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The expression pattern of two calcium binding proteins (CaBP), calbindin D28k (CB) and parvalbumin (PV), in the superior colliculus (SC) of the adult rabbit, as well as the morphology of the immunoreactive cells were examined. The study was performed on 12 rabbits. Coronal sections from postmortem SC were analyzed by light microscopy, and drawings of CaBP-labeled cells were obtained using a drawing tube. No previous information is available on either the CB/PV expression or the morphology of CB/PV positive cells in the SC of the adult rabbit. Therefore, in this study we show that CB neurons and neuropil form three main tiers: the first located within the stratum zonale (SZ) and the upper part of the stratum griseum superficiale (SGS), the second located within the stratum griseum intermedium (SGI), and the third, located within the medial and central areas of the stratum griseum profundum (SGP). In contrast to this layer labeling, almost no CB-positivity is found within the other collicular layers. On the other hand, the densest concentration of PV labeled cells and terminals is found within a single dense tier that spanned the ventral part of the startum griseum superficiale (SGS) and the dorsal part of the stratum opticum (SO). Anti-PV neurons are also scattered through the deeper layers below the dense tier. In contrast, almost no anti-PV labeled neurons or neuropil are found within the stratum zonale (SZ) and upper SGS. This distribution represents a new pattern of sublamination in the SC of this species. All the previously described cell types in other mammals are observed in the rabbit SC: marginal cells, horizontal cells, pyriform cells, narrow-field vertical cells, wide-field vertical cells, and stellate/multipolar cells. Detailed drawings of all these cellular types are represented to show their complete morphology. The results of this study indicate that both CB and PV are present in a variety of neurons, which present a number of homologies between mammals, but have a different location and/or distribution, according to the different species. These findings are thus relevant to better understand the organisation of the SC in mammals.
Anat Rec 2000 07 01
PMID:Calbindin D28k and parvalbumin immunoreactivity in the rabbit superior colliculus: an anatomical study. 1086 66

This article reviews the current knowledge of the early onset of the monoaminergic innervation in the developing cerebral cortex in humans and of changes in the distribution of tyrosine hydroxylase (TH) immunoreactivity in different neuronal populations of the developing telencephalon. The early genesis of the central monoaminergic neurons in mammals has led to postulations of a trophic role of monoamines in brain morphogenesis--especially in the cerebral cortex. The developmental effects of amines can be linked to the transient expression of different molecules linked to dopamine or serotonin neurotransmission. We present novel data on the immunocytochemistry of the vesicular monoamine transporter (VMAT2) and of the high-affinity serotonin transporter (SERT) in human fetuses. SERT is a marker of the serotoninergic axons and allows visualization of the serotonin afferents of the raphe in the human telencephalon. In addition, during a restricted time period corresponding to 12-14 postovulatory weeks, we found SERT-immunolabeled fibers in the rostral and caudal limbs of the internal capsule that do not correspond to serotoninergic fibers, but do coincide with the calbindin D28k-labeled thalamocortical fiber tracts. The present observations are correlated with findings in rodents, in which a transient expression of SERT is visible in the thalamocortical axons during early postnatal life. The function of this transporter has been shown to be important for the fine-tuning of cortical sensory maps during the critical period of development of these maps. Although the present observation does not allow ascertainment of which neurons transiently express SERT, it lends support to the notion that serotonin and serotonin uptake could have important developmental roles, during the formation of brain connections in humans, as they have in rodents.
Anat Rec 2002 Jun 01
PMID:Changing distribution of monoaminergic markers in the developing human cerebral cortex with special emphasis on the serotonin transporter. 1199 77

