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PubMed1999

BDNF attenuates retinal cell death caused by chemically induced hypoxia in rats.

PURPOSE: To investigate the neuroprotective effects of brain-derived neurotrophic factor (BDNF) against potassium cyanide (KCN)-induced retinal damage. METHODS: Rats were injected intravitreally with iodinated BDNF. Two days later, eyeballs were dissected into various parts, and the level of radioactivity in each part was measured. Retinal damage was induced by incubating rat eyeballs with 5 mM KCN. BDNF was injected intravitreally 2 days before KCN treatment, and subsequent morphometric analysis was carried out to evaluate the retinal cell damage. To elucidate the mechanisms of BDNF's neuroprotective effects, the intravitreal concentrations of amino acids and the expression of calretinin were investigated. RESULTS: Intravitreally injected BDNF was distributed evenly throughout the eyes, and the incorporation of iodinated BDNF into the retina was three times higher than in other ocular tissues. Immunohistochemical analysis demonstrated that exogenous BDNF diffused throughout the retina and was especially concentrated in the inner (INL) and outer nuclear layer. Morphometric analysis showed that the number of INL cells of the posterior area, 880 microm from the optic nerve head, was 190 +/- 4 with KCN treatment and 284 +/- 9 in control animals. Cell death appeared to be necrotic. When eyes injected with either phosphate-buffered saline (PBS) or BDNF were subjected to treatment with KCN, the number of INL cells was 186 +/- 5 in the PBS-treated controls and 253 +/- 8 in eyes treated with BDNF. Also, BDNF increased the number of calretinin-positive cells in the INL and reduced the KCN-induced elevation of intravitreal glutamate levels. CONCLUSIONS: BDNF injected intravitreally reaches the retina and attenuates the INL cell death caused by KCN-induced metabolic insult. The neuroprotective effects of BDNF are partly ascribed to the upregulation of a calcium-binding protein and the attenuation of glutamate release into the vitreous body.

Animals↗
PubMed1997

Orally projecting interneurones in the guinea-pig small intestine.

1. Orally projecting, cholinergic interneurones are important in mediating ascending excitatory reflexes in the small intestine. We have shown that there is just one major class of orally projecting interneurone, which we have characterized using retrograde labelling in organ culture, combined with immunohistochemistry, intracellular recording and dye filling. 2. Orally projecting interneurones, previously shown to be immunoreactive for choline acetyltransferase, tachykinins, enkephalin, calretinin and neurofilament protein triplet, have axons up to 14 mm long and are the only class of cells with orally directed axons more than 8.5 mm long. 3. They are all small Dogiel type I neurones with short dendrites, usually lamellar in form, and a single axon which sometimes bifurcates. Their axons give rise to short varicose collaterals in myenteric ganglia more than 3 mm oral to their cell bodies. 4. Orally projecting interneurones receive prominent fast excitatory post synaptic potentials (fast EPSPs). A major source of fast EPSPs is other ascending interneurones located further aborally. They also receive fast EPSPs from circumferential pathways. 5. In the stretched preparations used in this study, orally projecting interneurones were highly excitable, firing repeatedly to depolarizing current pulses and had negligible long after-hyperpolarizations following their action potentials. They did not receive measurable non-cholinergic slow excitatory synaptic inputs. 6. Ascending interneurones had a characteristic inflection in their membrane responses to depolarizing current pulses and their first action potential was typically delayed by approximately 30 ms. Under single electrode voltage clamp, ascending interneurones had a transient outward current when depolarized above -70 mV from more hyperpolarized holding potentials. Ascending interneurones also consistently showed marked inward rectification under both current clamp and voltage clamp conditions. 7. This class of cells has consistent morphological, neurochemical and electrophysiological characteristics and are important in mediating orally directed enteric reflexes.

Animals↗
PubMed1997

Calretinin neurons in human medial prefrontal cortex (areas 24a,b,c, 32', and 25).

