Biomedical subjects

S Shiosaka

Publications and source records attributed to S Shiosaka.

At least 55 records · Page 3Linked to original sources

PubMed1995

Ontogeny of neuropsin mRNA expression in the mouse brain.

In our previous study, we found a novel gene encoding a serine protease termed neuropsin (NP) which exhibited activity-dependent gene expression or repression in the mouse hippocampus (Chen et al., 1995). In the present study, we examined the ontogeny of NP mRNA expression by in situ hybridization in the brain. Weak hybridization signals were also observed in the choroid plexus at this stage in addition to neuronal labeling. The signals continued to show this localization pattern until postnatal day 12. After embryonic day 18, the number of hybridization signals localized in the neurons of the forebrain limbic area were more predominant than those in the lower brainstem. NP gene expression spread in the anterior olfactory nucleus, hippocampus, septal nuclei, diagonal band of Broca, amygdala and limbic cortex successively from early embryonic to adult stage, though signals in the other brain regions were gradually decreased after birth. Thus, the widespread localization and two types of expression pattern, constitutive or transient, suggest that NP is a multiple functional protein involved in development, neuronal plasticity and cerebrospinal fluid production.

Animals↗

PubMed1994

Basal magnocellular and pontine cholinergic neurons coexpress FGF receptor mRNA.

By in situ hybridization histochemistry, fibroblast growth factor receptor gene (flg)-expressing neurons were newly identified in the basal magnocellular nuclei (the vertical and horizontal limbs of the diagonal band, and Meynert's nucleus). The present study also confirmed flg localization in the laterodorsal tegmental nucleus and the pedunculopontine tegmental nucleus of the pons. Immuno- and in situ hybridization histochemistry on the same sections demonstrated that choline acetyltransferase and flg were colocalized in single neurons of the diagonal band, Meynert's nucleus and the pontine tegmental areas. The results suggest that a significant number of the basal magnocellular and a majority of the mesopontine cholinergic neurons are directly affected by fibroblast growth factors (FGF) via FGF receptor gene.

Animals↗

PubMed1994

Gene transfer and the expression of a foreign gene in vivo in post-mitotic neurons of the adult rat brain using the hemagglutinating virus of the Japan-liposome method.

Neurons in the adult rat brain were transfected in vivo with a simple plasmid that harbored the gene for beta-galactosidase from Escherichia coli under control of a chicken beta-actin promoter by use of the hemagglutinating virus of Japan (HVJ) and liposomes. Cells that expressed beta-galactosidase were detected only in the target area of the central nervous system for 10 days by light microscopic analysis. Since electron microscopic analysis revealed that the products of the histochemical reaction were predominantly associated with the nuclear membrane and the endoplasmic reticulum of positive cells, it appeared that the products were translated endogenously and had not been entrapped by endocytosis. Furthermore, the products were observed in typical neuronal cells with a large, round, and pale nuclei, and with direct axo-somatic and axo-dendritic synaptic contacts. This report suggests the possibility of introducing functionally significant genes into neurons in targeted areas of the adult central nervous system.

Animals↗

PubMed1994

Growth factors and extracellular signal-regulated kinases (mitogen-activated protein kinase) in the rat pineal gland.

The present study revealed that the pineal gland expressed basic fibroblast growth factor (bFGF) and FGF-receptor 1 (FGFR1/flg), suggesting that bFGF in the pineal gland acts in an autocrine or paracrine manner, which is mediated by FGFR1/flg. The present study also examined gene expression of the extracellular signal-regulated kinase (ERK) family (ERK1-3) which may be intracellular signal mediators of growth factors. ERK1 [mitogen-activated protein kinase (MAP-kinase)] was strongly expressed throughout the pineal gland, while expression of ERK2 and ERK3 was not found. These findings suggest the presence of a signal pathway from bFGF to ERK1 via FGFR1/flg in the pineal gland.

Animals↗

PubMed1994

Electron microscopic observation of bFGF immunoreactivity in the hippocampus.

Light and electron microscopic observation showed two types of neuronal immunostaining for basic fibroblast growth factor in the hippocampal CA2 subfield, where the densest immunoreactive neurons were localized in the brain. One neuronal type showed intense nuclear (eu- and heterochromatin) immunostaining but weak cytoplasmic immunostaining (N-type), and the other showed intense cytoplasmic but no or only faint nuclear immunoreactivity (C-type). The N-type also showed weak immunoreactivity in the perinuclear rough endoplasmic reticulum and contained bFGF mRNA as observed by in situ hybridization histochemistry, showed that this type can produces the bFGF protein. The N-type localized exclusively in the CA2 subfield. The C-type showed strong immunoreactivity on the rER, free ribosomes, and Golgi apparatus, although no clear evidence for bFGF production was observed. The multivesicular bodies, a pathway of endocytosis in hippocampal neurons showed apparent immunoreactivity under EM observation of both of types neurons (Parton et al. J. Cell Biol. 119: 123-137, 1992) suggesting a receptor-mediated type of incorporation of the bFGF.

