Cerebellar Weight Research Articles

Arbor vitae cerebelli: Fractal properties and their quantitative assessment by novel “contour scaling” fractal analysis method (an anatomical study)

BackgroundArbor vitae cerebelli (tree-like branching white matter of the cerebellum) has a complex spatial configuration that is challenging to assess using conventional morphometric methods. This study proposes a fractal approach to describe and quantify the anatomy of Arbor vitae cerebelli. For this purpose, a new “contour scaling” method for fractal analysis of cerebellar white matter was developed. Material and methodsThe cerebella of 100 cadavers (50 male and 50 female) who died from causes unrelated to brain pathology, aged 20–95 years, were examined. Mid-sagittal sections of the cerebellar vermis were studied. The fractal dimension values of the cerebellar white matter were determined using both the developed fractal analysis method and the conventional “box counting” method, along with measurements of non-fractal parameters including cerebellar weight, area and perimeter of the vermis cross-section, perimeter-to-area ratio, and circularity. ResultsConsidering the cerebellar white matter as a tree-like fractal, it was found to have 7 or 8 primary branches, which subdivide into 10-18 second-iteration branches, 19–38 third-iteration branches, and 34–53 fourth-iteration branches. Females more often had 8 primary branches compared to males, while males had a greater number of branches in the second to fourth iterations. The mean fractal (Hausdorff) dimension was 1.697 (1.721 in males, 1.674 in females, P = 0.01). The fractal dimension correlated most strongly with the perimeter and area of the vermis cross-section and had no significant relationship with age. ConclusionThe fractal (Hausdorff) dimension, determined using the novel “contour scaling” method, quantitatively assesses the degree of branching of the cerebellar white matter. An increase in the absolute size of the cerebellum leads to a higher degree of branching of its white matter and an increase in the number of its constitutive components – white matter branches and folia.

The Role of Vitamin C on ATPases Activities in Monosodium Glutamate-Induced Oxidative Stress in Rat Striatum and Cerebellum.

Monosodium glutamate (MSG) is a silent excitotoxin used as a flavour enhancer but exerts serious health hazards to consumers. MSG plays a role in neuronal function as the dominant excitatory neurotransmitter. It is transferred into the blood and ultimately increases brain glutamate levels, causing functional disruptions notably via oxidative stress. The study evaluated the toxic effect of high consumption of MSG and the modulatory role of vitamin C on ATPase activities in the striatum and cerebellum of male Wistar rats for five weeks. Rats were grouped into four (A-D): group A was fed with rat's show only; Group B was fed with diet containing 15% MSG; Group C was treated with vitamin C (200mg/kg b.wgt orally in 0.9% saline solution) only for 3weeks; and group D rats were fed with MSG and vitamin C. The findings show that MSG does not affect body and cerebellum weights but increases striatal weight. MSG increases the malondialdehyde (MDA) level and significantly decreases catalase (CAT) and superoxide dismutase (SOD) activities and glutathione (GSH) levels. MSG significantly impaired striatal and cerebellar ATPases activities (Na+/K+-, Ca2+-, Mg2+- and total ATPases). Vitamin C treatment abolishes MSG-induced oxidative stress and improves ATPase activities. The findings show that vitamin C has beneficial effects in improving the functions of membrane-bound ATPases against MSG toxicity in rat's striatum and cerebellum.

Developmental Ethanol Exposure Impacts Purkinje Cells but Not Microglia in the Young Adult Cerebellum.

Fetal alcohol spectrum disorders (FASD) caused by developmental ethanol exposure lead to cerebellar impairments, including motor problems, decreased cerebellar weight, and cell death. Alterations in the sole output of the cerebellar cortex, Purkinje cells, and central nervous system immune cells, microglia, have been reported in animal models of FASD. To determine how developmental ethanol exposure affects adult cerebellar microglia and Purkinje cells, we used a human third-trimester binge exposure model in which mice received ethanol or saline from postnatal (P) days 4-9. In adolescence, cerebellar cranial windows were implanted and mice were aged to young adulthood for examination of microglia and Purkinje cells in vivo with two-photon imaging or in fixed tissue. Ethanol had no effect on microglia density, morphology, dynamics, or injury response. However, Purkinje cell linear frequency was reduced by ethanol. Microglia-Purkinje cell interactions in the Purkinje Cell Layer were altered in females compared to males. Overall, developmental ethanol exposure had few effects on cerebellar microglia in young adulthood and Purkinje cells appeared to be more susceptible to its effects.

