Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Calbindin 2”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 847 records · Page 47Linked to original sources
PubMed1998

The value of anti-calretinin antibody in the differential diagnosis of normal and reactive mesothelia versus metastatic tumors in effusion cytology.

In effusion cytology the distinction of reactive mesothelia from metastatic carcinoma cells may be a diagnostic challenge. Immunocytochemistry using antibodies suitable to detect epithelial cells must be considered carefully due to limited sensitivity and specificity of these antibodies. Efficient results in histological differential diagnosis of malignant mesothelioma versus lung-adenocarcinoma applying a novel antiserum against the calcium binding protein calretinin inspirated us to investigate the value of anti-calretinin antibody in effusion cytology combined with an epithelial marker. Cytoslides prepared by cytocentrifugation from 42 malignant and 65 reactive effusion specimens were immunostained using antibodies against calretinin and the epithelial marker Ber-EP4. Positive immunoreaction for calretinin in normal and reactive mesothelial cells was noted in 93% of the cases, whereas immunoreaction for calretinin was completely negative in the metastatic cells in 95% of the malignant effusions. Metastatic carcinoma cells were detected with anti-Ber-EP4 in 83% of malignant effusions. Non-specific positive reactions for Ber-EP4 in single mesothelial cells were observed in 16% of all cases and, moreover, frequently with macrophages or neutrophilic granulocytes. Our results demonstrate high sensitivity and specificity of anti-calretinin antibody for mesothelial cells in effusion specimens. They support its application to improve the diagnostic reliability of epithelial markers, especially because anti-calretinin antibody could be helpful in the assessment of false positive and false negative reactions of epithelial markers.

Antibodies, Monoclonal↗
PubMedDate not supplied

Immunocytochemical localization of calretinin in the olfactory system of the adult lamprey, Lampetra fluviatilis.

The distribution of calretinin immunoreactive (CR-ir) structures in the adult lamprey (Lampetra fluviatilis) olfactory system was studied by using immunocytochemical techniques. In the olfactory epithelium, a subpopulation of olfactory receptor cells was CR-ir. In the olfactory bulbs, three different cell populations were observed. Large CR-ir cells (mitral cells) were located medially to the olfactory glomeruli and occasionally between them. In the inner cellular layer, two types of CR-ir perikarya were found: numerous small CR-ir cells (granule cells) and some medium-sized CR-ir cells (putative displaced periglomerular cells). In addition, different intensities of CR-ir fibers were present in particular rootlets of the olfactory nerves, as well as in particular glomeruli. The presence of CR-ir cells and fibers in all layers of the lamprey olfactory bulbs supports the idea that this protein is present in pathways underlying the processing of sensory information throughout evolution.

Animals↗
PubMedDate not supplied

The telencephalon of the frog Xenopus based on calretinin immunostaining and gene expression patterns.

To further understand the organization and evolution of the telencephalon, we analyzed in the frog Xenopus laevis the expression of the genes Distal-less-4 (Xdll-4, comparable to the mouse gene Dlx2) and GAD-67 (XGAD-67, expressed in GABAergic cells), and compared this with calretinin immunostaining and the cytoarchitecture of the telencephalon. Our results show that like in other vertebrates, the telencephalon of the frog Xenopus is divided into two major territories: a basal, subpallial region showing a high density of cells expressing Xdll-4 and XGAD-67, and a dorsal, pallial region showing only few, dispersed cells expressing these genes. The subpallial territory of the frog Xenopus includes the septum, the amphibian basal ganglia, some basal forebrain cholinergic cell groups and some amygdala nuclei. In the pallium of the frog Xenopus, medial, dorsal, lateral, and ventral parts could be distinguished, similar to those described in amniotes. In summary, the amphibian telencephalon shows a basic morphogenetic organization similar to that of amniotes, which suggests that this organization is common to the telencephalon of all tetrapods.

Animals↗
PubMedDate not supplied

Organization of the mouse dorsal thalamus based on topology, calretinin immnunostaining, and gene expression.

