The developmental expression of calcium-binding protein, calbindin D-28k (CB), during the first 6 months was studied in the canine hippocampus by immunohistochemistry. CB immunoreactivity appeared from on P0 in the dentate granule cells, mossy fibers in CA 3 area, CA 2 and CA 1 pyramidal cells, and interneurons in all regions. According to their morphology and location, these could represent the presumptive pyramidal cells and interneurons. From on P7, the CB immunoreactive pyramidal cells clearly distinguished and started to form two rows in CA 1 area as time progressed, while scattered multipolar neurons gradually decreased. CB immunoreactive cell processes increased in length up to P28. The adult-like distribution of CB immunoreactivity was established about P60. After P60, CB immunoreactivity appeared in dentate granule cells, mossy fibers in CA 3 area and pyramidal cells in CA 1 where formed two rows and CA 2 areas as well as in interneurons of the strata oriens and pyramidale. Taken together, developmental expression of CB in the canine hippocampus was summarized that CB imunoreactivity was observed in all regions on P0 and reached adult-like distribution about P60. These data also suggested the possibility of prenatal expression of CB on the basis of the staining pattern at P0.
Calbindins* ; Hippocampus* ; Immunohistochemistry ; Interneurons ; Neurons ; Pyramidal Cells

To understand the changes in expression of calcium binding proteins(CaBP) during the experimental focal ischemia, expression of two kinds of CaBP, paralvumin(PV) and calbindin D-28K(Calbindin), immunocytochemically, and activities of cytochrome oxidase(CO) and acetylcholinesterase(AchE), histochemically, in focal ischemic brain of the rat were investigated. Two groups of focal ischemic infarction were produced in Sprague Dawley rats(200-350 mg):Group I, Clip compression of left middle cerebral artery(MCA) for 5-10 mins and release;Group II, Electric coagulation of left MCA for 2-24 hrs. In the group I, CO activity and PV- and Calbindin-immunoreactivity(IR) were decreased in the left MCA territory, and decreased in number of PV- and Calbindin-IR neurons and degree of IR, but AchE activity was nearly same as that of control cortex. In the group II, decrease of CO and AchE activities, and marked increase of PV- and Calbindin IRs were noted on neuropil in the layers I through VI of ischemic region. Characteristically pyramidal cells, which did not express the both CaBPs in the control cortex, of layer V of ischemic cortex showed PV- and Calbindin Irs in the cell body and apical dendrite. These findings suggest that 1) PV- and Calbindin-IR neurons, mainly non-pyramidal cells, are more vulnerable than pyramidal cell to ischemic injury, 2) CaBP may have some roles in hypoxic neuronal injury, and 3) PV and Calbindin-immunocytochemistry can be used as useful technique in evaluation of experimental ischemia.
Animals ; Brain ; Calbindins ; Calcium ; Cytochromes ; Dendrites ; Infarction ; Ischemia* ; Neocortex* ; Neurons* ; Neuropil ; Pyramidal Cells ; Rats*

It is suggested that calbindin buffers the concentration of intracellular calcium as the calcium binding protein in the cell. In the neurodegerative disease such as Parkinsonian disease, Huntington 'disease, Alzheimer 'disease there is some change of calbindin. The calcium mediated neurotoxicity begins due to the decrease of calbindin gene in those disease. In this study the substantia nigra of the normal rat is immunostained with anti -calbindin antibody, the morphological characteristics and distribution of calbindin positive neurons are studied to confirm the suggestive neuroprotective role of calbindin in the Parkinsonian disease. In the substantia nigra tissues of rats, calbindin was immunostained in the cell body and cellular processes of the polygonal or ovoid neurons. The calbindin immumostained neurons were distributed mainly in the substantia nigra lateralis than substantia nigra compacta and have even distribution from cephalic section to caudal section. The degree of calbindin -immunostaining was similar from medial area to lateral area, from ventral area to dorsal area in the one section of substantia nigra. These results support the potentiative neuroprotective role of calbindin in the Parkinsonian disease.
Animals ; Buffers ; Calbindins* ; Calcium ; Carrier Proteins ; Immunohistochemistry ; Neurons* ; Rats* ; Substantia Nigra*