The calcium-binding protein calretinin (CR) is present in a subpopulation of local-circuit neurons in the mammalian cerebral cortex containing gamma-aminobutyric acid. This light microscopic investigation provides a detailed qualitative and quantitative morphological analysis of CR-immunoreactive (CR+) neurons in the medial prefrontal cortex (mPFC; areas 24a,b,c, 32', and 25) of the normal adult human. The morphology of CR+ neurons and their areal and laminar distributions were consistent across human mPFC. The principal organisational features of CR+ labelling were the marked laminar distribution of immunoreactive somata and the predominantly vertical orientation of labelled axon-like and dendritic processes. Several types of CR- neurons were present in layer 1, including horizontally aligned Cajal-Retzius cells. In layers 2-6, CR+ neurons displayed a variety of morphologies: bipolar cells (49% of CR+ population), vertically bitufted cells (35%), and horizontally bitufted cells (3.5%). These neuron types were mainly located in layer 2/upper layer 3, and their dendritic processes were commonly aspiny and sometimes highly beaded. Aspiny (8%) and sparsely spiny multipolar (5%) CR+ neurons were also found. The mean somatic profile diameter of CR+ cells was 11.6 +/- 0.3 microm (mean +/- S.D). CA+ puncta formed pericellular baskets around unlabelled circular somatic profiles in layers 2/3 and around unlabelled pyramidal-shaped somata in layers 5/6. The somatic sizes of these unlabelled cell populations were significantly different. Immunolabelled puncta were also found in close contact with CR+ somata. Cortical depth distribution histograms and laminar thickness measurements defined the proportions of the overall CR- cell population in each layer: 7% in layer 1, 78% in layers 2/3, 14% in layers 5/6, and 1% in the white matter. In the tangential plane, CR+ neurons were distributed uniformly at all levels of the cortex. By using stereological counting procedures on immunoreacted Nissl-stained sections, CR+ neurons were estimated to constitute a mean 8.0% (7.2-8.7%) of the total neuron population in each cortical area. These data are compared with similar information obtained for the mPFC in monkey and rat (Gabbott and Bacon [1996b] J. Comp. Neurol. 364:657-608; Gabbott et al., [1997] J. Comp. Neurol. 377:465-499). This study provides important morphological insights into a neurochemically distinct subclass of local-circuit inhibitory neurons in the human mPFC.

Adult↗
PubMed1996

Differential effects of excitatory amino acids on mesencephalic neurons expressing either calretinin or tyrosine hydroxylase in primary cultures.

In mesencephalic primary cultures derived from E14 rat embryos, calretinin- and tyrosine hydroxylase-immunoreactive neurons comprised 2% and 5% of the total cell population, respectively, at 6-7 days in vitro. The number of calretinin-immunoreactive neurons was unchanged after a 12- or 24-h exposure to 500 microM kainic acid (KA), but a 50% cell loss was detected after a 48-h exposure to KA. Tyrosine hydroxylase-immunoreactive neurons demonstrated a 50% and 67% cell loss at 24- and 48-h exposures to 500 microM KA. A 500 microM N-methyl-D-aspartic acid (NMDA) incubation for 24 h had no effect on calretinin-immunoreactive cell number, but did significantly reduce tyrosine hydroxylase-immunoreactive cell numbers by 26%. In tyrosine hydroxylase-immunoreactive cells, exposure to KA appeared to stimulate the retraction of the neuritic tree and to cause somatic swelling. In contrast, calretinin-immunoreactive neurons developed larger and more complex neuritic trees after a 24-h exposure to 500 microM KA but not NMDA. Immunohistochemical colocalization studies revealed that all tyrosine hydroxylase-immunoreactive and the majority of calretinin-immunoreactive neurons expressed the glutamate receptor subunits GluR2-R3. Very low levels of NMDAR1 receptor subunits were detected on cells in this culture and GluR4 receptor subunits were not detectable. Our experiments showed that glutamate receptors present in both calretinin- and tyrosine hydroxylase-immunoreactive cells were functional, since phosphorylated cAMP/Ca2+ response element-binding protein levels were increased in both cell types after 10 or 30 min exposures to 500 microM KA. The present results indicate that in the mesencephalic cultures tyrosine hydroxylase-immunoreactive cells are more vulnerable to KA excitotoxicity than calretinin-immunoreactive neurons.

Animals↗
PubMed1996

Distribution of the calcium-binding proteins parvalbumin and calretinin in the auditory brainstem of adult and developing rats.

