scholarly journals IMMUNOHISTOCHEMICAL STUDY OF CALBINDIN IN THE DEVELOPING CEREBELLUM OF THE RAT

2020 ◽  
Vol 18 (6) ◽  
pp. 692-697
Author(s):  
O. A. Karnyushko ◽  
◽  
S. M. Zimatkin ◽  
Keyword(s):  
Purkinje Cells ◽  
Calcium Binding ◽  
Age Groups ◽  
Physiological Role ◽  
Postnatal Period ◽  
Nerve Fibers ◽  
Adult Rats ◽  
White Rats ◽  
Calbindin D28k ◽  
Developing Cerebellum

Background. Calbindin is a calcium-binding protein that supports calcium homeostasis for the normal functioning of neurons. Purpose. To study the distribution of immunoreactivity of calbindin-D28K in the structures of the developing cerebellum of the rat.Material and methods. The study was performed on 16 outbred white rats of different age groups: 2-, 7-, 15-days (early postnatal period), 45-days (puberty period). The cerebellum samples were taken and fixed in zinc-ethanol-formaldehyde for immunohistochemistry. Calbindin-D28K immunoreactivity was determined on paraffin sections using primary polyclonal rabbit antibodies.Results. In the cerebellar cortex, calbindin immunoreactivity was detected on the 2nd day after development of Purkinje cells (PC) in their perikaryons, and by the 15th day in their dendrites and it did not change by the 45th day. In all terms of the study in PC, it was detected not only in the cytoplasm, but also in their nucleus. In the granular layer, calbindin immunoreactivity decreased in rats in postnatal ontogenesis, however, in adult rats, some neurons were moderately immunopositive. Among them, from the 15th day after birth, the calbindin-immunoreactive afferent nerve fibers running in the white matter were detected. There were no significant differences in the distribution of calbindin between the lobes of the cerebellum of different phylogenetic age. Conclusions. Considering that the expression of сalbindin-D28k is detected throughout the entire period of development of Purkinje cells, as well as its physiological role in maintaining the function and homeostasis of calcium in them, it can be concluded that сalbindin-D28k is a valuable marker for the morphofunctional characteristics of PC in the developing and adult cerebellum of rats in normal and pathological conditions.

scholarly journals Effects of Ascorbic Acid on Osteopontin Expression and Axonal Myelination in the Developing Cerebellum of Lead-Exposed Rat Pups

2019 ◽  
Vol 16 (6) ◽  
pp. 983 ◽  
Author(s):  
Sung Nam ◽  
Jin Seo ◽  
Sang-Soep Nahm ◽  
Byung-Joon Chang
Keyword(s):  
Ascorbic Acid ◽  
Purkinje Cells ◽  
Calcium Binding ◽  
Acid Treatment ◽  
Neuronal Development ◽  
Wire Mesh ◽  
Female Rats ◽  
Functional Protein ◽  
Nissl Staining ◽  
Developing Cerebellum

Osteopontin (OPN) is a multi-functional protein that binds to integrin and calcium-binding phosphoprotein. OPN is required for normal neuronal development and its axonal myelination. We studied the combined effect of lead (Pb) and ascorbic acid treatment on OPN expression in the developing cerebellum. We randomly divided pregnant female rats into three groups: control, Pb (lead acetate, 0.3%, drinking water), and Pb plus ascorbic acid (PA; ascorbic acid, 100 mg/kg, oral intubation) groups. The blood level of Pb was significantly increased, while ascorbic acid reduced Pb levels in the dams and pups. At postnatal day (PND) 21, results from Nissl staining and OPN immunohistochemistry demonstrated that OPN was detected in the Purkinje cell layer in the cerebellum. Ascorbic acid treatment mitigated Pb exposure-induced reduction in the number of intact Purkinje cells and OPN immunoreactive Purkinje cells in the cerebellum of pups. In addition, Pb-induced reduction in the number of oligodendrocytes and myelin-associated glycoprotein is associated with the malformation of the myelin sheath. Ascorbic acid provided protection from Pb-induced impairments. Pb-induced structural deficits in the cerebellum resulted in functional deterioration observed during locomotive tests (bar holding test and wire mesh ascending test), while ascorbic acid ameliorated these harmful effects. Present results suggest that the change of OPN is associated with myelination in the developing cerebellum. The results also demonstrated that exposure to Pb is harmful, while ascorbic acid treatment is beneficial.

