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N Shirasawa

Publications and source records attributed to N Shirasawa.

At least 19 recordsLinked to original sources
PubMed2000

Cytological characterization of a pituitary folliculo-stellate-like cell line, Tpit/F1, with special reference to adenosine triphosphate-mediated neuronal nitric oxide synthase expression and nitric oxide secretion.

An immortal nonhormone-producing cell line with a characteristic star-shaped morphology, named Tpit/F1, was derived from an anterior pituitary gland of a temperature-sensitive large T antigen transgenic mouse. To characterize Tpit/F1 cells, we performed cytological studies, which revealed that Tpit/F1 cells express the messenger RNAs of neruonal nitric oxide (NO) synthase, S-100 protein, basic fibroblast growth factor, and pituitary-restricted transcription factor. The Tpit/F1 cells response to pituitary adenylate cyclase-activating peptide comprised the stimulated secretion of interleukin-6. Furthermore, glucocorticoids stimulate glutamine synthase production by Tpit/F1 cells. Considering these cytological characteristics together with their morphology, we deduced that Tpit/F1 cells are derived from pituitary folliculo-stellate (FS) cells. Our cytophysiological analyses of Tpit/F1 cells revealed that intracellular Ca2+ increased dose dependently on ATP administration (0-100 microM), and that this effect did not require the presence of extracellular Ca2+ and was not abolished by treatment with gadolinium, a Ca2+ channel blocker. The ATP-induced increase in intracellular Ca2+ ([Ca2+]i) was completely abolished by treatment with the Ca2+-adenosine triphosphatase (Ca2+-ATPase) inhibitor thapsigargin, which suggests that ATP increases [Ca2+]i by mobilizing internally stored Ca2+ followed by an influx of Ca2+. Moreover, UTP was equipotent with ATP in causing the [Ca2+]i increase in Tpit/F1 cells. Also, the Ca2+ response was prevented by the phospholipase C inhibitor, U-73122, but not by its inactive analog, U-73343. From these results we therefore concluded that ATP acts on Tpit/F1 cells via P2Y2-purinoceptors. Interestingly, both neuronal nitric oxide synthase messenger RNA and NO secretion were increased by ATP administration (10 and 100 microM). These results suggest the biological significance of the topological colocalization of FS cells and endocrine cells. Namely, ATP is cosecreted with hormones from endocrine cells and stimulates NO production by FS cells, and the released NO may regulate neighboring endocrine cell and blood vessels.

Adenosine Triphosphate↗
PubMed1999

Glucocorticoids induce glutamine synthetase in folliculostellate cells of rat pituitary glands in vivo and in vitro.

Glutamine synthetase (GS) is a glucocorticoid-inducible enzyme that has a key role for glutamate metabolism in the central and peripheral nervous system. In this study GS activity was measured and the amount of immunoreactive GS (ir-GS) cells in the rat anterior pituitary gland was quantified as a function of age. In addition, the effects of GS inhibitors, glucocorticoid administration, and adrenalectomy on GS activity were examined. Some of the ir-GS cells were also immunoreactive for S100 protein (ir-S100) which is a known marker for folliculostellate cells (FS) in the anterior pituitary. FS cells expressing GS were first detected in 3-d-old rats, and this cell population, expressed as the immunostained cell area divided by a standard unit area, increased as a function of age. The percentages of FS cells also expressing GS were 0.2, 6.4, 25 and 74% at 3 d, 30 d, 60 d and 2 y of age, respectively. GS enzyme activity also increased in parallel with the increase of ir-GS cell population maturation. The subcutaneous injection of methionine sulphoximine, a GS and gamma-glutamylcysteine synthetase inhibitor, reduced pituitary GS activity by 83%, but increased the population of ir-GS cells 3.5-fold in 30-d-old rats. Buthionine sulphoximine, a specific inhibitor of y-glutamylcysteine synthetase, had little effect on GS activity or the ir-GS cell population. Neither methionine sulphoximine nor buthionine sulphoximine changed the population of ir-S100 protein cells (FS cells). Dexamethasone and hydrocortisone increased the population of ir-GS cells by 3.1 and 4.2-fold, respectively, within 12 h after administration. A significant increase of GS activity due to the injection of glucocorticoids was observed in the anterior pituitary, but not in the brain, retina or liver of immature rats. Adrenalectomy did not cause decrease of pituitary GS activity, and dexamethasone administration increased GS activity in both adrenalectomised and intact rats. In the monolayer culture of anterior pituitary cells, glucocorticoids increased GS activity by x 1.5, and methionine sulphoximine reduced the activity by over 94%. These results demonstrate that GS in folliculostellate cells is a glucocorticoid-inducible enzyme in vivo and in vitro, and that the age-dependent increase of GS activity is independent of endogenous adrenal glucocorticoids.

Adrenalectomy↗
PubMed1996

Immunoreactive luteinizing hormone (ir-LH) cells in the lung and stomach of chick embryos.