Parvalbumin (PV), calretinin (CR), and calbindin (CB) are calcium-binding proteins which are presumably involved in the regulation of the intracellular calcium concentration. Within the rat auditory system, CB is transiently expressed in several nuclei during the period of synapse refinement, indicating a specific function of CB during development, yet little is known in this regard about PV and CR. In order to gather more information about calcium-binding proteins during development, we analyzed the spatiotemporal distribution of PV and CR in the rat auditory brainstem using immunocytochemistry. In the adult, PV was heavily present in somata and neuropil of all nuclei and in fibers of all tracts. CR was found in somata of the cochlear nucleus and peripheral aspects of the inferior colliculus as well as in fibers extending into the superior olivary complex and the nuclei of the lateral lemniscus. The developmental expression of PV was characterized by a relatively late appearance in somata (at postnatal day 8), followed by a rapid increase to adult levels. In contrast, CR immunoreactivity was already strong two days before birth, yet the number and intensity of labeled neurons subsequently decreased and CR disappeared almost completely in the superior olivary complex, nuclei of the lateral lemniscus, and central aspects of the inferior colliculus. These data, together with those on CB, show that CR, CB, and PV are sequentially expressed during auditory brainstem development. They also suggest that the presence of the three proteins can be correlated with definite developmental stages.

Aging↗
PubMed1996

Calretinin-immunoreactive cells and fibers in the human amygdaloid complex.

Calretinin is a calcium-binding protein that colocalizes with GABA in the cerebral cortex and hippocampus of the rat and the monkey. In the present study, we investigated the distribution of calretinin-immunoreactive cells and fibers in the human amygdaloid complex. A conspicuous feature was the high density of calretinin neurons in the human amygdala. The highest densities of the calretinin-immunoreactive neurons were observed in the anterior cortical nucleus, accessory basal nucleus, amygdalohippocampal area, and in the nucleus of the lateral olfactory tract. The paralaminar nucleus, central nucleus, medial nucleus, and the periamygdaloid cortex contained the lowest densities of calretinin neurons. In most of the amygdaloid areas, the calretinin cells had the appearance of aspiny or sparsely spiny local circuit neurons. However, in the amygdalohippocampal area, we found also densely spined dendrites. The cortical areas and the central nucleus were characterized by intense neuropil labeling, while the deep nuclei contained a high density of calretinin-immunoreactive fibers and terminals. Calretinin immunoreactivity was also found in the intra-amygdaloid fiber bundles, stria terminalis, and in the ventral amygdalofugal pathway. This suggests that in addition to the local circuit neurons, calretinin immunoreactivity is also located in neurons that connect the amygdaloid complex with the other brain areas. The distribution and morphological characteristics of calretinin-immunoreactive neurons differed from those of another calcium-binding protein, parvalbumin, in the human amygdala (Sorvari et al. [1995] J. Comp. Neurol. 360:185-212). This suggests that these two calcium-binding proteins are located in different populations of neurons.

Adult↗
PubMed1996

Distribution of calretinin-immunoreactive septal axons in the normal and deafferented medial habenula of adult rats.

To characterize the neural circuitry and plasticity of the septohabenular pathway, the present study analyzes the distribution of calretinin-immunoreactive fibers within the normal and deafferented medial habenula (MHb) at the light and ultrastructural levels. In the adult rat, a dense plexus of calretinin-positive fibers was found throughout the entire MHb neuropil; these immunoreactive terminals formed asymmetric synaptic contacts with unstained dendritic profiles. Calretinin-positive axons that innervate the MHb originated from neurons of the ipsilateral posterior septum, specifically those of the nucleus septofimbrialis and the nucleus triangularis. Unilateral deafferentation of the MHb resulted in the complete loss of calretinin-immunostained fibers within the ipsilateral MHb after 7 days; no reduction was apparent on the contralateral side. Four weeks after unilateral MHb deafferentation, new calretinin-immunoreactive fibers were found confined to the caudal regions of the MHb, these axons again formed asymmetrical contacts with unstained dendritic profiles. No calretinin-positive axons, however, were found within the MHb at 4 weeks following bilateral deafferentation, thus suggesting that the source of these new fibers within the long-term deafferented MHb arises from the contralateral septal neurons. Supporting this idea, injections of biotinylated dextran amine into the 4-week deafferented MHb resulted in retrogradely labeled somata observed in the contralateral posterior septum. These data reveal that septal projections to the MHb, which are normally ipsilateral, respond to a unilateral deafferentation by extending contralateral fibers that cross the midline at the habenular commissure and reinnervate the caudal regions of the nucleus.

