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1.
Nam SM Kim YN Yoo DY Yi SS Kim W Hwang IK Seong JK Yoon YS 《Neurochemical research》2012,37(2):253-260
In this study, we investigated the differences in calbindin D-28k (CB), calretinin, (CR) and parvalbumin (PV) immunoreactivity
in the hippocampus of Zucker diabetic fatty (ZDF) rats and Zucker lean control (ZLC) rats. In addition, we observed the effects
of hypothyroidism on the levels of immunoreactivity of these proteins in ZDF rats. For this study, 7-week-old ZDF rats were
used, and methimazole treatment was continued for 5 weeks to induce hypothyroidism. The animals were sacrificed at 12 weeks
of age. ZDF rats showed increased blood glucose levels compared to those in ZLC rats. Methimazole intervention significantly
reduced total and free T3 levels, and it ameliorated the increase of blood glucose levels in ZDF rats. In ZLC rats, CB, CR,
and PV immunoreactivity was detected in regions of the hippocampus proper. In vehicle-treated ZDF rats, CB, CR, and PV immunoreactivity
was significantly decreased in the hippocampus. However, in the methimazole-treated rats, CB, CR, and PV immunoreactivity
was significantly increased compared to that in the vehicle-treated rats. These results suggest that hypothyroidism ameliorated
the diabetes-induced reduction of CB, CR, and PV immunoreactivity in the hippocampus. 相似文献
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3.
The distribution and morphology of neurons containing three calcium-binding proteins, calbindin D28K, calretinin, and parvalbumin in the adult rabbit visual cortex were studied. The calcium-binding proteins were identified using antibody immunocytochemistry. Calbindin D28K-immunoreactive (IR) neurons were located throughout the cortical layers with the highest density in layer V. However, calbindin D28K-IR neurons were rarely encountered in layer I. Calretinin-IR neurons were mainly located in layers II and III. Considerably lower densities of calretinin-IR neurons were observed in the other layers. Parvalbumin-IR neurons were predominantly located in layers III, IV, V, and VI. In layers I and II, parvalbumin-IR neurons were only rarely seen. The majority of the calbindin D28K-IR neurons were stellate, round or oval cells with multipolar dendrites. The majority of calretinin-IR neurons were vertical fusiform cells with long processes traveling perpendicularly to the pial surface. The morphology of the majority of parvalbumin-IR neurons was similar to that of calbindin D28K: stellate, round or oval with multipolar dendrites. These results indicate that these three different calcium-binding proteins are contained in specific layers and cells in the rabbit visual cortex. 相似文献
4.
5.
Distribution of Calretinin, Calbindin D28k, and Parvalbumin in Subcellular Fractions of Rat Cerebellum: Effects of Calcium 总被引:4,自引:0,他引:4
Abstract: The distribution of calretinin, calbindin D28k, and parvalbumin was examined in subcellular fractions prepared from rat cerebellum and analyzed by immunoblot. Calretinin was also quantified by radioimmunoassay. As expected, all three soluble, EF-hand calcium-binding proteins were predominantly localized in the cytosolic fraction. Calretinin and calbindin D28k were also detected in membrane fractions. Calretinin was more abundant in synaptic membrane than in microsomal fractions. The cerebellar microsomal fraction contained the greatest concentration of membrane-associated calbindin D28k. The association of calretinin and calbindin D28k with membrane fractions was decreased in samples prepared or incubated in low calcium. Quantification of calretinin in subcellular fractions of rat cerebellum revealed a greater amount of calretinin in cytosolic fractions prepared or incubated in low calcium and reduced amounts of calretinin in all membrane fractions incubated in low calcium with the exception of the mitochondrial fraction. These results imply that calretinin and calbindin D28k might have physiological target molecules that are associated with, or are components of, brain membranes. 相似文献
6.
