Fifteen years ago, I had the precious opportunity to write a monograph with two great neuroscientists, Professor Sir John Eccles and Professor J. Szentagothai. This monograph, The Cerebellum as a Neuronal Machine, summarizes there markable progress of cerebellar studies in the 1960s, and symbolizes the challenge of examining complex structures and functions of the brain at the level of neurons and synapses. While the neuronal network structure of the cerebellar cortex was analyzed in detail morphologically by Professor Szentagothai and electrophysiologically by Sir John, my contribution to this monograph was to establish the general inhibitory action of cerebellar Purkinje cells upon cerebellar and vestibular nuclear cells and to formulate a basic schema of organization through the cerebellar cortex, cerebellar nuclei, and brain stem structures. During the past fifteen years, there have evolved a number of new issues in studies of the cerebellum. In addition to the microelectrode, Golgi staining, and electron mi croscopy, which were the major tools in the 1960s, a number of new tools and experimental materials have been introduced to further advance analysis at neuronal and synaptic levels. These include tissue culture, brain slices, mutant and X-ray irradiated animals, marker techniques utilizing axonal flow, high-voltage electron microscopy, intracellular dye injection, immunohistochemistry, chemical microanalyses, and computer simulations. In the light of these advances, the first aim of this monograph is to review new data obtained during the past fifteen years and incorporate them into concepts of the cerebellar neuronal machine. Advances in the 1970s, though, are not merely an extension of studies in the 1960s, which were centered around the initial question of " H o w is the cerebellar machinery constructed?", but have been associated with an expansion of the scope of research issues, especially to the development of theoretical modeling. During the 1970s, a number of international symposia with appeal to theoreticians were held. In addition, there are two further questions to be considered: "How does the cerebellar machinery operate?" and "How does the cerebellar machinery control diverse system functions of the body?" The neuronal network theories of Marr and others thus attempted to clarify the operations of cerebellar circuitry on the basis of an assumption of a special type of synaptic plasticity in the cerebellar cortex. Being impressed by the close conformity of this assumption to the neuronal circuitry connections in the vestibulocerebellum, worked out in our lab oratory in the early 1970s, I was prompted to verify the assumption with experimental data. Fortunately, positive evidence for the plasticity assumption has now become avail able. These neuronal network models of the cerebellar cortex answer the second question above, "How does the cerebellar neuronal circuitry operate?" but do not reply to the third question, "What does the cerebellum do?" or "What roles does the cerebellum play in the diverse system functions of the body?" Addressing this particular question, I have tried to develop control system models of the cerebellum and formulated the basic view that the cerebellum acts as an element that endows a motor or autonomic system with three major capabilities— coordination, orthometria, and compensation— just as a modern computer endows an engineering system with multivariable, predictive, and adaptive-learning control capabilities. Although much inference and speculation still prevail, there has been a remarkable amount of progress in our understanding of structural-functional relationships in the cerebellum. The second aim of this monograph is to review the enormous amount of currently accumulated data with reference to the neuronal networks and control system structures of the cerebellum. In addressing the above second and third questions of "How does the cerebellar machinery operate and control diverse system functions of the body?" I concentrated on studies of the cerebellar flocculus, in the hope that this region would represent a prototype of cerebellar control functions. As the specific relationship of the flocculus with vestibulo-ocular reflexes rapidly became clear, I proceeded to formulate the flocculus hypothesis of the vestibulo-ocular reflex control, and devoted all of my subsequent efforts to experimental verification of this hypothesis. I have attempted to converge all of the presently available approaches to the examination of a single system, i.e., the area in the rabbit flocculus that controls the vestibulo-ocular reflex. Although there are diverse opinions concerning the validity of the hypothesis, I believe that there is sufficient justification for maintaining the flocculus hypothesis and expanding it to a general hypothesis of neural control. The third and main objective of this monograph is to examine involvement of the cerebellum in diverse system functions of the body from this particular point of view. Thus, this monograph attempts not only to provide a comprehensive review of recent experimental data in a descriptive form but also to examine them in the light of con structive models of the cerebellum. The latter effort has met with difficulties, because there are still large gaps in our knowledge of both experimental facts and theories. Therefore, this volume is not a textbook-like introduction of established concepts but, rather, represents an intermediate stage of our advance toward understanding the operation of the cerebellar machinery and its role in the neural control of system functions of the body. Although this aspect of the monograph may be biased, I hope readers will understand my intent in writing it.The difficulties presented are certainly not specific to studies of the cerebellum; they are common to studies of the mechanism so fany brain function. I believe that a clear picture of the current status of our knowledge will help us to formulate perspectives of the future development of studies of the cerebellum and neural control.
Central nervous system myelomatosis is uncommon and is associated with a particularly poor prognosis. PET images, from a 53-year-old man referred to a fully digital F-FDG PET for relapsed multiple myeloma, revealed high F-FDG uptakes located in the cortex and sulci of the right central area and within the meningeal envelopes of the cerebellum, the trigeminal nerves, and on the spinal canal. These particular uptakes gave evidence of a central nervous system myelomatosis subsequently confirmed by plasma cells documented in cerebrospinal fluid. Such interesting images could be obtained owing to the potential of high-resolution images provided by fully digital F-FDG PET.
Purpose To evaluate whether patients with neurofibromatosis type 1 (NF1)-a multisystem neurodevelopmental disorder with myriad imaging manifestations, including focal transient myelin vacuolization within the deep gray nuclei, brainstem, and cerebellum-exhibit differences in cortical and subcortical structures, particularly in subcortical regions where these abnormalities manifest. Materials and Methods In this retrospective study, by using clinically obtained three-dimensional T1-weighted MR images and established image analysis methods, 10 intracranial volume-corrected subcortical and 34 cortical regions of interest (ROIs) were quantitatively assessed in 32 patients with NF1 and 245 age- and sex-matched healthy control subjects. By using linear models, ROI cortical thicknesses and volumes were compared between patients with NF1 and control subjects, as a function of age. With hierarchic cluster analysis and partial correlations, differences in the pattern of association between cortical and subcortical ROI volumes in patients with NF1 and control subjects were also evaluated. Results Patients with NF1 exhibited larger subcortical volumes and thicker cortices of select regions, particularly the hippocampi, amygdalae, cerebellar white matter, ventral diencephalon, thalami, and occipital cortices. For the thalami and pallida and 22 cortical ROIs in patients with NF1, a significant inverse association between volume and age was found, suggesting that volumes decrease with increasing age. Moreover, compared with those in control subjects, ROIs in patients with NF1 exhibited a distinct pattern of clustering and partial correlations. Discussion Neurofibromatosis type 1 is characterized by larger subcortical volumes and thicker cortices of select structures. Most apparent within the hippocampi, amygdalae, cerebellar white matter, ventral diencephalon, thalami and occipital cortices, these neurofibromatosis type 1-associated volumetric changes may, in part, be age dependent.
Cerebellar ataxia is a frequent and often disabling syndrome severely impairing motor functioning and quality of life. Patients suffer from reduced mobility, and restricted autonomy, experiencing an even lower quality of life than, e.g., stroke survivors. Aminopyridines have been demonstrated viable for the symptomatic treatment of certain forms of cerebellar ataxia. This article will give an outline of the present pharmacotherapy of different cerebellar disorders. As a current key-therapy for the treatment of downbeat nystagmus 4-aminopyridine (4-AP) is suggested for the treatment of downbeat nystagmus (5-10 mg Twice a day [TID]), a frequent type of persisting nystagmus , due to a compromise of the vestibulo-cerebellum. Studies with animals have demonstrated, that a nonselective blockage of voltage-gated potassium channels (mainly Kv1.5) increases Purkinje-cell (PC) excitability. In episodic ataxia type 2 (EA2), which is frequently caused by mutations of the PQ-calcium channel, the efficacy of 4-AP (5-10 mg TID) has been shown in a randomized controlled trial (RCT). 4-AP was well tolerated in the recommended dosages. 4-AP was also effective in elevating symptoms in cerebellar gait ataxia of different etiologies (2 case series). A new treatment option for cerebellar disease is the amino-acid acetyl-DL-leucine, which has significantly improved cerebellar symptoms in three case series. There are on-going randomized controlled trials for cerebellar ataxia (acetyl-DL-leucine vs placebo; ALCAT), cerebellar gait disorders (SR-form of 4-AP vs placebo; FACEG) and EA2 (sustained-release/SR-form of 4-AP vs acetazolamide vs placebo; EAT2TREAT), which will provide new insights into the pharmacological treatment of cerebellar disorders.