To better understand the organization and evolution of the dorsal thalamus, we have made a first approach to analyze the possible histogenetic compartments of the mammalian dorsal thalamus using mouse embryos. For that, we have analyzed the expression of the proneural gene Math4a and the protein calretinin. Our results suggest the existence of rostrodorsal, caudoventral, and ventral compartments in the embryonic dorsal thalamus of the mouse, which partly parallel the dorsoventral histogenetic tiers postulated in the dorsal thalamus of sauropsids. The rostrodorsal compartment of the mouse dorsal thalamus is characterized by expression of Math4a, and it appears to include sensory and motor thalamic nuclei projecting to the dorsal pallium (isocortex). This compartment appears equivalent to the lemnothalamus proposed by Butler in tetrapods based on hodological grounds. The caudoventral and ventral compartments of the mouse dorsal thalamus lack expression of Math4a in the mantle, but they are characterized by several populations of calretinin-immunorective neurons that show projections to the claustroamygdaloid region in the ventrolateral pallium. More studies will be needed to analyze if the compartments proposed in this study represent true histogenetic units, and to find homologous developmental fields in all vertebrates.

Animals↗
PubMed1997

Calretinin-containing neurons in rat cerebellar granule cell cultures.

Using an antiserum against calretinin, a calcium-binding protein, we discovered two distinct neuronal cell types that stain intensely in enriched cerebellar granule cells. One neuronal cell type resembles unipolar brush cells, whereas the other resembles Lugaro cells. During early culture times, these calretinin-positive neurons are most numerous but represent less than one percent of the total neuronal population. In cultured cells, calretinin mRNA levels peak at day three in vitro, followed by a rapid decline to undetectable levels by day six in vitro. However, calretinin-immunoreactive neurons are observed up to 29 days in vitro. Excitotoxic concentrations of glutamate receptor agonists failed to elicit an excitotoxic response on the intensely staining calretinin-positive neurons, whereas greater than 95% of the cerebellar granule cells were susceptible to the excitotoxic actions of the glutamate receptor agonists. To distinguish between the two possibilities that calretinin-positive neurons either do not express glutamate receptors or they are not susceptible to the excitotoxic effects of glutamate receptor agonists, we performed immunocytochemistry using glutamate receptor antibodies to detect the presence of receptor protein. We found that the AMPA/kainate glutamate receptor (GluR2R3) colocalized with calretinin, suggesting that calretinin-immunoreactive neurons express the AMPA/kainate receptor; cerebellar granule cells, which are known to express this receptor, were also immunoreactive for the GluR2R3 receptor.

Animals↗
PubMed2000

Development of the auditory brainstem of birds: comparison between barn owls and chickens.

Birds have proved to be extremely useful models for the study of hearing function. In particular, chickens and barn owls have been widely used by a number of researchers to study diverse aspects of auditory function. These studies have benefited from the advantages offered by each of these two species, including differences of auditory specialization. Direct comparisons between chickens and barn owls become complicated when the degree of auditory specialization and their modes of development are brought into consideration. In this article we review the available literature on the development of the auditory brainstem of chickens and barn owls in the context of such differences. In addition, we present a time line constructed on the basis of common stages of structural differentiation, rather than chronological time. We suggest that such a time line should be considered when discussing comparative data between these two species. Such an approach should facilitate the interpretation of similarities and differences observed in the developmental processes of the auditory system of chickens and barn owls.

Animals↗
PubMed2001

Early bilateral deafening prevents calretinin up-regulation in the dorsal cortex of the inferior colliculus of aged CBA/CaJ mice.

This study was conducted to test the hypothesis that age-related calretinin (CR) up-regulation seen in the dorsal cortex of the inferior colliculus (ICdc) of old hearing CBA mice is dependent upon neural activity within the auditory pathway. We tested this hypothesis by bilaterally deafening young CBA/CaJ mice with kanamycin, and then aging them until 24 months. This manipulation mimics the lack of sound-evoked auditory activity experienced by old C57BL/6J mice, who are deaf and do not show CR up-regulation with age. Cell counts revealed that the density of CR+ cells in the ICdc of old hearing CBA mice was statistically different from old deafened CBA mice raised under identical conditions. Old hearing CBAs possessed an average of 27.54 more CR+ cells/100 microm2 than old deafened CBAs. When old deafened CBAs were compared to young hearing CBAs, young hearing C57s, and old deaf C57s, there was no significant difference in mean CR+ cell density in ICdc. Thus, only the old normal hearing CBAs showed an increase in CR+ cells with age, supporting the hypothesis that CR up-regulation depends upon sound-evoked activity. Moreover, these results demonstrate that up-regulation of CR expression was not simply due to a mouse strain difference.

