The hippocampus makes new memories and is involved in mental cognition, and the hippocampal dentate gyrus (DG) is critical because neurogenesis, which occurs throughout life, occurs in the DG. We observed the differentiation of neuroblasts into mature neurons (granule cells) in the DG of C57BL/6 mice at various early postnatal (P) ages: P1, P7, P14, and P21 using doublecortin (DCX) immunohistochemistry (IHC) for neuroblasts and calbindin D-28k (CB) IHC for granule cells. DCX-positive cells decreased in the DG with age; however, CB+ cells increased over time. At P1, DCX and CB double-labeled (DCX+CB+) cells were scattered throughout the DG. At P7, DCX+CB+ cells (about 92% of CB+ cells) were seen only in the granule cell layer (GCL) of the dorsal blade. At P14, DCX+CB+ cells (about 66% of CB+ cells) were found in the lower half of the GCL of both blades. In contrast, at P21, about 18% of CB+ cells were DCX+CB+ cells, and they were mainly located only in the subgranular zone of the DG. These results suggest that the developmental pattern of DCX+CB+ cells changes with time in the early postnatal stages.
Animals ; Calcium-Binding Protein, Vitamin D-Dependent ; Cognition ; Dentate Gyrus ; Hippocampus ; Immunohistochemistry ; Mice ; Neurogenesis ; Neurons

OBJECTIVES: We attempted to compartmentalize the periaqueductal gray (PAG) of the rabbit in terms of the different distribution patterns between NADPH-diaphorase (NADPHd)- and calbindin D28K (CB)-positive neurons. METHODS: Immunohistochemistry and immunofluorescent labelling for CB and histochemistry for NADPHd were carried out on coronally-sectioned midbrain slices of the rabbit. RESULTS: NADPHd-positive neurons were selectively localized in the dorsolateral (DL), the middle one-third of the lateral (L), the dorsal half of the ventrolateral (VLd) PAG, and the supraoculomotor cap nucleus (Su3C). Clusters of CB-immunoreactive perikarya marked the dorsal half of DL (DLd), Su3C, the ventral one-third of L, and the ventral half of the ventrolateral (VLv) PAG. Double labelling for NADPHd and CB revealed that two markers labelled different neuronal groups in DLd and Su3C subdivisions. CONCLUSION: The present data suggest that NADPHd and CB can be regarded as reliable neurochemical markers to reveal the longitudinally-columnar organization within the PAG and to subdivide each columnar area.
Calcium-Binding Protein, Vitamin D-Dependent ; Immunohistochemistry ; Mesencephalon ; Neurons ; Periaqueductal Gray

The distributions of calretinin (CR)- and parvalbumin (PV)-immunoreactive neurons in the main olfactory bulb (MOB) of the goat were examined in this study. As in other animals, the goat MOB has a characteristic laminar structure with laminar types and distribution patterns in each layer. CR-immunoreaction was observed in all layers of the MOB, except for the olfactory nerve layer. Most of CR-immunoreactive neurons were observed in the glomerular and granule cell layers. Relatively small number of CR-immunoreactive neurons was detected in other layers. These CR-immunoreactive neurons were interneurons. PV-immunoreaction was detected in all layers. In contrast to CR, olfactory nerve bundles were immunostained with PV. Most of PV-immunoreactive neurons were distributed in the glomerular and granule cell layers. PV-immunoreactive neurons were interneurons. This result suggests that CR and PV may play important roles in the olfactory signal modulation through interneurons in the goat MOB.
Animals ; Calcium-Binding Protein, Vitamin D-Dependent ; Calcium-Binding Proteins ; Goats ; Immunohistochemistry ; Interneurons ; Neurons ; Olfactory Bulb ; Olfactory Nerve ; Smell