In the present study, we investigated changes in glutamate decarboxylase 65 (GAD65) and GAD67 immunoreactivity and protein
levels in the main olfactory bulb (MOB) after 5 min of transient forebrain ischemia in gerbils. GAD65 immunoreactivity in
the sham-operated group was shown in neurons and neuropil except for the somata of granule cells. GAD65 immunoreactivity was
increased in neurons in the external plexiform layer 60 days after ischemia, and in mitral cells 30 and 60 days after ischemia.
GAD67 immunoreactivity in the sham-operated group was shown in periglomerular cells, neuron in the external plexiform layer
and granule cells with neuropil. GAD67 immunoreactivity in periglomerular cells was increased 10, 45 and 60 days after ischemia.
GAD67 immunoreactivity in neurons in the external plexiform layer was increased 10 and 15 days after ischemia. Mitral cells
showed strong GAD67 immunoreactivity 10 days after ischemia. However, GAD67 immunoreactivity in the granule cells was not
changed with time after ischemia. In Western blot analysis for GAD65 and GAD67 protein levels in the ischemic gerbil MOB,
GAD65 level was not changed after ischemia; GAD67 level was increased 10 days after ischemia. These results suggest that transient
ischemia causes changes in GAD65 and GAD67 immunoreactivity in the gerbil MOB, and this change may induce a malfunction in
olfaction after an ischemic insult.
Ki-Yeon Yoo and In Koo Hwang equally contributed to this article. 相似文献
7.
Age-Dependent Changes in Calretinin Immunoreactivity and its Protein Level in the Gerbil Hippocampus
Choong Hyun Lee In Koo Hwang Jung Hoon Choi Ki-Yeon Yoo Ok Kyu Park Sung-Oh Huh Yun Lyul Lee Hyung-Cheul Shin Moo-Ho Won 《Neurochemical research》2010,35(1):122-129
Calretinin (CR)-immunoreactive interneurons are well known as the interneuron specific interneurons in the hippocampus. CR-immunoreactive
neurons form cellular network and regulate the activity of other GABAergic inhibitory interneurons in the hippocampus. In
the present study, we investigated age-related changes in CR-immunoreactive neurons and protein levels in the gerbil hippocampus
during normal aging. In all subregions of the gerbil hippocampus, the number of CR-immunoreactive neurons was significantly
decreased in the postnatal month 6 (PM 6) group compared to that in the PM 1 group. Thereafter, CR-immunoreactive neurons
were decreased with age. In addition, the number of CR-immunoreactive cells in the subgranular zone were significantly decreased
in the PM 6 group. We also observed that CR protein levels were decreased gradually with age. These results indicate that
both CR immunoreactivity and its protein level were decreased with age in the gerbil hippocampus during normal aging. 相似文献
8.
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Laminar Distribution of Putative Neurotransmitter Amino Acids and Ligand Binding Sites in the Dog Olfactory Bulb 总被引:1,自引:1,他引:1
Coronal sections of frozen dog olfactory bulb have been dissected into four anatomically distinct layers. The laminar distribution of ten amino acids, the dipeptide carnosine, and nine [3H]ligand binding sites in these layers was determined. GABA and tyrosine levels were highest in the mitral cell-granule cell layer, and glutamate levels were slightly elevated in the glomerular layer. The distributions of all other amino acids did not show significant differences across the layers. Carnosine was predominantly localized in the fiber and glomerular layers. With the exception of quinuclidinyl benzilate, the [3H]ligand binding sites showed more discrete distributions. Muscimol, diazepam, kainic acid, and spiroperidol binding were predominantly localized in the mitral cell-granule cell layer, where clonidine binding was at a minimum. Dihydromorphine binding was high in both the fiber and the mitral cell-granule cell layers. Carnosine binding was maximal in the glomerular layer. The implications of these observations with regard to biochemical and neurophysiological data are discussed. 相似文献
10.
