Calbindin-D28K has been implicated in the regulation of neuronal cell death. Previously, we demonstrated that calbindin-D28K prevents staurosporine (STS)-induced caspase activation and subsequent apoptosis in a neuronal cell line. However, the role of calbindin-D28K in STS-induced activation of calpain and necrotic cell death was not identified. Staurosporine induced the elevation of intracellular calcium after 1 hr of treatment. Overexpression of calbindin-D28K and presence of a calcium chelator, BAPTA, prevented the increase of calcium in STS-treated cells. Cleavage of Bax by calpain was prevented by the overexpressed calbindin-D28K. Permeabilization of the plasma membrane, a factor in necrosis, as well as apoptotic change of the nucleolus induced by STS, was prevented by calbindin-D28K. Thus, our study suggests that calbindin-D28K may exert its protective functions by preventing calpain activation in necrotic cell death, in addition to its effect on the caspase-apoptosis pathway.
Apoptosis ; Calbindin 1* ; Calcium ; Calpain ; Cell Death ; Cell Line ; Cell Membrane ; Membranes* ; Necrosis ; Neurons ; Staurosporine

The present study was aimed at investigating the expression of calbindin-D28k (CaBP-D28k) in human fallopian tube, which were collected from 33 childbearing age women undergoing abdominal hysterectomy with adnexectomy for benign disease in the pelvic cavity. These women had normal menstrual cycle and history of normal pregnancy. Isthmus, ampullary and umbrella segments of fallopian tubes were respectively collected. These specimens were divided into 6 groups based on their menstrual cycles: early-proliferative stage (n=6), mid-proliferative stage (n=5), late-proliferative stage (n=5), early-secretory stage (n=7), mid-secretory stage (n=5) and late-secretory stage (n=5). The expressions of CaBP-D28k protein and mRNA in fallopian tubes were determined by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) methods. Positive expressions of CaBP-D28k protein and mRNA were observed in human fallopian tubes. There was no significant difference in the expression of CaBP-D28k protein among the isthmus, ampulla and umbrella segments in the same phase of menstrual cycle (P>0.05). However, in the menstrual cycle, the expression level of CaBP-D28k protein in the epithelium was the lowest during the early- and mid-proliferative stages and increased in both the late-proliferative and early-secretory stages (P<0.05), and then decreased in the mid- and late-secretory stages (P<0.05). The expressed CaBP-D28k protein was disposed to gobbets or dispersed sheets in cytoplasm in the early- and mid- proliferative stages, and showed concentrated granules on the top of cells in the late-proliferative and early-, mid-secretory stages. Then in the late-secretory stage redistribution renewed as in the early- and mid-proliferative stages. The CaBP-D28k mRNA obviously increased in the late-proliferative and early-secretory stages (P<0.05). These findings indicate that the expressions of CaBP-D28k protein and mRNA exist in human fallopian tubes and exhibit a cyclic change.
Calbindin 1 ; metabolism ; Fallopian Tubes ; metabolism ; Female ; Gene Expression ; Humans ; Menstrual Cycle

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*

Voluntary running is known to dramatically increase the cell proliferation and neurogenesis in the dentate gyrus of the adult mouse hippocampus. However, it is crucial to realize that adding excitatory neurons could result in serious maladaptive outcomes for hippocampal circuit function. To investigate the response of mature granule cells on the increase of cell proliferation during voluntary running, we investigated the temporal change of calbindin-D28k (a marker for mature granule cells) using immunohistochemistry during voluntary running with upregulated neurogenesis. By using immunohistochemsitry for Ki-67 and doublecortin (DCX), we observed that the cell proliferation and differentiation of granule cells increased at 1 week of voluntary running. We found that, at 6 weeks of voluntary running, the cell proliferation and differentiation of granule cells returned to sedentary control levels. On the other hand, calbindin-D28k immunoreactivity decreased in the granular cell layer of the dentate gyrus and CA3 region of hippocampus after 1 week of voluntary running. At 6 weeks of voluntary running, the density of the calbindin-D28k in the granular cell layer and CA3 region was returned to the sedentary control level. These results demonstrate that the cell proliferation and differentiation are increased at early point of voluntary running, and the granule cell activity in the dentate gyrus is temporally changed for response to the increase of cell proliferation and differentiation during voluntary running.
Adult ; Animals ; Calbindin 1* ; Cell Proliferation ; Dentate Gyrus* ; Hand ; Hippocampus ; Humans ; Immunohistochemistry ; Mice* ; Neurogenesis ; Neurons ; Running*