Amino Acid Sequence↗

PubMed1993

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.

Animals↗

PubMed1992

Localization of basic fibroblast growth factor-like immunoreactivity in the rat brain.

The immunohistochemical localization of basic fibroblast growth factor (bFGF) was studied in the adult rat brain, using a specific antibody against a synthetic bFGF fragment (the N-terminal 12 residues). Widespread but uneven regional localization of bFGF-like immunoreactive neurons and fibers was observed. Ependymal cells were also stained. The immunoreactive neurons were found in the cerebral cortex, olfactory bulb, septum, basal magnocellular nuclei, thalamus, hypothalamus, globus pallidus, hippocampus, amygdala, red nucleus, central gray of the midbrain, cerebellum, dorsal tegmental area, reticular formation, cranial motor nuclei and spinal cord. Immunoreactive fiber bundles and nerve terminals were also detected. These results indicate that bFGF is produced by or present in a specific neuronal cell population of the central nervous system.

Amino Acid Sequence↗

PubMed1992

Basic fibroblast growth factor-like immunoreactivity in the trigeminal proprioceptive and motor systems.

Basic fibroblast growth factor (bFGF) isolated from the brain and pituitary, has been shown to induce cell divisions in a variety of cell types. It also acts as a potent stimulator of angiogenesis, and it is important in the survival of several types of cultured neurons. Despite considerable information on the functions of bFGF, there is incomplete knowledge about the ways in which it reaches remote tissues and its subcellular localization in the adult brain. Here we report our findings that a certain population of neurons with free ribosomes and rough endoplasmic reticulum immunoreactive for bFGF in the mesencephalic nucleus of the trigeminal nerve sends proprioceptive fibers to muscle spindles in the masseter muscle, and immunoreactive axons to the trigeminal motor nucleus to form synapses with the bFGF-containing motoneurons whose axons further constitute myoneural junctions in the periphery. Moreover, some bFGF neurons contain electron dense immunoreaction deposits in the euchromatin but not in the heterochromatin of the nucleus. These findings suggest that endogenous bFGF is transported within nerve processes and functions in mature neuronal circuits subserving the masseteric reflex arcs, and that bFGF is produced in free ribosomes and/or rough endoplasmic reticulum and is transported into the genetically active euchromatin as well.

Animals↗

PubMed1992

Increase of basic fibroblast growth factor immunoreactivity and its mRNA level in rat brain following transient forebrain ischemia.

We examined the time course of basic fibroblast growth factor (bFGF) immunoreactivity and its mRNA level mainly in the hippocampus after transient forebrain ischemia using immunohistochemistry, enzyme immunoassay (EIA), Western blot analysis and in situ hybridization. Neuronal death in the hippocampal CA1 subfield was observed 72 h after 20 min of ischemia. The number of bFGF-immunoreactive(IR) cells increased 48 h-5 days after ischemia in all hippocampal regions. At 10 and 30 days, the bFGF-IR cells in the CA1 subfield had further increased in numbers and altered their morphology, enlarging and turning into typical reactive astrocytes with the advancing neuronal death in that area. In contrast, the number of bFGF-IR cells in other hippocampal regions had decreased 30 days after ischemia. The EIA study showed a drastic increase in bFGF levels in the hippocampus 48 h after ischemia (150% of that in normal rat) which was followed by further increases. In Western blot analysis, three immunoreactive bands whose molecular weights correspond to 18, 22 and 24 kDa were observed in normal rat and ischemia increased all their immunoreactivities. In the in situ hybridization study of the hippocampus, bFGF mRNA positive cells were observed in the CA1 subfield in which many bFGF-IR cells existed after ischemia. These data demonstrate that transient forebrain ischemia leads to an early and strong induction of bFGF synthesis in astrocytes, suggesting that the role of bFGF is related to the function of the reactive astrocytes which appear following brain injury.

Animals↗

PubMed1992

Attempts to make models for Alzheimer's disease.

