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PubMed2003

Chronological alterations of calbindin D-28k immunoreactivity in the gerbil main olfactory bulb after ischemic insult.

We investigated spatial and temporal alterations of calbindin D-28k (CB) immunoreactivity in the gerbil main olfactory bulb after transient ischemia-reperfusion. In sham-operated animals, CB-immunoreactive (IR) neurons were found in the periglomerular layer, external plexiform layer and granule cell layer. At 1-4 days after ischemic insult, the number of CB-IR neurons significantly increased. This result suggests that the increased CB may buffer the intracellular calcium at an early time point after the ischemic insult. In contrast, 10-30 days after the ischemic insult, the number of CB-IR neurons significantly decreased as compared to sham-operated animals. This result suggests that a malfunction in olfactory process may have occurred in the olfactory bulb at a later time point after the ischemic insult.

Animals↗
PubMed2003

Purkinje-like cells in the cochlear nucleus of the Common Tree Shrew (Tupaia glis) identified by calbindin immunohistochemistry.

The dorsal cochlear nucleus (DCN) of Tree Shrews (Tupaia glis; n=2) was examined by calbindin (CB) immunohistochemistry for the presence of Purkinje-like cells (PLCs), detected previously in only four different mammals. We found up to eight CB-immunoreactive PLCs in the left and right DCN, and a few axons, likely of PLC origin, that appeared to leave the DCN. These findings suggest that PLCs may have a wider distribution through mammalian species, and may represent more than just misrouted cells.

Animals↗
PubMed1999

Calbindin 28kD and parvalbumin immunoreactive neurons receive different patterns of synaptic input in the cat superior colliculus.

Recent evidence suggests that neurons containing the calcium binding proteins calbindin 28kD (CB) and parvalbumin (PV) have differing distributions which match respectively the distribution of W and Y retinal ganglion cell inputs to the cat superior colliculus (SC). In this study we have used electron microscope immunocytochemistry to study directly the synaptic inputs to neurons containing CB and PV. Aspiration lesions of areas 17-18 of visual cortex were made 4 days prior to sacrifice in order to identify degenerating cortical terminals (CT). Retinal terminals (RTs) were identified by their characteristic morphology including large round synaptic vesicles and pale mitochondria. We photographed RTs and CTs that were in contact with immunoreactive profiles sampled in both the superficial gray and optic layers (ol) of SC. CB immunoreactive (ir) dendrites were usually of small to medium caliber and were found to receive synaptic input from RTs. These RTs were all small profiles forming a single synaptic contact with asymmetric densifications. CBir profiles also received other synaptic input, including from terminals with dark mitochondria that contained flattened synaptic vesicles (F profiles). No CBir dendrites were found to receive CT input even though degenerating CTs were found in the vicinity of CBir profiles. By contrast, both RT and CT were found to contact PVir dendrites. RT terminals contacting PVir dendrites were both small and larger profiles with round synaptic vesicles and asymmetric synaptic densifications. CT were undergoing electron dense degeneration but still sometimes formed asymmetric synaptic densifications with PV neurons. PV cells also received F profile synaptic input. We conclude that CB neurons receive small RT synapses that are probably of W origin, while PV neurons receive both RT and CT synapses which are likely related to the Y pathway.

Animals↗
PubMed2003

Expression of human Calbindin-D(9k) correlated with age, vitamin D receptor and blood calcium level in the gastrointestinal tissues.

OBJECTIVE: Calbindin-D(9k) (CaBP-9k) has been thought to be one of the important factors of calcium absorption and metabolism in the intestine. The understanding of CaBP-9k role in the gastrointestinal tissues is important in human in relation with aging and gender, and whether there is any relationship with mineral homeostasis and risk for the development of age-related calcium disorders such as osteoporosis. DESIGN AND METHODS: The expression of CaBP-9k and vitamin D receptor (VDR) was analyzed in the human gastrointestinal tissues with different ages and gender. In addition, the correlation among CaBP-9k expression, age and calcium concentration in blood was elucidated in these tissues. RESULTS: A significant increase in the expression of CaBP-9k mRNA was observed in the bulb and 2(nd) portion of duodenum over age in both men and women. However, no significant age-associated change in VDR mRNA level was detected in the duodenum. The concentration of blood calcium level decreased with age in both men and women. CONCLUSION: The duodenal CaBP-9k may be not involved in calcium absorption in these tissues. The functional significance of the increase in the intestinal CaBP-9k expression with age is not currently clear and requires further investigation.