The study aimed to compare the whole-brain gray matter volume (GMV) and white matter volume (WMV) difference between primary angle closure glaucoma (PACG) patients and health controls (HCs) using a voxel-based morphometry method. A total of 27 patients with PACG (17 males and 10 females) and 27 HCs (17 males and 10 females), closely matched for age and education, were enrolled in the study. All subjects underwent magnetic resonance imaging (MRI) scans. The MRI data were processed using SPM8 software in voxel-based morphometry 8 toolbox. The relationship between the mean GMV values of brain regions and the clinical features including psychological testing and mean retinal nerve fiber layer (RNFL) thickness in PACG groups were analyzed by using Pearson correlation. Compared with HCs, PACG patients showed significantly decreased GMV values in the left cerebellum posterior lobe (CPL), right extra-nuclear, and right superior temporal gyrus. In contrast, PACG patients showed significantly increased GMV values in the left CPL, right CPL, right superior temporal gyrus, right thalamus and right insula (P<0.01). Moreover, in the PACG group, the left mean RNFL showed a positive correlation with the mean GMV values of the left CPL (r=0.719; P<0.001) and the right mean RNFL showed a positive correlation with the mean GMV values of the left CPL (r=0.721; P<0.001). The Hamilton depression score showed a positive correlation with the mean GMV values of right insula (r=0.897; P<0.001). Our results demonstrated that PACG patients showed altered brain structure in various regions related to visuomotor function, thalamocortical pathway, and emotion function, which might provide a useful informations to understanding the anatomy neural mechanisms of deficit in vision loss and depression in PACG.
To further knowledge of the physiology of opioid receptors in birds, the structure and expression of the μ-, δ-, and κ-opioid receptor genes were studied in a peregrine falcon ( Falco peregrinus), a snowy owl ( Bubo scandiacus), and a blue-fronted Amazon parrot ( Amazona aestiva). Tissue samples were obtained from birds that had been euthanatized for poor release prognosis or medical reasons. Samples were taken from the brain (telencephalon, thalamus, pituitary gland, cerebellum, pons, medulla oblongata, mesencephalon), the spinal cord and dorsal root ganglions, and plantar foot skin. Messenger RNA was recovered, and reverse transcription polymerase chain reaction (RT-PCR) was performed to generate complementary DNA (cDNA) sequences. Gene structures were documented by directly comparing cDNA sequences with recently published genomic sequences for the peregrine falcon and the blue-fronted Amazon parrot or by comparisons with genomic sequences of related species for the snowy owl. Structurally, the avian μ-opioid receptor messenger RNA (mRNA) species were complex, displaying differential splicing, alternative stop codons, and multiple polyadenylation signals. In comparison, the structure of the avian κ-receptor mRNA was relatively simple. In contrast to what is seen in humans, the avian δ-receptor mRNA structure was found to be complex, demonstrating novel 3-prime coding and noncoding exons not identified in mammals. The role of the δ-opioid receptor merits further investigation in avian species.
To systematically study somatic variants arising during development in the human brain across a spectrum of neurodegenerative disorders. In this study we developed a pipeline to identify somatic variants from exome sequencing data in 1461 diseased and control human brains. Eighty-eight percent of the DNA samples were extracted from the cerebellum. Identified somatic variants were validated by targeted amplicon sequencing and/or PyroMark® Q24. We observed somatic coding variants present in >10% of sampled cells in at least 1% of brains. The mutational signature of the detected variants showed a predominance of C>T variants most consistent with arising from DNA mismatch repair, occurred frequently in genes that are highly expressed within the central nervous system, and with a minimum somatic mutation rate of 4.25 × 10 per base pair per individual. These findings provide proof-of-principle that deleterious somatic variants can affect sizeable brain regions in at least 1% of the population, and thus have the potential to contribute to the pathogenesis of common neurodegenerative diseases.
Twenty-two feral rock pigeons ( Columba livia) from 10 counties in California with ataxia, torticollis, and difficulty standing and flying were admitted to rehabilitation centers in late winter and spring of 2017 and died or were euthanized. Common necropsy findings included thin body condition, generalized deep red discoloration of organs, and hemorrhagic subcutaneous neck tissues. Meningoencephalitis was observed microscopically in 16 pigeons, and 3 also had protozoal schizonts in the brain. The most consistently affected regions of the brain were cerebellum and brainstem. Skeletal muscles, and less frequently the heart, contained large intrasarcoplasmic bradyzoites typically without inflammation. Fifteen of the 22 birds tested positive using pan- Sarcocystis polymerase chain reaction. The sequence of the amplicon was most closely related to S. calchasi, and the 8 subtyped sequences had 100% homology with S. calchasi. This investigation demonstrated the transcontinental and North American spread of S. calchasi causing a disease outbreak in free-ranging rock pigeons, thus warranting increased surveillance in susceptible native columbids.
Primary vitreoretinal lymphoma (PVRL) can be a diagnostic challenge and commonly presents as a partially steroid-responsive vitritis or as subretinal cream-colored infiltrates. The authors present a patient with PVRL who initially presented with bilateral vitritis; however, after two non-diagnostic vitrectomy specimens and two unremarkable brain MRIs, she was lost to follow-up. She presented 2.5 years later with a white plaque on the posterior capsule of her left intraocular lens, though the vitreous cavity was free of infiltrate. Repeat biopsy revealed diffuse large B-cell lymphoma, and brain MRI demonstrated an enhancing lesion of the cerebellum, consistent with primary central nervous system lymphoma.
Functional connectivity is frequently derived from fMRI data to reduce a complex image of the brain to a graph, or "functional connectome". Often shortest-path algorithms are used to characterize and compare functional connectomes. Previous work on the identification and measurement of semi-metric (shortest circuitous) pathways in the functional connectome has discovered cross-sectional differences in major depressive disorder (MDD), autism spectrum disorder (ASD), and Alzheimer's disease. However, while measurements of shortest path length have been analyzed in functional connectomes, less work has been done to investigate the composition of the pathways themselves, or whether the edges composing pathways differ between individuals. Developments in this area would help us understand how pathways might be organized in mental disorders, and if a consistent pattern can be found. Furthermore, studies in structural brain connectivity and other real-world graphs suggest that shortest pathways may not be as important in functional connectivity studies as previously assumed. In light of this, we present a novel measurement of the consistency of pathways across functional connectomes, and an algorithm for improvement by selecting the most frequently occurring "normative pathways" from the k shortest paths, instead of just the shortest path. We also look at this algorithm's effect on various graph measurements, using randomized matrix simulations to support the efficacy of this method and demonstrate our algorithm on the resting-state fMRI (rs-fMRI) of a group of 34 adolescent control participants. Additionally, a comparison of normative pathways is made with a group of 82 age-matched participants, diagnosed with MDD, and in doing so we find the normative pathways that are most disrupted. Our results, which are carried out with estimates of connectivity derived from correlation, partial correlation, and normalized mutual information connectomes, suggest disruption to the default mode, affective, and ventral attention networks. Normative pathways, especially with partial correlation, make greater use of critical anatomical pathways through the striatum, cingulum, and the cerebellum. In summary, MDD is characterized by a disruption of normative pathways of the ventral attention network, increases in alternative pathways in the frontoparietal network in MDD, and a mixture of both in the default mode network. Additionally, within- and between-groups findings depend on the estimate of connectivity.
Transient Cerebral Arteriopathy (TCA) is one of the main causes of childhood stroke. Here we present an unusual case of Arterial Ischemic Stroke (AIS) caused by a TCA of posterior flow and originally located in the right thalamus. The detection of enterovirus in the cerebrospinal fluid allowed us to suppose a probable post infectious etiology. The course of symptoms was self-limited and the child had a complete clinical recovery after five days. A new ischemic lesion on the antero-inferior paravermian region of the left cerebellum was revealed by a following brain Magnetic Resonance Imaging (MRI) three months later and these findings were reported by further brain MRI control performed after 15 months. Comparing follow up Magnetic Resonance Angiography (MRA) with previous High Resolution Vessel Wall Magnetic Resonance Imaging (HRMI), we found a vessel narrowing at the level of the Posterior Inferior Cerebellar Artery that might explain the arteriopathy process. In conclusion, clinical and radiological course allow us to speculate that this multifocal cerebral arteriopathy might be a transient lesion due to enterovirus infection. To our knowledge, there are only three articles describing TCA enterovirus-related, and brain MRA was performed in only one case; in addition, no one with the involvement of the posterior circulation.