Profound reductions in cortical acetylcholine levels together with degeneration of cholinergic neurons in the basal forebrain have been reported in patients with Alzheimer's disease. A similar loss of the cholinergic neurons of the basal forebrain and impairment of learning and memory occur in animals injected with a nerve growth factor-diphtheria toxin conjugate, suggesting that this animal model is suitable to analyze cholinergic roles on learning and memory processes, and also the pathogenesis of Alzheimer's disease. In addition, animal models constructed by electrolytic or neurotoxic lesioning of the basal magnocellular nucleus, and models made by transgenetic technology were described.

Alzheimer Disease↗

PubMed1991

Specific cholinergic destruction in the basal magnocellular nucleus and impaired passive avoidance behavior of rodents.

A nerve growth factor (NGF)-diphtheria toxin conjugate (NGDT) was found to selectively abolish or depress the activity of NGF receptor-bearing cholinergic neurons of the basal magnocellular nucleus (BMN). Bilateral cortical injections of NGDT impaired the retention of passive avoidance behavior in mice. A memory deficit was also revealed when cortical injections of NGDT were administered after the acquisition of a passive avoidance response. Thus, retrograde destruction of BMN cholinergic neurons by the cortical injection of NGDT interfered with both learning and memory processes. The animal model outlined here should be useful in analyzing the pathogenesis of Alzheimer's disease and the functions of the cholinergic system in the BMN.

Animals↗

PubMed1991

Localization of basic FGF-like immunoreactivity in the hypothalamo-hypophyseal neuroendocrine axis.

We examined the localization of basic fibroblast growth factor (basic FGF) in the adult rat brain by immunohistochemical and Western blotting analysis using a specific antibody against a synthetic basic FGF fragment (N-terminal 12 residues). The antibody did not cross-react with acidic FGF. Basic FGF-like immunoreactivity was located exclusively in the neuronal elements and had very heterogeneous distribution. Immunoreactive cell bodies were observed in the paraventricular, supraoptic and circular nuclei of the hypothalamus. Numerous immunoreactive neuronal processes originating from these basic FGF-positive cells extended lateroventrally and then caudally to the internal layer of the median eminence. In addition, the neurohypophysis contained a significant number of basic FGF-like immunoreactive fibers. Western-blotting analysis revealed that the hypothalamus and the hypophysis contained a main band of basic FGF immunoreactive with an apparent molecular weight of 17 kDa. These results show that the hypothalamo-hypophyseal neuroendocrine pathway contains basic FGF.

Amino Acid Sequence↗

PubMed1991

Neurotensin and neuromedin N elevate the cytosolic calcium concentration via transiently appearing neurotensin binding sites in cultured rat cortex cells.

Through assessment of the changes in the intracellular free-calcium concentration ([Ca2+]i), which was measured using the calcium sensitive dye, fura-2, the character of the neurotensin (NT) binding sites which appeared transiently during the early ontogenetic stage in the rat cerebral cortex was analyzed in primary cultures of cerebral cortex cells from neonatal rats. NT (1-1000 nM) elevated [Ca2+]i of the cells even when extracellular calcium was chelated with 1 mM ethylene glycol-bis(beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA). These findings suggest that the transiently appearing NT-binding sites in the cortex are receptors for NT and that some of the NT-induced increase in [Ca2+]i is due to mobilization from the intracellular calcium store. Further application of NT after 10 min washing caused an increase in [Ca2+]i again. This is in contrast to the findings for cortical slices from adult rats and mRNA-injected oocytes; desensitization due to NT was of long duration and further application of NT failed to activate the neurons which had responded the first time to NT. These facts suggest that the character of the NT-binding sites in the cerebral cortex differs between neonatal and adult rats. In addition, we showed that neuromedin N had a similar property to NT as to mobilization of [Ca2+]i and acted only on NT-responsive cells, suggesting the interaction between NT and neuromedin N at the postsynaptic level via the same receptor.

Amino Acid Sequence↗

PubMed1991

A sex-specific cytochrome P-450(F-1) colocalized with various neuropeptides in the paraventricular and supraoptic nuclei of female rats.

Numerous cells containing P-450(F-1) were detected in the magnocellular and parvocellular neurons of the paraventricular nucleus of the hypothalamus. Electron microscopic analysis of immunoreactive neurons has shown that P-450(F-1) immunoreactivity is present on the Golgi apparatus and rough endoplasmic reticulum. In the paraventricular nucleus, the P-450(F-1)-positive magnocellular neurons frequently contained oxytocin and some of them also contained CRF. Vasopressin was colocalized with P-450(F-1), but these neurons did not express CRF. In the supraoptic nucleus, P-450(F-1) was colocalized with oxytocin or CRF in single neurons, but not with vasopressin. No cells exhibiting the colocalization of both P-450(F-1) and somatostatin were observed in these nuclei. The results of the present study concerning colocalization of P-450 and peptides suggest that P-450(F-1) is involved in the hypothalamo-hypophyseal neuroendocrine function in the female rat.