Calbindin D-28K (CALB) is one of the calcium-binding proteins which is assumed to be buffering, transport of Ca2+, and regulation of various enzyme systems. In the spinal cord, a subpopulation of calbindin-immunoreactive neurons located in the ventral portion of lamina VII, medial to the motoneuron column, has recently been proposed to be Renshaw cells (RCs), that mediate recurrent inhibition of spinal alpha-motoneurons, based on the anatomical location. In this study, we have performed to investigate the correlation between RCs containing high levels of CALB and motoneurons in the ventral horn of lumbar spinal cord of the ataxic pogo mice, that characterized by a failures of interlimb coordination, and prolonged excessive tone of hindlimb extensor muscles. We have shown that CALB immunoreactive RCs was significantly decreased in the ventral horn of lumbar spinal cord of the ataxic pogo mice (p.0.05), when compared with the control mice. Whereas, CALB immunoreactivity expression levels were no difference in the dorsal horn. Furthermore, CALB protein was significantly decreased in the lumbar spinal cord of the ataxic pogo mice (p.0.01). However, there were no difference in the cervical and thoracic spinal cord of the between control and pogo mice. These results suggest that motoneurons of ventral horn of the lumbar spinal cord might be more excited state, results in the decreased CALB immunoreactive RCs have not mediated a motoneuron excitability, in the atxic mice, pogo.
Animals ; Calcium-Binding Protein, Vitamin D-Dependent ; Calcium-Binding Proteins ; Hindlimb ; Horns ; Mice ; Muscles ; Neurons ; Spinal Cord

Alzheimer's disease (AD) is a neurodegenerative disease that proceeds with the age-dependent neuronal loss, an irreversible event which causes severe cognitive and psychiatric devastations. In the present study, we investigated whether the compound, AAD-2004 [2-hydroxy-5-[2-(4-trifluoromethylphenyl)-ethylaminobenzoic acid] which has anti-oxidant and anti-inflammatory properties, is beneficial for the brain of Tg-betaCTF99/B6 mice, a murine AD model that was recently developed to display age-dependent neuronal loss and neuritic atrophy in the brain. Administration of AAD-2004 in Tg-betaCTF99/B6 mice from 10 months to 18 months of age completely repressed the accumulation of lipid peroxidation in the brain. AAD-2004 markedly suppressed neuronal loss and neuritic atrophy, and partially reversed depleted expression of calbindin in the brain of Tg-beta-CTF99/B6. These results suggest that AAD-2004 affords neurodegeneration in the brain of AD mouse model.
Alzheimer Disease ; Animals ; Aspirin ; Atrophy ; Brain ; Calcium-Binding Protein, Vitamin D-Dependent ; Lipid Peroxidation ; Mice ; Neurodegenerative Diseases ; Neurons

BACKGROUND: It is important to differentiate between schwannomas and neurofibromas for the cases in which the histopathologic features overlap. Depending on the tumor type, surgeons can decide on a treatment method and whether to preserve or sacrifice the nerve; the possibility of malignant transformation in the case of neurofibromas also needs to be considered. METHODS: We studied 101 cases of schwannoma and 103 cases of neurofibroma. All the hematoxylin and eosin slides for these cases were reviewed, and tissue microarrays were prepared from the representative areas. Immunohistochemical analysis was performed using antibodies for S-100 protein, calretinin, CD56 and CD34. RESULTS: All the tumors except 3 neurofibromas were positive for the S-100 protein. Calretinin was found in 26.7% of the schwannomas (27/101), but it was not found in any of the neurofibromas. CD56 was positive in 77.2% of the schwannomas (78/101) and in 9.8% of the neurofibromas (10/102). CD34 was positive in 42.5% of the schwannomas (43/101) and in 80.2% of the neurofibromas (81/101). Statistically, calretinin was significantly specific for schwannomas (p<0.001) and CD56 was also sensitive for these tumors (p<0.001). On the other hand, a CD34 expression seemed highly sensitive (p<0.001) for neurofibromas. CONCLUSIONS: We concluded that combined immunohistochemical analysis for calretinin, CD56, and CD34 may be very useful for differentiating schwannomas from neurofibromas.
Antibodies ; Calcium-Binding Protein, Vitamin D-Dependent ; Diagnosis, Differential ; Eosine Yellowish-(YS) ; Hand ; Hematoxylin ; Immunohistochemistry ; Neurilemmoma ; Neurofibroma ; S100 Proteins