This study examined and compared the immunocytochemical distribution of the two calcium-binding proteins calbindin D-28k and parvalbumin in the sensory and motor cortex of the cat. In this experimental animal, calcium-binding protein calbindin D-28k immunoreactive neurons were mainly found many pyramidal cells distributed in layers 2 and 3 of the two cortical areas. Calbindin D-28k neuropil labeling was heaviest in layers 1 to 3. In contrast to parvalbumin, we found only minor differences in distribution, size and mor-phology of calbindin D-28k cell body or neuropil staining in the two cortical areas. Parvalbumin- immunoreactive cells were in all layers cortex except layer 1 and reached their peak density in the middle layers. The two cortical areas differed markedly in number, cell size and morphology of immunoreactive cells. Parvalbumin positive cells were more than twice as numerous in the two cortical areas compared to the calbindin D-28k positive cells. Calbindin D-28k and parvalbumin-immunoreactive somata were round, oval, spindle and polygonal in shape, and the positive neurons were unipolar, bipolar,multipolar and horizontal in shape. The diameters of the somata of the two positive neurons were 15~20 micrometer. Also, two positive dendrites were considerably densely arrayed in arborization.
Animals ; Calbindins* ; Calcium-Binding Proteins ; Cats* ; Cell Count ; Dendrites ; Motor Cortex* ; Neurons ; Neuropil ; Pyramidal Cells

We have studied the distribution of calcium-binding proteins parvalbumin (PV -IR) and calbindin D-28k (CB -IR) immunoacitivity in the mongrel dog olfactory bulb using monoclonal antibodies and the avidin -biotin -immunoperoxidase method. The possible coexistence of both markers was determined by segmental histochemical and immunohistochemical double labelling of the same section. In the olfactory bulb of mongrel dog, PV-IR and CB-IR were mainly located in the external plexiform layer, and a few scattered in the glomerular layer, mitral cell layer, and glanule cell layer in the order of thier existence. In addition, three neuronal types were observed in the glomerular layer and external plexiform layer border. Horizontal cells and vertical cells of Cajal were also observed after both PV-IR and CB-IR labeling. Distict groups of PV-IR and CB-IR, differing in size, shape, dendritic branching pattern, and staining intensity, were distinguished in the all layers of the olfactory bulb. Specific neuronal populations were positive for both PV-IR and CB-IR markes. No cell colocalized both stains in the mongrel dog olfactory bulb. The number of PV-IR were more than abundant in olfactory bulb compared to the CB-IR cells. The PV-IR and CB-IR postitive cell somata were round, oval, spindle and polygonal in shape, and they were unipolar, bipolar and multipolar in type. The diameters of the somata of the PV-IR and CB-IR neurons were 20~40 micro meter, respectively. Also dendrites of the these neurons were densely arrayed in arborization.
Animals ; Antibodies, Monoclonal ; Avidin ; Calbindins* ; Calcium-Binding Proteins ; Coloring Agents ; Dendrites ; Dogs* ; Immunohistochemistry ; Neurons* ; Olfactory Bulb*

We have examined the ontogeny of parvalbumin and calbindin D-28k immunoreactivities in the canine anterior cingulate cortex from the day of birth (P0) through P180. At P7, parvalbumin immunoreactivity appears firstly in layer VI multipolar cells. The parvalbumin immunoreactivity in GABAergic interneurons appears to follow an 'inside-out' gradient of radial mergence and reaches an adult-like pattern by the end of the 6th postnatal month. Immunoreactivity is limited mainly to developing nonpyramidal cells, whereas pyramid-like parvalbumin immunoreactive cells are transiently observed in layer V from the P14 to the P90. The developmental pattern of calbindin D-28k immunoreactivity differs from that of parvalbumin immunoreactivity. Calbindin D-28k immunoreactivity develops firstly in layer V pyramidal cells from P0, which continues through the third postnatal month. Calbindin D-28k immunoreactive interneurons are located in the infragranular layers and white matter at P0 and increase in both the supragranular and infragranular layers by P14. This is followed by an adult-like pattern at the P180. These data suggested that parvalbumin and calbindin D-28k may play a role in protecting immature neurons from intracelluar calcium influx during postnatal development.
Animals ; Calbindins* ; Calcium ; Dogs ; Gyrus Cinguli* ; Immunohistochemistry ; Interneurons ; Neurons ; Parturition ; Pyramidal Cells*