Assessment of Protective Roles of Aqueous Seed Extract of Sour-Sop (Annona muricata) in Cerebellum Following Cadmium-induced Neurotoxicity of Adult Wistar Rats

Cadmium, a deleterious heavy metal that pervades the environment, has the potential to accumulate in the body and cause health complications. In the domain of traditional medicine, medicinal plants have been employed to combat the toxicity of heavy metals and treating of many diseases in traditional medicine.
 Aim: To access the protective impact of Annona muricata seed extract on cadmium-triggered neurotoxicity in the cerebellum of albino wistarrat.
 Place of Study: Department of Anatomy, Faculty of Basic Medical Sciences Olabisi Onabanjo University, Ogun State between June 2022 and June 2023
 METHODOLOGY: After a period of 14 days of acclimatization, 40 healthy male wistar rats were randomly allocated to four groups, group1(2ml/kg of distilled water), group2 cadmium only(2g/kg SC), group3 (2g/kg cadmium + 100mg/kg extract of Annona muricata) and group4(2g/kg cadmium + 200mg/kg extract of Annona muricata) The rats were subjected to cadmium subcutaneously followed by the oral administration of aqueous seed extract of Annona Muricata for fifteen consecutive days.
 Results: The impact of the extract on antioxidant enzymes activities of the cerebellum, cerebellar weight, as well as the histology of the cerebellum were scrutinized. The data revealed that the aqueous seed extract of Annona muricata, in a dose-dependent manner, augmented the levels of superoxide dismutase (SOD) (P=0.005) and catalase (CAT) (P=0.027) in contrast to the cadmium-only group. The relative weight of the cerebellum exhibited a significant increasein the treated groups compared to the cadmium-only group.
 The histology of the cerebellum delineated pathological changes arising from the exposure to cadmium, while Annona muricata brought about regenerative changes.
 Conclusion: In summary, the study posits that owing to the presence of phytochemicals in Annona muricata aqueous seed extract, it was efficacious in mitigating the neurotoxicity induced by cadmium in the cerebellum of male rats.

Volumetric calculation of cerebellum in Akkaraman sheep and Hair goat using Cavalieri’s principle

The aim of this study is to calculate the cerebellum volume in Akkaraman sheep and Hair goat using the Cavalieri's principle. Cerebellum of 12 healthy 1-2 year old male animals (6 sheep and 6 goats) were used in the study. The weight of the cerebellum separated from the rhombencephalon was calculated with a sensitive electronic balance, and its volume was calculated with Archimedes' principle. In the volume measurement to be made with Cavalieri's principle, the cerebellum was blocked with 8% agar to prevent tissue loss during slicing. The stained sections were scanned with a horizontal scanner at 600 dpi resolution. Gray matter and white matter volume were calculated separately by dropping the point counting grid on cross sections in ImageJ software. Cerebellum weight was calculated as 11.6 gr in sheep and 12.55 gr in goats. The ratio of cerebellum weight to total brain weight was calculated as 0.10 in both species. It was observed that the cerebellum weighs an average of 9.8% of the total brain weight in sheep. In the goat, it was observed that the cerebellum constituted 10.11% of the brain. Gray matter and white matter volumes were calculated as 6.75 ml and 3.36 ml in sheep, respectively. In the goat, the gray matter and white matter volumes were measured as 6.80 ml and 3.82 ml, respectively. Total cerebellum volume was found to be 10.14 ml and 10.65 ml in sheep and goats, respectively. In sheep, 65.55% of the cerebellum volume consisted of gray matter and 33.08% of white matter. In goats, 63.88% of the cerebellum consisted of gray matter and 35.85% of white matter. No statistical difference was observed in the volume measurement results obtained in both species (p>0.05). In recent years, there has been an increase in neurodegenerative disease models in farm animals. These diseases can cause changes in the volume of the cerebellum. In this context, it is thought that the volume values obtained from healthy sheep and goat cerebellum in the current study will be important for future studies

Effects of RNA M6A demethylase ALKBH5 gene deficiency on morphology and function of cerebellum in aged mice