Adult↗
PubMed2002

Decrease in calbindin content significantly alters LTP but not NMDA receptor and calcium channel properties.

The contribution of the cytosolic calcium binding protein calbindin D(28K) (CaBP) to the synaptic plasticity was investigated in hippocampal CA1 area of wild-type and antisense transgenic CaBP-deficient mice. We showed that long-term potentiation (LTP) induced by tetanic stimulation in CaBP-deficient mice was impaired. The fundamental biophysical properties of NMDA receptors and their number were not modified in CaBP-deficient mice. We also demonstrated that the physiological properties of calcium channels were identical between genotypes. An insufficient Ca(2+) entry through NMDA receptors or calcium channels, or a decrease in NMDA receptor density are unlikely to explain this impairment of LTP. Interestingly, we showed that the loss of LTP was not prevented by glycine but was restored in the presence of a low concentration of the NMDA receptor antagonist D-APV (5 microM) and of the calcium chelator BAPTA-AM (5 microM). Moreover, we observed a loss of LTP in the wild-type mice when the postsynaptic tetanic-induced [Ca(2+)](i) rise is excessively increased. Conversely, a weaker tetanus stimulation allowed LTP induction and maintenance in CaBP-deficient mice. These results suggest that a higher cytosol [Ca(2+)](i), due to the decrease of CaBP expression may impair LTP induction and maintenance mechanisms without affecting the mechanisms of calcium entry. Thus, CaBP plays a critical role in long term synaptic plasticity by limiting the elevation of calcium rise in the cytosol to some appropriate spatio-temporal pattern.

Animals↗
PubMed1990

Immunocytochemical demonstration of the calcium-binding proteins calbindin-D 28k and parvalbumin in the subiculum, hippocampus and dentate area of the domestic pig.

The distribution of the calcium-binding proteins calbindin-D 28k (CaBP) and parvalbumin (PV) in the hippocampal region of the domestic pig was demonstrated by immunocytochemistry. Scattered CaBP-immunoreactive cell bodies were present in the subiculum, stratum oriens, pyramidal cell layer and stratum radiatum of the hippocampal regio superior and inferior, and the outer plexiform layer and outer hilar cell layer of the dentate hilus. Other cell bodies and bundles of stained fibers were present in stratum moleculare of regio superior and inferior, and in the outer third of the molecular layer of the fascia dentata. Terminal-like CaBP-immunoreactivity was seen in the subiculum and around cell bodies in the pyramidal cell layer of regio superior and inferior and the dentate granular cell layer. Scattered PV-immunoreactive cell bodies were present in stratum oriens and the pyramidal cell layer of regio superior and inferior, and in the outer plexiform layer and outer hilar cell layer of the dentate hilus. Terminal-like PV-immunoreactivity surrounded the cell bodies in the pyramidal cell layer of regio superior and inferior and in the dentate granular cell layer. The distribution of CaBP and PV in the pig hippocampus is compared to that of other more commonly used experimental animals. Whereas the distribution of PV-immunoreactivity in the pig hippocampus appears identical to that of the rat hippocampus, the distribution of CaBP-immunoreactivity in the pig hippocampus differs markedly from that of the rat hippocampus, the most prominent feature being a lack of CaBP-immunoreactivity in the granule cells, mossy fibers and pyramidal cells in the pig. The functional implications of calcium-binding proteins in the brain are discussed.

Animals↗
PubMedDate not supplied

Genetic regulation of placental function: a quantitative in situ hybridization study of calcium binding protein (calbindin-D9k) and calcium ATPase mRNAs in sheep placenta.