The maturation pattern of the calcium binding proteins parvalbumin (PV) and calbindin-D28K (CB) from the day of birth, postnatal day 0 (P0) to 30 days (P5, P10, P15, P20, P30) and adult was studied in the rat amygdala using immunohistochemistry. PV and CB immunoreactivities in the amygdala of the rat showed very different patterns during postnatal development. The first PV-immunoreactive neurons appeared in the cortical amygdaloid nuclei and the basolateral amygdaloid nucleus at P5, and then in the lateral amygdaloid nucleus and the basomedial amygdaloid nucleus at P10. Adult patterns of PV-immunoreactive neurons were reached at P20. In contrast, CB-immunoreactive neurons were already found at birth in all amygdaloid nuclei except the intercalated nucleus. The intensity and number of immunoreactivity of CB-containing neurons increased during the first 10 days of postnatal life but dramatically decreased at P15. Mature patterns CB-immunoreactive neurons were achieved at P20. These two calcium binding proteins exhibited a non-homogeneous distribution in the adult amygdala, PV-immunoreactive neurons were mainly localized in the basolateral nuclear group but not in the medial amygdaloid nucleus, the cental amygdaloid nucleus and the intercalated nucleus. In contrast, CB-immunoreactive neurons were distributed in almost all amygdaloid nuclei except the intercalated nucleus. The present results showing different postnatal maturation patterns such as time of appearance, the number and distribution of immunoreactive cells suggest that PV and CB may play a different functional role during the postnatal development of the amygdala.
Adult ; Amygdala ; Animals ; Calbindin 1 ; Calbindins* ; Calcium-Binding Proteins ; Humans ; Immunohistochemistry ; Neurons* ; Parturition ; Rats*

Calbindin D(28k),a calcium binding protein,is found in various tissues,including some cells in the distal nephron.It plays an important role in the regulation of calcium reabsorption. We previously reported the expression of calbindin D(28k) in adult rat kidney.However,the exact time of expression during differentiation in the embryonic kidney is not known.During development,intercalated cells are deleted from the medullary collecting duct by two distinct mechanisms.However,the reason for the different modes of cell death is not known.As calbindin is reported to protect cells against apoptosis,we examined the expression of calbindin D(28k) in the developing rat kidney.Kidneys from 16-,17-,18-and 20-day-old fetuses and 1-,3-,5-,7-,14-and 21-day-old pups and adult Sprague awley rats were processed for immunohistochemistry using a monoclonal antibody against calbindin D(28k) .Intercalated cells were identified by immunostaining for H+ -ATPase and by electron microscopy.Calbindin D(28k) immunoreactivity first appeared in subpopulations of cells in the connecting tubule and medullary collecting duct in the 17-day-old fetus.In the connecting tubule,calbindin D(28k) was expressed only in H+ -ATPase negative connecting tubule cells,and there was no labeling of intercalated cells.In the medullary collecting duct,calbindin D(28k) immunostaining was observed in a few cells with apical H+ -ATPase,characteristic of type A intercalated cells.The numbers of calbindin D(28k) -positive type A intercalated cells increased from day 18 of gestation.In contrast,there was little or no calbindin D(28k) immunoreactivity in the type B intercalated cells or principal cells.During the first two weeks after birth,calbindin D(28k) -positive type A intercalated cells were lost from the terminal part of the medullary collecting duct by simple extrusion. After two weeks,calbindin D(28k) immunostaining decreased in the type A intercalated cells throughout the medullary collecting duct.However,the immunoreactivity of calbindin D(28k) in the cortical collecting duct was increased in some of the type A intercalated cells and the adult pattern was observed in 21-day-old pups.Thus,we propose that the different expression of calbindin D(28k) in type A and type B intercalated cells may be responsible-at least partly-for the different modes of cell death demonstrated in these cells during kidney development.
Adult ; Animals ; Calbindins* ; Calcium ; Cell Death ; Fetus ; Humans ; Immunohistochemistry ; Kidney* ; Rats*