Cerebellar Purkinje cells (PCs) play a crucial role in motor functions and their progressive degeneration is closely associated with spinocerebellar ataxias. Although immunohistochemical (IHC) analysis can provide a valuable tool for understanding the pathophysiology of PC disorders, the method validation of IHC analysis with cerebellar tissue specimens is unclear. Here we present an optimized and validated IHC method using antibodies to calbindin D28k, a specific PC marker in the cerebellum. To achieve the desired sensitivity, specificity, and reproducibility, we modified IHC analysis procedures for cerebellar tissues. We found that the sensitivity of staining varies depending on the commercial source of primary antibody. In addition, we showed that a biotin-free signal amplification method using a horseradish peroxidase polymer-conjugated secondary antibody increases both the sensitivity and specificity of ICH analysis. Furthermore, we demonstrated that dye filtration using a 0.22 micrometer filter eliminates or minimizes nonspecific staining while preserving the analytical sensitivity. These results suggest that our protocol can be adapted for future investigations aiming to understand the pathophysiology of cerebellar PC disorders and to evaluate the efficacy of therapeutic strategies for treating these diseases.
Antibodies ; Calcium-Binding Protein, Vitamin D-Dependent ; Cerebellum ; Filtration ; Horseradish Peroxidase ; Purkinje Cells ; Sensitivity and Specificity ; Spinocerebellar Ataxias

Nitric oxide (NO) modulates the activities of various channels and receptors to participate in the regulation of neuronal intracellular Ca2+ levels. Ca2+ binding protein (CaBP) expression may also be altered by NO. Accordingly, we examined expression changes in calbindin-D28k, calretinin, and parvalbumin in the cerebral cortex and hippocampal region of neuronal NO synthase knockout(-/-) (nNOS-/-) mice using immunohistochemistry. For the first time, we demonstrate that the expression of CaBPs is specifically altered in the cerebral cortex and hippocampal region of nNOS-/- mice and that their expression changed according to neuronal type. As changes in CaBP expression can influence temporal and spatial intracellular Ca2+ levels, it appears that NO may be involved in various functions, such as modulating neuronal Ca2+ homeostasis, regulating synaptic transmission, and neuroprotection, by influencing the expression of CaBPs. Therefore, these results suggest another mechanism by which NO participates in the regulation of neuronal Ca2+ homeostasis. However, the exact mechanisms of this regulation and its functional significance require further investigation.
Animals ; Calcium ; Calcium-Binding Protein, Vitamin D-Dependent ; Calcium-Binding Proteins ; Carrier Proteins ; Cerebral Cortex ; Homeostasis ; Immunohistochemistry ; Mice ; Neurons ; Nitric Oxide ; Nitric Oxide Synthase ; Synaptic Transmission

Nitric Oxide (NO) actively participates in the regulation of neuronal intracellular Ca2+ levels by modulating the activity of various channels and receptors. To test the possibility that modulation of Ca2+ buffer protein expression level by NO participates in this regulatory effect, we examined expression of calbindin-D28k, calretinin, and parvalbumin in the cerebellum of neuronal NO synthase knock-out (nNOS(-/-)) mice using immunohistochemistry. We observed that in the cerebellar cortex of the nNOS(-/-) mice, expression of calbindin-D28k and parvalbumin were significantly increased while expression of calretinin was significantly decreased. These results suggest another mechanism by which NO can participate in the regulation of Ca2+ homeostasis.
Animals ; Calcium ; Calcium-Binding Protein, Vitamin D-Dependent ; Calcium-Binding Proteins ; Cerebellar Cortex ; Cerebellum ; Homeostasis ; Immunohistochemistry ; Mice ; Neurons ; Nitric Oxide ; Nitric Oxide Synthase

To understand the neurochemical properties of the gastric myenteric plexus of ruminants, the expression patterns of calbindin D-28k (CB), calretinin (CR), substance P (SP) and calcitonin gene-related peptide (CGRP) were explored in the Korean native goat. In gastric myenteric plexus, CB and SP immunoreactivity were observed in round- or ovalshaped neurons. CR and CGRP immunoreactivity were detected only in the nerve fibers. This immunohistochemical localization of CB, CR, CGRP and SP in the myenteric plexus of the goat stomach exhibited species-specific patterns. These findings suggest that these substances may be directly or indirectly related to the gastric functions of the goat stomach.
Animals ; Calcitonin Gene-Related Peptide ; Calcium-Binding Protein, Vitamin D-Dependent/metabolism ; Calcium-Binding Proteins/*metabolism ; Goats/*metabolism ; Immunohistochemistry/veterinary ; Myenteric Plexus/*metabolism ; Stomach/*innervation ; Substance P/metabolism