scholarly journals Early Subset of Cerebellar Nuclei Neurons Derived from Mesencephalon in Mice

2017 ◽  
Author(s):  
Maryam Rahimi-Balaei ◽  
Xiaodan Jiao ◽  
Fiona E. Parkinson ◽  
Behzad Yeganeh ◽  
Hassan Marzban
Keyword(s):  
Neural Crest ◽  
Purkinje Cells ◽  
Ventricular Zone ◽  
Cerebellar Nuclei ◽  
Nerve Fibers ◽  
Neurotrophin Receptor ◽  
Cerebellar Development ◽  
Developing Cerebellum ◽  
Rhombic Lip ◽  
Migratory Pathway

ABSTRACTDuring cerebellar development, cerebellar nuclei (CN) neurons and Purkinje cells are the earliest born among the different neuronal subtypes. Purkinje cells are the sole output of the cerebellar cortex and project to the CN. The CN represents the main output of the cerebellum, which is generated from the rhombic lip and the ventricular zone. We used immunohistochemistry, embryonic cultures, dye tracers and in situ hybridization to examine the origin of a new subset of CN neurons from the mesencephalon during early cerebellar development. Our results show that a subset of CN neurons, which are immunopositive for α-synuclein (SNCA) and Otx2, originate from the mesencephalon and cross the isthmus toward the rostral end of the nuclear transitory zone. Double immunostaining of the SNCA with Otx2 or p75 neurotrophin receptor (p75ntr) indicates that these cells are derived from neural crest cells. We also showed that this population of neurons with nerve fibers terminates at the subpial surface of putative lobules VI/VII. The SNCA+/Otx2+/p75+ cells, which divide the cerebellar primordium into rosterodorsal and caudoventral compartments, show increased cleaved caspase-3 activation, which suggests temporary presence of these cells due to apoptosis. These results strongly suggest that early CN neurons originate from the mesencephalic neural crest population and cross the isthmus to contribute as a subset of the CN. Their temporary presence in the nuclear transitory zone suggests that these neurons/fibers play a regulatory role as a signaling center to attract early afferent pioneer axons and provide neuronal migratory pathway during early cerebellar development.Significance StatementDuring cerebellar development two germinal zones are involved in cerebellar neurogenesis: the rhombic lip and the ventricular zone, which are located in the developing cerebellum itself. Our findings indicate that a subset of cerebellar nuclei neurons have an external origin, the mesencephalon, and they are the earliest born neurons that enter to the developing cerebellum. In this study, we focused on the origin of these cells and traced their migratory pathway from the mesencephalon while crossing the isthmus, followed them when they entered to the developing cerebellum. We also demonstrated their potential role on later born cells during cerebellar development.

scholarly journals Loss of dysferlin or myoferlin results in differential defects in excitation–contraction coupling in mouse skeletal muscle

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
David Y. Barefield ◽  
Jordan J. Sell ◽  
Ibrahim Tahtah ◽  
Samuel D. Kearns ◽  
Elizabeth M. McNally ◽  
...  
Keyword(s):  
Ryanodine Receptor ◽  
Calcium Binding ◽  
Physiological Role ◽  
Muscle Disease ◽  
Calcium Handling ◽  
Membrane Repair ◽  
Tubule Formation ◽  
Ec Coupling ◽  
Associated Proteins ◽  
Receptor Clusters

AbstractMuscular dystrophies are disorders characterized by progressive muscle loss and weakness that are both genotypically and phenotypically heterogenous. Progression of muscle disease arises from impaired regeneration, plasma membrane instability, defective membrane repair, and calcium mishandling. The ferlin protein family, including dysferlin and myoferlin, are calcium-binding, membrane-associated proteins that regulate membrane fusion, trafficking, and tubule formation. Mice lacking dysferlin (Dysf), myoferlin (Myof), and both dysferlin and myoferlin (Fer) on an isogenic inbred 129 background were previously demonstrated that loss of both dysferlin and myoferlin resulted in more severe muscle disease than loss of either gene alone. Furthermore, Fer mice had disordered triad organization with visibly malformed transverse tubules and sarcoplasmic reticulum, suggesting distinct roles of dysferlin and myoferlin. To assess the physiological role of disorganized triads, we now assessed excitation contraction (EC) coupling in these models. We identified differential abnormalities in EC coupling and ryanodine receptor disruption in flexor digitorum brevis myofibers isolated from ferlin mutant mice. We found that loss of dysferlin alone preserved sensitivity for EC coupling and was associated with larger ryanodine receptor clusters compared to wildtype myofibers. Loss of myoferlin alone or together with a loss of dysferlin reduced sensitivity for EC coupling, and produced disorganized and smaller ryanodine receptor cluster size compared to wildtype myofibers. These data reveal impaired EC coupling in Myof and Fer myofibers and slightly potentiated EC coupling in Dysf myofibers. Despite high homology, dysferlin and myoferlin have differential roles in regulating sarcotubular formation and maintenance resulting in unique impairments in calcium handling properties.