The calcium requirement of the ovine fetus increases progressively throughout pregnancy. The 9-kDa calcium binding protein (calbindin-D9k; 9CBP) is considered to be a reliable marker for epithelia mediating calcium transport. This quantitative in situ hybridization study shows that the levels of 9CBP mRNA show a pregnancy stage-related increase which correlates with fetal calcium demand only in maternal endometrial gland and fetal interplacentomal trophoblast epithelia. Levels of 9CBP mRNA in the placentome, which has by far the greater area of maternofetal contact, show no changes during pregnancy. mRNA for the CaATPase enzyme, a second requirement for calcium transport, is shown to be present in epithelia in interplacentomal and placentomal regions but shows no change in concentration as pregnancy progresses. Results with the 9CBP and CaATPase mRNAs confirm our recent immunocytochemical results with ruminant placenta and indicate the basis for a cellular calcium transport system analogous to that in the enterocyte. The interplacentomal trophoblast system appears to be eminently suitable for investigations of details of the cellular mechanism and control of epithelial calcium transport.

Animals↗
PubMed2003

Neurochemical heterogeneity of the thalamic reticular and perireticular nuclei in developing rabbits: patterns of calbindin expression.

The thalamic reticular nucleus (TRN) forms an essential part of the circuits that link the thalamus to the cortex, whereas the perireticular thalamic nucleus (PRN) consists of scattered neurons that are located in the internal capsule, in close relation to the TRN. A common feature of these nuclei in different species is the immunoreactivity for some calcium binding proteins with a developmental pattern of expression. In the present study, sections from rabbits at different ages were examined to determine the calbindin (CB) expression in the developing TRN and PRN at the first stages of development. These CB-expressing cells constitute an important subpopulation of neurons in the caudal half of the developing TRN. In the adult, there are still positive CB somata in the middle and caudal halves of the nucleus. In the PRN, where the developmental pattern of CB expression has not been described before, the number of CB perireticular cells decreases progressively. Our results, together with previous data in the rabbit suggest the existence of remarkable neurochemical heterogeneity in the TRN and PRN of the rabbit.

Animals↗
PubMed1996

A chronological study of the expression of glial fibrillary acidic protein and calbindin-D28 k by reactive astrocytes in the electrically lesioned rat brain.

Immunoreactivity of neuronal and glial marker proteins of reactive astrocytes around the electrically damaged pyramidal layer and stratum radiatum of the hippocampal CA1 region and corpus callosum was chronologically studied in electrically lesioned rat brains. A monoclonal antibody against calbindin-D28 k (CD28-Ab) and a polyclonal antibody against glial fibrillary acidic protein (GFAP-Ab) were used for immunostaining. Immunoreactivity of CD28 and GFAP in the reactive astrocytes was detected in brains 1-6 weeks post-lesion but not in non-lesioned brains. The number of immunohistochemically stained reactive astrocytes around the electrically damaged areas were counted and then compared with the number of those in the same areas of non-lesioned brains. The number of CD28- and GFAP-immunoreactive astrocytes began to increase around the lesion from 1-3 weeks following lesion in the pyramidal layer of the hippocampal CA1 region and from 1-4 weeks following lesion in the stratum radiatum of the hippocampal CA1 region and corpus callosum. These immunoreactive astrocytes could be observed for 6 weeks (the maximum survival time studies) in all areas of the lesioned brains studied. The increase in the number of reactive astrocytes might have been induced by the stimulatory effects of neurotrophic factors, or growth factors, produced around the lesioned site. The constancy in the number of reactive astrocytes after 3 and 4 weeks in the lesioned areas may have been due to the termination of the initial phase of the repair process, i.e. space-filling. Reactive astrocytes which were stained by GFAP-Ab were separated into two groups, based on the presence of CD28, i.e. CD28-positive and CD28-negative reactive astrocytes. The presence of CD28 might confer certain functions via calcium-mediated mechanisms on CD28-positive astrocytes in addition to the constructive role mediated by GFAP.

Animals↗
PubMed2004

Changes in the expression of calbindin D-28k in the gerbil hippocampus following seizure.