Animals↗
PubMed1997

Calretinin in the thalamic reticular nucleus of the rat: distribution and relationship with ipsilateral and contralateral efferents.

In order to investigate the existence of anatomical subdivisions within the thalamic reticular nucleus (Rt), the distribution of reticular neurons expressing the calcium binding protein calretinin was investigated in the rat by means of immunocytochemistry. Calretinin immunoreactive (Cr-ir) neurons were mainly distributed in the lateral and ventral regions, and along the medial border of the Rt rostral pole. Caudal to the rostral pole, many neurons were Cr-ir in the more dorsal part of the rostral two-thirds (the "dorsal cap") of the Rt. Fewer Cr-ir neurons were present more caudally along the lateral and medial borders, and in the caudalmost part of the nucleus, related to the acoustic thalamus. The distribution of Cr-ir neurons in the rostral Rt was compared with that of neurons projecting to the ipsilateral and contralateral anterior, intralaminar, midline, and mediodorsal nuclei, or to the contralateral rostral Rt. The retrograde transport of Fluorogold revealed a remarkably precise topography of the rostral Rt: different reticular areas were found to project to different thalamic nuclei, or to different rostrocaudal or mediolateral portions of the same thalamic nucleus, with a limited degree of overlap. The double-labeling experiments demonstrated that the reticular neurons projecting to the ipsilateral anterodorsal, midline, mediodorsal, and anterior intralaminar nuclei frequently expressed calretinin; by contrast, the majority of the reticular commissural neurons did not express the protein, with the exception of neurons projecting to the contralateral mediodorsal and midline nuclei. The ipsilaterally projecting calretinin-positive neurons were frequently located along the medial edge of the rostral pole and in the dorsal cap of the nucleus, segregated from the commissural calretinin-negative neurons. The combined analysis of calretinin expression patterns and tract tracing data provided further insight in the anatomical organization of the thalamic reticular nucleus, suggesting a different neurophysiological role for the ipsilaterally vs. the contralaterally projecting reticular neurons in the modulation of the synaptic activity of the dorsal thalamus.

Animals↗
PubMed1998

A novel disruption of cortical development in p35(-/-) mice distinct from reeler.

The p35/cdk5 neuronal-specific kinase complex has been shown to play an important role in the laminar configuration of cortical neurons. Mice lacking either p35 or cdk5 exhibit a disrupted cortical lamination pattern. We showed previously that instead of the normal "inside-out" layering pattern of cortical neurons, cortical neurons are layered from "outside-in" in p35 mutant mice. To gain insight into the mechanisms that underlie these defects, we examined the organization of landmark structures formed during cortical development and the migratory behavior of p35(-/-) cortical neurons by using bromodeoxyuridine labeling. In the present study, we show that reelin localization in the marginal zone is normal in p35 mutant mice. Furthermore, the preplate splits into the marginal zone and subplate properly, a developmental event that fails to occur in reeler mice. Finally, the migration of the earliest born cortical plate neurons is normal in p35 mutant mice; cortical neurons subsequently generated remain underneath these neurons. These data suggest that the p35/cdk5 kinase is required for cortical plate neurons to migrate past preexisting neurons and take up superficial positions to constitute the inside-outside layering order of cortical lamination.

Animals↗
PubMed1998

Calretinin in pretecto- and olivocerebellar projections in the chick: immunohistochemical and experimental study.

Calretinin (CaR) is a calcium-binding protein that is distributed extensively in the central nervous system. It is localized in the cell bodies and neurites of specific neuronal populations and serves, therefore, as a reliable anatomical marker. Some components of the pretectocerebellar projection, which connects specific pretectal nuclei to caudal cerebellar folia, are concerned with the cerebellar control of visual reflexes. We investigated the distribution of pretectocerebellar-projecting neurons in relation to cells that show CaR immunoreactivity. Cells that project to the cerebellar cortex in the diencephalic primary visual nuclei and in other grisea, like the nucleus spiriformis medialis and the nucleus dorsofrontalis, colocalized with those that appeared to be immunolabeled intensely with anti-CaR antiserum. To explore the hypothesis of a common developmental origin of these pretectal cerebellopetal neurons, we also investigated the development of CaR-immunopositive cells in the chick pretectum and the arrival of their fibers in the cerebellum, from 10 days of incubation (stage 36) to posthatching stages. Finally, we analyzed the source of CaR+ climbing fibers and found a subpopulation of CaR+ cells in the inferior olivary nucleus. On the whole, these results suggest that there is a common developmental origin of pretectal cerebellopetal neurons, some of which share the property of CaR expression. The functional significance of this correlation needs to be investigated.