Objective: To investigate the effects of RNA m6A demethylase ALKBH5 gene deficiency on cerebellar morphology and function in the aged mice, and to explore the role of ALKBH5 in cerebellar degeneration. Methods: Western blot was performed to detect the protein level of ALKBH5 in the cerebellum of wild-type mice of various ages. The expression of NeuN, Calbindin-D28K, MAP2, GFAP and other proteins in the cerebella of middle-aged (12-month-old) and aged (18-month-old) wild-type mice and ALKBH5-/- mice was examined using immunohistochemistry. The balance beam test and gait analysis were performed to test the balance ability and motor coordination of the mice. Results: With aging of the mice, the expression of ALKBH5 in the cerebellum increased gradually in an age-dependent manner. In the aged mice, but not middle-aged mice, the body weight, whole brain weight and cerebellum weight of ALKBH5-/- mice decreased by 15%, 10% and 21%, respectively (P<0.05). The expression of ALKBH5 in the Purkinje cells was much higher than that in other types of neural cells. Correspondingly, ALKBH5-deficiency caused 40% reduction in the number of Purkinje cells, as well as the length and density of neuronal dendrites in the aged mice (P<0.01). In addition, the time for the aged ALKBH5-/- mice to pass the balance beam was 70% longer than that of the wild type mice of the same age, with unstable gaits (P<0.01). Conclusions: Gene deficiency of RNA m6A demethylase ALKBH5 causes cerebellar atrophy, Purkinje neuron loss and damage in the aged mice. These changes eventually affect mice's motor coordination and balance ability. These results suggest that imbalanced RNA m6A methylation may lead to neurodegenerative lesions in the cerebellum of mice.

Perinatal Protein Restriction Impacts Nuclear O-GalNAc Glycosylation in Cells of Liver and Brain Structures of the Rat

BackgroundPost-translational modifications are key factors in the modulation of nuclear protein functions controlling cell physiology and an individual’s health. ObjectivesThis study examined the influence of protein restriction during the perinatal period on the nuclear O-N-acetylgalactosamine (O-GalNAc) glycosylation of cells from the liver and parts of the brain in the rat. MethodsPregnant Wistar rats were divided into 2 groups on day 14 of pregnancy and fed ad libitum 1 of 2 isocaloric diets containing 24% (well-fed) or 8% (protein-restricted diet) casein until the end of the experiment. Male pups were studied after weaning at 30 d of life. Animals and their organ/tissues (liver, cerebral cortex, cerebellum and hippocampus) were weighed. Cell nuclei were purified, and the presence in nucleus and cytoplasm of all factors required for the initiation of O-GalNAc glycan biosynthesis, i.e., the sugar donor (UDP-GalNAc), enzyme activity (ppGalNAc-transferase) and the glycosylation product (O-GalNAc glycans), were evaluated by western blotting, fluorescent microscopy, enzyme activity, enzyme-lectin sorbent assay and mass spectrometry. ResultsThe perinatal protein deficit reduced progeny weight, as well as the cerebral cortex and cerebellum weight. UDP-GalNAc levels in the cytoplasm and nuclei of the liver, the cerebral cortex, cerebellum, or hippocampus were not affected by the perinatal dietary protein deficits. However, this deficiency affected the ppGalNAc-transferase activity localized in the cerebral cortex and hippocampus cytoplasm as well as in the liver nucleus, thus reducing the “writing” ppGalNAc-transferase activity of O-GalNAc glycans. In addition, liver nucleoplasm from protein-restricted offspring revealed a significant reduction in the expression of O-GalNAc glycans on important nuclear proteins. ConclusionsOur results report an association between the consumption of a protein-restricted diet by the dam and her progeny with the modulation in the offspring’ liver nuclei O-GalNAc glycosylation, which may ultimately regulate nuclear protein functions.