Previous studies have reported that calbindin D-28k (CB), a calcium-binding protein, containing neurons in the hippocampus play an important role in hippocampal excitability in epilepsy, because CB modulates the free calcium ion during seizure. Hence, in the present study, we investigated changes of CB expression in the hippocampus and its association in the Mongolian gerbil to identify roles of CB in epileptogenesis. CB immunoreactivity in the hippocampus was significantly lower in the pre-seizure group of seizure sensitive (SS) gerbils as compared with those seen in the seizure resistant (SR) gerbils. The distribution of CB immunoreactivity in the hippocampus showed significant difference after seizure on-set in SS gerbils. CB immunoreactivity in the hippocampal CA1, CA2 areas, and subiculum was lowest at 3h after seizure on-set; thereafter, the immunoreactivity became to increase to 12h after seizure on-set. Mossy fibers, Schaffer collaterals and dentate granule cells showed the highest CB immunoreactivity at 3h after seizure on-set; thereafter, the immunoreactivity became to decrease. In the case of the intrinsic and output connections of the hippocampus, a rapid decrease of CB serves an inhibitory function, which regulates the seizure activity and output signals from the hippocampus.

Animals↗
PubMed2002

Stimulation of calbindin-D(9k) mRNA expression in the rat uterus by octyl-phenol, nonylphenol and bisphenol.

Quantification of estrogen-induced changes in the expression levels of endogenous genes such as pS2 and vitellogenin could be an assay to detect estrogenicity of chemicals. Considering its regulation by estrogen, in the present study, we hypothesize that the calbindin-D(9k) (CaBP-9k) gene has the possibility as a biomarker for estrogenic response of the environmental estrogens. We analyzed the time- and dose-dependent CaBP-9k mRNA expression in the immature rats by 3-day injection of 17beta-estradiol (E2) and alkylphenol acid [octyl-phenol (OP) and nonylphenol (NP)] and bisphenol A(BPA)) which are environmentally persistent and reported to have some estrogenic activity in experimental test systems. The expression of CaBP-9k mRNA was compared with uterotropic response of the compounds. A significant increase in CaBP-9k mRNA expression was observed when treated with 1000 mg/kg body weight (BW) per day of OP (18-fold versus control), NP (17-fold versus control) and BPA (6-fold versus control) for 3 days in dot blot assays. Using Northern blot analysis, a more dramatic increase of CaBP-9k mRNA expression was observed when treated with 1000 mg/kg BW per day of OP (132-fold versus control) and NP (113-fold versus control) for 3 days. Treatment with 10 and 100 mg/kg BW per day of NP and 100 mg/kg BW per day of OP for 3 days induced a small but significant increase in CaBP-9k mRNA expression. As expected, a single dose of E2 (40 microg/kg BW per day) for 3 days induced a significant increase in CaBP-9k mRNA expression as revealed by dot (15-fold versus control) or Northern blot assay (102-fold versus control). In a time response experiment using Northern blot assay, a significant increase in CaBP-9k mRNA expression was observed as early as 3 h, peaked at 6 h and continued until 72 h after treatment with 1000 mg/kg BW per day of OP, NP, and 48 h after treatment with 1000 mg/kg BW per day of BPA. A similar time-dependent response was observed when assessed by dot blot assay. Uterotropic response of the compounds was determined and compared with CaBP-9k mRNA expression. The alkylphenolic compounds induced a significant increase in the uterine wet weight at 1000 mg/kg BW per day of OP and NP, not BPA. A strong correlation between in vivo uterotropic assay and CaBP-9k mRNA expression assay was observed. In order to investigate the possible mechanisms by which the compounds regulate CaBP-9k mRNA expression, we studied the effect of the compound on the ERalpha mRNA level using total RNA from the treated rats. The alkylphenolic compounds as well as E2 stimulate the expression of ERalpha mRNA in a similar pattern to that of CaBP-9k mRNA in terms of dose- and time-dependent response. Strong regulation of CaBP-9k mRNA expression by E2 and the environmental estrogens and its correlation with in vivo uterotropic assay suggest that CaBP-9k gene can be used as a biomarker gene for assaying estrogenicity of putative estrogenic compounds.

Animals↗
PubMed2003

Mouse calbindin-D(9k) gene expression in the uterus during late pregnancy and lactation.