Principal neurons encode information by varying their firing rate and patterns precisely fine-tuned through GABAergic interneurons. Dysregulation of inhibition can lead to neuropsychiatric disorders, yet little is known about the molecular basis underlying inhibitory control. Here, we find that excessive GABA release from basket cells (BCs) attenuates the firing frequency of Purkinje neurons (PNs) in the cerebellum of Fragile X Mental Retardation 1 (Fmr1) knockout (KO) mice, a model of Fragile X Syndrome (FXS) with abrogated expression of the Fragile X Mental Retardation Protein (FMRP). This over-inhibition originates from increased excitability and Ca transients in the presynaptic terminals, where Kv1.2 potassium channels are downregulated. By paired patch-clamp recordings, we further demonstrate that acutely introducing an N-terminal fragment of FMRP into BCs normalizes GABA release in the Fmr1-KO synapses. Conversely, direct injection of an inhibitory FMRP antibody into BCs, or membrane depolarization of BCs, enhances GABA release in the wild type synapses, leading to abnormal inhibitory transmission comparable to the Fmr1-KO neurons. We discover that the N-terminus of FMRP directly binds to a phosphorylated serine motif on the C-terminus of Kv1.2; and that loss of this interaction in BCs exaggerates GABA release, compromising the firing activity of PNs and thus the output from the cerebellar circuitry. An allosteric Kv1.2 agonist, docosahexaenoic acid, rectifies the dysregulated inhibition in vitro as well as acoustic startle reflex and social interaction in vivo of the Fmr1-KO mice. Our results unravel a novel molecular locus for targeted intervention of FXS and perhaps autism.
Previous studies reported the delayed recovery group after circadian rhythm disruption in mice showed higher quinpiroleinduced locomotor activity. This study aimed to compare not only Protein Kinase C (PKC) activities in frontal, striatal, hippocampus and cerebellum, but also relative PKC activity ratios among brain regions according to recovery of circadian rhythm. The circadian rhythm disruption protocol was applied to eight-week-old twenty male Institute Cancer Research mice. The circadian rhythm recovery patterns were collected through motor activities measured by Mlog system. Depressive and manic proneness were examined by forced swim test and quinpirole-induced open field test respectively. Enzyme-linked immunosorbent assay was employed to measure PKC activities. The delayed recovery group presented greater locomotor activities than the early recovery group (p=0.033). The delayed recovery group had significantly lower frontal PKC activity than the other (p=0.041). The former showed lower frontal/cerebellar PKC activity ratio (p=0.047) but higher striatal/frontal (p=0.038) and hippocampal/frontal (p=0.007) PKC activities ratios than the latter. These findings support potential mechanism of delayed recovery after circadian disruption in bipolar animal model could be an alteration of relative PKC activities among mood regulation related brain regions. It is required to investigate the PKC downstream signaling related to the delayed recovery pattern.
Many viruses depend on the extensive membranous network of the endoplasmic reticulum (ER) for their translation, replication, and packaging. Certain membrane modifications of the ER can be a trigger for ER stress, as well as the accumulation of viral protein in the ER by viral infection. Then, unfolded protein response (UPR) is activated to alleviate the stress. Zika virus (ZIKV) is a mosquito-borne flavivirus and its infection causes microcephaly in newborns and serious neurological complications in adults. Here, we investigated ER stress and the regulating model of UPR in ZIKV-infected neural cells in vitro and in vivo. Mice deficient in type I and II IFN receptors were infected with ZIKV via intraperitoneal injection and the nervous tissues of the mice were assayed at 5 days post-infection. The expression of phospho-IRE1, XBP1, and ATF6 which were the key markers of ER stress were analyzed by immunohistochemistry assay in vivo. Additionally, the nuclear localization of XBP1s and ATF6n were analyzed by immunohistofluorescence. Furthermore, two representative neural cells, neuroblastoma cell line (SK-N-SH) and astrocytoma cell line (CCF-STTG1), were selected to verify the ER stress in vitro. The expression of BIP, phospho-elF2α, phospho-IRE1, and ATF6 were analyzed through western blot and the nuclear localization of XBP1s was performed by confocal immunofluorescence microscopy. RT-qPCR was also used to quantify the mRNA level of the UPR downstream genes in vitro and in vivo. ZIKV infection significantly upregulated the expression of ER stress markers in vitro and in vivo. Phospho-IRE1 and XBP1 expression significantly increased in the cerebellum and mesocephalon, while ATF6 expression significantly increased in the mesocephalon. ATF6n and XBP1s were translocated into the cell nucleus. The levels of BIP, ATF6, phospho-elf2α, and spliced xbp1 also significantly increased in vitro. Furthermore, the downstream genes of UPR were detected to investigate the regulating model of the UPR during ZIKV infection in vitro and in vivo. The transcriptional levels of atf4, gadd34, chop, and edem-1 in vivo and that of gadd34 and chop in vitro significantly increased. Findings in this study demonstrated that ZIKV infection activates ER stress in neural cells. The results offer clues to further study the mechanism of neuropathogenesis caused by ZIKV infection.
Discriminating strokes in patients with acute dizziness/vertigo is challenging especially when other symptoms and signs of central nervous involvements are not evident. Despite the developments in imaging technology over the decades, a significant proportion of acute strokes may escape detection on imaging especially during the acute phase or when the lesions are small. Thus, small strokes causing isolated dizziness/vertigo would have a higher chance of misdiagnosis in the emergency department. Even though several diagnostic algorithms have been advanced for acute vascular vertigo, we still await more comprehensive and sophisticated ones that can also be applied to transient vestibular symptoms due to vascular compromise. In this respect, vascular and perfusion imaging would be informative. Application of artificial intelligence and tele-consultation may be future perspectives for real-time decision in acute dizziness and vertigo. Several new constellations of ocular motor and vestibular findings have been added to the strokes involving the brainstem and cerebellum. Defining these characteristics would help understanding the function of central vestibular structures and allow more accurate localization of the strokes involving these structures.
The dorsal mesial frontal cortex contains the supplementary motor area (SMA) and the pre-supplementary motor area (pre-SMA), which play an important role in action and cognition. Evidence from cytoarchitectonic, stimulation, and functional studies suggests structural and functional divergence between the two subregions. However, a microstructural map of these areas obtained in a representative sample of brains in a stereotaxic reference space is still lacking. In the present study we show that the dorsal mesial frontal motor cortex comprises two microstructurally different brain regions: area SMA and area pre-SMA. Area-specific cytoarchitectonic patterns were studied in serial histological sections stained for cell bodies of ten human postmortem brains. Borders of the two cortical areas were identified using image analysis and statistical features. The 3D reconstructed areas were transferred to a common reference space, and probabilistic maps were calculated by superimposing the individual maps. A coordinate-based meta-analysis of functional imaging data was subsequently performed using the two probabilistic maps as microstructurally defined seed regions. It revealed that areas SMA and pre-SMA were strongly co-activated with areas in precentral, supramarginal and superior frontal gyri, Rolandic operculum, thalamus, putamen and cerebellum. Both areas were related to motor functions, but area pre-SMA was involved in more complex processes such as learning, cognitive processes and perception. The here described subsequent analyses led to converging evidence supporting the microstructural, and functional segregation of areas SMA and pre-SMA, and maps will be made available to the scientific community to further elucidate the microstructural substrates of motor and cognitive control.
In recent yearsestradiol has emerged as a potential regulator of transient receptor potential vanilloid (TRPV) cationic channels in the peripheral tissues and sensory neurons, however, its analogous role in theCNS is poorly understood. TRPV channels modulate Ca signalling, neurotransmission and behaviour, andexpression of these ion channels and estrogen receptors show a great degree of overlap in different brain regions. Herein, we probe if TRPV1-6 genes contain estrogen receptor-binding sites and if their expression in different brain regions is modulated during estrous cycle. Bioinformatics analysis of the mouse TRPV1-6 gene sequences showed presence of putative functional estrogen response element in their promoter regions. Using qRT-PCR, TRPV1-6 mRNA expression was observed in the olfactory bulb, cortex, hypothalamus, hippocampus, brainstem, and cerebellum of mouse. In these regions, compared to estrus, metestrus, and diestrus, reduced levels of TRPV1 and TRPV5 but elevated TRPV2 and TRPV6 mRNA levels were observed during proestrus. Lower levels of TRPV3 and TRPV4 mRNAs were seen during estrus but higher expression of TRPV3 during metestrus and diestrus, and TRPV4 during proestrus was observed. Estradiol seems to regulate TRPV1/TRPV5 and TRPV2/TRPV6 mRNA expression in opposite manner. Except TRPV4 mRNA expression in the hippocampus and TRPV6 expression in the olfactory bulb, hippocampus and brainstem, expression of other member of TRPV subfamily in distinct brain regions of male mice was comparable to those in metestrus and diestrus mice. We suggest that the circulating levels of estradiol during the estrous cycle may differentially regulate the activity of TRPV1-6 ion channels in the brain.
