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A Wanaka

Publications and source records attributed to A Wanaka.

At least 73 records · Page 4Linked to original sources

Localization of trkB mRNA in postnatal brain development.

We investigated the localization of trkB mRNA, which encodes a putative component of high-affinity brain-derived neurotrophic factor (BDNF) or the neurotrophin-3 (NT-3) receptor, in the postnatal rat brain by in situ hybridization histochemistry. At birth, trkB mRNA was strongly expressed in various regions with the thalamus and cerebral cortex showing the strongest expression. As the rat grows, expression generally persisted or declined in most regions with the exception of the hippocampus where trkB mRNA expression increased during postnatal development. In the adult brain, trkB mRNA was detected in the olfactory system, cerebral cortex, hippocampal formation, amygdala, and cerebellar cortex. These findings, together with the developmental profiles of BDNF and NT-3 mRNA expressions, suggest that trkB product (gp145trkB) mainly transduces NT-3 signals early in the postnatal period, and BDNF signals later in the period.

Aging↗

MFH-1, a new member of the fork head domain family, is expressed in developing mesenchyme.

We have isolated a novel mouse gene, MFH-1 (mesenchyme fork head 1) that is related to the Drosophila fork head and rat HNF3 genes. MFH-1 encodes a distinct fork head domain that is classified into a distinct subfamily. A recombinant MFH-1 protein could bind to the HNF3 binding site. MFH-1 is expressed temporally in developing embryos, first in the non-notochordal mesoderm and later in areas of mesenchymal condensation in the trunk, head, and limbs. Our results suggest that MFH-1 might be involved in the formation of special mesenchymal tissues.

Amino Acid Sequence↗

Developmentally regulated expression of pleiotrophin, a novel heparin binding growth factor, in the nervous system of the rat.

Pleiotrophin (PTN) is a newly identified heparin-binding growth factor which is closely related to the retinoic acid-inducible MK protein. PTN is expressed at high levels in perinatal brain and promotes neurite outgrowth from embryonic brain neurons and mitogenesis in fibroblasts, suggesting that it may play an important role in the development of the nervous system. We have used in situ hybridization to examine PTN expression in the developing and adult rat nervous systems. During embryogenesis, PTN mRNA is primarily expressed by neuroglial progenitor cells in the subependymal layer of the central nervous system (CNS), whereas during the perinatal period high levels of PTN transcripts are found in neurons as well as glial elements (astrocytes and oligodendrocytes). In the adult brain, PTN expression is markedly decreased relative to early postnatal brain and, in contrast to the neuronal and glial expression observed in young animals, is confined to specific neuronal subpopulations (especially hippocampal CA1-3 regions, cerebral cortex laminae II-IV). PTN is also expressed in the developing spinal cord and eye. In the peripheral nervous system (PNS), PTN mRNA is present in ganglionic neurons during embryogenesis. In adult ganglia, however, PTN expression becomes localized to the satellite cells of the ganglia. The developmental pattern of PTN expression in the CNS and the 'switch' in expression from neurons to satellite cells in the PNS suggests that it has important functions not only in the developing nervous system, but also in the adult CNS and PNS and that the functions performed by this growth factor change during ontogeny. We have also found that levels of PTN mRNA are dramatically but transiently elevated in neurons of the hippocampus, piriform cortex and parietal cortex following a chemically induced seizure, indicating that neuronal PTN mRNA expression is increased by intense physiological stimuli and may play a role in the response to these stimuli.

Aging↗

Upregulation of fibroblast growth factor-receptor messenger RNA expression in rat brain following transient forebrain ischemia.

Recently, we demonstrated that transient forebrain ischemia in rats leads to an early and strong induction of basic fibroblast growth factor (bFGF) synthesis in astrocytes in the injured brain regions. In this study, in order to clarify the targets of such raised endogenous bFGF levels, the messenger RNA (mRNA) expression of its receptors (flg and bek) in the hippocampus following transient forebrain ischemia induced by four-vessel occlusion for 20 min was investigated using an in situ hybridization technique. Transient forebrain ischemia induced an increase in the number of flg mRNA-positive cells from an early stage (24 h after ischemia) in the hippocampal CA1 subfield where delayed neuronal death occurred later (48-72 h after ischemia). This increase became more marked with the progression of neuronal death and was still evident in the same area 30 days later. The time course of the appearance and distribution pattern of flg mRNA-positive cells in the CA1 subfield were quite similar to those of bFGF mRNA-positive cells. On the other hand, in situ hybridization for bek mRNA showed only slight and transient (observed 72 h and 5 days after ischemia) increases in the number of mRNA-positive cells in the CA1 subfield following ischemia. The use of in situ hybridization and glial fibrillary acidic protein immunohistochemistry in combination demonstrated that the cells in the CA1 subfield that exhibited ischemia-induced flg or bek mRNA expression were astrocytes. These data indicate that transient forebrain ischemia induces upregulation of fibroblast growth factor-receptor expression, accompanied by increased bFGF expression in astrocytes, and suggest that the increased astrocytic bFGF levels in injured brain regions act on the astrocytes via autocrine systems and are involved in the development and maintenance of astrocytosis.

