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Biomedical subjects

M Kawata

Publications and source records attributed to M Kawata.

At least 73 records · Page 4Linked to original sources
PubMed1998

Vulnerability to aging in the rat serotonergic system.

Distributional changes of serotonergic fibers associated with aging were demonstrated immunohistochemically. Old rat brains were morphologically characterized by the presence of peculiar features of serotonergic fibers not found in the young adult brain. In 24-month-old rats, these aberrant serotonergic fibers were subdivided into two groups according to morphological alterations: type 1 fibers consisting of thin fibers with moderately enlarged varicosities, and type 2 fibers consisting of much thicker fibers that have even larger varicosities and a tortuous course. These two types of fibers were distributed differentially in the forebrain. Type 1 fibers were found mainly in the striatum and frontoparietal cortex, whereas type 2 fibers were found in the posterior cingulate cortex and dentate gyrus of the hippocampal formation. Both types of aberrant fibers were seen in amygdala, frontoparietal cortex, hypothalamus, and thalamus. In 36-month-old rats, more highly degenerating arborizations were detected, and these aberrant ramifications were classified as follows based on shape as: type 3 fibers consisting of highly arborized thin fibers with a larger number of larger varicosities, and type 4 fibers consisting of thick fibers with abundant larger varicosities. Distributional difference indicated that type 1 fibers progress into type 3 fibers, whereas type 2 develop into type 4 fibers. These findings suggest the possibility that one set of pathological fibers emanate from the dorsal raphe nucleus and the other from the median raphe. Moreover, both two sets of serotonergic fibers show age-related aberrations in their morphology over same time course.

Aging↗
PubMed1998

In vivo time course of morphological changes and DNA degradation during the degeneration of castration-induced apoptotic prostate cells.

The in vivo time course of the morphological changes and DNA degradation in castration-induced apoptotic prostate cells was studied from the earliest to the latest stage of the degeneration process. To study this problem, we first induced apoptotic prostate cells in rats by castration for 3 days and then promptly and continuously blocked the death of healthy prostatic cells in the castrated rats by in vivo testosterone replacement. Because testosterone replacement could not stop the irreversible lysis of already damaged prostate cells, apoptotic cells at different stages of the degeneration process were eliminated sequentially from the prostate after the healthy prostate cells had been protected. Prostate cells at the earliest stage of apoptosis at the time when the castrated rats received testosterone replacement disappeared last. By tracing the morphological and DNA degradation of apoptotic cells after hormone treatment, we estimated the time course of prostate cell death from the early to the final stage. In the morphological evolution of apoptotic prostate cells, the clumping of nuclear chromatin, the degeneration of cytoplasm and the involution of the cell surface occurred and progressed simultaneously, resulting in the rapid formation of apoptotic bodies that were gradually digested by other cells. The DNA ladders of apoptotic cells were progressively cleaved into a mononucleosomal subunit that was further degraded at an additional site, generating a heterogeneous population of small nucleotides. The final digestion of DNA fragments occurred within the apoptotic bodies. The whole course of prostate cell death after castration took about 44 h.

Animals↗
PubMed1998

Steroid hormones and their receptors in the brain.

Steroid hormones regulate several important functions of the brain by altering the expression of particular genes through their receptors. First in this paper the localization of glucocorticoid receptor immunoreactivity and mRNA in the brain was examined. Second biphasic effects of glucocorticoid on the hippocampus was described and particular emphasis was given on the apoptosis. Third the significance of estrogen receptor in the sexually dimorphic areas was discussed. These results suggest that steroids modulate the gene expression along with the alteration of cell structures in a different manner in a tissue-specific pattern.

Animals↗
PubMed1998

The effects of oestrogen and progesterone on serotonin and its metabolite in the lateral septum, medial preoptic area and ventromedial hypothalamic nucleus of female rats.