The effect of two second-generation antidepressants escitalopram and venlafaxine on the activity of brain and liver cytochrome P450 2D (CYP2D) involved in the metabolism of psychotropics and neurotransmitters was determined in the chronic mild stress (CMS) model of depression. Escitalopram or venlafaxine (10 mg/kg ip/day each) were administered to control and CMS rats for 5 weeks. The activity of CYP2D was studied by measurement of the rate of bufuralol 1'-hydroxylation in microsomes derived from the liver or different brain structures. The obtained results indicate that CMS and the studied antidepressants had different effects on the CYP2D activity depending on the location of the enzyme. In the brain, CMS produced an increase in the CYP2D activity in the hippocampus. Chronic escitalopram or venlafaxine had no effect on the CYP2D activity in the brain of nonstressed rats, however, the antidepressants increased the enzyme activity in the frontal cortex, hypothalamus and cerebellum of stressed animals. In the liver, CMS did not affect the CYP2D activity, while chronic escitalopram or venlafaxine significantly decreased the CYP2D activity and protein level in nonstressed and stressed rats. We conclude that: 1) CMS stimulates the CYP2D activity in the hippocampus and triggers the stimulatory effect of antidepressants on CYP2D in other brain structures; 2) the local brain metabolism of CYP2D substrates (neurosteroids, neurotransmitters, psychotropics) may be enhanced by CMS and/or antidepressants; 3) in contrast to the brain, the liver metabolism of CYP2D substrates may be slower during long-term treatment with escitalopram or venlafaxine.
The aims of this study were to examine both static functional connectivity (FC) and dynamic FC alterations in motor execution regions after stroke and to investigate whether the altered static or dynamic FC was associated with the clinical behaviors in stroke patients. Seventy-six stroke patients and 55 healthy controls (HC) were recruited. Static FC and dynamic FC maps were computed based on the seeds of six core regions in motor execution network. Correlation analyses were performed between static or dynamic FC and clinical behavioral scores in stroke patients. Compared with the HC, the stroke patients had significantly higher static FC between the seeds and the precentral or postcentral gyrus, frontal gyrus, inferior parietal lobule, thalamus and insula, and lower static FC between the seeds and the cerebellum and middle temporal gyrus. There were significant differences in dynamic FC between the seeds and precuneus, calcarine gyrus, insula, inferior parietal lobule, precentral gyrus, and middle temporal, frontal or occipital gyrus between the stroke patients and HC. Furthermore, a significant negative correlation was found between the Fugl-Meyer assessment scores and dynamic FC of the ipsilesional primary motor cortex and contralesional precentral gyrus in patients. The current study shows that the patterns of both static FC and dynamic FC changed after stroke, and correlation between motor function and temporal variability in the FC of the precentral gyrus was significant in stroke patients. Our findings indicate that dynamic FC might be a potential indicator for evaluating motor function after stroke.
The aim was to investigate normal patterns of brain metabolism determined by F-fluorodeoxyglucose positron emission tomography (FDG PET)/MRI during childhood and adolescence. A retrospective analysis was carried out on all paediatric patients who underwent FDG PET/MRI at our institution between March 2016 and August 2017. Exclusion criteria were neurological disease, central nervous system metastases, previous chemotherapy/radiotherapy, general anaesthesia/sedation and medications suspected to affect cerebral metabolism. Standardized uptake value (SUV)mean and SUVmax were calculated for 12 brain grey matter regions. Subgroup analysis of childhood (≤10 years old) and adolescence (≥11 years old) was also carried out. From 492 PET/MRI scans, 28 patients (11 children, 17 adolescents) were deemed representative of normal brain metabolism. SUVmean and SUVmax increased with age in all regions. The highest rates of increasing SUVmean were in the thalamus, basal ganglia, frontal lobes, insula and occipital lobes. Higher SUVmean was found in the right frontal, right lateral temporal, right temporal pole, right cingulate/paracingulate, right thalamus, left occipital, left basal ganglia, left insula and left cerebellum compared with the contralateral side. This SUVmean asymmetry was present in both childhood and adolescence in the majority of regions. The highest rates of increasing SUVmax with age were in the occipital lobes, frontal lobes, thalamus and central region. There was no asymmetry in SUVmax in the majority of regions. This FDG PET/MRI study shows that normal brain metabolism measured by SUVmean and SUVmax increases with age in all regions, proceeding at different rates between distinct anatomical sites. Our results suggest that there is mild asymmetry in SUVmean, but mostly symmetric SUVmax during normal development.
Maternal health and nutrition during the perinatal period is the predominant factor influencing the functional development of the brain. Maternal malnutrition during the perinatal period causes retardation of brain development. The current study investigates the role of Astaxanthin (AsX) in spatial learning and memory and BDNF in perinatally undernourished Wistar rats. The albino wistar rats were perinatally undernourished and administered with different dosages of AsX. The spatial learning and memory performance and BDNF level were assessed. Data were collected and analysed. The % Correct choice during the acquisition phase, performance at the end of the acquisition phase and the mean BDNF level at the Hippocampus, Cerebellum, and Cerebral cortex showed significant decline (P<0.001) in the PUN group and significantly high (P<0.001) in the PUNA2 group compared to the control. However, the mean RME and mean WME during different days of the acquisition phase were significantly high (P<0.001) in the PUN group and insignificant (P>0.05) in PUNA2 compared to the control. The results showed that AsX effectively modulated the cognitive deficit that occurred in perinatally undernourished rats. This can be attributed to BDNF upregulation as evidenced by the significant increase of the BDNF level.
Liponeurocytoma is a rare benign tumor of the central nervous system, which develops mainly in adult patients within the posterior fossa. The WHO has categorized this entity in their last classification of 2016 as a benign grade II tumor. Histopathological characteristics contain neuronal and variable astrocytic differentiation with foci of lipomatous distinction. Up to now only a few case reports and case series are published and the knowledge of this tumor is still very limited. General treatment guidelines do not exist. Aim of this study was to analyze the literature to create treatment guidelines. PRISMA (i.e., Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines were followed to query existing online databases between January 1, 1978 and May 15, 2018. Systematic review of the literature revealed 73 cases (40 female, 33 male) of a liponeurocytoma whereof 59 cases (80.8%) occurred in the posterior fossa. Ki-67/MIB-1 proliferation index was described in 58 cases showing a mean value of 3.73% ± 4.01. Follow-up was performed in 49 cases with a median length of 52.02 ± 50.52 months, showing tumor recurrence in 14 cases (28.57%). Tumor recurrence was observed in only one case (1/12, 8.33%) after adjuvant radiotherapy, while tumor recurrence was seen in 13/29 patients (44.83%), who did not receive adjuvant radiotherapy. Liponeurocytomas are rare benign tumors, occurring mainly in the cerebellum. Therapy of choice is surgery. Postoperative radiotherapy seems to lower the risk of tumor recurrence and should be offered to the patient.
Locomotion in vertebrates relies on motor circuits in the spinal cord receiving inputs from the hindbrain to execute motor commands while dynamically integrating proprioceptive sensory feedback. The spatial organization of the neuronal networks driving locomotion in the hindbrain and role of inhibition has not been extensively investigated. Here, we mapped neuronal activity with single-cell resolution in the hindbrain of restrained transgenic Tg(HuC:GCaMP5G) zebrafish larvae swimming in response to whole-field visual motion. We combined large-scale population calcium imaging in the hindbrain with simultaneous high-speed recording of the moving tail in animals where specific markers label glycinergic inhibitory neurons. We identified cells whose activity preferentially correlates with the visual stimulus or motor activity and used brain registration to compare data across individual larvae. We then morphed calcium imaging data onto the zebrafish brain atlas to compare with known transgenic markers. We report cells localized in the cerebellum whose activity is shut off by the onset of the visual stimulus, suggesting these cells may be constitutively active and silenced during sensorimotor processing. Finally, we discover that the activity of a medial stripe of glycinergic neurons in the domain of expression of the transcription factor engrailed1b is highly correlated with the onset of locomotion. Our efforts provide a high-resolution, open-access dataset for the community by comparing our functional map of the hindbrain to existing open-access atlases and enabling further investigation of this population's role in locomotion.