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Differential expression of two members of FGF receptor gene family, FGFR-1 and FGFR-2 mRNA, in the adult rat central nervous system.

The fibroblast growth factor (FGF) receptor gene family now contains four members that encode homologous receptor-tyrosine kinases (RTKs) to each other. We have demonstrated that one of the members, FGFR-1 (also termed as flg), is expressed in the widespread but specific neuronal populations in the adult rat central nervous system (CNS). In the present study, we examined the expression pattern of another member, FGFR-2 (also termed as bek) and compared it with that of FGFR-1. In contrast with FGFR-1, we detected FGFR-2 expression primarily in the fiber tracts of the adult rat CNS, suggesting that the oligodendrocytes are the main sites of the FGFR-2 expression. These findings indicate that FGF may regulate neurons and glial cells through different subtypes of its cognate receptor.

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Co-localization of basic fibroblast growth factor-like immunoreactivity and its receptor mRNA in the rat spinal cord and the dorsal root ganglion.

In the present study, we examined the localizations of basic fibroblast growth factor-like immunoreactivity (bFGF-LI) and its receptor mRNA in the spinal cord and the dorsal root ganglion of the rat. Anti-bFGF peptide antibody and cRNA probe were employed to visualize the localizations of bFGF-LI and FGF receptor (FGF-R) mRNA, respectively. In the spinal cord, we observed that a number of neurons including the motor neurons and interneurons were positive for both substances. In the dorsal root ganglion (DRG), the large neurons preferentially showed co-localization of bFGF-LI and FGF-R mRNA, while the small neurons were not always positive for both. Given the fact that FGF-R is a membrane-spanning protein, these findings suggest the following two possibilities: (1) bFGF acts on the neurons of the spinal cord and the DRG in an autocrine and/or paracrine manner; (2) FGF-R mRNA-positive neurons take up bFGF from innervating neurons and/or surrounding glias in a receptor-mediated fashion.

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Localization of mRNA for c-kit receptor and its ligand in the brain of adult rats: an analysis using in situ hybridization histochemistry.

Localization of mRNA for the c-kit receptor and its ligand (Sl factor) in the brain of adult rats was studied using in situ hybridization histochemistry. The mRNA for the c-kit receptor was detected in the forebrain, the lower brain stem and the cerebellum. In the forebrain, the c-kit mRNA signals were detected in the olfactory bulb, the caudate-putamen, throughout the superficial cortex, the accumbens nucleus, the nucleus of vertical limb diagonal band, the bed nucleus of anterior commissure, Ammon's horn, the entopeduncular nucleus, the subthalamic nucleus, the dorsal raphe nucleus, the parasubiculum, the presubiculum, the ventricular nucleus of lateral lemniscus, and the entorhinal cortex. In the lower brain stem, the signals were detected in the inferior colliculus, the spinal vestibular nucleus, the spinal tract nucleus of trigeminal nerve, and the pyramidal tract. In the cerebellum, the signals were detected in the molecular layer of the cortex and cerebellar nuclei. By contrast, the signals of mRNA for Sl factor were detected in the forebrain and the cerebellum. In the forebrain, the signals were detected in the olfactory bulb, the endopiriform nucleus, the septohippocampal nucleus, the habenular nuclei, and most of the thalamic nuclei. In the cerebellum, the signals were detected in Purkinje cells. Several pairs of structures were found in which mRNA of either the c-kit receptor or the Sl factor was expressed and between which the synaptic connection had been reported, suggesting that the interaction between the c-kit receptor and the Sl factor may play some roles in the development of such synaptic connections.

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Specific expressions of Fyn and Lyn, lymphocyte antigen receptor-associated tyrosine kinases, in the central nervous system.

The Src-like protein-tyrosine kinases Fyn and Lyn are expressed in lymphocytes. Fyn is expressed in T cells at elevated levels and is associated with the T cell antigen receptor complex, whereas Lyn is expressed in B cells and is associated with membrane-bound immunoglobulin. Thus, these kinases are suggested to participate in antigen-mediated signal transduction in lymphocytes. Previous report showed that fyn was also expressed in brain, but its cellular distribution was not examined. Expression of Lyn in neural tissues was not previously reported. Here we report that both fyn and lyn are expressed in discrete regions of the brain. To throw light on their functions in the brain, we investigated their expressions during brain ontogenesis in mice. In situ hybridization analysis showed that Fyn mRNA was specifically expressed in neurons of embryos and newborn mice. In adult animals, fyn mRNA was expressed in oligodendrocytes as well as neurons. In contrast, the expression of lyn mRNA was relatively low in brains of embryos and newborn mice, but in adults the transcript was specifically expressed in the granular layer of the cerebellum. Therefore, the Fyn and Lyn kinases may regulate distinct functions of specific cells during brain development. The specific expressions of Fyn and Lyn in both lymphatic and neural tissues could suggest common signalling mechanisms in the immune system and central nervous system.