Brain plasticity, in relation to new adult mammalian neurons generated in the subgranular zone of the hippocampus, has been well described. However, the functional outcome of new adult olfactory neurons born in the subventricular zone of the lateral ventricles is not clearly defined, as manipulating neurogenesis through various methods has given inconsistent and conflicting results in lab mice. Several small rodent species, including Peromyscus leucopus, display seasonal (photoperiodic) brain plasticity in brain volume, hippocampal function, and hippocampus-dependent behaviors; plasticity in the olfactory system of photoperiodic rodents remains largely uninvestigated. We exposed adult male P. leucopus to long day lengths (LD) and short day lengths (SD) for 10 to 15 weeks and then examined olfactory bulb cell proliferation and survival using the thymidine analog BrdU, olfactory bulb granule cell morphology using Golgi-Cox staining, and behavioral investigation of same-sex conspecific urine. SD mice did not differ from LD counterparts in granular cell morphology of the dendrites or in dendritic spine density. Although there were no differences due to photoperiod in habituation to water odor, SD mice rapidly habituated to male urine, whereas LD mice did not. In addition, short day induced changes in olfactory behavior were associated with increased neurogenesis in the caudal plexiform and granule cell layers of the olfactory bulb, an area known to preferentially respond to water-soluble odorants. Taken together, these data demonstrate that photoperiod, without altering olfactory bulb neuronal morphology, alters olfactory bulb neurogenesis and olfactory behavior in Peromyscus leucopus. 相似文献
11.
Calbindin-D28k is expressed in osteoblastic cells and suppresses their apoptosis by inhibiting caspase-3 activity 总被引:5,自引:0,他引:5
Bellido T Huening M Raval-Pandya M Manolagas SC Christakos S 《The Journal of biological chemistry》2000,275(34):26328-26332
The rate of osteoblast apoptosis is a critical determinant of the rate of bone formation. Because the calcium-binding protein calbindin-D(28k) has anti-apoptotic properties in neuronal cells and lymphocytes, we searched for the presence of this protein in osteoblastic cells and investigated whether it can modify their response to proapoptotic signals. Calbindin-D(28K) was expressed at low levels in several osteoblastic cell lines and at high levels in primary cultures of murine osteoblastic cells. Transient transfection of rat calbindin-D(28k) cDNA blocked tumor necrosis factor alpha (TNFalpha)-induced apoptosis in osteoblastic MC3T3-E1 cells, as determined by cell viability and nuclear morphology of cells cotransfected with the green fluorescent protein targeted to the nucleus, whereas transfection of the empty vector had no effect. Calbindin-D(28k) levels in several stably transfected MC3T3-E1 lines were directly related to protection from TNFalpha-induced apoptosis. Purified rat calbindin-D(28k) markedly reduced the activity of caspase-3, a critical molecule for the degradation phase of apoptosis, in a cell-free assay. In addition, cell extracts from MC3T3-E1 cells expressing high levels of calbindin-D(28k) decreased caspase-3 activity, compared with extracts from vector-transfected cells. This effect was apparently unrelated to the calcium binding properties of calbindin, as chelation of calcium by EGTA or addition of other calcium-binding proteins such as calbindin-D(9k), S100, calmodulin, and osteocalcin, did not affect caspase-3 activity. Last, calbindin-D(28k) interacts with the active form of caspase-3 as demonstrated by a GST pull-down assay. These results demonstrate that calbindin-D(28k) is a biosynthetic product of osteoblasts with a role in the regulation of apoptosis. They also reveal that the antiapoptotic properties of calbindin-D(28k) may result not only from calcium buffering but also from the ability of the protein to interact with and to inhibit caspase-3 activity, a property that is independent of its calcium binding capability. 相似文献
12.