Calbindin-D(9k) (CaBP-9k) is a cytosolic calcium binding protein mainly expressed in duodenum, placenta and uterus. In order to understand the expression pattern and regulation of uterine CaBP-9k gene, the expression of CaBP-9k mRNA and its regulation by estrogen (E2) and progesterone (P4) were investigated in the mouse uterus during late pregnancy (from day 12 to 18) and lactation. The expression levels of uterine CaBP-9k, estrogen receptor alpha (ERalpha) and progesterone receptor (PR) mRNAs were measured by Northern blot analysis. The expression levels of mouse uterine CaBP-9k mRNA gradually increased from pregnancy day 16 (P16), peaked at P18 (6.0-fold vs. P12) and declined at birth and during lactation. The expression levels of ERalpha and PR mRNAs indicated a similar fluctuation as CaBP-9k mRNA, suggesting the role of sex steroids/receptors in the regulation of CaBP-9k gene. To investigate effect of steroid hormone on CaBP-9k mRNA expression, three groups of animals were injected (s.c) with steroid hormone antagonists (RU486, tamoxifen, and ICI182780), respectively. RU486, a P4 antagonist, induced a significant decrease in CaBP-9k mRNA expression at 48 (3.2-fold) and 72 h (3.8-fold). However, tamoxifen and ICI182780, E2 antagonists, had no effect on CaBP-9k mRNA expression. Combined treatment with RU486 and ICI182780 did not further decrease the expression level of CaBP-9k mRNA when compared with RU486 treatment at 48 and 72 h. In addition, the treatment with RU40555, a glucocorticoid/progesterone antagonist, resulted in a decrease at 48 and 72 h following treatment. These results indicate that E2 is not likely involved in the regulation of CaBP-9k gene in the mouse uterus during late pregnancy and lactation. In conclusion, the present results suggest that P4, not E2 is a key regulator of CaBP-9k mRNA expression during late pregnancy and lactation.

Animals↗
PubMed2002

Long-term changes in calbindin D(28K) immunoreactivity in the rat hippocampus after cardiac arrest.

Calbindin D(28K) (CB) expression was analyzed in the rat hippocampus following 10-min-cardiac arrest-induced ischemia within a year after reperfusion. In rats examined 3 days after ischemia, CB immunoreactivity disappeared completely from CA1 pyramidal neurons and from most CA2 pyramids. In the stratum granulosum of the dentate gyrus, mossy fibers, and hippocampal interneurons, CB immunoreactivity was preserved, although staining was somewhat paler than that in control rats. A similar pattern of CB immunoreactivity was found in rats sacrificed 14 days and 1 month after cardiac arrest. From the 14th postischemic day, neuronal loss in the stratum pyramidale of CA1 but not in that of CA2 became apparent. The reappearance of CB immunoreactivity in CA1 and CA2 pyramidal neurons was noticed 6 months after ischemia, and the pattern was identical to that observed in animals sacrificed 12 months after the ictus. The prolonged loss and delayed reappearance of CB immunoreactivity in the hippocampus demonstrate that ischemia may induce long-term disturbances of protein expression, which may in turn result in impairment of hippocampal functioning.

Animals↗
PubMed2002

Age-related change of calbindin D-28k immunoreactive neurons in the rat main olfactory bulb.

We examined the aged-related changes of calbindin D-28k (CB)-immunoreactive (IR) neurons in the rat main olfactory bulb (MOB). The localization of CB-IR neurons was found to be almost entirely restricted to the glomerular layer. However, a few CB-IR neurons were observed in the granular layer. Prior to the 6th postnatal month (PM 6), the CB-IR neurons had long processes, and the population of CB-IR neurons had increased significantly compared to PM 12 and 24. At PM 12, CB-IR neurons showed a tendency to be smaller and have fewer dendrites than at guess. This phenomenon became remarkable at PM 24. The distinct reduction of CB-IR dendrites in glomeruli may be due to age-related functional restrictions. Increased calcium levels in the MOB neurons may be induced to allow cytotoxic event detection in the MOB neurons, and unbalanced Ca(2+)/Mg(2+)-ATPase may also induce aging-related MOB morphological changes.

Aging↗
PubMed1999

Early social isolation decreases the expression of calbindin D-28k in rat cerebellar Purkinje cells.