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Expression of FGF receptor gene in rat development.

We examined the expression of FGF-receptor (FGF-R) mRNA during rat development with in situ hybridization histochemistry. Embryonic tissues (E9, E12, E14, E17) and postnatal neural tissues (P1, P7, P14, adult) were examined. We detected significant levels of FGF-R mRNA in various tissues at different developmental stages. As postulated by previous studies using other methods, FGF-R gene expression was observed primarily in mesoderm- and neuroectoderm-derived tissues. In the nervous system, the pattern of gene expression was developmentally regulated; in embryos, FGF-R mRNA was mainly detected in the ependymal layer of the central nervous system (CNS). Postnatally, FGF-R transcripts were observed in specific neuronal populations, such as hippocampal neurons. FGF-R mRNA was also found in sensory systems such as trigeminal and dorsal root ganglia in late stage embryos; however, FGF-R mRNA decreased in the postnatal period. FGF-R mRNA expression was modulated in the developing retina: FGF-R messages were observed in the pigment epithelium and neuroblast layer at embryonic stages; in the postnatal period, they were found in the ganglion cell and inner granular layer. In non-neuronal embryonic tissues, a wide variety of organs expressed FGF-R message. Particularly, the prevertebral column, bone, kidney and skin showed high levels of expression. These observations reinforce the idea that FGF exerts effects on the development of various tissues.

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Developmental study of nerve growth factor receptor mRNA expression in the postnatal rat cerebellum.

Nerve growth factor receptor (NGFR) mRNA expression was examined in the developing rat cerebellum with in situ hybridization histochemistry. The probe used in this study was the 57-mer oligonucleotide complementary to the rat NGFR cDNA (bases 798-855). The specificity of the hybridization was determined with a 'sense' probe. From the age of postnatal day 0 (PND 0), hybridization signals were detected in the external granule cell layer, Purkinje cell layer, and meningeal cells of the cerebellum. Until PND 10, the external granular cell layer consistently expressed high levels of NGFR message, with the densest hybridization signal observed in the proliferative zone. NGFR mRNA level in this layer rapidly decreased at later ages and was barely detectable after PND 16. In the Purkinje cell layer beyond PND 4, the hybridization signal became more intense, reached a peak level around PND 10, and then gradually decreased. NGFR mRNA was detected until PND 30 in this layer. Meningeal cells expressed NGFR message from PND 0 to PND 8. These results suggest that NGF may be involved in cerebellar development as a mediator of morphogenesis and/or synaptogenesis.

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Localization of FGF receptor mRNA in the adult rat central nervous system by in situ hybridization.

Fibroblast growth factor receptor (FGF-R) mRNA expression was examined in the adult rat CNS. Northern blot analysis showed a distinct 4.3 kb transcript in various CNS regions. In situ hybridization revealed widely distributed, but specific, populations of cells that express FGF-R mRNA. The most intense hybridization signals were observed in the hippocampus and in the pontine cholinergic neurons. The limbic system and brainstem nuclei, including motor nuclei, showed robust labeling. Cerebellar granule cells and spinal cord neurons were positive for FGF-R mRNA. The distribution of FGF-R mRNA differed significantly from that of NGF receptor mRNA; particularly, no hybridization signal was detected in basal forebrain cholinergic neurons. These results strongly suggest that FGF or FGF-like molecules may exert effects on specific neuronal populations in the mature CNS.

Amino Acid Sequence↗

Immunocytochemical analysis of [Met5]enkephalin-Arg6-Gly7-Leu8 immunoreactive structures in the rat superior cervical ganglion.