Afferent Pathways↗
PubMed1999

Spiny calretinin-immunoreactive neurons in the hilus and CA3 region of the rat hippocampus: local axon circuits, synaptic connections, and glutamic acid decarboxylase 65/67 mRNA expression.

We have used the Golgi method and Golgi electron microscopic techniques to analyze the axonal arborization and efferent connections of spiny calretinin-immunoreactive neurons in the CA3 region and hilus of the rat hippocampal formation. In the hilus, the axons of spiny calretinin-immunoreactive neurons sent out numerous collaterals that arborized in the hilar region and the molecular layer. In the CA3 region, these axons extended mainly to the stratum radiatum and pyramidal layer but also to the stratum oriens and stratum lacunosum-moleculare. Axonal varicosities were distributed widely throughout the axonal collaterals. Electron microscopic studies revealed that the axon terminals of spiny calretinin-immunoreactive neurons established synaptic contacts mainly with dendritic shafts. We next analyzed the expression of glutamic acid decarboxylase (GAD65/67) mRNAs in spiny nonpyramidal neurons that were identified by calretinin immunoreactivity. We found that spiny calretinin-positive neurons in the CA3 region and hilus of the rat hippocampal formation expressed the two isoforms of GAD: GAD65 and GAD67 mRNAs. These findings show that the spiny calretinin-immunoreactive neurons of hippocampus give rise to local axonal arborizations, suggesting that they are inhibitory.

Animals↗
PubMed1999

Immunochemical localization of calretinin in the retina of the turbot (Psetta maxima) during development.

The expression of the calcium-binding protein calretinin was analysed by immunohistochemistry techniques in the retina of turbot (Psetta maxima) from embryonic to juvenile stages. Calretinin immunoreactivity was first detected in retinae from newly hatched larvae, in which the anlage of the inner plexiform layer and a subset of amacrine and ganglion cells displayed a faint immunolabelling. First appearance of photoreceptors during larval life coincided with an increase in the intensity of the labelling. During subsequent larval development, the expression of calretinin affected distinctive retinal components. The inner plexiform layer, optic fiber layer, and a population of amacrine and ganglion cells were invariably labelled. Occasional bipolar cells were labelled at the end of the larval period. By metamorphosis, calretinin is sequentially expressed in horizontal cells, and bipolar immunoreactive cells become numerous. The pattern of calretinin immunoreactivity of the inner plexiform layer changes from the larval to juvenile period. In all cases, calretinin immunoreactivity exhibited variations between the peripheral retina, which contains the most recently differentiated retinal components, and the remainder of the differentiated retina. Our results suggest that the progressive expression of calretinin in the turbot retina appears associated with some degree of neuronal differentiation. Once the definitive pattern of calretinin immunoreactivity is established in the turbot retina, both similarities and differences with the calretinin location in the retina of other vertebrates can be demonstrated.

Animals↗
PubMed1999

Laminar boundaries persist in the hippocampal dentate molecular layer of the mutant Shaking Rat Kawasaki despite aberrant granule cell migration.