Ca2+ binding proteins (CaBPs), calbindin-D28k (calbindin) and calretinin, are thought to contribute to the regulation of intracellular Ca2+ in many neuronal populations and perhaps more importantly, signal functional modulation in neuronal activity. In the present experiments, light microscopic immunohistochemistry revealed that the immunoreactivity of calbindin and calretinin was contained in varicose axons in the posterior pituitary. The dual labeling study with confocal microscopy demonstrated that calbindin immunoreactivity was present in the terminals of both oxytocin (OXT) and arginine-vasopressin (AVP) neurons. However, calretinin immunoreactivity was exclusively seen in the OXT terminals. Moreover, the dual labeling study showed that most calretinin-positive terminals contained calbindin immunoreactivity, demonstrating the colocalization of calbindin and calretinin in the same OXT nerve terminals. By electron microscopy, calbindin and calretinin immunoreactivities were seen in the neurosecretory axons and nerve terminals. These immunoreactive nerve terminals were seen to contain more clear microvesicles than dense-core neurosecretory granules. This immunoelectron microscopic observation suggests that both calbindin and calretinin localize preferentially in the active zone of the nerve terminals, which usually face the perivascular space around fenestrated capillaries. In spite of similar localization of calbindin and calretinin within the posterior pituitary, Western blot analysis showed some differences between the two CaBPs. Calbindin was present mostly in the soluble fraction with little in the insoluble fraction, but a substantial portion of calretinin was present in both the insoluble and soluble fractions. Moreover, dehydration induced by drinking 2% NaCl solution and deprivation of drinking water increased calretinin levels in the posterior pituitary as compared with control, but the calbindin level was not changed. The present findings demonstrate that calbindin and calretinin colocalize in the active zones of OXT nerve terminals, but only calretinin is upregulated with dehydration, suggesting different physiological role of calbindin and calretinin in the nerve terminals. 相似文献
13.
Calbindin-D9k (CaBP9k) is a vitamin D-dependent, calcium binding protein first identified in the cytoplasm of the intestinal epithelial cell. Using biotin-streptavidin immunohistochemistry, CaBP9k was localized to the maternal caruncular epithelium, fetal chorionic epithelium, and trophoblastic binucleated cells of the bovine placenta. Within the maternal epithelium the intensity of staining increases from second trimester pregnancies to term pregnancies, indicating a higher intracellular concentration of CaBP9k in the epithelium at term. Luminal and glandular epithelium of the non-caruncular endometrium also stained positively for CaBP9k in all stages of pregnancy observed. No CaBP9k was identified within the stroma or myometrium of the pregnant cow uterus. The increased level of CaBP9k in the caruncular epithelium during the last trimester is hypothesized to be in response to the rising demand for calcium to aid in the mineralization of the fetal skeleton. CaBP9k may play a role in enhancing calcium transport across the placenta in cattle. 相似文献
14.
Jung Hoon Choi Ki-Yeon Yoo Ok Kyu Park Choong Hyun Lee Sung Koo Kim In Koo Hwang Yun Lyul Lee Hyung-Cheul Shin Moo-Ho Won 《Cellular and molecular neurobiology》2010,30(6):929-938
Neurogenesis occurs during the embryonic stage and throughout life. Brain injuries such as ischemic insults enhance cell proliferation
in some areas of the brain. We examined proliferation of newly generated cells in each layer of the gerbil main olfactory
bulb (MOB) after 5 min of transient cerebral ischemia using bromodeoxyuridine (BrdU) immunohistochemistry. Ischemia-related
neuronal death in the MOB was not detected using Fluoro-Jade B histofluorescence and TUNEL staining. Many BrdU-positive (+) cells were found in the rostral migratory stream in control and ischemic MOBs. Significant increase of BrdU+ cells was observed in the granule cell layer (GCL) and glomerular layer (GL) from 15 days post-ischemia, and BrdU+ cells were very much higher than those of the control group 30 days post-ischemia. At this time point after ischemia/reperfusion,
a few BrdU+ cells in the GL and GCL were co-localized with calretinin+ cells, and many BrdU+ cells expressed doublecortin, a marker of immature neurons. These results indicate that cell proliferation is increased in
the GCL and GL without apparent neuronal loss from 15 days after transient cerebral ischemia in gerbils. 相似文献
15.