Characterization of the enteric neurons is vital for understanding their physiological role. We have used single and dual label fluorescence and peroxidase-based immunohistochemistry in myenteric and submucosal whole mounts from the rat small intestine to evaluate the morphology and distribution of enteric neurons immunoreactive for the following phenotypic antigens: neuronal nitric oxide synthase (NOS), neurokinin-1 receptor (NK-1R), calretinin (Calr), calbindin (Cal), and neurofilament-M (NF-M). NOS-immunoreactive neurons had Dogiel type I morphology, were abundant in the myenteric plexus compared to the submucosal plexus, and never coexpressed NK-1R immunoreactivity. NK-1R- and Calr-immunoreactive neurons had Dogiel type II morphology and were distributed comparably in both plexuses. NK-1R and Calr-immunoreactivity were coexpressed in many of the same neurons. Calbindin-immunoreactive neurons exhibited four distinct morphologies: small and large Dogiel type II neurons, Dogiel type I neurons, and small elongated neurons. These neurons were significantly fewer in number in the myenteric plexus compared to the submucosal plexus. Neurofilament-M-immunoreactive neurons had three morphologies, Dogiel type II neurons, small Dogiel type II neurons, and a less common subpopulation of small, elongated, multipolar neurons. These neurons were also fewer in number in the myenteric plexus compared to the submucosal plexus. The distribution of these phenotypic markers may assist future work that elucidates the functional activities of these enteric neurons such as control of intestinal motility and adaptation to the entry of gastric contents.
Anat Rec A Discov Mol Cell Evol Biol 2003 Mar
PMID:Morphology and distribution of nitric oxide synthase-, neurokinin-1 receptor-, calretinin-, calbindin-, and neurofilament-M-immunoreactive neurons in the myenteric and submucosal plexuses of the rat small intestine. 1255 37

Cells in the epithelial rest of Malassez (ERM cells) express calbindin D28k (CB); however, the hormonal regulation of CB in ERM cells remains to be elucidated. We investigated the immunohistochemical localization of CB and 1,25-dihydroxyvitamin D3 receptor (VDR) during root formation of mouse molar teeth in order to clarify whether the expression of CB in ERM cells is dependent on vitamin D. At the early stage of root formation (postnatal (PN) days 10-14), both CB- and VDR-immunoreactive cells were observed intermittently along the root surface. In the apical portion, almost all CB-immunoreactive cells showed VDR immunoreactivity; however, VDR-immunoreactive cells in the most apical portion were immunonegative for CB. In the middle and cervical portions, the distributions of the two proteins were completely different. At the late stage of root formation (PN28d) and in adult animals, CB immunoreactivity was distributed in cells found along the acellular cementum at the bifurcation region, as well as between the dentin and cellular cementum in the apical portion (although these lacked immunoreactivity for VDR). The present results indicate that CB expression in newly disrupted cells from Hertwig's epithelial root sheath occurs in a vitamin-D dependent manner, whereas the expression of CB in mature ERM cells may be independent of vitamin D.
Anat Rec A Discov Mol Cell Evol Biol 2003 Aug
PMID:Immunolocalization of calbindin D28k and vitamin D receptor during root formation of murine molar teeth. 1284 6

In small laboratory animals, such as guinea pigs, immunoreactivity for the calcium-binding protein calbindin (CALB) can be used to distinguish functionally different classes of myenteric neurones. The rumen of sheep is a highly specialized gastrointestinal region, and the control of its functions requires specific intrinsic innervation patterns. The aim of this study was to neurochemically identify and characterize CALB-positive myenteric neurones of the ovine rumen. Therefore, we performed quadruple immunohistochemistry against CALB, substance P (SP), vasoactive intestinal peptide (VIP), and nitric oxide synthase (NOS) using whole-mount preparations of the ruminal myenteric plexus. On average, 3 +/- 2 and 1 +/- 0.4 myenteric neurones/ganglion were CALB-immunoreactive in suckling lambs and adult sheep, respectively. These neurones had Dogiel type-I morphology. Most of them (89.2% +/- 8.7% and 71.7% +/-44.8% in suckling lambs and adult sheep, respectively) did not colocalize any of the other antigenes. Since it has been shown in previous studies that ruminal myenteric neurones are immunoreactive for either choline acetyltransferase (ChAT) or NOS, we defined neurones which were CALB-positive and NOS-negative as CALB/ChAT. The other CALB-positive neurones were encoded CALB/NOS/+/-VIP (10.3% +/- 9.3% and 26.7% +/- 46.2% in suckling lambs and adult sheep, respectively) or CALB/ChAT/SP (0.5% +/- 1.0% and 1.7% +/- 1.9% in suckling lambs and adult sheep, respectively). We used cryostat sections of the ruminal wall to analyze the projections of the CALB-positive neurones. CALB-immunoreactive somata were exclusively located within the myenteric plexus. CALB-immunoreactive nerve fibers were found primarily in the lamina propria of the ruminal papillae. We conclude that CALB-positive myenteric neurones within the ovine rumen project to the epithelium; however, their functional role remains to be investigated.
Anat Rec A Discov Mol Cell Evol Biol 2004 Jun
PMID:Calbindin-immunoreactive neurones in the ovine rumen. 1516 40