Calbindin-D28k, a vitamin D-dependent calcium binding protein, plays a cardinal role in transport of calcium in kidney. Previous studies have demonstrated calbindin-D28k immunoreactivity in the distal nephron of mammalian kidney. However, it is well known that in most species including rat and human, there is a gradual transition from the distal convoluted tubule to the cortical collecting duct, and that the connecting segment do no tclearly demarcated, because of intermingling of distal convoluted tubule cells, connecting tubule cells, principal cells and at least two configurations of intercalated cells. In this study, to identify the cell types of calbindin-D28k-positive cells in distal nephron of rat kidney, we used double immunostaining with an antibody against calbindin-D28k and antibodies against thiazide sensitive Na+/Cl- cotransporter for distal convoluted tubule or H+-ATPase for intercalated cells. In the distal convoluted tubule, most of the distal convoluted tuble cells were calbindin-D28k-positive, whereas the intercalated cells were calbindin-D28k-negative. In the connecting tubule, 68% of the cells were calbindin-D28k-positive, and about 97% of the positive cells were connecting tubule cells and only 3% of them were intercalated cells. In the cortical collecting duct, and outer medullary collecting duct of outer stripe and inner stripe, only 8.6%, 11.8% and 4.4% of cells were weak positive for calbindin-D28k respectively. These weak positive cells in the collecting duct are mainly identified as intercalated cells. These findings indicate that calbindin-D28k is involved in not only transcellular transport of calcium but also processes regulating intracellular calcium in rat kidney.
Animals ; Antibodies ; Calbindin 1 ; Calcium* ; Carrier Proteins* ; Humans ; Immunohistochemistry ; Kidney* ; Nephrons* ; Rats* ; Transcytosis ; Vitamins

The circling mice with tmie gene mutation are known as an animal deafness model, which showed hyperactive circling movement. Recently, the reinvestigation of circling mouse was performed to check the inner ear pathology as a main lesion of early hearing loss. In this trial, the inner ear organs were not so damaged to cause the hearing deficit of circling (cir/cir) mouse at 18 postnatal day (P18) though auditory brainstem response data indicated hearing loss of cir/cir mice at P18. Thus, another mechanism may be correlated with the early hearing loss of cir/cir mice at P18. Hearing loss in the early life can disrupt the ascending and descending information to inferior colliculus (IC) as integration site. There were many reports that hearing loss could result in the changes in Ca²⁺ concentration by either cochlear ablation or genetic defect. However, little was known to be reported about the correlation between the pathology of IC and Ca²⁺ changes in circling mice. Therefore, the present study investigated the distribution of calcium-binding proteins (CaBPs), calbindin-D28k, parvalbumin, and calretinin immunoreactivity (IR) in the IC to compare among wild-type (+/+), heterozygous (+/cir), and homozygous (cir/cir) mice by immunohistochemistry. The decreases of CaBPs IR in cir/cir were statistically significant in the neurons as well as neuropil of IC. Thus, this study proposed overall distributional alteration of CaBPs IR in the IC caused by early hearing defect and might be helpful to elucidate the pathology of central auditory disorder related with Ca²⁺ metabolism.
Animals ; Calbindin 1* ; Calbindin 2* ; Calcium-Binding Proteins ; Deafness ; Ear, Inner ; Evoked Potentials, Auditory, Brain Stem ; Hearing ; Hearing Loss ; Immunohistochemistry ; Inferior Colliculi* ; Metabolism ; Mice* ; Neurons ; Neuropil ; Parvalbumins ; Pathology