Jung Hoon Choi Ki-Yeon Yoo Choong Hyun Lee Ok Kyu Park Bing Chun Yan Hua Li In Koo Hwang Jeong Ho Park Sung Koo Kim Moo-Ho Won 《Neurochemical research》2010,35(10):1599-1608
In the present study, we investigated age-related differences in neuronal progenitors in the gerbil main olfactory bulb (MOB)
using doublecortin (DCX), a marker for neuronal progenitors which differentiate into neurons in the brain. No difference in
the number of neuronal nuclei (NeuN)-immunoreactive neurons was found in the MOB at variously aged gerbils. At postnatal month
(PM) 1, DCX immunoreaction was detected in all layers of the MOB except for the olfactory nerve layer. At this time point,
DCX-immunoreactive cells (neuronal progenitors) were very abundant; however, they did not have fully developed-processes.
From PM 3, the number of DCX-immunoreactive neuronal progenitors was decreased with age. At PM 6, DCX-immunoreactive cells
showed very well-developed processes. In western blot analysis, DCX protein level in the MOB was highest at PM 1. Thereafter,
levels of DCX protein were decreased with age. In the subventricular zone of the lateral ventricle, the number of Ki-67-immunoractive
cells (proliferating cells) was also significantly decreased with age. In addition, increases of α-synuclein-immunoreactive
structures were observed in the MOB with age. These results suggest that decrease in DCX-immunoreactive neuronal progenitors
and its protein levels in the MOB with age may be associated with reduction of cell proliferation in the SVZ and with an increase
in α-synuclein in the MOB. 相似文献
16.
Choong Hyun Lee In Koo Hwang Ki-Yeon Yoo Jung Hoon Choi Ok Kyu Park Jae-Chul Lee Young-Gil Jeong In Se Lee Moo-Ho Won 《Cellular and molecular neurobiology》2009,29(5):665-672
The hippocampus is associated with learning and memory function and shows neurochemical changes in aging processes. Calbindin
D-28k (CB) binds calcium ion with a fast association rate. We examined age-related changes in CB immunoreactivity and its
protein level in the gerbil hippocampus during normal aging. In the hippocampal CA1 region (CA1) and CA2, CB immunoreaction
was found in some neurons in the stratum pyramidale (SP) at postnatal month 1 (PM 1). CB immunoreactivity in neurons was markedly
increased at PM 3. Thereafter, CB immunoreactivity was decreased with time: CB-immunoreactive (+) neurons were fewest at PM 24. In the CA3, a few CB+ neurons were found only in the SP at PM 1 and in the stratum radiatum at PM 18 and 24. In addition, mossy fibers were stained
with CB at PM 1. CB immunoreactivity in mossy fibers was markedly increased at PM 3, thereafter it was decreased with time.
In the dentate gyrus, many granule cells (GC) in the granule cell layer were stained with CB at PM 1. CB immunoreactivity
in GC was markedly increased at PM 3, thereafter CB immunoreactivity was decreased with time. In Western blot analysis, CB
protein level in the gerbil hippocampus was highest at PM 3, thereafter CB protein levels were decreased with time. This result
indicates that CB in the gerbil hippocampus is abundant at PM 3 and is decreased with age. 相似文献
17.
18.
Chemical and Surgical Lesions of Rat Olfactory Bulb: Changes in Thyrotropin-Releasing Hormone and Other Systems 总被引:1,自引:0,他引:1
N. A. Sharif 《Journal of neurochemistry》1988,50(2):388-394
Stereotaxic injection of kainic acid (15 micrograms) into rat olfactory bulbs was accompanied by a 53% (n = 4; p less than 0.02) depletion of endogenous thyrotropin-releasing hormone (TRH) as compared to sham-operated controls 2 weeks postlesion. TRH levels remained unaltered in three other caudal regions. Bulbar kainate lesions produced a 58% (n = 5; p less than 0.001) decrease in TRH receptor binding capacity without affecting the receptor affinity. Kainate lesions also reduced bulbar muscarinic and benzodiazepine receptors by 60% and 48%, respectively. Again, no changes in TRH receptors were apparent in six other brain areas after bulbar kainate treatment. Injection of the dopaminergic neurotoxin, 6-hydroxydopamine (8 micrograms), into rat bulbs decreased TRH receptors by 35% (n = 4; p less than 0.05) 1 week postlesion. One month after surgical bulbectomy, TRH and TRH receptor levels in a number of brain areas were unaltered compared to those of control animals. These studies suggest that TRH in the olfactory bulb originates intrinsically and may be produced predominantly for local use. Secondly, TRH receptors in the bulb appear to be postsynaptically localized on intrinsic neurons, although a small proportion are also associated with presynaptic elements of dopaminergic noradrenergic neurons. Bulbar TRH receptors exhibited nanomolar affinity and a pharmacological selectivity akin to that of the pituitary gland and other brain regions. 相似文献
19.