The effects of sex steroid hormones on serotonin and its metabolite, 5-hydroxyindole-3-acetic acid (5-HIAA) in the lateral septal nucleus (LS), the medial preoptic area (MPA) and the ventromedial nucleus of the hypothalamus (VMH) of female rats were investigated, using immunohistochemistry and high-performance liquid chromatography (HPLC). Female rats were divided into three groups: ovariectomized rats (OVX group); OVX-rats treated with estradiol benzoate alone (E2 group); and OVX-rats treated with E2 plus progesterone (E2 + P group). We analysed the density of serotonin-immunoreactive fibres with a computer-assisted image analysis system, and measured the tissue concentrations of serotonin and 5-HIAA. Many serotonin-immunoreactive fibres were observed in the LS, MPA and VMH in all three groups. The density of serotonin-immunoreactive fibres in the MPA and VMH was significantly lower in the E2 and E2+P groups compared to the OVX group, whereas the LS showed no detectable differences among the three groups. In the HPLC study, the concentrations of serotonin in the MPA and VMH of the E2 and E2+P groups were significantly lower than that in the OVX group. There was no significant difference in the concentration of serotonin in the LS. The concentration of 5-HIAA and the ratio of 5-HIAA/serotonin in the LS, MPA and VMH showed no significant differences among the OVX, E2 and E2+P groups. The present results suggest that E2 priming for sexual behaviour can affect the serotonergic system by decreasing serotonin content, but not the turnover rate, in the MPA and VMH of female rats.

Animals↗
PubMed1998

Detection of epithelial ovarian-cancer-associated antigens involved in immune complexes by monoclonal antibodies.

To detect antigenic molecules involved in immune complexes (ICs) in patients with epithelial ovarian cancer, we developed several monoclonal antibodies (Mabs) by hybridoma technology from mice immunized with ovarian cancer tissues. Hybridoma supernatants were differentially screened with a panel of ICs purified from ascites of patients with ovarian cancer and with ICs from pooled normal human sera by the enzyme-linked immunosorbent assay (ELISA). From about 6,000 supernatants screened in 6 fusions, 4 Mabs showing preferential binding to the ascitic ICs were selected. Their antibody specificity was further examined with serum-free conditioned media of various cancer cell lines by an ELISA inhibition assay. The reactivity of 3 Mabs was inhibited by the media of epithelial ovarian cancer cell lines, but not by any of the non-ovarian cell lines tested, suggesting that the target antigens were derived from ovarian cancer cells. Furthermore, ICs detected by one Mab, designated 2F11, were elevated in the sera from 18 of 42 (42.9%) patients with epithelial ovarian cancer, but not in those from 39 patients with benign ovarian tumors or from 29 healthy individuals (with the exception of 1 serum), using sandwich ELISA with protein A as the capture reagent.

Adult↗
PubMed1997

Adrenalectomy-induced granule cell death is predicated on the disappearance of glucocorticoid receptor immunoreactivity in the rat hippocampal granule cell layer.

In this study, we observed the changes of glucocorticoid receptor (GR)-immunoreactivity (ir) and cell death in the rat hippocampal granule cell layer at various periods after adrenalectomy (ADX). Our results revealed that all of the rats shortly after ADX showed a rapid loss of GR-ir and subsequent appearance of degenerating cells in the granule cell layer. One month after ADX, however, about 80% of the rats displayed a restoration of GR-ir and the absence of degenerating cells in the granule cell layer, and this phenomenon was successively noted for 6 months. Hippocampal structural destruction 3 and 6 months after ADX was found in about 20% of the rats with loss of GR-ir in the granule cell layer; the ADX rats with even weak GR-ir in this area had a normal hippocampus. The treatment of rats with synthetic GR agonist, dexamethasone, immediately after ADX prevented the loss of GR-ir and significantly reduced the number of degenerating cells in the granule cell layer. Our results clarified that granule cell death after ADX was necessarily accompanied by the disappearance of GR-ir in the granule cell layer, suggesting that ADX-induced granule cell death is predicated on the loss of GR-ir and that the presence of GR-ir in this area may be important for granule cell survival.

Adrenalectomy↗
PubMed1997

Differential expression of estrogen receptor mRNA and protein in the female rat preoptic area.

The expression of estrogen receptor (ER) mRNA and ER protein in the medial preoptic area of ovariectomized rat was investigated at both cellular and regional levels using non-isotopic in situ hybridization and immunohistochemistry. ER mRNA was localized in the cytoplasm, while both liganded and unliganded forms of the ER protein were confined to the nucleus. Furthermore, ER mRNA containing cells were evenly distributed throughout the medial preoptic area, showing a homogeneous staining pattern compared to that of ER protein. ER immunoreactive cells were highly distributed in the medial, moderately in the lateral aspect of the medial preoptic area, showing a heterogeneous staining pattern with strongly and weakly labeled cells. These results suggest that ER protein levels are controlled by cellular posttranscriptional mechanisms.