Indirect immunofluorescence and immunoelectron microscopy were employed to analyze the enkephalinergic systems in the rat superior cervical ganglion (SCG). These systems were identified using specific antiserum against [Met5]Enkephalin-Arg6-Gly7-Leu8 (ENK-8), a peptide which is derived only from proenkephalin A. Abundant ENK-8 like immunoreactive (ENK-8-LI) neurons and fibers were observed in the SCG, but their distribution patterns were heterogenous; ENK-8-LI neurons were localized preferentially in the caudal two-thirds of the SCG, while immunoreactive fibers were found to be distributed more densely in the rostral one-third than in the remaining part of the SCG. Most of the ENK-8-LI neurons were large and had ultrastructural features resembling those of principal cells, some were identified electron microscopically as small intensely fluorescent (SIF) cells. ENK-8-LI fibers were varicose in appearance and surrounded the perikarya of neurons. Since most of these fibers were not detected after experimental decentralization of the SCG and since ENK-8-LI terminals were seen to contain small lucent vesicles, most of the former were thought to be preganglionic fibers. Immunoreactive fibers mainly formed synaptic contacts with the dendrites of non-immunoreactive principal cells, but a small proportion of ENK-8-LI principal cells also received synaptic input from them. Occasionally, immunoreactive fibers formed synapses with the processes or the soma of both ENK-8-LI and non-immunoreactive SIF cells. On the basis of these findings, we conclude that: (1) preganglionic ENK-8-LI fibers terminate mainly on the principal cells, which are devoid of ENK-8-LI structures; (2) the majority of ENK-8-LI neurons are principal cells, while the remainder are SIF cells; (3) inputs to these cells mainly involve structures lacking ENK-8 immunoreactivity; and (4) there are, however, a small number of ENK-8-LI preganglionic fibers which terminate on ENK-8-LI principal cells and SIF cells.

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Immunocytochemical localization of beta-adrenergic receptors in the rat brain.

The localization of beta-adrenergic receptors (beta-AdR) in the rat brain was examined immunocytochemically with antibody against affinity-purified beta 2-receptors. This antibody was confirmed to cross-react with beta 1-AdR. beta-AdR were widely but unevenly distributed in the rat brain. In the forebrain and diencephalon, cells with beta-AdR were abundant in the cerebral cortex, hippocampus, and various hypothalamic nuclei other than the ventromedial hypothalamic nucleus and the suprachiasmatic nucleus. Few or no cells with beta-AdR were found in the thalamus except in the lateral and medial geniculate nucleus, habenular nucleus and paraventricular thalamic nucleus. The striatum, globus pallidus, and accumbens nucleus also contained few beta-AdR. In the lower brainstem, areas related to auditory function, such as the inferior colliculus, the nucleus of the lateral lemniscus and the cochlear nuclei, as well as areas related to cerebellar function, such as the reticulotegmental nucleus of the pons, the pontine nuclei, the lateral reticular nucleus and the inferior olive, were rich in beta-AdR. Considerable immunoreactivity was also found in the parabrachial nucleus and the nucleus of the solitary tract.

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Presence of catecholaminergic axon-terminals containing beta-adrenergic receptor in the periventricular zone of the rat hypothalamus.

The present study, using a light microscopic double-immunofluorescence method, has revealed the presence of fibers containing both tyrosine hydroxylase- and beta-adrenergic receptor-like immunoreactivities in the rat hypothalamic periventricular zone. Subsequent immunoelectron microscopic analysis demonstrated that these belong to axon terminals. These findings suggest that presynaptic beta-adrenergic receptor is present in catecholaminergic terminals.

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Localization of glycine and beta-adrenergic receptors in the rat brain.

The localization of glycine (GR) and adrenergic receptors (AdR) was examined in the rat brain using a monoclonal antibody against the affinity purified glycine receptor and a polyclonal antibody against purified beta2-AdR. GR were concentrated in the lower brainstem, whereas no immunoreactivity was observed in the diencephalon and forebrain except in a few diencephalic nuclei. The highest density was found in the cranial motor nuclei, reticular formation, parabrachial area, dorsal and ventral cochlear nuclei, and dorsal and ventral tegmental nuclei. On the other hand, AdR were widely but unevenly distributed in the rat brain. The present study further showed the presence of AdR in catecholaminergic terminals in the hypothalamus.

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Localization of glycine receptors in the rat central nervous system: an immunocytochemical analysis using monoclonal antibody.

The localization of glycine receptors was immunocytochemically examined in the rat brain using a monoclonal antibody against the affinity-purified glycine receptor. Glycine receptors were concentrated in the lower brainstem, whereas no immunoreactivity was observed in the diencephalon and forebrain except in a few diencephalic nuclei. The highest density of receptors was found in the cranial motor nuclei, reticular formation, parabrachial area, dorsal and ventral cochlear nuclei, and dorsal and ventral tegmental nuclei. Differences were observed in the distribution of immunoreactive elements in the various brain regions. In the cerebellar cortex, the immunoreactivity was exclusively seen along the dendrites of the Purkinje cells. On the other hand, glycine receptors were detected on the cellular membrane of the soma of the cochlear nuclei, trigeminal motor nucleus, parabrachial area, lateral reticular nucleus, dorsal nucleus of the lateral lemniscus, cerebellar nuclei, trigeminal spinal nucleus, anterior horn and reticular formation. In other regions, the receptors were evenly distributed throughout the neuropil.

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