The cerebellum plays an important role in depression. Cerebro-cerebellar circuits have been found to show aberrance in bipolar disorder (BD) and major depressive disorder (MDD). However, whether the cerebro-cerebellar connectivity contributes equally to the pathologic mechanisms of BD and MDD remains unknown. We recruited 33 patients with MDD, 32 patients with BD, and 43 healthy controls (HC). We selected six seed regions (three per hemisphere) in the cerebrum, corresponding to the affective, cognitive control, and default mode networks, to establish cerebro-cerebellar functional connectivity maps. Relative to the HC, both the BD and MDD patients exhibited weaker negative connectivity between the right subgenual anterior cingulate cortex and the cerebellar vermis IV_V (p = 0.03, p = 0.001) and weaker positive connectivity between the left precuneus and the left cerebellar lobule IX (p = 0.043, p = 0.000). Moreover, the MDD patients showed weaker positive connectivity in the left precuneus-left cerebellar lobule IX circuit than the BD patients (p = 0.049). In addition, the BD patients showed weaker positive connectivity in the right dorsolateral prefrontal cortex-left cerebellar lobule Crus Ι circuit compared to the HC (p = 0.002) or the MDD patients (p = 0.013). Receiver operating characteristic curves analyses showed that the altered cerebro-cerebellar connectivities could be used to distinguish the patients from the HC with relatively high accuracy. Our findings suggested that differences in connectivity of cerebro-cerebellar circuits, which are involved in affective or cognitive functioning, significantly contributed to BD and MDD.
The meninges are pivotal in protecting the brain against traumatic brain injury (TBI), an ongoing issue in most mainstream sports. Improved understanding of TBI biomechanics and pathophysiology is desirable to improve preventative measures, such as protective helmets, and advance our TBI diagnostic/prognostic capabilities. This study mechanically characterised the porcine meninges by performing uniaxial tensile testing on the dura mater (DM) tissue adjacent to the frontal, parietal, temporal, and occipital lobes of the cerebellum and superior sagittal sinus region of the DM. Mechanical characterisation revealed a significantly higher elastic modulus for the superior sagittal sinus region when compared to other regions in the DM. The superior sagittal sinus and parietal regions of the DM also displayed local mechanical anisotropy. Further, fatigue was noted in the DM following ten preconditioning cycles, which could have important implications in the context of repetitive TBI. To further understand differences in regional mechanical properties, regional variations in protein content (collagen I, collagen III, fibronectin and elastin) were examined by immunoblot analysis. The superior sagittal sinus was found to have significantly higher collagen I, elastin, and fibronectin content. The frontal region was also identified to have significantly higher collagen I and fibronectin content while the temporal region had increased elastin and fibronectin content. Regional differences in the mechanical and biochemical properties along with regional tissue thickness differences within the DM reveal that the tissue is a non-homogeneous structure. In particular, the potentially influential role of the superior sagittal sinus in TBI biomechanics warrants further investigation. This study addresses the lack of regional mechanical analysis of the cortical meninges, particularly the dura mater (DM), with accompanying biochemical analysis. To mechanically characterise the stiffness of the DM by region, uniaxial tensile testing was carried out on the DM tissue adjacent to the frontal, parietal, temporal and occipital lobes along with the DM tissue associated with the superior sagittal sinus. To the best of the authors' knowledge, the work presented here identifies, for the first time, the heterogeneous nature of the DM's mechanical stiffness by region. In particular, this study identifies the significant difference in the stiffness of the DM tissue associated with the superior sagittal sinus when compared to the other DM regions. Constitutive modelling was carried out on the regional mechanical testing data for implementation in Finite Element models with improved biofidelity. This work also presents the first biochemical analysis of the collagen I and III, elastin, and fibronectin content within DM tissue by region, providing useful insights into the accompanying macro-scale biomechanical data.
Bilirubin neurotoxicity has been studied for decades and has been shown to affect various mechanisms via significant modulation of gene expression. This suggests that vital regulatory mechanisms of gene expression, such as epigenetic mechanisms, could play a role in bilirubin neurotoxicity. Histone acetylation has recently received attention in the CNS due to its role in gene modulation for numerous biological processes, such as synaptic plasticity, learning, memory, development and differentiation. Aberrant epigenetic regulation of gene expression in psychiatric and neurodegenerative disorders has also been described. In this work, we followed the levels of histone 3 lysine 14 acetylation (H3K14Ac) in the cerebellum (Cll) of the developing (2, 9, 17 days after the birth) and adult Gunn rat, the natural model for neonatal hyperbilirubinemia and kernicterus. We observed an age-specific alteration of the H3K14Ac in the hyperbilirubinemic animals. The GeneOntology analysis of the H3K14Ac linked chromatin revealed that almost 45% of H3K14Ac ChiP-Seq TSS-promoter genes were involved in CNS development including maturation and differentiation, morphogenesis, dendritogenesis, and migration. These data suggest that the hallmark Cll hypoplasia in the Gunn rat occurs also via epigenetically controlled mechanisms during the maturation of this brain structure, unraveling a novel aspect of the bilirubin-induced neurotoxicity.
To investigate chemotherapy dosage-related cognitive impairment and its neural mechanisms in breast cancer (BC) patients. Twenty-eight breast cancer patients after each chemotherapy cycle and matched 29 healthy control subjects underwent structural magnetic resonance imaging. Voxel-based morphometry analysis was performed to compare group differences in the gray matter for the whole brain. Furthermore, mediation analysis was conducted to explore the role of brain structures in chemotherapy dosage-related cognitive impairment. Voxel-based morphometry analysis was performed in gray matter for the whole brain of BC patients after chemotherapy. The results revealed that the gray matter density in the left inferior frontal gyrus, right middle frontal gyrus, right fusiform area, and bilateral cerebellum was decreased in the BC patients compared to controls. The number of chemotherapy cycles was negatively associated with general cognitive capacity, verbal fluency and digit span performance in the BC patients. In addition, decreased gray matter density in the right middle frontal gyrus could mediate the chemotherapy dosage effects on verbal fluency performance. These findings indicate that the dose-response relationship between chemotherapy and cognitive impairment may depend on the decreases in gray matter density of the frontal cortical structures.
Immune-mediated cerebellar ataxias (IMCAs), a clinical entity reported for the first time in the 1980s, include gluten ataxia (GA), paraneoplastic cerebellar degenerations (PCDs), anti-glutamate decarboxylase 65 (GAD) antibody-associated cerebellar ataxia, post-infectious cerebellitis, and opsoclonus myoclonus syndrome (OMS). These IMCAs share common features with regard to therapeutic approaches. When certain factors trigger immune processes, elimination of the antigen(s) becomes a priority: e.g., gluten-free diet in GA and surgical excision of the primary tumor in PCDs. Furthermore, various immunotherapeutic modalities (e.g., steroids, immunoglobulins, plasmapheresis, immunosuppressants, rituximab) should be considered alone or in combination to prevent the progression of the IMCAs. There is no evidence of significant differences in terms of response and prognosis among the various types of immunotherapies. Treatment introduced at an early stage, when CAs or cerebellar atrophy is mild, is associated with better prognosis. Preservation of the "cerebellar reserve" is necessary for improvement of CAs and resilience of the cerebellar networks. In this regard, we emphasize the therapeutic principle of "Time is Cerebellum" in IMCAs.
Diffusion tensor histology holds great promise for quantitative characterization of structural connectivity in mouse models of neurological and psychiatric conditions. There has been extensive study in both the clinical and preclinical domains on the complex tradeoffs between the spatial resolution, the number of samples in diffusion q-space, scan time, and the reliability of the resultant data. We describe here a method for accelerating the acquisition of diffusion MRI data to support quantitative connectivity measurements in the whole mouse brain using compressed sensing (CS). The use of CS allows substantial increase in spatial resolution and/or reduction in scan time. Compared to the fully sampled results at the same scan time, the subtle anatomical details of the brain, such as cortical layers, dentate gyrus, and cerebellum, were better visualized using CS due to the higher spatial resolution. Compared to the fully sampled results at the same spatial resolution, the scalar diffusion metrics, including fractional anisotropy (FA) and mean diffusivity (MD), showed consistently low error across the whole brain (< 6.0%) even with 8.0 times acceleration. The node properties of connectivity (strength, cluster coefficient, eigenvector centrality, and local efficiency) demonstrated correlation of better than 95.0% between accelerated and fully sampled connectomes. The acceleration will enable routine application of this technology to a wide range of mouse models of neurologic diseases.
Cognitive and social capacities require postnatal experience, yet the pathways by which experience guides development are unknown. Here we show that the normal development of motor and nonmotor capacities requires cerebellar activity. Using chemogenetic perturbation of molecular layer interneurons to attenuate cerebellar output in mice, we found that activity of posterior regions in juvenile life modulates adult expression of eyeblink conditioning (paravermal lobule VI, crus I), reversal learning (lobule VI), persistive behavior and novelty-seeking (lobule VII), and social preference (crus I/II). Perturbation in adult life altered only a subset of phenotypes. Both adult and juvenile disruption left gait metrics largely unaffected. Contributions to phenotypes increased with the amount of lobule inactivated. Using an anterograde transsynaptic tracer, we found that posterior cerebellum made strong connections with prelimbic, orbitofrontal, and anterior cingulate cortex. These findings provide anatomical substrates for the clinical observation that cerebellar injury increases the risk of autism.