Mesenchymal Stem Cell Transplantation Ameliorates Ara-C-Induced Motor Deficits in a Mouse Model of Cerebellar Ataxia

This study investigated the therapeutic effects of transplanting human mesenchymal stem cells (hMSCs) into wild-type mice that were intraperitoneally administered cytosine arabinoside (Ara-C) to develop cerebellar ataxia (CA) during the first three postnatal days. hMSCs were intrathecally injected into 10-week-old mice once or thrice at 4-week intervals. Compared to the nontreated mice, the hMSC-treated mice showed improved motor and balance coordination, as measured using the rotarod, open-field, and ataxic scoring assessments, and increased protein levels in Purkinje and cerebellar granule cells, as measured using calbindin and NeuN protein markers. Multiple hMSC injections preserved Ara-C-induced cerebellar neuronal loss and improved cerebellar weight. Furthermore, the hMSC implantation significantly elevated the levels of neurotrophic factors, including brain-derived and glial cell line-derived neurotrophic factors, and suppressed TNF-α-, IL-1β-, and iNOS-mediated proinflammatory responses. Collectively, our results demonstrate that hMSCs exhibit therapeutic potential for Ara-C-induced CA by protecting neurons through the stimulation of neurotrophic factors and inhibition of cerebellar inflammatory responses, which can improve motor behavior and alleviate ataxia-related neuropathology. In summary, this study suggests that hMSC administration, particularly multiple treatments, can effectively treat ataxia-related symptoms with cerebellar toxicity.

Heat shock protein amplification improves cerebellar myelination in the Npc1nih mouse model.

Niemann-Pick disease type C (NPC) is a rare prematurely fatal lysosomal lipid storage disease with limited therapeutic options. The prominent neuropathological hallmarks include hypomyelination and cerebellar atrophy. We previously demonstrated the efficacy of recombinant human heat shock protein 70 (rhHSP70) in preclinical models of the disease. It reduced glycosphingolipid levels in the central nervous system (CNS), improving cerebellar myelination and improved behavioural phenotypes in Npc1nih (Npc1-/-) mice. Furthermore, treatment with arimoclomol, a well-characterised HSP amplifier, attenuated lysosomal storage in NPC patient fibroblasts and improved neurological symptoms in Npc1-/- mice. Taken together, these findings prompted the investigation of the effects of HSP amplification on CNS myelination. We administered bimoclomol daily or rhHSP70 6 times per week to Npc1-/- (BALB/cNctr-Npc1m1N/J, also named Npc1nih) mice by intraperitoneal injection from P7 through P34 to investigate the impact on CNS myelination. The Src-kinase inhibitor saracatinib was administered with/without bimoclomol twice daily to explore the contribution of Fyn kinase to bimoclomol's effects. Treatment with either bimoclomol or rhHSP70 improved myelination and increased the numbers of mature oligodendrocytes (OLs) as well as the ratio of active-to-inactive forms of phosphorylated Fyn kinase in the cerebellum of Npc1-/- mice. Additionally, treatment with bimoclomol preserved cerebellar weight, an effect that was abrogated when co-administered with saracatinib, an inhibitor of Fyn kinase. Bimoclomol-treated mice also exhibited increased numbers of immature OLs within the cortex. These data increase our understanding of the mechanisms by which HSP70 regulates myelination and provide further support for the clinical development of HSP-amplifying therapies in the treatment of NPC. Funding for this study was provided by Orphazyme A/S (Copenhagen, Denmark) and a Pathfinder Award from The Wellcome Trust.

Effects of Passiflora foetida on the Number of Purkinje Cells in the Cerebellum of Albino Rats

Permot (Passiflora foetida) is one type of plant that has been used as a mosquito coil. Besides being effective at killing Aedes aegypti mosquitoes, mosquito coils made from permot leaf extract are also effective at killing German cockroaches and house flies. Until now there have been no reports stating the side effects of using permot leaf extract in humans. This study was designed to demonstrate the harmful side effects of inhalation of smoked mosquito coils made from permot leaf extract on the cerebellar cortex of adult male rats. Thirty-five adult male rats were used in this study. Rats were divided into 7 groups, namely group 1 as a negative control (without exposure), group 2 as a positive control (transfluthrin exposure 3000 ppm), groups 3 to 7 were treatment groups of permot leaf extract with graded doses (500 ppm, 1000 ppm, 2000 ppm, 3000 ppm, and 4000 ppm), and was treated for 12 weeks. All rats were sacrificed in a timely manner and cerebellar specimens were taken, prepared and observed using a light microscope. Previously, rats were weighed periodically to determine the effect of toxicity. The results showed that exposure to mosquito coil smoke made from 3000 ppm Transflutrin and 4000 ppm permot leaf extract caused weight loss in rats. Likewise on the weight of the rat cerebellum. The effect of decreasing the number of Purkinje cells also occurred in the group exposed to mosquito coils made from Transflutrin 3000 ppm and permot leaf extract 4000 ppm. Therefore, the safe and effective dose of mosquito coils made from permot leaves is less than 4000 ppm, because exposure to doses of more than 4000 ppm can cause degenerative changes in the cerebellar cortex.