The migration of macrophages and lymphocytes that produce cytokines such as tumor necrosis factor-alpha (TNF-alpha) causes beta-cell death, leading to type 1 diabetes. Similarly, in type 2 diabetes, the adipocyte-derived cytokines including TNF-alpha are elevated in the circulation, causing inflammation and insulin resistance. Thus, the studies described in this article using TNF-alpha are relevant to furthering our understanding of the pathogenesis of diabetes mellitus. We used RINr1046-38 (RIN) insulin-producing beta-cells, which constitutively express calbindin-D(28k), to characterize the effect of TNF-alpha on apoptosis, replication, insulin release, and gene and protein expression. Western blots of TNF-alpha-treated RIN cells revealed a decrease in calbindin-D(28k). By ELISA, TNF-alpha-treated beta-cells had 47% less calbindin-D(28k) than controls. In association with the decline in calbindin-D(28k), TNF-alpha treatment of RIN cells led to a 73% greater increase in changes in intracellular calcium concentration (Delta[Ca(2+)](i)) in TNF-alpha-treated cells as compared to that in control RIN cells upon treatment with 50 mM KCl; caused a greater increase in the [Ca(2+)](i) following the addition of 5.5 microM ionomycin; increased by more than threefold the apoptotic rate, expressed as the percentage of TUNEL-positive nuclei to total nuclei; decreased the rate of cell replication by 36%; and increased and decreased selectively the expression of specific genes as determined by microarray analysis. The subcellular localizations of Bcl-2, an antiapoptotic protein, and Bax, a proapoptotic protein, within RIN cells were altered with TNF-alpha treatment such that the two were colocalized with mitochondria in the perinuclear region. We conclude that the proapoptotic action of TNF-alpha on beta-cells is manifested via decreased expression of calbindin-D(28k) and is mediated at least in part by [Ca(2+)](i).
Anat Rec A Discov Mol Cell Evol Biol 2005 Oct
PMID:Tumor necrosis factor-alpha-induced changes in insulin-producing beta-cells. 1611 68

The cytoarchitecture of the cerebral cortex in mammals has been traditionally investigated using Nissl, Golgi, or myelin stains and there are few comparative studies on the relationships between neuronal morphology and neurochemical specialization. Most available studies on neuronal subtypes identified by their molecular and morphologic characteristics have been performed in species commonly used in laboratory research such as the rat, mouse, cat, and macaque monkey, as well as in autopsic human brain specimens. A number of cellular markers, such as neurotransmitters, structural proteins, and calcium-buffering proteins, display a highly specific distribution in distinct classes of neocortical neurons in a large number of mammalian species. In this article, we present an overview of the morphologic characteristics and distribution of three calcium-binding proteins, parvalbumin, calbindin, and calretinin, and of a component of the neuronal cytoskeleton, nonphosphorylated neurofilament protein in the neocortex of various species, representative of the major subdivisions of mammals. The distribution of these neurochemical markers defined several species- and order-specific patterns that permit assessment of the degree to which neuronal morphomolecular specialization, as well as the regional and laminar distribution of distinct cell types in the neocortex, represents derived or ancestral features. In spite of the remarkable diversity in morphologic and cellular organization that occurred during mammalian neocortical evolution, such patterns identified several associations among taxa that closely match their phylogenetic relationships.
Anat Rec A Discov Mol Cell Evol Biol 2005 Nov
PMID:Morphomolecular neuronal phenotypes in the neocortex reflect phylogenetic relationships among certain mammalian orders. 1621 36