Cerebellar hypoplasia is a rare disorder of cerebellar formation in which the cerebellum is not completely developed, smaller than it should be, or completely absent. The prevalence of cerebellar hypoplasia at birth is unknown, and little is known about epidemiological risk factors. Using data from the National Birth Defects Prevention Study (NBDPS), a population-based, case-control study, we analyzed clinical features and potential risk factors for nonsyndromic cerebellar hypoplasia. The NBDPS included pregnancies with estimated delivery dates from 1997-2011. We described clinical features of cerebellar hypoplasia cases from the study area. We explored risk factors for cerebellar hypoplasia (case characteristics, demographics, pregnancy characteristics, maternal health conditions, maternal medication use, and maternal behavioral exposures) by comparing cases to non-malformed live born control infants. We calculated crude odds ratios (ORs) and 95% confidence intervals using logistic regression models. We identified 87 eligible cerebellar hypoplasia cases and 55 mothers who participated in the NBDPS. There were no differences in clinical features between interviewed and non-interviewed cases. Cerebellar hypoplasia cases were more likely than controls to be from a multiple pregnancy, be born preterm, and have low birth weight. Cerebellar hypoplasia cases were more likely to be born in or after 2005, as opposed to earlier in NBDPS. We found elevated ORs that were not statistically significant for maternal use of vasoactive medications, non-Hispanic black mothers, and mothers with a history of hypertension. Although unadjusted, our findings from a large, population-based study can contribute to new hypotheses regarding the etiology of cerebellar hypoplasia.
The aim of this study was to compare volumetric parameters in the abnormal and normal posterior fossa using the Virtual Organ Computer-aided AnaLysis VOCAL technique to determine whether fetuses with an abnormal posterior fossa have different volumes. A prospective study was conducted on 17 fetuses with an abnormal posterior fossa including, Dandy Walker malformation (n = 6), vermian hypoplasia (n = 3), mega cisterna magna (n = 8) and 99 healthy control fetuses from 20 to 34 weeks' gestation. Measurement of the fetal cisterna magna and cerebellar volume was performed in the standard transcerebellar plane through the VOCAL method. To establish the correlation of volumes with gestational age, polynomial regression analysis was performed. For comparison between groups, univariate ANCOVA was performed using gestational age as a covariate. The reliability was analyzed by the intraclass correlation coefficient (ICC). Cerebellar volume and cisterna magna volume was correlated with gestational age. Posterior fossa volume was significantly larger in Dandy Walker malformation (DWM) (p < 0.0001) and mega cisterna magna (p < 0.0001) in comparison to the control group. In vermian hypoplasia group, cisterna magna volume does not seem to expand (p = 0.298). Cerebellar volume does not seem to change in subgroups when the influence of gestational age is discarded (p = 0.09). The ratio of cerebellar volume to the cisterna magna volume decreases significantly in abnormal fetuses (p < 0.0001). Good intraobserver and interobserver reliability was found for both cerebellum and cisterna magna measurements. Volume analysis may have a role in discrimination of different posterior fossa pathologies.
Autoimmune epilepsy (AE) is becoming increasingly recognized as a potentially reversible cause of frequent or medically intractable seizures and cognitive deterioration. We describe various presentations of autoimmune encephalopathy which have specifically presented with seizure and describe reported imaging findings. This is organized as a review of the more common autoantibodies which can specifically precipitate seizure according to the intracellular or extracellular location of the targeted antigen. For each antibody, we illustrate their pathophysiology, characteristic clinical presentations with typical effective treatments and prognoses and imaging findings on MRI and PET/CT exams. Parenchymal involvement is variable with the limbic structures typically affected; however, non-limbic cortex, cerebellum, brainstem and basal ganglia can also be involved. In the acute setting, affected regions typically demonstrate T hyperintensity with mild mass effect from edema and increased F-FDG uptake. Chronically involved parenchyma will often undergo atrophy and demonstrate decreased metabolism; mesial temporal sclerosis (MTS) is often the end result when the limbic system is involved. Without treatment, long term effects from AE range from ongoing cognitive dysfunction and refractory seizures to death. Familiarity with AE may prompt appropriate antibody screening, particularly in cases of refractory seizure disorders. Early investigation and proper management of AE cases may help to prevent parenchymal and neurologic deterioration in these patients.
Due to concerns for enhanced absorption of manganese (Mn) from drinking water compared to diet, bioavailability of Mn from drinking water remains a major data gap in understanding Mn kinetics. In this study, PBPK models for adult rats and humans were updated with a drinking water exposure route and were used to assess the homeostatic control of Mn uptake, excretion and tissue kinetics between the two different ingestion modes. Drinking water model parameters were estimated from tissue kinetic data from a drinking water study in rats. The published study included a 10 ppm-Mn diet with additional Mn added to drinking water to give a total ingested Mn dose equivalent to that from a 200 ppm diet. The 200 ppm diet and equivalent mixed drinking water/diet exposures provided Mn concentrations for brain (striatum, olfactory bulb, and cerebellum), liver and bone after 7 and 61 days of Mn exposure. Modeling of these data sets indicated that (1) the oral Mn bioavailability is similar for diet or drinking water and (2) homeostatic control of gut uptake of Mn occurs with either drinking water or dietary ingestion. This updated description for absorption and distribution of Mn from gut was added to a human Mn-PBPK model to simulate Mn exposure from multiple routes of exposure (i.e. dietary intake, drinking water, and inhalation). This increases the utility of the Mn PBPK model by allowing for the simulation of multiple Mn exposure scenarios, including variable daily food and drinking water exposures in a human population.
Gait impairment during complex walking in older adults is thought to result from a progressive failure to compensate for deteriorating peripheral inputs by central neural processes. It is the primary hypothesis of this paper that failure of higher cerebral adaptations may already be present in middle-aged adults who do not present observable gait impairments. We therefore compared metabolic brain activity during steering of gait (i.e., complex locomotion) and straight walking (i.e., simple locomotion) in young and middle-aged individuals. Cerebral distribution of [18F]-fluorodeoxyglucose, a marker of brain synaptic activity, was assessed during over ground straight walking and steering of gait using positron emission tomography in seven young adults (aged 24±3) and seven middle-aged adults (aged 59±3). Brain regions involved in steering of gait (posterior parietal cortex, superior frontal gyrus, and cerebellum) are retained in middle-age. However, despite similar walking performance, there are age-related differences in the distribution of [ 18F]-FDG during steering: middle-aged adults have (i) increased activation of precentral and fusiform gyri, (ii) reduced deactivation of multisensory cortices (inferior frontal, postcentral, fusiform gyri), and (iii) reduced activation of the middle frontal gyrus and cuneus. Our results suggest that pre-clinical decline in central sensorimotor processing in middle-age is observable during complex walking.
To evaluate the prevalence of anti-AQP4 antibody in serum and CSF samples from patients being investigated for possible neuromyelitis optica spectrum disorder (NMOSD) referred to the PathWest State reference laboratory using a sensitive cell-based assay (CBA). NMOSD is an inflammatory CNS disease distinct from MS, which is relatively rare in Western countries. A proportion of patients with NMOSD have detectable serum IgG antibodies that target the water channel aquaporin-4 (AQP4-IgG), but the frequency varies in different populations studied and according to the assay method employed. Sera or CSF from a diagnostic cohort of 196 consecutive patients with possible NMOSD which had previously been screened by indirect immunofluorescence (IIF) on primate cerebellum were re-tested for AQP4-IgG reactivity to the M1 and M23 isoforms of AQP4 using a commercial CBA. A control group of 205 patients with definite MS was also included in the study. Of the 196 patients, only 5 sera were AQP4-IgG positive, representing 2.6% of patients in the diagnostic cohort. All 5 AQP4-IgG positive patients fulfilled the 2015 revised diagnostic criteria for NMOSD and were females of varied ethnic origins, 4 of whom had longitudinally extensive transverse myelitis. The CBA confirmed AQP4-IgG positivity in the four patients previously reported as positive by IIF, and an additional patient with NMOSD who had previously been diagnosed as MS was also identified. None of the 205 MS sera were AQP4-IgG positive. Our study confirms the utility and greater reliability of the M1/M23 CBA for detecting AQP4-IgG in patients with possible NMOSD, and indicates a prevalence of seropositive NMOSD in the Western Australian population similar to that in other Western populations.