It has been reported that new apical anion exchanger perndrin, encoded by the pendred syndrome (PDS/pds, Slc26A4) gene, was expressed in the AE1-negative intercalated cells of rat and mouse kidneys. The purpose of this study was performed that expression of pendrin in the subtypes of intercalated cells in human kidney. The normal human renal tissues obtained from nephrotomized kidneys for renal cell carcinoma were fixed in periodate-lysine-paraformalde-hyde, and processed for immunohistochemistry. Subtypes of intercalated cells were identified by using antibodies for H(+)-ATPase and AE1, and connecting tubule cells and principal cells of collecting duct were identified using antibodies for calbindin D28K and AQP2, respectively. In human kidney, pendrin was expressed in the apical domain of AE1-negative intercalated cells including type B cells with diffuse and/or basolateal H(+)-ATPase, non A-non B (non -A/B) type intercalated cells with apical H(+)-ATPase and bipolar type of intercalated cells with apical and basolateral H(+)-ATPase. The AQP2-positive principal cells of cortical collecting duct were also had apical pendrin immunoreactivity. However, there was no pendrin immunoreactivity in AE1-positive type A intercalated cells, calbindin D28K-positive connecting tubule cells, and AQP2-positive medullary collecting duct. These results suggest that pendrin is an apical anion exchanger not only in the AE1-negative intercalated cells (type B, non-A/B and bipolar cells) but also in the principal cells of cortical collecting duct, and has an essential role in HCO3-secretion in human kidney.
Animals ; Antibodies ; B-Lymphocytes ; Calbindin 1 ; Calbindins ; Carcinoma, Renal Cell ; Humans* ; Immunohistochemistry ; Kidney* ; Mice ; Proton-Translocating ATPases ; Rats

Dopamine plays an important role in cognitive functions including decision making, attention, learning and memory in the anterior cingulate cortex (ACC). However, little is known about dopamine receptors (DAR) expression patterns in ACC neurons, especially GABAergic interneurons. The aim of the present study was to investigate the expression of the most abundant DAR subtypes, D1 receptors (D1Rs) and D2 receptors (D2Rs), in major types of GABAergic interneurons in rat ACC, including parvalbumin (PV)-, calretinin (CR)-, and calbindin D-28k (CB)-containing interneurons. Double immunofluorescence staining and confocal scanning were used to detect protein expression in rat brain sections. The results showed a high proportion of PV-containing interneurons express D1Rs and D2Rs, while a low proportion of CR-positive interneurons express D1Rs and D2Rs. D1R- and D2R-expressing PV interneurons are more prevalently distributed in deep layers than superficial layers of ACC. Moreover, we found the proportion of D2Rs expressed in CR cells is much greater than that of D1Rs. These regional and interneuron type-specific differences of D1Rs and D2Rs indicate functionally distinct roles for dopamine in modulating ACC activities via stimulating D1Rs and D2Rs.
Animals ; Calbindin 1 ; physiology ; Calbindin 2 ; physiology ; Calcium-Binding Proteins ; physiology ; Dopamine ; physiology ; Gyrus Cinguli ; cytology ; Interneurons ; physiology ; Parvalbumins ; physiology ; Rats ; Receptors, Dopamine D1 ; physiology ; Receptors, Dopamine D2 ; physiology

BACKGROUNDWe investigated the co-expression of calbindin-D28k (CB), calretinin (CR) and parvalbumin (PV, a combination of the three is referred to as CaBPs) with gamma-aminobutyric acid (GABA) or glycine in neurons of the rat medullary dorsal horn (MDH).