Aging↗
PubMed2002

Hypoplasia of spiral and Scarpa's ganglion cells in GABA(A) receptor beta(3) subunit knockout mice.

This study documents morphologic alterations in the spiral ganglion and Scarpa's ganglion from gamma-aminobutyric acid A (GABA(A)) receptor beta(3) subunit null mutant mice. The ganglion cells of the mutant mice were hypoplastic in hematoylin&eosin-stained sections. Hypoplasia was observed at every location of the spiral ganglion and Scarpa's ganglion except the apical cochlear turn. Calretinin immunostaining demonstrated a selective hypoplasia of calretinin-negative cells at every location of spiral and Scarpa's ganglion cells, while the soma area of calretinin-positive cells was not affected by the gene deletion. Meanwhile, in the spiral ganglion of both wild type and knockout mice, there were apical to basal gradients in the soma size and the proportion of calretinin-positive cells. The absence of statistically significant hypoplasia in hematoylin&eosin sections through the apical turn of the cochlea can be explained by the relatively higher proportion of calretinin-positive ganglion cells, which were unaffected by the gene deletion. These findings suggest that GABA(A) receptor isoforms containing the beta(3) subunit may play an important role in the development and differentiation of non-calyceal terminals of Scarpa's ganglion cells and type II and smaller type I spiral ganglion cells.

Animals↗
PubMed2003

The effects of early bilateral deafening on calretinin expression in the dorsal cochlear nucleus of aged CBA/CaJ mice.

The aim of this study was to test the hypothesis that calretinin (CR) levels in the aged mouse auditory brainstem depend upon hearing ability. Old animals with good hearing, and thus higher sound-evoked activity levels, were predicted to have higher levels of CR immunoreactivity than old animals with hearing loss. CR immunoreactivity was analyzed in the deep layer (layer III) of the dorsal cochlear nucleus (DCN) in CBA/CaJ mice that were bilaterally deafened at 3 months of age with kanamycin, and then aged until 24 months. This manipulation partially mimics the lack of sound-evoked auditory activity experienced by old C57BL/6J mice, who are deaf at 24 months of age (but show residual hearing at 15 months) and have lower levels of CR immunoreactivity than old CBA mice with normal hearing [Hear. Res. 158 (2001) 131]. Cell counts revealed that the density of CR+ cells in DCN layer III of the deafened CBA mice was statistically different from old intact CBA mice raised under identical conditions. Old deafened CBAs showed a decline of 47% in the mean density of CR+ cells compared to old hearing CBAs, thus supporting the hypothesis. Interestingly, while there tended to be fewer CR+ cells in the old deaf C57s as compared to young C57s and young and old CBAs with normal hearing, the difference was not statistically significant. It is possible that the residual hearing of C57 mice at 15 months may provide sufficient auditory input to maintain CR at levels higher than CBA mice that are deafened completely at 3 months of age, and are profoundly deaf for a much longer time (21 months).

Aging↗
PubMed1998

Establishment of conditionally immortalized rat utricular epithelial cell lines using a retrovirus-mediated gene transfer technique.

Supporting cells in the inner ear sensory epithelium are most likely hair cell progenitors. In an effort to establish an in vitro model system of hair cell differentiation, we developed immortalized epithelial cell lines by transferring the tsA58 allele of the SV40 large T antigen oncogene into neonatal rat utricular supporting cells using a retrovirus. The established cell lines have been stably maintained continuously for more than 25 passages and display many features similar to primary supporting cells. They grow in patches and assume a polygonal morphology. Immunocytochemical characterization of the established cell lines reveals that these cells can be labeled by epithelial cell markers, but not by fibroblast, glial or neuronal markers. The immortalized cells grow rapidly in serum medium at permissive temperature, but the majority cease proliferation when cultured in serum free medium at non-permissive temperature. These cells respond to mitogenic growth factors including bFGF, EGF and TGF-alpha and express growth factor receptors in a manner similar to the primary supporting cells. Furthermore, we find that the cells undergo a morphological differentiation when cultured in serum free medium at non-permissive temperature in the presence of bFGF. Under these conditions, the cells shrink in size, become elongated, and express early hair cell markers such as calretinin and calmodulin. The utricular epithelial cell line we have established may potentially provide an invaluable system for studying hair cell differentiation and regeneration.