scholarly journals Immunohistochemical localization of calbindin-D28k in the kidney and cerebellum of the porcupines (Hystrix cristata)

2012 ◽  
Vol 48 (No. 12) ◽  
pp. 369-372
Author(s):  
S. Timurkaan ◽  
A. Aydin ◽  
M. Kaban
Keyword(s):  
Purkinje Cells ◽  
Immunohistochemical Localization ◽  
Regulatory Mechanisms ◽  
Distal Nephron ◽  
Intracellular Protein ◽  
High Affinity ◽  
Hystrix Cristata ◽  
Calbindin D28k ◽  
Convoluted Tubules

The localization of calbindin in the kidney and cerebellum of Hystrix cristata was investigated immu­nohistochemically using an antiserum against the 28k Da calbindin of chicken duodenum. Calbindin-D28k is an intracellular protein with a high affinity for calcium. This protein is exclusively localized in the distal convoluted tubules of the kidney and in Purkinje cells of the cerebellum. Functionaly, calbindin-D28k is supposed to be involved in the regulation of the reabsorption of calcium in the distal nephron, though the exact regulatory mechanisms are not yet known.

Development of sensitivity to cAMP-inducing hormones in the rat stomach

1984 ◽  
Vol 247 (3) ◽  
pp. G231-G239
Author(s):  
C. Gespach ◽  
Y. Cherel ◽  
G. Rosselin
Keyword(s):  
Biological Activities ◽  
Physiological Role ◽  
H2 Receptor Antagonist ◽  
Adult Rats ◽  
Gastric Glands ◽  
Camp Generation ◽  
Final Maturation ◽  
Noncompetitive Inhibitor ◽  
Regulatory Properties

Development of cAMP responses to secretin, pancreatic glucagon, and histamine was measured in gastric glands of fetal (day 20), postnatal (days 1-30), and adult rats (day 65). cAMP stimulation by these hormones was already detected on day 20 of gestation. cAMP generation showed biphasic variations during the 1st days of life and at the onset of weaning (day 20). Anticipated weaning at day 14 triggered precocious maturation (efficacies) of the cAMP-generating systems sensitive to secretin, glucagon, and histamine without changing the potencies of the hormones. During development, the general characteristics (potency and pharmacological or regulatory properties) of the receptor-cAMP systems studied were comparable with those evidenced in adult rats. At days 5, 20, and 65, vasoactive intestinal peptide and the peptide having N-terminal histidine and C-terminal isoleucine amide (PHI) were about 100 times less potent than secretin (EC50 = 1.5 X 10(-9) M secretin). The histamine action could be blocked by the competitive H2-receptor antagonist cimetidine (70-100% inhibition) as well as by the noncompetitive inhibitor somatostatin (37-62% inhibition). The data indicate that these regulatory hormones (secretin, glucagon(s), histamine, and somatostatin) might have a direct effect on gastric glands and may modulate their biological activities (metabolism, differentiation, proliferation, and exocrine and endocrine secretions) from the neonatal period in rats. The important physiological role of weaning on the final maturation of the cAMP-generating systems in rat gastric glands is underlined.