Animals↗

PubMed1991

Catecholamine and acetylcholine in the rat cerebral cortex with special reference to pathogenetic mechanisms of Alzheimer's disease.

A direct synapse between catecholamine fibers and neuropeptide Y-containing neurons is demonstrated in rat cerebral cortex using an immunohistochemical double-staining method under the electron microscope. A new method to produce a selective reduction in cholinergic neurons in the basal forebrain without damage to the non-cholinergic neurons, passing fibers or other cholinergic systems, is described. This animal model seems to be very useful to analyze the pathogenesis of Alzheimer's disease and to examine the function of cholinergic neurons of the basal forebrain.

Acetylcholine↗

PubMed1990

Long-term survival of grafted cells, dopamine synthesis/release, synaptic connections, and functional recovery after transplantation of fetal nigral cells in rats with unilateral 6-OHDA lesions in the nigrostriatal dopamine pathway.

In animal models of hemi-Parkinson's disease, survival of grafted nigral cells, their synaptic connections, dopamine (DA) synthesis/release, and recovery from motor disturbances were investigated, and these were compared among 3 groups of animals raised for 3 months, 1 year and 2 years after the transplantation. Fetal nigral DAergic cell suspensions were transplanted in the ipsilateral caudate nucleus of rats with unilateral 6-OHDA lesions in the nigrostriatal DA pathway. Motor disturbances, assessed by methamphetamine-induced rotation, recovered partly in the 2nd week, significantly in the 4th week after the grafting, and remained stable thereafter. Many tyrosine hydroxylase (TH)-positive cells were detected along the grafting tracks. The number of TH-positive cells was similar in the 3 groups of animals. These TH-positive cells made synaptic connections in the host caudate. By in vivo microdialysis measurement, extracellular DA, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) around the grafted sites recovered to 30-100% of those of controls. No significant differences were observed in the concentration of DA, DOPAC and HVA among 3 groups of animals. They also responded to methamphetamine loading though the magnitudes were smaller. Using a TH cDNA probe, TH-positive cells were found to express TH mRNA in in situ hybridization-autoradiographic analysis. Data indicate that grafted fetal DAergic cells survive, synthesize and release DA, make synaptic connections in the host brain and ameliorate motor disturbances for over 2 years. There were no differences in these parameters among the 3 groups of animals, and no untoward side effects were observed even at 2 years after the grafting. Thus it was confirmed that the grafting of neuronal cells into the brain is a promising approach to restore disturbed function.

Animals↗

PubMed1990

Histamine acts directly on calcitonin gene-related peptide- and substance P-containing trigeminal ganglion neurons as assessed by calcium influx and immunocytochemistry.

Primary cultures of rat trigeminal ganglion cells were exposed to histamine, and the intracellular free-calcium concentration, [Ca2+]i, was measured by the calcium-sensitive dye fura-2. Histamine increased the [Ca2+]i of the neurons. Pretreatment of the cells with histamine H1-receptor blocker, or removal of extracellular calcium, abolished the response, however, the response was not altered by pretreatment with H2-blocker. Immunocytochemical analysis showed that these cultured cells that responded to histamine identically showed substance P- or calcitonin gene-related peptide-like immunoreactivity.

Animals↗

PubMed1990

Different ontogenetic profiles of cells expressing prepro-neurotensin/neuromedin N mRNA in the rat posterior cingulate cortex and the hippocampal formation.

The ontogeny of the expression of prepro-neurotensin/neuromedin N messenger RNA (prepro-NT/NN mRNA) in the rat posterior cingulate cortex (retrosplenial cortex) and the hippocampal formation was investigated using in situ hybridization histochemistry. In the primordium of the posterior cingulate cortex and the hippocampal formation, prepro-NT/NN mRNA was first expressed on embryonic day 17, and was found in the subiculum, layers II-III in areas 29a and 29b, and layer VI in the posterior cingulate cortex at birth. Expression was also observed in the CA1 field. In the adult rat, the expression of prepro-NT/NN mRNA was reduced in the posterior cingulate cortex, and only a few positive cells were seen here. However, the CA1 field and the subiculum still contained numerous positive cells.

Animals↗