Animals↗
PubMed1997

S100beta promotes the extension of microtubule associated protein2 (MAP2)-immunoreactive neurites retracted after colchicine treatment in rat spinal cord culture.

S100beta, a glial derived calcium-binding protein with neurotrophic activity in the central nervous system, stimulates neurite extension of fetal raphe, cortex, spinal cord, and dorsal root ganglion neurons. The effects of S100beta on neurite length and microtubule associated protein2 (MAP2) immunoreactivity (IR) after microtubule disruption with colchicine were investigated in primary rat spinal cord culture. The incubation with S100beta (20 ng/ml) for 3 h after exposure to colchicine (10(-6) M) for 30 min altered the distribution of MAP2-IR. The length of MAP2-IR neurites increased by 65% compared to that in colchicine treatment alone. MAP2-IR intensity in the cell body was reduced by 26% compared to that in colchicine treatment alone. These results indicate that neurites shrink when the microtubular cytoskeletal system is disrupted and S100beta rapidly promotes re-assembly and/or stabilization.

Animals↗
PubMed1997

The in vivo time course for elimination of adrenalectomy-induced apoptotic profiles from the granule cell layer of the rat hippocampus.

Although apoptotic cellular degeneration has been reported to be extremely rapid with the use of in vitro models, the time needed to clear apoptotic neurons in the in vivo brain is unknown. In this study we used a simple morphological approach to solve this problem. Four days after adrenalectomy (ADX), all of the operated rats morphologically displayed hippocampal granule cell apoptosis that was prevented completely by corticosterone replacement immediately after ADX. Therefore, we intravenously injected the rats with corticosterone 4 d after ADX and subsequently maintained them on corticosterone replacement in saline drinking water. This corticosterone replacement could protect healthy granule cells promptly and continuously against hormone-deficient apoptosis, because the normal glucocorticoid receptor immunoreactivity within the granule cell nuclei, which disappeared after ADX, was identified 1 hr after corticosterone replacement was started, and this effect persisted for several days. However, this corticosterone treatment could not prevent the irreversible apoptosis of the already degenerated granule cells at various stages of the same progressive apoptotic process. Then we successively traced the disappearance of apoptotic granule cells throughout the hippocampus at different time points by Nissl and silver staining. Given that the apoptotic cells at the earliest stage of the degenerating process when the ADX rats received corticosterone injection were the last to disappear, the period from corticosterone injection until the disappearance of the last degenerating debris of apoptotic cells was taken to represent the time course for elimination of apoptotic neurons in vivo. We discovered that the elimination of apoptotic granule cells took 72 hr.

Adrenalectomy↗
PubMed1997

Short inverted repeats function as hotspots of intermolecular recombination giving rise to oligomers of deleted plastid DNAs (ptDNAs).

We have determined the nucleotide sequences around the junction points of oligomeric-deleted ptDNAs possessing a head-to-head or tail-to-tail configuration from long-term cultured cell lines and albino plants. It was shown that DNA rearrangement occurred by direct fusion of deleted ptDNAs in an inverted orientation, which was linked by an asymmetrical sequence of 254-698 bp derived from either of the ptDNAs joined. It is notable that inverted repeats of 7-14 bp flank the asymmetrical sequences at each of the junction points. These features of the DNA sequence around the junction points are commonly observed in oligomeric ptDNA with a large-scale deletion regardless of the cell lines employed. It is suggested that the short inverted repeats are involved in the intermolecular recombination of ptDNA.

Base Sequence↗
PubMed1997

Chronic loss of glucocorticoids following adrenalectomy down-regulates the expression of glucocorticoid receptor mRNA in the rat forebrain.