This studies were examined and compared the immunohistochemical distribution of the two calcium -binding proteins calbindin D -28K and parvalbumin positive neurons in the brain stem and spinal cord after transection of spinal cord in rats. In this experiment, calbindin D -28K immunoreactive neurons were mainly found in many pyramidal cells distributed in the brain stem and spinal cord of rats. Calbindin D -28K neuropil labeling was strongly noted in brain stem and in spinal all segments of the spinal cord. In contrast to parvalbumin, little differences were found in distribution, size and morphology of calbindin D -28K cell body or neuropil staining in the brain stem and spinal cord. Parvalbumin immunoreactive cells were demonstrated in all lamina of the gray matter of the spinal cord. These immunoreactive cells had the most high density in the layer I and II dorsal horn and several nuclei of the ventral horn of the all segments of the spinal cord. These immunoreactive cells between the brain stem and spinal cord were quite different markedly in number, cell size and morphology The number of parvalbumin positive cells were more than twice in the brain stem and spinal cord compared to the calbindin D -28K positive cells. Calbindin D -28K and parvalbumin -immunoreactive somata were round, oval, spindle and polygonal in shape, and the positive neurons were unipolar, bipolar, multipolar and horizontal in shape. The diameters of the somata of the two positive neurons were 30 ~40 micrometer, respectively. Also dendrites of two positive neurons were densely arrayed in arborization.
Animals ; Brain Stem* ; Brain* ; Calbindins* ; Calcium ; Cell Count ; Dendrites ; Horns ; Neurons* ; Neuropil ; Pyramidal Cells ; Rats* ; Spinal Cord*

This study was examined and compared the immunocytochemical distribution of the two calcium-binding proteins calbindin D-28K and parvalbumin immunreactive neurons in the medulla oblongata and spinal cord after transection of spinal cord in rats. In this experiment, calbindin D-28K immnunoreactive neurons were mainly found in many pyramidal cells distributed medulla oblongata and spina1 cord of rats. Parvalbumin immunoreactive cells were demonstrated in all lamina of the gray matter of the spinal cord. These immunoreactive cells had the most high density in the severa1 nuclei of the ventra1 horn of the all segments of the spina1 cord. Calbindin D-28K neuropil labeling was strongly noted in spina1 all segments of the spinal cord. In contrast parva1bumin immunoreactive, little differences were found in distribution, size and morphology of calbindin D-28K cell body or neuropil staining in the spinal cord. The number of parvalbumin immunoreactive cells were more than twice in the medulla oblongata and spinal cord compared to the calbindin D-28K immunoreactive cells. Calbindin D-28K and parvalbumin-immmoreactive somata were round, ova1, spind1e and polygona1 in shape, and the immunoreactive neurons were unipolar, bipolar, multipolar and horizontal in shape. The diameters of the somata of the two immunoreactive neurons were 40 ~50 micrometer, respectively. Also dendrites of two immunoreactive neurons were densely arrayed in network. These results suggest that CB-IR and PV-IR most high density in the of the VII~X layers in the ventra1 horn of the all segments of the spina1 cord.
Animals ; Calbindins* ; Calcium-Binding Proteins ; Dendrites ; Horns* ; Medulla Oblongata* ; Neurons* ; Neuropil ; Pyramidal Cells ; Rats* ; Spinal Cord Injuries* ; Spinal Cord*