Alleles↗
PubMed1999

Developmental expression of calretinin-immunoreactivity in the thalamic eminence of the fetal mouse.

An investigation of the developmental expression of calretinin immunoreactivity and mRNA expression was carried out in the developing mouse diencephalon. Attention was focused on the thalamic eminence, which is a prominent structure previously described in the thalamus of the fetal mammalian diencephalon and adult lower vertebrates. Calretinin-positive staining was first observed in the thalamic eminence beginning at embryonic day 11. In situ hybridization histochemistry confirmed the presence of calretinin mRNA in the thalamic eminence. During subsequent embryonic development calretinin expression was very intense in neurons in the thalamic eminence though embryonic day 17, and thereafter, was increasingly difficult to distinguish. By postnatal day 0 the thalamic eminence was no longer discernable. Additional neurons within the murine diencephalon also expressed calretinin positive cell bodies and, or neuronal processes, including the stria medullaris, the habenular commissure and the paraventricular thalamic nucleus. It is possible that the thalamic eminence may form during development in order to act as an organizing center for the diencephalon.

Animals↗
PubMed1999

Distribution of calretinin-like immunoreactivity in the brain of Rana esculenta.

The distribution of calretinin-like immunoreactivity has been analyzed in the brain of Rana esculenta. Several neurons of nuclei belonging to sensory pathways, subhabenular area and left habenula were immunopositive. Immunoreactivity was present in fibers of motor and sensory pathways, thalamus, tegmentum and isthmus. The immunolabeling pattern partially overlapped that previously described in the rat. However, in comparison with the rat, fewer cells and fibers were immunoreactive and there were less positive brain nuclei. especially in the pallium, septum and striatum, that were totally negative. Taking into consideration that these regions are rather simple in the frog, the presence of calretinin seems to be consistent with the degree of complexity of brain areas and segregation of different nuclei.

Animals↗
PubMed2002

A selective decrease in the relative density of parvalbumin-immunoreactive neurons in the hippocampus in schizophrenia.

Neuropathological studies have demonstrated deficits of GABAergic interneurons in the hippocampus in schizophrenia. and selective deficits in some GABAergic sub-populations defined by calcium-binding proteins (CBPs) have been reported in the cortex in schizophrenia. In the present study, the relative densities of cells immunoreactive for the CBPs parvalbumnin (PV) and calretinin (CR) were determined in hippocampal tissue sections taken from patients with schizophrenia, bipolar disorder and major depression and from matched control subjects (15 per group). No significant difference in the density of CR-immunoreactive neurons was found between subject groups. Relative to normal controls, schizophrenic patients showed a significant and profound deficit in the relative density of PV-immunoreactive neurons in all hippocampal sub-fields. These reductions were more apparent in male than female schizophrenic patients, and were unrelated to antipsychotic drug treatment, age or duration of illness. The density of PV-immunoreactive neurons did not differ significantly from controls in the depression group, although a trend toward decreased relative density of PV-immunoreactive neurons was apparent in bipolar disorder that reached significance in one sub-field. The findings provide further evidence to support a profound and selective abnormality of a sub-population of GABAergic neurons in the hippocampus in schizophrenia.

Bipolar Disorder↗
PubMed2003

Miliary pulmonary metastases from a clinically occult pleural mesothelioma.