In the negative feedback model, loss of endogenous glucocorticoids up-regulates the expression of glucocorticoid receptor mRNA. To elucidate further the effect of chronic lack of glucocorticoids on the expression of glucocorticoid receptor mRNA and protein, in situ hybridization and immunohistochemical methods were used to examine the long-term alteration of glucocorticoid receptor mRNA and its immunoreactivity in the forebrain of adrenalectomized rats. Constant lack of glucocorticoids resulted in marked decrease in the expression of glucocorticoid receptor mRNA and disappearance of glucocorticoid receptor immunoreactivity in many forebrain structures. In particular, in the suprapyramidal blade of the hippocampal granule cell layer and cerebral cortex, many cells showed almost no glucocorticoid receptor mRNA signals. These results suggest that long-term loss of endogenous glucocorticoids down-regulates the levels of glucocorticoid receptor mRNA, leading to reduction in the synthesis of glucocorticoid receptors in the rat forebrain. Therefore, the presence of endogenous glucocorticoids is vital to the continued expression of glucocorticoid receptor mRNA.

Adrenal Glands↗
PubMed1997

Astrocytic lineage analysis by detection of GFAP promoter activity in vitro.

To survey the emergence and onset of differentiation of the astrocytic lineage in the developing mouse cerebral wall, the promoter activity of a 2.5 kb 5'-flanking region of glial fibrillary acidic protein (GFAP) was measured in individual developing brain cells using a retrovirus-mediated gene transfer system. We identified precursors for astrocytes in primary culture of embryonic mouse cerebral wall cells by detection of GFAP promoter activity, which was detected approximately 3 days prior to the appearance of GFAP immunoreactivity. Since retroviruses only integrate into the chromosomes of actively proliferating cells, cells detected by this method should have been mitotically active at the time of retroviral infection on day 15 postfertilization (E15). Furthermore, we observed that cells activating GFAP promoter were located near the ventricular surface of cultured cerebral wall slices as a cluster of spherical cells. These results demonstrate that precursor cells for astrocytes exist within the germinative zone of developing cerebral wall, and that these cells are mitotically active on day E15, which is a late stage of neuronal production period in the mouse cerebral wall. The morphology, location and mitotic activity of these cells suggest that they are unlikely to be cells that have been transformed from radial glial cells.

Animals↗
PubMed1997

Vaginal atresia with transverse septum in a cat.

A six and half-year-old nulliparous mixed breed cat which had the complaints of vomiting, abdominal distention and depression was presented to the Veterinary Teaching Hospital of Osaka Prefecture University. She was suspected of pyometra by clinical signs and tests. By laparotomy, it was clarified that both uterine horns and vagina showed distension by the accumulation of secretions, and the vagina ended blindly leaving a tough connective tissue at the border between cranial and caudal part of the vagina. Postoperative contrast-radiograph of the remaining vagina proved it had no persistence of the hymen. From these findings, the condition was diagnosed as a feline atresia vaginalis with the transverse vaginal septum which is caused by the embryonic failure of canalization of the paramesonephric duct between the end of the Müllerian duct and the urogenital sinus.

Animals↗
PubMed1997

Localization of heat shock protein in osteoarthritic cartilage.

In osteoarthritic (OA) chondrocytes, expression of 70 kD heat shock protein (HSP70) is enhanced under stress. The HSP70 level is also related to OA severity in humans. In the present study, specimens of OA cartilage were obtained from 14 surgically treated female patients. Their disease severity was evaluated according to Mankin's histological/histochemical grading scores. The ratio of HSP70-positive cells was examined by using immunohistochemistry. As OA became more severe, the ratio of HSP70-positive cells increased in deeper areas from the surface, and there was a positive correlation between the grade and the ratio. Distribution of cells expressing HSP70 mRNA detected by in situ hybridization was similar to the distribution of positive cells in the immunohistochemistry. It was presumed that HSP70 plays a role in cell protection from stress, and that mechanical and biological factors which are related to the enhancement of HSP70 expression, have a considerable influence on OA progressions.

Aged↗
PubMed1997

Assessment of intravascular ultrasound-bearing balloon catheter-guided percutaneous transluminal coronary angioplasty and stenting.