METHODSImmunofluorescence histochemical double-staining for CaBPs and GABA or glycine was performed on the sections from rat MDH.

RESULTSCB-, CR-, PV-, GABA- and glycine-like immunoreactive (LI) neurons were differentially observed in all layers of the MDH, but particularly in lamina II. Neurons that exhibited immunoreactivity for both CaBPs and GABA or glycine were also observed mainly in lamina II. A few of them were found in laminae I and III. The percentages of neurons which co-expressed CB/GABA or CB/glycine out of the total numbers of CB- and GABA-LI neurons or CB- and glycine-LI neurons were 5.3% and 12.1% or 4.1% and 10.0%, respectively. The ratios of CR/GABA or CR/glycine co-existing neurons out of the total numbers of CR- and GABA-LI neurons or CR- and glycine-LI neurons were 5.8% and 7.6% or 4.4% and 7.1%, respectively. The rates of PV/GABA or PV/glycine co-localized neurons out of the total numbers of PV- and GABA-LI neurons or PV- and glycine-LI neurons were 11.1% and 5.1% or 9.9% and 5.1%, respectively.

CONCLUSIONThe results indicate that some neurons in the MDH contain both CaBPs and GABA or glycine.

Animals ; Calbindin 1 ; Calbindin 2 ; Calbindins ; Calcium-Binding Proteins ; analysis ; Fluorescent Antibody Technique ; Glycine ; analysis ; Immunohistochemistry ; Medulla Oblongata ; cytology ; Parvalbumins ; analysis ; Posterior Horn Cells ; chemistry ; Rats ; S100 Calcium Binding Protein G ; analysis ; gamma-Aminobutyric Acid ; analysis

This study was designed to identify expression of calcium-binding proteins and synaptic reorganizations of dentate mossy fibers in hippocampal sclerosis of human temporal lobe epilepsy. Hippocampal neuronal density was quantitively analyzed in temporal lobe epilepsy group (n=50) to investigate the degree of hippocampal sclerosis and it was compared with that of autopsy control (n=3). To verify the distribution of calcium-binding proteins in neurons of epileptic hippocampi, the parvalbumin (PV)-immunoreactive and calbindin-D28K (CB)-immunoreactive neurons were quantitively analyzed in each area of Ammon's horn by immunohistochemical stain. Also, to clarify synaptic reorganizations of the dentate mossy fibers, a part of each hippocampus was examined under light microscopy and transmission electron microscopy using Timm sulphide silver method. In epileptic hippocampi, severity of hippocampal sclerosis (HS) was graded four, which consisted of 3 cases with no HS, 6 mild HS, 12 moderate HS, and 29 severe HS. The hippocampal neuronal loss was most prominent in CA1, followed by CA4 and CA2. Expression of calcium-binding proteins was more prevalent in CA2 of all groups. The proportion of PV-immunoreactive neurons in CA1 and CA4 significantly increased in the moderate and severe HS group, whereas the proportion of CB-immunoreactive neurons did not correlated with the severity of HS. Timm granules were noted in inner molecular supragranular layer of dentate gyrus of epileptic hippocampi and they tended to increase in proportion along with the severity of hippocampal sclerosis. Transmission electron microscopy showed that supragranular Timm granules corresponded to synaptic terminals of mossy fibers. These results suggest that parvalbumin appears to have more protective effect against neuronal loss and that mossy fiber synaptic reorganization seems to play a major role in pathogenesis of hippocampal sclerosis of human temporal lobe epilepsy.
Autopsy ; Calbindin 1 ; Calcium* ; Calcium-Binding Proteins* ; Dentate Gyrus ; Epilepsy, Temporal Lobe* ; Hippocampus ; Humans ; Microscopy ; Microscopy, Electron, Transmission ; Nerve Fibers, Myelinated ; Neurons ; Presynaptic Terminals ; Sclerosis* ; Silver ; Temporal Lobe*