Early developmental alcohol exposure alters behavioral outcomes following adolescent re-exposure in a rat model.

Prenatal alcohol exposure alters brain development, affecting cognitive, motor, and emotional domains, and potentially leading to greater alcohol intake during adolescence. The present study investigated whether early alcohol exposure modifies vulnerability to behavioral alterations associated with adolescent alcohol exposure in a rodent model. Sprague-Dawley rats received ethanol or sham intubations during two developmental periods: (1) the third trimester equivalent of brain development in humans (postnatal days [PD] 4-9) and (2) adolescence (PD 28-42). Both exposures resulted in blood alcohol concentrations around 200 mg/dl. Subjects were tested in the open field (PD 45-48) and on hippocampal and prefrontal cortical (PFC) dependent tasks: the Morris water maze (PD 52-58) and trace fear conditioning (PD 63-64). Neonatal alcohol exposure reduced forebrain and cerebellar weight, increased open-field activity, and slowed acquisition of trace fear conditioning. Adolescent alcohol exposure did not disrupt learning or significantly induce gross brain pathology, suggesting that 200 mg/dl/day of ethanol disrupts cognitive development during the 3rd trimester equivalent, but not during adolescence. Interestingly, females exposed to alcohol only during adolescence exhibited an increased conditioned fear response and more rapid habituation of locomotor activity in the open field, suggesting alterations in emotional responding. Moreover, subjects exposed to a combination of neonatal and adolescent alcohol exposure spent significantly more time in the center of the open field chamber than other groups. Similarly, males exposed to the combination exhibited less thigmotaxis in the Morris water maze. These results indicate that combined exposure to alcohol during these two critical periods reduces anxiety-related behaviors and/or increases risk taking in a sex-dependent manner, suggesting that prenatal alcohol exposure may affect risk for emotional consequences of adolescent alcohol exposure.

Cadmium and Mercury Exposure: Oxidative, Neurobehavioural and Histological Alterations to the Cerebellum of Wistar Rats

Background: The unprecedented increase in metal exposure has been aided by modern industrialization and anthropogenic activities. Cadmium and mercury are recognized as two of the most common heavy metals with destructive impacts on most organ systems. The present study was designed to investigate and improve existing literature on the possible deleterious effects of cadmium and mercury exposure.&#x0D; Methods: Adult Wistar rats were treated with cadmium chloride (5 mg/kg/day) and mercury chloride (4 mg/kg/day) for 14 days. Body, brain and cerebellar weights, motor deficits, antioxidant and lipid peroxidation activities as well as histological alterations to the cerebellum were evaluated at the end of the experiment.&#x0D; Results: Findings showed a significant reduction in body and brain weights, dysregulation of antioxidant enzymes activity and impaired locomotion and exploratory activity in treated rats. Also, an increase in lipid peroxidation and degeneration of Purkinje cells of the cerebellum was observed in treated rats.&#x0D; Conclusion: Overall, these results corroborate previous findings that cadmium and mercury induce deleterious effects on the cerebellum and central nervous system. In addition, this study helps to provide an anatomical perspective and information on the exact cerebellar changes induced by cadmium and mercury in Wistar rats.

Exploration of Aqueous Phyllanthus amarus Leaf Extract as a Protective Agent in Mercury Chloride-Exposed Wistar Rats: A Neurobehavioural Study