To investigate the usefulness of an intravascular ultrasound (IVUS)-bearing balloon catheter, we analyzed results in 17 patients with 19 lesions who underwent percutaneous transluminal coronary angioplasty (PTCA) with IVUS-bearing balloon catheters (group A) and 25 patients with 28 lesions who underwent PTCA without IVUS (group B). The patients had angina pectoris with or without old myocardial infarction. The success rate (less than 50% residual stenosis on coronary angiography) in group A was 17/17 patients (100%) and 19/19 lesions (100%), and that in group B was 21/25 patients (84%) and 24/28 lesions (86%) (NS). The percent stenosis, by coronary angiography, in groups A and B was 71.3% and 76.8% before PTCA (NS) and 4.8% and 22.3% in groups A and B after PTCA (P = 0.005). Complication rates in the two groups were not significantly different, but non-Q myocardial infarction was observed in one patient with a type C lesion (according to the American Heart Association/American College of Cardiology [AHA/ACC] classification) in group A. Postdilation of Wiktor stents was determined by IVUS in four patients. These results suggest that the IVUS-bearing balloon catheter is useful to achieve larger lumen size for optimal PTCA.

Adult↗
PubMed1996

Neuro-glial neurotrophic interaction in the S-100 beta retarded mutant mouse (Polydactyly Nagoya). III. Transplantation study.

The hippocampus and caudo-dorsal cortex of the homozygote of polydactyly mutant mouse (Polydactyly Nagoya, Pdn/Pdn) were markedly reduced in S-100 beta positive astrocytes and serotonergic fibers as compared to the heterozygote (Pdn/+) and wild type (+/+) [39]. The Pdn/Pdn mice die within 2 days after birth, so it is impossible to examine postnatal changes. To demonstrate the developmental change of Pdn/Pdn hippocampal tissue, we transplanted hippocampal pieces of neonatal Pdn/Pdn and +/+ mice into the right and left hippocampus of the same adult +/+ mice, respectively, and immunocytochemically examined them. Two weeks after transplantation, +/+ hippocampal tissue contained a large number of glial fibrillary acidic protein (GFAP) and S-100 beta positive astrocytes and a number of serotonergic fibers. While Pdn/Pdn hippocampal tissue contained numerous GFAP positive astrocytes, S-100 beta positive astrocytes and serotonergic fibers were not observed. Two months after transplantation, GFAP and S-100 beta were expressed in the Pdn/Pdn hippocampal tissue similar to the +/+ tissue. Serotonergic fibers were distributed in the +/+ tissue, while no serotonergic fibers were observed in the Pdn/Pdn transplant tissue. In contrast, no difference was observed in the tyrosine hydroxylase positive fibers between Pdn/Pdn and +/+ grafts. The expression of 5-HT1A receptor-like immunoreactivity was higher in the +/+ tissue than that of Pdn/Pdn tissue. The present results suggest that the expression of S-100 beta in the astrocytes of early stage of transplantation is a critical for fiber ingrowth of serotonergic neurons and expressions of 5-HT1A receptor.

Animals↗
PubMed1996

The alteration of glucocorticoid receptor-immunoreactivity in the rat forebrain following short-term and long-term adrenalectomy.

To examine the effect of short-term and long-term adrenalectomy (ADX) on the glucocorticoid receptor (GR) expression, we performed an immunohistochemical study on the rat forebrain. One day after ADX, the GR-immunoreactivity significantly decreased or disappeared in most forebrain structures, while relatively strong GR-immunoreactivity was still found within the hypothalamus especially in the arcuate nucleus (ARC) and the parvocellular paraventricular nucleus (PVN). Two weeks following ADX, GR-immunoreactive cells disappeared in many structures of the forebrain including most parts of hypothalamus while moderate GR-immunoreactivity was still observable in the ARC and PVN. More than 3 months after ADX, the rats still survived when they received replacement of corticosterone during the first 2 weeks following the operation. Moderate GR-immunoreactivity in the ARC and PVN of the hypothalamus was exhibited whereas no immunoreactive cells remained in the cerebral cortex, thalamus and other forebrain structures when these animals showed obvious cell death in the granule cells of the dentate gyrus, identified with the silver impregnation method for degenerating cells. Massive cell loss in this hippocampal region is an indicator of a complete ADX, in addition to the blood corticosterone level. These results demonstrate topographic differences of GR expression in the rat forebrain after ADX with only continuous immunoreactivity in the ARC and PVN of the hypothalamus, suggesting that some neurons in the ARC and PVN could keep active GR probably in order to maintain their survival after removing the adrenal gland.

Adrenalectomy↗