The well-organized cerebellum is an ideal model to investigate the developmental appearance and localization of pre- and postsynaptic structures. One of the synaptic proteins abundant in the central nervous system and localized in presynaptic vesicle membranes is the synaptic vesicle protein 2 (SV2). SV2 was shown to be involved in priming and modulating synaptic vesicles and having an effect in epileptic diseases. So far there are no data available describing the developmental localization of this protein in the cerebellum. We followed the expression pattern of SV2 and compared it with the expression of the neuronal calcium-binding protein Calbindin and the AMPA glutamate receptor subunits 2/3 (GluR 2/3), both shown to be early expressed in the developing chick cerebellum predominantly in Purkinje cells. We detected the expression of SV2 in presynaptic terminals (mainly from climbing and mossy fibers) as soon as they are formed at embryonic day 16 in the inner molecular layer. Purkinje cells express Calbindin and GluR 2/3 in the soma and postsynaptically in the primary dendrites at this stage. With ongoing development, the pattern of SV2 expression follows the development of Purkinje cell dendrites in the molecular layer, suggesting a synaptic refinement of labeled climbing and later parallel fibers.
Anat Rec (Hoboken) 2008 May
PMID:Expression of SV2 in the developing chick cerebellum: comparison with Calbindin and AMPA glutamate receptors 2/3. 1838 60

Squirrels are highly visual mammals with an expanded cortical visual system and a number of well-differentiated architectonic fields. To describe and delimit cortical fields, subdivisions of cortex were reconstructed from serial brain sections cut in the coronal, sagittal, or horizontal planes. Architectonic characteristics of cortical areas were visualized after brain sections were processed with immunohistochemical and histochemical procedures for revealing parvalbumin, calbindin, neurofilament protein, vesicle glutamate transporter 2, limbic-associated membrane protein, synaptic zinc, cytochrome oxidase, myelin or Nissl substance. In general, these different procedures revealed similar boundaries between areas, suggesting that functionally relevant borders were being detected. The results allowed a more precise demarcation of previously identified areas as well as the identification of areas that had not been previously described. Primary sensory cortical areas were characterized by sparse zinc staining of layer 4, as thalamocortical terminations lack zinc, as well as by layer 4 terminations rich in parvalbumin and vesicle glutamate transporter 2. Primary areas also expressed higher levels of cytochrome oxidase and myelin. Primary motor cortex was associated with large SMI-32 labeled pyramidal cells in layers 3 and 5. Our proposed organization of cortex in gray squirrels includes both similarities and differences to the proposed of cortex in other rodents such as mice and rats. The presence of a number of well-differentiated cortical areas in squirrels may serve as a guide to the identification of homologous fields in other rodents, as well as a useful guide in further studies of cortical organization and function.
Anat Rec (Hoboken) 2008 Oct
PMID:Architectonic subdivisions of neocortex in the gray squirrel (Sciurus carolinensis). 1878 Feb 99

A natural defect in rat cerebellum postnatal development has been found in the fissura prima, consisting in various complex configurations of the cerebellar layers. We investigated the genesis of fissure malformations through immunoreactions for PCNA, GFAP, GABAA alpha6, and calbindin to label proliferating cells of the external granular layer (egl), radial glial fibers, mature granule cells, and Purkinje cells, respectively. Results on critical stages of rat postnatal development provided interesting evidences on how the malformation develops in fissures prima and secunda. Early (postnatal day 10) at the site of malformation, the Bergmann radial glia was often retracted and showed distortions and irregular running. The interruption of GFAP-positive radial glial fibers could fit in with the presence of clusters of PCNA-unlabeled cells in the sites of fusion of the egl; the clusters of cells are granule cells since their soma is labeled by GABAA alpha6. Moreover, an altered migration of granule cell precursors to the internal granular layer was evident which, in turn, affected Purkinje cell differentiation and the growth of their dendrites. In summary, the changed relationship among glial fiber morphology, granule cell migration, and Purkinje cell differentiation suggests how the Bergmann glial fibers have a basic role in the foliation process, being the driving physical force in directing migration of the granule cells at the base of fissure.
Anat Rec (Hoboken) 2010 Mar
PMID:Postnatal development of the central nervous system: anomalies in the formation of cerebellum fissures. 2009 88


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