Mercury (Hg) exposure is known to damage vital components of the central nervous system primarily through oxidative stress. Although Phyllanthus amarus (PA) is reported to be rich in antioxidants, there is a dearth of relevant research evidence to show its protective activity against Mercuric chloride (HgCl2)-induced neurotoxicity. Accordingly, this study was designed to investigate the possible protective activity of aqueous Phyllanthus amarus leaf extract (APALE) in HgCl2-exposed rats using standard neurobehavioral assessments. Thirty adult Wistar rats were randomized into six groups of five rats each viz: A: Control, distilled water; B: HgCl2 (4 mg/kg body wt.); C: APALE (200 mg/kg body wt.) + HgCl2 (4 mg/kg); D: APALE (400 mg/kg body wt.) + HgCl2 (4 mg/kg); E: APALE (200 mg/kg body wt.); F: APALE (400 mg/kg body wt.). All administrations were through an orogastric tube for 14 days. Thereafter, neurobehavioral tests (open field, novel object and Y-maze) were done and experimental rats were sacrificed by cervical dislocation. Body, brain and cerebellar weights were examined across all groups. HgCl2 exposure significantly (p&lt;0.0.05) decreased whole body weight across experimental groups. Similarly, a reduction in brain and cerebellar weight was observed. Significant neurobehavioral alterations to locomotion, discrimination index and cognition/memory, induced by HgCl2, were observed across experimental groups. However, protection against the deleterious effects induced by HgCl2 was observed following pretreatment with APALE. Conclusively, these findings show that APALE demonstrated potent protective activity against HgCl2-induced neurotoxicity and may be useful in the management of Hg toxicity and other related disorders.

Size anomaly and alteration of GABAergic enzymes expressions in cerebellum of a valproic acid mouse model of autism

Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder characterized by social communication deficit and repetitive behaviour. In the past few years, increasing clinical evidence has shown that the cerebellum may contribute to the neuropathology of ASD. However, studies in the mechanism for the involvement of the cerebellum in autism remained speculative. Although some have suggested the possibility of a change of glutamate decarboxylases in the cerebellum of autistic patients, this remains controversial and is limited to the alteration in transcriptional level. This study aimed to investigate the cerebellar structure and determine the expression of rate-limiting GABAergic enzymes in GABA signalling of the autism cerebellum. Pregnant C57BL/6 J mice were intraperitoneally injected with a dosage of 500 mg/kg valproic acid (VPA) on embryonic day 10.5 for autistic behavioural induction. This study found that early prenatal exposure to VPA led to tail deformation and decreased cerebellar weight and size. Early adult mouse models with autistic behaviour showed reduced expression of both isoforms of glutamate decarboxylases (GAD) 65 and 67 in the cerebellum. Also, protein expressions of cerebellar type 1 GABA transporter (GAT-1) and GABA transaminase (GABAT) were reduced in VPA mice. It indicated that abnormal GABA production, recycling, and metabolism could alter the excitatory-inhibitory balance in the autistic cerebellum. Thus, our findings provide supporting evidence that cerebellum impairment could be an etiology of environmentally induced autism. Changes in cerebellar structure and the altered GABAergic enzymes in the cerebellum provide targets for future therapeutic studies in idiopathic autism.

Impact of cisplatin administration on cerebellar cortical structure and locomotor activity of infantile and juvenile albino rats: the role of oxidative stress.

The central neurotoxicity of cisplatin (CisPt) has always raised questions especially during development, but few studies are available. Hence, this work was designed to assess the CisPt's impacts on the postnatal rat cerebellum via evaluation of locomotor activity, histological and immunohistochemical studies, and to focus on cerebellar oxidative stress-related alterations. Eighty newborn pups were divided into 2 equal experimental groups: the control group was kept without any treatment and CisPt-treated group received a single subcutaneous injection of CisPt (5μg /g b.w.) in their nape at PD10. Ten rats at PD11, PD17, and PD30 ages were weighed, then deeply anesthetized and sacrificed. For locomotor assessment, 20 pups were divided equally into control and CisPt-treated groups and tested at PD11-13, PD15-17, and PD28-30 ages. CisPt-treated rats suffered from decreased motor activity and showed decreased body and cerebellar weights, reduced levels of enzymatic antioxidants (SOD and CAT), and non-enzymatic antioxidant defense (GSH), and increase of lipid peroxidation marker (MDA). Histopathologically, CisPt sowed deleterious changes within cerebellar cortical layers in the form of vacuolations, decreased thickness, and hemorrhage (in PD17), while Purkinje cells exhibited profound degenerative changes in the form of swelling, disrupted arrangement, distortion, and nuclear shrinkage. In CisPt-treated rats, GFAP demonstrated upregulated, hypertrophied, and branched Bergmann glial fibers and reactive astrogliosis. Immuno-localization of Ki-67-positive cells revealed defective migration associated with decreased proliferation in early ages in addition to glial proliferation in PD30. In conclusion, CisPt causes oxidative stress-related deleterious effects on structure of developing cerebellar cortex and affects locomotor activity.