Mesothelioma is a rare neoplasm of the serosal membranes. Signs and symptoms of a pleural effusion typically herald discovery of the tumor. We report a case of miliary metastatic mesothelioma involving both lungs in a 54-year-old man who presented with right-sided chest discomfort, numerous pulmonary nodules detected by computed tomography of the chest, and absent pleural effusion. Immunohistochemical and electron microscopy studies performed on wedge biopsies of parenchymal pulmonary nodules led to the diagnosis of metastatic mesothelioma. Subsequent pleural evaluation and biopsy of pleural thickening noted at a site of prior chest wall trauma identified the primary neoplasm and confirmed the diagnosis as malignant epithelioid mesothelioma. The histologic appearance of discohesive epithelioid cells in a distinctly myxoid background was the clue in this case leading to the consideration of metastatic mesothelioma and a thorough immunohistochemical evaluation of the tumor. This case shows that mesothelioma may metastasize throughout the lungs in a miliary pattern and the metastases may be clinically detected before the primary pleural tumor. Metastatic mesothelioma is a consideration for metastatic pulmonary tumors of unknown origin.

Biomarkers, Tumor↗
PubMed1998

Two-laser dual-immunofluorescence confocal laser scanning microscopy using Cy2- and Cy5-conjugated secondary antibodies: unequivocal detection of co-localization of neuronal markers.

The ability of the confocal laser scanning microscope (CLSM) to visualize in one focal plane the fluorescence associated with multiple markers renders this instrument extremely valuable for the study of co-localization of various markers in the somata and cellular processes of neurons. In the present protocol we deal with the question whether or not co-localization exists in neurons of two different neuronal markers. The conventionally used method towards answering this type of question is double-immunofluorescence microscopy. Fundamental to this approach, independent from whether the preparations are observed in a normal fluorescence microscope or in a CLSM, is that each of the applied fluorescent labels should not chemically interact with the other label or inadvertently be visible through the illumination/filter setup designed for the other fluorophore. In the field of double-label CLSM, three types of approach are distinguished: the single-laser, two-color approach, the two-laser, two-color approach, and the time-resolved approach (Brismar and Ulfhake, 1997). Each type of approach has its own advantages and disadvantages. In the instrument in our institute (a Zeiss LSM 410), combinations of fluorophores like fluorescein isothiocyanate (FITC) and tetramethyl rhodamine isothiocyanate (TRITC) are less useful, since TRITC produces a detectable signal in the FITC illumination/filter setup. Instead of experimenting with filter sets we have chosen to take two measures to eliminate this problem. Our first measure is to use fluorophores whose absorption/emission spectra overlap as little as possible. We have selected among the recently developed carbocyanine fluorophores one fluorescing in the visible range (Cy2) (green, in the same range as FITC and with much better resistance to fading than FITC; cf. Härtig et al., 1996), and another fluorescing in the near infrared range (Cy5, infrared; cf. Mesce et al., 1993). Our second measure to ensure excellent signal separation is the adoption of a two-laser, two-color approach. Co-localization of the calcium binding protein, calretinin, and a neurotransmitter, gamma-aminobutyric acid (GABA), in interneurons in the entorhinal cortex and the hippocampus of the rat was used as the principal test model. We compare the above two-laser, two-color approach with a single-laser, two-color CLSM approach using as markers Cy2 and the red fluorophore, Texas Red (physical characteristics resembling TRITC). In this paper considerable attention is paid to control experiments to verify the reliability of the staining procedure. The results show that our two-laser, two-color CLSM approach produces a complete and unambiguous separation of the fluorescent labels, Cy2 and Cy5. We are currently using this method to determine the degree of co-localization of neurochemical substances in CNS neurons.

Antibodies↗
PubMed2002

The distribution of P2X3 purine receptor subunits in the guinea pig enteric nervous system.