The present report provides the first detailed description of the hippocampus in the Shaking Rat Kawasaki (SRK) mutant by using a panel of antibody markers to delineate its laminar organization. The mutant was characterised at postnatal day 21 by severe malformations of both neuronal position and orientation, the most striking of which was the presence of a rounded central granule cell mass in the dentate gyrus rather than the normal V-shaped granule cell layer. Despite this finding, the SRK dentate gyrus not only retained a cell-sparse molecular layer (thinner but similar in gross appearance to that of control littermates), but the sharp laminar boundary between its inner and outer parts was as clearly marked by IM1 and OM4 antibody staining as it was in the normal dentate gyrus. These immunocytochemical data suggest that the entorhinal terminal field of the dentate gyrus may be relatively normal in the mutant, despite entorhinal afferents appearing to take an abnormal trajectory after they fail to cross the hippocampal fissure. Laminar malformations included disruption of the SRK pyramidal cell layer, with spreading of the CA3 mossy fibre projection to an ectopic infrapyramidal position, radial displacement of CA1 pyramids, and transposition of a hitherto unremarked longitudinal fibre bundle immunoreactive for calretinin from its normal position in the stratum lacunosum-moleculare of field CA2 to an alvear position in SRK. The SRK malformations were very like but not identical to those seen in the mouse reeler mutant, suggesting similar underlying developmental mechanisms.

Animals↗
PubMed1999

Development of layer I of the human cerebral cortex after midgestation: architectonic findings, immunocytochemical identification of neurons and glia, and in situ labeling of apoptotic cells.

The development of layer I was studied in the human frontal cortex from 21 weeks of gestation (GW) to 2.5 postnatal months in series of adjacent sections processed for thionin staining, Bodian silver staining, and immunocytochemical labeling of neurons and glia. In addition, the terminal dUTP nick-end labeling (TUNEL) method was used to label in situ DNA fragmentation. A progressive decrease of cell density and the disappearance of the subpial granular layer (SGL) appeared as distinctive developmental features of human layer I, consistently with previous investigations. The neuronal antigen microtubule-associated protein2 was found to label preferentially Cajal-Retzius cells and dendritic processes extending from the cortical plate. At midgestation, the calcium binding protein calretinin stained in the marginal zone numerous neurons, including the Cajal-Retzius cells and their processes. Calretinin-immunoreactive neurons decreased during the subsequent maturation: such decline was abrupt in the SGL, whereas bipolar calretinin-immunopositive cells accumulated in the inner marginal zone to be presumably incorporated into the cortical plate. Cajal-Retzius cells expressed calretinin throughout the examined developmental stages. The glial antigen vimentin was already expressed at midgestation, and vimentin immunopositivity decreased progressively in cell bodies and fibers of layer I during development. Glial fibrillary acidic protein-positive elements gradually matured, and the positive cell bodies displayed the features of mature astrocytes at the end of gestation. Moreover, a decrease of free glial cells was observed in layer I, suggesting their progressive incorporation into the cortical plate. TUNEL-positive cells were detected at midgestation in the marginal zone, and they were concentrated in the SGL until its disappearance; their number decreased dramatically throughout layer I after 30 gestational weeks. TUNEL-positive nuclei or regressive changes were not detected in Cajal-Retzius cells throughout the examined developmental stages. Thus, our data point out that naturally occurring cell death is an active mechanism contributing to the disappearance of the SGL but not to the subsequent developmental reshaping of human layer I, in which, instead, migratory phenomena should play a major role. In addition, our findings argue against a disappearance of Cajal-Retzius cells due to regressive processes.

Apoptosis↗
PubMed1999

Development of calretinin immunoreactivity in the brainstem auditory nuclei of the barn owl (Tyto alba).

The early development of calretinin immunoreactivity (CR-IR) was described in the auditory nuclei of the brainstem of the barn owl. CR-IR was first observed in the auditory hindbrain at embryonic day (E17) and a day later (E18) in the inferior colliculus. In each of the auditory nuclei studied, CR-IR did not develop homogeneously, but began in the regions that map high best frequencies in the adult barn owl. In the hindbrain, CR-IR was first observed in the rostromedial regions of the cochlear nucleus magnocellularis and the nucleus laminaris, and in the dorsal regions of the nucleus angularis and in the nucleus of the lateral lemniscus. In the inferior colliculus, CR-IR began in the ventral region of the central core. The edge of these gradients moved along the future tonotopic axes during the development of all nuclei studied, until adult patterns of CR-IR were achieved about a week after hatching.

Animals↗
PubMed2000

Pattern of calretinin immunoreactivity in the main olfactory system and the vomeronasal system of the tree shrew, Tupaia belangeri.