Choline supplementation prevents the effects of bilirubin on cerebellar-mediated behavior in choline-restricted Gunn rat pups.

Background:Bilirubin is produced by the breakdown of hemoglobin, and is normally catabolized and excreted. Neurotoxic accumulation of serum bilirubin often occurs in premature infants. The homozygous Gunn rat lacks uridine diphosphate-glucuronosyltransferase-1A1 (UGT1A1), the enzyme needed to biotransform bilirubin. This rodent model of hyperbilirubinemia emulates many aspects of bilirubin toxicity observed in the human infant. We demonstrate that choline supplementation in early postnatal development is neuroprotective in the choline restricted Gunn rat, when hyperbilirubinemia is induced on postnatal day 5.Methods:We first compared behaviors and cerebellar weight of pups born to dams consuming regular rat chow to those of dams consuming choline-restricted diets. Secondly, we measured behaviors and cerebellar weights of pups born to choline-restricted dams, reared on a choline-restricted diet, supplemented with or without choline and treated with or without sulfadimethoxine (SDMX).Results:A choline-restricted diet did not change the behavioral outcomes, but cerebellar weight was reduced in the choline-restricted group regardless of genotype or SDMX administration. SDMX induced behavioral deficits in jj pups, and choline supplementation improved most behavioral effects and cerebellar weight in SDMX-treated jj rats.Conclusion:These results suggest that choline may be used as a safe and effective neuroprotective intervention against hyperbilirubinemia in the choline deficient premature infant.

Perinatal exposure to nonylphenol promotes proliferation of granule cell precursors in offspring cerebellum: Involvement of the activation of Notch2 signaling

Nonylphenol (NP), a widely diffused persistent organic pollutant (POP), has been shown to impair cerebellar development and cause cerebellum-dependent behavioral and motor deficits. The precise proliferation of granule cell precursors (GCPs), the source of granular cells (GCs), is required for normal development of cerebellum. Thus, we established an animal model of perinatal exposure to NP, investigated the effect of NP exposure on the cerebellar GCPs proliferation, and explored the potential mechanism involved. Our results showed that perinatal exposure to NP increased cerebellar weight, area, and internal granular cell layer (IGL) thickness in offspring rats. Perinatal exposure to NP also resulted in the GCPs hyperproliferation in the external granular layer (EGL) of the developing cerebellum, which may underlie the above-mentioned cerebellar alterations. However, our results suggested that perinatal exposure to NP had no effects on the length of GCPs proliferation. Meanwhile, perinatal exposure to NP also increased the activation of Notch2 signaling, the regulator of GCPs proliferation. In conclusion, our results supported the idea that exposure to NP caused the hyperproliferation of GCPs in the developing cerebellum. Furthermore, our study also provided the evidence that the activation of Notch2 signaling may be involved in the GCPs hyperproliferation.

Neurobehavioural changes and morphological study of cerebellar purkinje cells in kaolin induced hydrocephalus.

Cerebellar abnormalities are commonly associated with hydrocephalus. However, the effect of hydrocephalus on the otherwise normal cerebellum has been largely neglected. This study assesses the morphological changes in the Purkinje cells in relation to cerebellar dysfunction observed in juvenile hydrocephalic rats. Fifty-five three-week old albino Wistar rats were used, hydrocephalus was induced by intracisternal injection of kaolin (n = 35) and others served as controls (n = 20). Body weight measurements, hanging wire, negative geotaxis, and open field tests were carried out at the onset and then weekly for 4weeks, rats were killed, and their cerebella processed for Hematoxylin and Eosin, Cresyl violet and Golgi staining. Qualitative and quantitative studies were carried out; quantitative data were analyzed using two-way ANOVA and independent T tests at p < 0.05. Hydrocephalic rats weighed less than controls (p = 0.0247) but their cerebellar weights were comparable. The hydrocephalic rats had a consistently shorter latency to fall in the hanging wire test (F(4,112) = 18.63; p < 0.0001), longer latency to turn in the negative geotaxis test (F(4,112) = 22.2; p < 0.0001), and decreased horizontal (F(4,112) = 4.172, p = 0.0035) and vertical movements (F(4,112) = 4.397; p = 0.0024) in the open field test than controls throughout the 4weeks post-induction. Cellular compression in the granular layer, swelling of Purkinje cells with vacuolations, reduced dendritic arborization and increased number of pyknotic Purkinje cells were observed in hydrocephalic rats. Hydrocephalus caused functional and morphological changes in the cerebellar cortex. Purkinje cell loss, a major pathological feature of hydrocephalus, may be responsible for some of the motor deficits observed in this condition.