One of the pivotal events in neural development is compartmentalization, wherein the neural tissue divides into domains and undergoes functional differentiation. For example, midbrain-hindbrain boundary (MHB) formation and subsequent isthmus development are key steps in cerebellar development. Although several regulatory mechanisms are known to underlie this event, little is known about cellular behaviors. In this study, to examine the cellular dynamics around the MHB region, we performed confocal time-lapse imaging in zebrafish embryos to track cell populations in the neural tube via 4D analysis. We used a transgenic line wherein enhanced green fluorescent protein (EGFP) expression is driven by the gastrulation brain homeobox 2 (gbx2) enhancer, which is involved in MHB maintenance. 4D time-lapse imaging of 5-20 h revealed a novel pattern in cell migration: a dynamic ventrocaudally directed migration from the MHB region toward the hindbrain. Furthermore, in the hindbrain region, these EGFP-positive cells altered their shapes and extended the axons. Immunohistochemical analysis and retrograde labeling showed that these cells in the hindbrain were in the process of neuronal differentiation, including reticulospinal neurons. These results revealed the dynamic and two-step behavior and possible fate of the cell population, which are linked to brain compartmentalization, leading to a deeper understanding of brain development and formation of neuronal circuits.
While the mouse has been a productive model for inner ear studies, a lack of highly specific genes and tools has presented challenges. The absence of definitive otic lineage markers and tools is limiting in vitro studies of otic development, where innate cellular heterogeneity and disorganization increase the reliance on lineage-specific markers. To address this challenge in mice and embryonic stem (ES) cells, we targeted the lineage-specific otic gene Fbxo2 with a multicistronic reporter cassette (Venus/Hygro/CreER = VHC). In otic organoids derived from ES cells, Fbxo2 specifically delineates otic progenitors and inner ear sensory epithelia. In mice, Venus expression and CreER activity reveal a cochlear developmental gradient, label the prosensory lineage, show enrichment in a subset of type I vestibular hair cells, and expose strong expression in adult cerebellar granule cells. We provide a toolbox of multiple spectrally distinct reporter combinations for studies that require use of fluorescent reporters, hygromycin selection, and conditional Cre-mediated recombination.
Strictly controlled dendrite patterning underlies precise neural connection. Dendrite self-avoidance is a crucial system preventing self-crossing and clumping of dendrites. Although many cell-surface molecules that regulate self-avoidance have been identified, the signaling pathway that orchestrates it remains poorly understood, particularly in mammals. Here, we demonstrate that the LKB1-SIK kinase pathway plays a pivotal role in the self-avoidance of Purkinje cell (PC) dendrites by ensuring dendritic localization of Robo2, a regulator of self-avoidance. LKB1 is activated in developing PCs, and PC-specific deletion of LKB1 severely disrupts the self-avoidance of PC dendrites without affecting gross morphology. SIK1 and SIK2, downstream kinases of LKB1, mediate LKB1-dependent dendrite self-avoidance. Furthermore, loss of LKB1 leads to significantly decreased Robo2 levels in the dendrite but not in the cell body. Finally, restoration of dendritic Robo2 level via overexpression largely rescues the self-avoidance defect in LKB1-deficient PCs. These findings reveal an LKB1-pathway-mediated developmental program that establishes dendrite self-avoidance.
The aim of this study was to evaluate the clinical utility of arterial spin labeling (ASL) magnetic resonance imaging (MRI) for the detection of cerebellar hypoperfusion in patients with spinocerebellar degeneration (SCD). Regional cerebral blood flow (CBF) were obtained from ASL and I-IMP single-photon emission computed tomography (SPECT) images by volume-of-interest analysis in patients with SCD (n = 16). Regional CBF were also measured by ASL in age-matched controls (n = 19) and by SPECT in separate controls (n = 17). The cerebellar CBF values were normalized to the CBF values for the whole gray matter (nCBF) in ASL and SPECT. The mean cerebellar nCBF measured by ASL was lower in patients with SCD (0.70 ± 0.09) than in the controls (0.91 ± 0.05) (p < 0.001), which was consistent with the comparison using SPECT (0.82 ± 0.05 vs. 0.98 ± 0.05, p < 0.001). The cerebellar nCBF measured by ASL significantly correlated with that determined by SPECT in patients (r = 0.56, p < 0.001). ASL imaging showed decreased cerebellar blood flow, which correlated with that measured by SPECT, in patients with SCD. These findings suggest the clinical utility of noninvasive MRI with ASL for detecting cerebellar hypoperfusion in addition to atrophy, which would aid the diagnosis of SCD.
Purpose We applied voxelwise apparent diffusion coefficient (ADC) histogram analysis in addition to structural magnetic resonance imaging (MRI) findings and patients' age for differentiation of intraaxial posterior fossa tumors involving the fourth ventricle. Participants and methods Pretreatment MRIs of 74 patients with intraaxial brain neoplasm involving the fourth ventricle, from January 1, 2004 to December 31, 2015, were reviewed. The tumor solid components were segmented and voxelwise ADC histogram variables were determined. Histogram-driven variables, structural MRI findings, and patient age were combined to devise a differential diagnosis algorithm. Results The most common neoplasms were ependymomas ( n = 21), medulloblastoma ( n = 17), and pilocytic astrocytomas ( n = 13). Medulloblastomas followed by atypical teratoid/rhabdoid tumors had the lowest ADC histogram percentile values; whereas pilocytic astrocytomas and choroid plexus papillomas had the highest ADC histogram percentile values. In a multivariable multinominal regression analysis, the ADC 10th percentile value from voxelwise histogram was the only independent predictor of tumor type ( p < 0.001). In separate binary logistic regression analyses, the 10th percentile ADC value, tumor morphology, enhancement pattern, extension into Luschka/Magendie foramina, and patient age were predictors of different tumor types. Combining these variables, we devised a stepwise diagnostic model yielding 71% to 82% sensitivity, 91% to 95% specificity, 75% to 78% positive predictive value, and 89% to 95% negative predictive value for differentiation of ependymoma, medulloblastoma, and pilocytic astrocytoma. Conclusion We have shown how the addition of quantitative voxelwise ADC histogram analysis of the tumor solid component to structural findings and patient age can help with accurate differentiation of intraaxial posterior fossa neoplasms involving the fourth ventricle based on pretreatment MRI.
Surface protein dynamics dictate synaptic connectivity and function in neuronal circuits. , a gene disrupted by copy number variations (CNVs) in neurodevelopmental disorders, including autism spectrum, was previously shown to regulate the surface expression of ASTN1 in glial-guided neuronal migration. Here, we demonstrate that ASTN2 binds to and regulates the surface expression of multiple synaptic proteins in postmigratory neurons by endocytosis, resulting in modulation of synaptic activity. In cerebellar Purkinje cells (PCs), by immunogold electron microscopy, ASTN2 localizes primarily to endocytic and autophagocytic vesicles in the cell soma and in subsets of dendritic spines. Overexpression of ASTN2 in PCs, but not of ASTN2 lacking the FNIII domain, recurrently disrupted by CNVs in patients, including in a family presented here, increases inhibitory and excitatory postsynaptic activity and reduces levels of ASTN2 binding partners. Our data suggest a fundamental role for ASTN2 in dynamic regulation of surface proteins by endocytic trafficking and protein degradation.
OBJECTIVE To determine brain region affinity for and retention of gadolinium in dogs after administration of gadodiamide and whether formalin fixation affects quantification. ANIMALS 14 healthy dogs. PROCEDURES 13 dogs received gadodiamide (range, 0.006 to 0.1 mmol/kg, IV); 1 control dog received a placebo. Dogs received gadodiamide 3 to 7 days (n = 8) or 9 hours (5) before euthanasia and sample collection. Brain regions were analyzed with inductively coupled mass spectrometry (ICP-MS) and transmission electron microscopy. Associations between dose, time to euthanasia, and gadolinium retention quantities (before and after fixation in 5 dogs) were evaluated. RESULTS Gadolinium retention was seen in all brain regions at all doses, except for the control dog. Exposure 3 to 7 days before euthanasia resulted in 1.7 to 162.5 ng of gadolinium/g of brain tissue (dose-dependent effect), with cerebellum, parietal lobe, and brainstem affinity. Exposure 9 hours before euthanasia resulted in 67.3 to 1,216.4 ng of gadolinium/g of brain tissue without dose dependency. Transmission electron microscopy revealed gadolinium in examined tissues. Fixation did not affect quantification in samples immersed for up to 69 days. CONCLUSIONS AND CLINICAL RELEVANCE Gadodiamide exposure resulted in gadolinium retention in the brain of healthy dogs. Cerebellum, parietal lobe, and brainstem affinity was detected with dose dependency only in dogs exposed 3 to 7 days before euthanasia. Fixation had no effect on quantification when tissues were immersed for up to 69 days. Physiologic mechanisms for gadolinium retention remained unclear. The importance of gadolinium retention requires further investigation.