The main objective of this study was to analyze the effects of early social isolation on the expression of calbindin D-28k (CAD-28k) in cerebellar Purkinje cells. Sprague-Dawley rats were reared either in isolation (IC) or socially housed conditions (SC) from postnatal days 18-32. Half of the rats of each group were sacrificed at 32 days of age and the cerebella processed for immunocytochemical labeling against CAD-28k. The remaining IC animals were housed under standard conditions for re-socialization between postnatal days 32 and 62. The results indicate that Purkinje cells of rats reared under early isolation show a marked loss of CAD-28k immunoreactivity, and that this deficiency is recovered by later social interaction, although the thickness of the molecular layer remains reduced.

Animals↗
PubMed1998

Distribution of calbindin D-28k and parvalbumin neurons and fibers in the rat basal ganglia.

This review deals with the distribution of immunoreactivity for calbindin D-28k (CB) and parvalbumin (PV) in the different nuclei of the rodent basal ganglia analyzed with the data available after the use of single and double antigen procedures applied to single sections. These findings reveal that CB and PV are distributed according to a highly heterogeneous pattern in the caudate putamen complex (CPu), globus pallidus (GP), entopeduncular nucleus (EP), subthalamic nucleus (STh) and substantia nigra (SN) of the rat. In each basal ganglia structure, the two calcium-binding proteins label different neuronal subsets. Therefore, the use of CB and PV immunohistochemistry may be considered as an excellent tool to define distinct chemoarchitectonic and functional domains within the complex organization of the basal ganglia. Double immunohistochemical methods are also useful to illustrate the relationships between the different chemical subdivisions of the CPu, GP, EP, STh and SN and the chemically characterized connections with each other and with other forebrain and brainstem structures. However, specific rules should be followed when combining single and double immunostaining procedures, and the results of such studies must be evaluated with caution. When they are used properly, these methods can reveal hitherto unknown principles of organization of the basal ganglia and thus shed new light on the anatomical and functional organization of this set of subcortical structures involved in the control of motor behavior.

Animals↗
PubMed2000

Calcification processes in the chick otoconia and calcium binding proteins: patterns of tetracycline incorporation and calbindin-D28K distribution.

In order to clarify the otoconia formation and turnover, tetracycline, an antibiotic that precipitates at calcifying fronts and serves as a fluorescent marker, was injected into eggs at different stages of chick embryonic development, as well as into postnatal chicken and into adult animals. The changes in the intensity, location patterns and time course of fluorescent labelling in each examined stage in the otolithic organs was studied. The presence and distribution of calbindin (CB)-D28K, one of the calcium-binding proteins constantly found in the mammalian and chicken cochlea and also in otolithic membrane of some adult mammals, was studied. Results in embryonal stages, postnatal and adult animals allow us to postulate that otoliths are mainly produced during the embryonal phase, but they may also be produced throughout the whole life span. Results also indicate that otoconia are dynamic structures which undergo turnover. The correspondence between the patterns of CB-D28K immunoreactivity and tetracycline fluorescence may indicate that CB-D28K participates in the formation of otoconia.

Aging↗
PubMed2001

Onset of calbindin-D 28K and parvalbumin expression in the lateral geniculate complex and olivary pretectal nucleus during postnatal development of the rat.

The onset and distribution of calbindin (CB) and parvalbumin (PV) immunoreactivity were investigated in the lateral geniculate nuclear complex and the olivary pretectal nucleus (OPT) in developing rats. CB expression occurred early (before eye-opening) in the relay neurons of the intergeniculate leaflet, parvocellular portion of the ventral lateral geniculate nucleus and OPT relating to ambient vision mediated by W-like retinal ganglion cells. On the contrary, PV expression occurred late (after eye-opening) in the relay neurons of the magnocellular portion of the ventral lateral geniculate nucleus (VLGMC) and OPT relating to focal vision mediated by Y-like retinal ganglion cells. A unilateral eye enucleating experiment indicated that the VLGMC and OPT received dense input from PV-positive Y-like retinal ganglion cells. The results show the different onsets of CB and PV expressions in the retino-recipient thalamic and pretectal nuclei receiving inputs from different kinds of retinal ganglion cells.

Aging↗