Adenosine 5'-triphosphate (ATP) excites 70-90% of enteric neurons through P2X type purine receptors, and is likely to be an enteric neurotransmitter. Recent studies indicate that the P2X2 subunit is expressed by specific subgroups of enteric neurons, and that there are enteric neurons that are responsive to ATP but lack this subunit. In the present work, we have investigated whether the P2X3 subunit is similarly localised to specific subgroups of neurons, and whether these are different from the P2X2 subunit-expressing neurons. The P2X3 subunit was localised by immunohistochemistry to nerve cells of the myenteric ganglia of the stomach, small and large intestines, and nerve cells of the submucosal ganglia in the small and large intestines of the guinea pig. All immunoreactivity was absorbed with the P2X3 receptor peptide against which the antiserum was raised. In myenteric ganglia of the ileum, P2X3 receptor immunoreactivity was in calretinin, enkephalin and nitric oxide synthase (NOS)-immunoreactive neurons. In submucosal ganglia, all calretinin-immunoreactive nerve cells were P2X3 receptor immunoreactive. In the submucosal ganglia of the ileum, 13 +/- 3% of neuropeptide Y (NPY)-immunoreactive neurons were also P2X3 receptor immunoreactive, whereas in the distal colon, almost all NPY-expressing nerve cells were P2X3 receptor immunoreactive. The localisation of the P2X3 subunit was largely distinct from that of the P2X2 subunit, although both subunits occur in some NOS neurons, where P2X2 and P2X3 subunits may form heteromeric receptors. Unlike the P2X2 subunit, the P2X3 subunit is not expressed in intrinsic sensory neurons in the ileum. It is concluded that the P2X3 receptor subunit is expressed in specific functional groups of neurons; the major types are excitatory and inhibitory muscle motor neurons, ascending interneurons and cholinergic secretomotor neurons.

Animals↗
PubMedDate not supplied

Most calretinin-containing amacrine cells in the rabbit retina co-localize glycine.

Calretinin-containing retinal amacrine cells are heterogeneous with regard to their neurochemical properties. In the rabbit retina, about 90% of them contain glycine, as evidenced in the present study by double-label immunocytochemistry. In a previous report, we showed that a small population of amacrine cells contains both gamma-aminobutyric acid and calretinin. In this study, we further identified this cell population by means of known secondary markers. However, none of the markers we tested (choline acetyltransferase, serotonin accumulation, NADPH-diaphorase, vasoactive intestinal polypeptide) co-localized with calretinin. A small population (1%) of the cells in the ganglion cell layer contains both calretinin and glycine. Since calretinin-positive cells in the ganglion cell layer have been identified as ganglion cells based on soma size and presence of calretinin-positive axons in the optic nerve fiber layer, this population may represent a class of ganglion cell which contains glycine. Our results, together with those of other studies, suggest that calretinin is not a general marker of any of the well-known amacrine cell types in the mammalian retina. Rather, calretinin, just as other calcium-binding proteins, is distributed in a species-specific manner. At the same time it appears that, as shown for horizontal cells, one or more of the major buffer-type calcium-binding proteins of the EF-hand family is present in most of the retinal amacrine cells.

Animals↗
PubMed1995

Conformational changes and calcium binding by calretinin and its recombinant fragments containing different sets of EF hand motifs.

Four recombinant fragments, representing different sets of EF-hand motifs of rat calretinin (CR) (I-II, I-III, III-VI, IV-VI), were prepared, and their Ca2+ -induced conformational changes were compared with those of full-length recombinant CR. All fragments were able to bind calcium ions as shown by 45Ca2+ overlay method on nitrocellulose and fluorescence measurements. The intrinsic tryptophan fluorescence intensity (FI) of apo-CR reversibly increased about 3-fold upon addition of calcium, indicating a change of conformation. The FI of fragments I-II (Trp 25) and I-III (Trp 25 and 116) increased about 1.4-fold on calcium binding, but that of fragment III-VI (Trp 116) increased 3.5-fold. Calcium titration of CR monitored by Trp fluorescence intensity showed that recombinant CR and some fragments bound Ca2+ with high affinity (Kd below 0.4 microM) and with high cooperativity. An apparent Hill coefficient for Ca2+ -induced fluorescence changes in CR was about 3.7. CR bound to organomercurial-agarose Cys 101 and 266 did not form cystine. The fluorescence intensities of cysteine-linked fluorescent probes 5-iodoacetamidofluorescein and N-(1-pyreneiodoacetamide) were increased approximately 1.3-fold upon calcium binding by CR. These data indicate that CR binds Ca2+ with high affinity and cooperativity and that this binding induces a change of conformation that involves the interaction of different parts of the molecule. Taken together, our results suggest that CR works as an on/off switch within a narrow range of free Ca2+ by interacting with as yet unidentified targets.

Animals↗