Functional and molecular characterization of NHE3 expression during ontogeny in rat jejunal epithelium

1997 ◽  
Vol 273 (6) ◽  
pp. C1937-C1946 ◽  
Author(s):  
James F. Collins ◽  
Hua Xu ◽  
Pawel R. Kiela ◽  
Jiamin Zeng ◽  
Fayez K. Ghishan
Keyword(s):  
Northern Blot ◽  
Polyclonal Antibodies ◽  
Membrane Vesicle ◽  
Age Groups ◽  
Fold Increase ◽  
Jejunal Mucosa ◽  
Mrna Levels ◽  
Adult Rats ◽  
Intestinal Brush Border Membrane ◽  
Protein Levels

Ontogenic changes occur in intestinal brush-border membrane vesicle (BBMV) Na+/H+exchange activity. The present studies were designed to investigate ontogenic changes in Na+/H+exchanger (NHE) isoform 3 in rat jejunum. pH-dependent Na+ uptake was assayed in four age groups of rats in the presence of 0, 50, or 800 μM HOE-694, a specific NHE inhibitor with differential sensitivities for NHE2 [inhibition constant ( K i) = 5 μM in PS120 fibroblasts] and NHE3 ( K i = 650 μM). Results showed that NHE2 and NHE3 contribute to basal BBMV uptake at all ages. Uptake levels were highest in 6-wk-old rats, lower in adult rats, and lowest in 2-wk-old (suckling) and 3-wk-old (weanling) rats. NHE3 contribution ranged from 92% at 6 wk of age to 59% at 2 and 3 wk of age. NHE3 inhibition by 800 μM HOE-694 was 38–45%. Statistical analysis showed that HOE-694 had a significant effect at both concentrations at all ages and that differences were present between all ages except 2- and 3-wk rats (at all HOE-694 concentrations). Northern blot analyses of jejunal mucosa showed lowest NHE3 mRNA levels in 2-wk animals and higher levels in all other age groups. Polyclonal antibodies were developed against an NHE3 COOH-terminal fusion protein, and antiserum was characterized with NHE3-transfected PS120 cells and by immunohistochemistry. Western blot analyses showed lowest protein levels in 2-wk animals and higher levels in the other ages. Suckling rats were subcutaneously injected with methylprednisone (MP) for 2 days and killed 1 day later. Northern blot analyses showed a twofold increase in NHE3 mRNA expression with MP treatment. Immunoblot analyses showed a 2.5-fold increase in NHE3 immunoreactive protein levels with MP injection. Overall, these data suggest that NHE3 is regulated during ontogeny and that ontogenic changes are most apparent around the time of weaning. Furthermore, the data suggest that NHE3 is regulated at transcriptional and posttranscriptional levels during mammalian intestinal development.

Dissimilar Anxiety-like Behavior in Prepubertal and Young Adult Female Rats on Acute Exposure to Aluminium

2021 ◽  
Vol 22 ◽  
Author(s):  
Trina Sengupta ◽  
Sutirtha Ghosh ◽  
Archana Gaur T. ◽  
Prasunpriya Nayak
Keyword(s):  
Body Weight ◽  
Young Adult ◽  
Adult Female ◽  
Developmental Stages ◽  
Elevated Plus Maze ◽  
Age Groups ◽  
Female Rats ◽  
Acute Exposure ◽  
Adult Rats ◽  
Plus Maze

Background: Puberty is a developmental transition in which an estrogenic surge occurs, mediating the release of xenoestrogens, like aluminium. Aluminium’s effect on anxiety in rodents at the different developmental stages is inconsistent. Aims: This study aimed at investigating the effect of the metalloestrogenic property of aluminium on anxiety-like behavioral changes in prepubertal and young adult female rats. Objective: Considering this aim, our objective was to evaluate the anxiety-like behavior by the elevated plus maze in prepubertal and young adult female rats with or without acute exposure to aluminium. Methods: To address this property of aluminium, 5mg/Kg body weight (Al-5) and 10 mg/Kg body weight (Al-10) of aluminium was administered intraperitoneally to female rats at two developmental stages, prepubertal (PP; n = 8 for each dose) and young adult (YA; n = 6 for each dose) for two weeks. Post-treatment, three days behavioral assessment of the rats was done employing elevated plus maze. Results: Reduced escape latency was seen in Al-5, Al-10 pre-pubertal rats, and Al-5 young-adult rats on day 3. A significant reduction in open arm time was seen in the Al-5 young-adult rats. Aluminium treatment in the pre-pubertal rats reduced their head dipping and grooming. Reduced sniffing, head dipping, and stretch-attended posture in the treated young-adult female rats showed that they had impaired risk-taking tendency. Conclusion: Differential effect on the anxiety-like behavior in the pre-pubertal and young-adult female rats might be due to the metalloestrogenic property of aluminium, acting differently on the two age groups.

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