Effects of Phosphatidylserine Source of Docosahexaenoic Acid on Cerebellar Development in Preterm Pigs.

Preterm birth, a major contributor to infant mortality and morbidity, impairs development of the cerebellum, the brain region involved in cognitive processing and motor function. Previously, we showed that at term-equivalent age, preterm pigs that received formula supplemented with docosahexaenoic acid (DHA) esterified to phosphatidylserine (PS) had cerebellar weights similar to those of newborn term pigs and were heavier than control preterm pigs. However, whether PS-DHA promotes the development of specific cerebellar cell populations or enhances key developmental processes remains unknown. Here we investigated the effects of the PS-DHA on development of the cerebellum in preterm pigs delivered via caesarean section and reared for ten days on a milk replacer with either PS-DHA (experimental group) or sunflower oil (control group). Upon necropsy, key cerebellar populations were analyzed using immunohistochemistry. Consumption of PS-DHA was associated with the expansion of undifferentiated granule cell precursors and increased proliferation in the external granule cell layer (EGL). Preterm pigs that received PS-DHA also had significantly fewer apoptotic cells in the internal granule cell layer (IGL) that contains differentiated granule neurons. PS-DHA did not affect the number of differentiating granule cells in the inner EGL, thickness of the inner EGL, density of Purkinje cells, or Bergmann glial fibers, or diameter of Purkinje cells. Thus, PS-DHA may support cerebellar development in preterm subjects by enhancing proliferation of granule cells, a process specifically inhibited by preterm birth, and increasing the survival of granule cells in the IGL. These findings suggest that PS-DHA is a promising candidate for clinical studies directed at enhancing brain development.

Behavioral, histopathological, and biochemical evaluations on the effects of cinnamaldehyde, naloxone, and their combination in morphine-induced cerebellar toxicity

Long-term morphine use for therapeutic approaches may lead to serious side effects. Several studies have suggested opioid antagonist and antioxidant therapy for reducing adverse effects of morphine. Cinnamaldehyde has a potent anti-oxidant property. In this study, separate and combined effects of cinnamaldehyde and naloxone (an opioid receptor antagonist) on behavioral changes and cerebellar histological and biochemical outcomes were investigated after long-term morphine administration. Seventy-eight rats were divided into two major morphine-treated and morphine-untreated groups. Morphine-treated group was subdivided into seven subgroups for receiving vehicle, normal saline, cinnamaldehyde (1.25, 5, and 20 mg/kg), naloxone, and cinnamaldehyde plus naloxone before morphine. Morphine-untreated group was subdivided into six subgroups and treated with vehicle, cinnamaldehyde (1.25, 5, and 20 mg/kg), naloxone, and their combination. Chemical compounds were administered for 28 consecutive days. Behavioral tests including footprint, rotarod, and beam balance tests were employed. Histopathological and biochemical alterations of cerebellum were determined. Body and cerebellum weights, stride width, time spent on the rotarod, Purkinje cell number, thickness of molecular and granular layers, superoxide dismutase (SOD), and total antioxidant capacity (TAC) decreased as a result of administrating morphine. Morphine increased beam transverse time, malondealdehyde (MDA), tumor necrosis factor-α (TNF-α), and caspase-3 levels. Histopathological changes such as cellular vacuolation and loss were also produced as a result of treatment with morphine. Cinnamaldehyde, naloxone, and their combination treatments improved all the above-mentioned alterations induced by morphine. We concluded that cinnamaldehyde produced a neuroprotective effect through anti-oxidant, anti-inflammatory, apoptotic, and probably naloxone-sensitive opioid receptor interaction mechanisms.