Calbindin-D(28k) controls [Ca(2+)](i) and insulin release. Evidence obtained from calbindin-d(28k) knockout mice and beta cell lines 总被引:3,自引:0,他引:3
Sooy K Schermerhorn T Noda M Surana M Rhoten WB Meyer M Fleischer N Sharp GW Christakos S 《The Journal of biological chemistry》1999,274(48):34343-34349
The role of the calcium-binding protein, calbindin-D(28k) in potassium/depolarization-stimulated increases in the cytosolic free Ca(2+) concentration ([Ca(2+)](i)) and insulin release was investigated in pancreatic islets from calbindin-D(28k) nullmutant mice (knockouts; KO) or wild type mice and beta cell lines stably transfected and overexpressing calbindin. Using single islets from KO mice and stimulation with 45 mM KCl, the peak of [Ca(2+)](i) was 3.5-fold greater in islets from KO mice compared with wild type islets (p < 0.01) and [Ca(2+)](i) remained higher during the plateau phase. In addition to the increase in [Ca(2+)](i) in response to KCl there was also a significant increase in insulin release in islets isolated from KO mice. Evidence for modulation by calbindin of [Ca(2+)](i) and insulin release was also noted using beta cell lines. Rat calbindin was stably expressed in betaTC-3 and betaHC-13 cells. In response to depolarizing concentrations of K(+), insulin release was decreased by 45-47% in calbindin expressing betaTC cells and was decreased by 70-80% in calbindin expressing betaHC cells compared with insulin release from vector transfected betaTC or betaHC cells (p < 0.01). In addition, the K(+)-stimulated intracellular calcium peak was markedly inhibited in calbindin expressing betaHC cells compared with vector transfected cells (225 nM versus 1,100 nM, respectively). Buffering of the depolarization-induced rise in [Ca(2+)](i) was also observed in calbindin expressing betaTC cells. In summary, our findings, using both isolated islets from calbindin-D(28k) KO mice and beta cell lines, establish a role for calbindin in the modulation of depolarization-stimulated insulin release and suggest that calbindin can control the rate of insulin release via regulation of [Ca(2+)](i). 相似文献
20.
Hippocampal interneurons are local circuit neurons which are responsible for inhibitory activity in the hippocampus. Parvalbumin
(PV) is one of useful markers for GABAergic interneurons, not for principle cells, in the hippocampus. In the present study,
we investigated age-related changes in PV immunoreactive neurons and protein levels in the gerbil hippocampus during normal
aging. PV immunoreactive neurons were detected in all hippocampal subregions of all groups. PV immunoreactive neurons, which
innervated principal neurons, were non-pyramidal neurons in the hippocampal CA1-3 regions, and were polymorphic neurons in
the dentate gyrus. In the hippocampal CA1 region, the number of PV immunoreactive neurons was significantly reduced in the
postnatal month 3 (PM 3) group, which was sustained by PM 18, and, at PM 24, the number of PV immunoreactive neurons was significantly
decreased. In the CA2/3 region and dentate gyrus, the number of PV immunoreactive neurons was significantly decreased at PM
6: Thereafter, the number of PV immunoreactive neurons was sustained until PM 24. In addition, changes in PV protein levels
in the gerbil hippocampus were similar to immunohistochemical changes during normal aging: PV protein levels were significantly
decreased with age by PM 6: Thereafter, PV protein levels were sustained by PM 24. These results suggest that PV immunoreactive
interneurons were decreased in the hippocampus with age in gerbils. 相似文献