The distribution of the calcium-binding protein calretinin was studied in peripheral and central parts of the main olfactory system (MOS) and the vomeronasal system (VNS) of adult tree shrew Tupaia belangeri. The calretinin immunoreaction was carried out with a peroxidase-coupled polyclonal antibody. In the VNS, complete labeling of all receptor cells and vomeronasal nerve fibers was observed, whereas only a subset of the somata and dendrites of receptor cells and of the olfactory nerve fibers of the MOS was immunoreactive. From the immunoreactive dendritic clubs of vomeronasal receptor cells, calretinin-labeled structures, presumably clumps of microvilli, arose that terminated within immunopositive portions of the mucus. In the main olfactory bulb, the neuropil of some of the glomeruli was immunoreactive. All periglomerular and many mitral cells were labeled. The external plexiform layer was subdivided into a faintly immunoreactive superficial half and a strongly immunoreactive deep half. Immunoreactive basal dendrites of mitral cells could be followed into either the deep half or the superficial half. In the laminated internal granular layer, a subset of immunopositive granule cells extended dendrites into the external plexiform layer. Mitral cells and granule cells with dendrites ascending to different levels of the external plexiform layer may represent functional subclasses. In the accessory olfactory bulb, all vomeronasal nerve fibers, glomeruli, and mitral/tufted cells were labeled, whereas immunoreactive periglomerular cells and internal granule cells were only scattered. In Tupaia, calretinin immunoreactivity is a more general property of the primary projecting neurons of the VNS than of the MOS and possibly indicates the involvement of calretinin in the perception of certain of the olfactory qualities.

Animals↗
PubMed2000

Cytologic differential diagnosis among reactive mesothelial cells, malignant mesothelioma, and adenocarcinoma: utility of combined E-cadherin and calretinin immunostaining.

BACKGROUND: The differential diagnosis between reactive mesothelial cells (RMs), malignant mesotheliomas (MMs), and adenocarcinomas (ACs) is often difficult in cytologic specimens, and the utility of various immunohistochemical markers have been explored. Because recent immunohistologic studies have suggested that E-cadherin (E-cad) and calretinin (Cal) may be useful markers for epithelial and mesothelial differentiations, respectively, the authors investigated their utility in cytologic diagnosis. METHODS: In this retrospective study, immunostaining was performed on smears retrieved from Papanicolaou-stained slides of effusions using the labeled streptavidin-biotin method. Sixteen cases of RM, 9 cases of MM, and 52 cases of AC from various sites, including 13 pulmonary primaries, were examined with primary antibodies against E-cad and Cal. RESULTS: The positive rates for E-cad and Cal, respectively, were as follows: RM, 0/16 (0%) and 16/16 (100%); MM, 9/9 (100%) and 8/8 (100%); and AC, 45/52 (86.5%) and 0/51 (0%). The E-cad expression by neoplastic cells was strongest in the intercellular junctions, and poorly differentiated neoplastic cells in the single cell form showed the weakest expression. CONCLUSIONS: In contrast to the results of previous immunohistochemical studies, the current study indicates that MMs constantly express E-cad, whereas RMs lack its expression in cytologic specimens, which would be useful in the differential diagnosis between the two. On the other hand, E-cad expression is not reliable for distinguishing AC from MM. The Cal expression can be a very useful marker for the distinction between AC and the mesothelial lineage. The combined immunostaining for E-cad and Cal has utility in differential diagnosis among RM, MM, and AC.

Adenocarcinoma↗
PubMed2000

Development of the anatomical alteration of the cerebellar fissura prima.

The development of the naturally occurring malformation of the cerebellar fissura prima was monitored in rats starting from 4 days of life to the adulthood. The first sign of the malformation was evident at 10 days of life and consisted of an interruption of the pia mater and the fusion of the external granular layers on the two sides of the fissura. Later, nests of apparently mature granule cells could be seen to be encircled by cells of the external granular layer and to be connected to the granule cell layer by thin bridges of cells. Calretinin immunoreactive fibers followed the bridges of cells to reach the ectopic masses of cells. Towards the end of histogenesis and in adult animals, brush cells and Golgi cells were present in the ectopic masses of granule cells. The latter appeared to contribute to the formation of normal glomeruli, as in the orthotopic granule cell layer. In addition, bundles of parallel fibers crossed the boundary between the molecular layers on the two side of the fissure, thus suggesting that parallel fibers can contact Purkinje cells of the opposite folium.

Animals↗