Parallel fibers in the molecular layer of the vertebrate cerebellum mediate slow spike conduction in the transverse plane. In contrast, electrophysiological recordings have indicated that rapid spike conduction exists between the lateral regions of the cerebellar cortex of the red-ear pond turtle (Trachemys Scripta). The anatomical basis for this commissure is now examined in that species using neuronal tracing techniques. Fluorescently tagged dextrans and lipophilic carbocyanine dyes placed in one lateral edge of this non-foliated cortex are transported across the midline of living brains in vitro and along the axonal membranes of fixed tissues, respectively. Surprisingly, the labeled commissural axons traversed the cortex within the Purkinje cell layer, and not in the white matter of the molecular layer or the white matter below the granule cell layer. Unlike thin parallel fibers that exhibit characteristic varicosities, this commissure is composed of smooth axons of large diameter that also extend beyond the cerebellar cortex via the cerebellar peduncles. Double labeling with Myelin Basic Protein antibody demonstrated that these commissural axons are ensheathed with myelin. In contrast to this transverse pathway, an orthogonal myelinated tract was observed along the cerebellar midline. The connections of this transverse commissure with the lateral cerebellum, the vestibular nuclear complex and the cochlear vestibular ganglia indicate that this commissure plays a role in bilateral vestibular connectivity. This article is protected by copyright. All rights reserved.
The putative cache (Ca channel and chemotaxis receptor) domain containing 1 (CACHD1) protein has predicted structural similarities to members of the α2δ voltage-gated Ca channel (VGCC) auxiliary subunit family. CACHD1 mRNA and protein were highly expressed in the male mammalian CNS, in particular in the thalamus, hippocampus and cerebellum, with a broadly similar tissue distribution to Ca3 subunits, in particular, Ca3.1. In expression studies, CACHD1 increased cell-surface localization of Ca3.1 and these proteins were in close proximity at the cell surface consistent with the formation of CACHD1-Ca3.1 complexes. In functional electrophysiological studies, co-expression of human CACHD1 with Ca3.1, Ca3.2 and Ca3.3 caused a significant increase in peak current density and corresponding increases in maximal conductance. By contrast, α2δ-1 had no effect on peak current density or maximal conductance in either Ca3.1, Ca3.2 or Ca3.3. Comparison of CACHD1-mediated increases in Ca3.1 current density and gating currents revealed an increase in channel open probability. In hippocampal neurons from male and female E19 rats, CACHD1 overexpression increased Ca3-mediated action potential (AP) firing frequency and neuronal excitability. These data suggest that CACHD1 is structurally an α2δ-like protein that functionally modulates Ca3 voltage-gated calcium channel activity.This is the first study to characterise the CACHD1 protein. CACHD1 is widely expressed in the CNS, in particular in the thalamus, hippocampus and cerebellum. CACHD1 distribution is similar to that of low-voltage-activated (Ca3, T-type) calcium channels, in particular to Ca3.1, a protein which regulates neuronal excitability and is a potential therapeutic target in conditions such as epilepsy and pain. CACHD1 is structurally a α2δ-like protein that functionally increases Ca3 calcium current. CACHD1 increases the presence of Ca3.1 at the cell surface, forms complexes with Ca3.1 at the cell-surface and causes an increase in channel open probability. In hippocampal neurons, CACHD1 causes increases in neuronal firing. Thus, CACHD1 represents a novel protein that modulates Ca3 activity.
Soft actuators have played an indispensable role in generating compliant motions of soft robots. Among the various soft actuators explored for soft robotic applications, dielectric elastomer actuators (DEAs) have caught the eye with their intriguing attributes similar to biological muscles. However, the control challenge of DEAs due to their strong nonlinear behaviors has hindered the development of DEA-based soft robots. To overcome the control challenge, this paper proposes a bioinspired control approach of DEAs. A three-dimensional muscle-like DEA, capable of large forces and giant deformation, is fabricated and adopted as the control platform. To facilitate the controller design, the dynamic model of the DEA is developed through experimental analysis, which takes electromechanical coupling, viscoelastic effects and dynamics uncertainties into consideration. Motivated by the proprioception of the biological muscles, the self-sensing capability of the actuator is explored and exhibits good accuracy. Thus the self-sensing of the actuator is utilized to provide the sensory feedback in the control loop without the need of additional external sensors. Inspired from the role of the cerebellum in motor learning, a cerebellum model articulation nonlinear controller is proposed to compensate the dynamics uncertainties and to provide motion correction. Finally, the effectiveness of the proposed control approach is verified by both the simulation and the experiments.
The aging process clearly increases the demand for antioxidant protection, especially in the brain, involving that provided by α-tocopherol (αT). However, little is known about the age-related changes in brain αT levels and the influencing effect of gender on it, in human or murine samples as well. Accordingly, the aim of the current study was to detect age-, gender- and region-specific changes in αT concentrations in mouse brain tissue and to assess the influencing effect of plasma αT levels on it. Female and male C57BL/6 mice at the ages of 6, 16 and 66 weeks (n = 9 in each group) were applied. αT levels were determined with high performance liquid chromatography (HPLC) from the striatum, cortex, hippocampus, cerebellum, brainstem and from plasma samples. A detailed validation process was carried out for the applied HPLC method as well. The results demonstrated that brain αT levels significantly increased in the striatum, cortex, and hippocampus with aging in both genders, but in a more pronounced way in females with an increasing magnitude of this difference. In case of the cerebellum, a moderate elevation could be detected only in females, whereas in case of the brainstem there was no significant change in αT level. With regard to plasma samples, no clear trend could be identified. The current study is the first to present age-dependent gender-specific changes in αT level in certain brain regions of the C57Bl/6 mouse strain, and may provide meaningful information for future therapeutic studies targeting aging-related processes.
We describe the phenomenon of crossed cerebellar diaschisis (CCD) in four subjects diagnosed with Alzheimer's disease (AD) according to the National Institute on Aging -Alzheimer Association (NIA-AA) criteria, in combination with 18F-FDG PET and 11C-PiB PET imaging. 18F-FDG PET showed a pattern of cerebral metabolism with relative decrease most prominent in the frontal-parietal cortex of the left hemisphere and crossed hypometabolism of the right cerebellum. 11C-PiB PET showed symmetrical amyloid accumulation, but a lower relative tracer delivery (a surrogate of relative cerebral blood flow) in the left hemisphere. CCD is the phenomenon of unilateral cerebellar hypometabolism as a remote effect of supratentorial dysfunction of the brain in the contralateral hemisphere. The mechanism implies the involvement of the cortico-ponto-cerebellar fibers. The pathophysiology is thought to have a functional or reversible basis but can also reflect in secondary morphologic change. CCD is a well-recognized phenomenon, since the development of new imaging techniques, although scarcely described in neurodegenerative dementias. To our knowledge this is the first report describing CCD in AD subjects with documentation of both 18F-FDG PET and 11C-PiB PET imaging. CCD in our subjects was explained on a functional basis due to neurodegenerative pathology in the left hemisphere. There was no structural lesion and the symmetric amyloid accumulation did not correspond with the unilateral metabolic impairment. This suggests that CCD might be caused by non-amyloid neurodegeneration. The pathophysiological mechanism, clinical relevance and therapeutic implications of CCD and the role of the cerebellum in AD need further investigation.
The neural histaminergic system innervates the cerebellum, with a high density of fibers in the vermis and flocculus. The cerebellum participates in motor functions, but the role of the histaminergic system in this function is unclear. In the present study, we investigated the effects of intracerebellar histamine injections and H1, H2 and H3 receptor antagonist injections (chlorpheniramine, ranitidine, and thioperamide, respectively) and H4 receptor agonist (VUF-8430) on locomotor and exploratory behaviors in mice. The cerebellar vermis of male mice was implanted with guide cannula. After three days of recovery,the animals received microinjections of saline or histamine (experiment1), saline or chlorpheniramine (experiment 2), saline or ranitidine(experiment 3), saline or thioperamide (experiment 4), and saline or VUF-8430 (experiment 5) in different concentrations. Five minutes postinjection,the open field test was performed. The data were analyzed using one-way ANOVA and Duncan's post hoc test. The microinjections of histamine, ranitidine or thioperamide did not lead any behavioral effects at the used doses. In contrast, animals that received chlorpheniramine at the highest dose (0.16 nmol) and VUF-8430 at the highest dose (1.48 nmol)were more active in the open field apparatus, with an increase in the number of crossed quadrants, number of rearings and time spent in the central area of the arena, suggesting that chlorpheniramine and VUF-8430 modulates locomotor and exploratory behaviors in mice.