Luteinizing hormone (LH) immunoreactivity was detected in the lung and stomach of chick embryos by the immunoperoxidase staining technique using specific antiserum to chicken LH. Immunoreactive LH (ir-LH) cells first appeared in the primordial cells of the epithelial layer of lung bud and foregut as well as of Rathke s pouch in the 3-day-old embryo, Hamburger and Hamilton stage 21. Ir-LH cells increased in number with advancing age of embryos in the lung, stomach, and pituitary gland. In the lung of 7-day-old embryos, stage 31, the ir-LH cells were distributed in the epithelium of primary, secondary, and tertiary bronchi, and their shapes were pseudostratified columnar, simple columnar, and simple cuboidal, depending on their sites in the intrapulmonary airway. Ir-LH cells were more numerous in the median part than in the lateral part of the lung, and the population in the epithelial layer of entobronchi of the secondary bronchi was 4 times higher than that in ectobronchi and laterobronchi of the secondary bronchi and in the primary bronchi. The immunoreactive products were found, either in the entire cell or in the apical part, facing the lumina of bronchi. In the stomach, ir-LH cells were found in the epithelial layer of gastric glands. No ir-LH cells were observed in interstitial regions, which consisted of mesenchymal cells and blood vessels, in the lung and stomach tissues. With advancing age, ir-LH cells changed their shapes to flat or squamous, coincident with the formation of parabronchi. Other pituitary hormones were not observed immunohistochemically in either the lung or stomach before hatching. Preabsorption of the antiserum against avian LH with the purified chicken LH or the extract of pituitaries from 10-day-old embryos completely destroyed the immunoreactivity to the cells in the lung and the pituitary. A single band of the immunoreaction products, whose molecular weight was around 25 K daltons, was shown by the immunostaining of nitrocellulose membrane transblotted after sodium dodecylsulphate-polyacrylamide gel electrophoresis of the purified pituitary LH, extracts of pituitaries from 10-day-old embryos, and the extracts of lungs from 7-, 10-, and 14-day-old chick embryos. These results demonstrated that ir-LH cells are present in extrapituitary tissues, and may play an important role during the development of chick embryonic lung and stomach.

Animals↗
PubMed1991

Immunohistochemistry of the pituitary pars distalis of the musk shrew, Suncus murinus.

Immunocharacteristics of the pituitary pars distalis cell types of the musk shrew, Suncus murinus, were studied by the unlabeled antibody enzyme technique, using peroxidase-antiperoxidase or avidin-biotin-peroxidase complex. The thyrotropin (TSH)-, gonadotropin (GTH)-, corticotropin (ACTH)-, prolactin (PRL)-, and growth hormone (GH)-secreting cells of the PD were identified on the basis of their immunoreactivity with different heterologous antisera. The TSH cells showed specific immunoreactivity with antisera against human (h) TSH beta and rat (r) TSH beta. Cells showing immunoreactivity with the antisera against hLH beta and ovine (o) LH beta were designated as GTH cells as no immunoreactivity was observed with antisera against hFSH beta and oFSH beta. The ACTH cells as well as the cells of the pars intermedia were revealed by anti-ACTH1-24 and anti-ACTH1-10 sera. Whereas the PRL cells were recognized by their immunoreactivity with antisera against hPRL and oPRL, the GH cells were identified with anti-hGH, anti-oGH, and anti-bovine (b) GH sera. TSH and GTH, TSH and ACTH, GTH and ACTH, ACTH and GH, ACTH and PRL, and GH and PRL cells were visualized in the same section using the dual immunoperoxidase technique. Comparison of the immunohistochemically identified cells with those described histochemically reveals several discrepancies, which expose the limitations of the latter techniques identifying adenohypophysial cells.

Adrenocorticotropic Hormone↗
PubMed1990

Cytoplasmic accumulations in rat primary brain cell cultures following treatment with E-64, a thiol protease inhibitor.

The present communication deals with morphological effects of E-64, a thiol protease inhibitor, on primary cultures dissociated from 15-day-old fetal rat brain. Brain cells were treated with different concentrations of E-64 varying from 0.1 to 50 micrograms/ml medium. Numerous cytoplasmic accumulations appeared in neuronal cells, astrocytes and oligodendrocytes exposed to a concentration of 1.0 micrograms/ml of E-64 or higher, and after withdrawal of E-64, the accumulations disappeared. These accumulations were immunocytochemically stained with the antibody against cathepsin B, a marker of lysosomal activity, and the staining patterns found with acid phosphatase were similar to those found with cathepsin B. It is hypothesized that the cytoplasmic accumulations were lysosomes and that the action of E-64 was reversible. Use of thiol protease inhibitors on primary brain cell culture may be an appropriate model system for studying protein storage diseases in neuronal cells.

Animals↗
PubMed1987

Appearance of the cyst- or ductule-like structures and their role in the restoration of the rat pituitary autograft.

The anterior pituitary glands of 31-day-old male rats were autotransplanted under renal capsule. At the 3rd and 5th day after transplantation, the autografts were observed by electron microscopy and by immunohistochemistry using antiserum against S-100 protein, which serves as marker of folliculo-stellate (FS) cells. On the 3rd day, a large of the graft was replaced with loose connective tissue in which the FS cells were remarkable in number. The FS cells encircled a small number of granular cells and formed cyst- or ductule-like structures with neighboring FS cells. Together, FS cells and the encircled granular cells formed a basement membrane surrounding the entire ductule. Throughout the regenerating gland, many such ductules were found. On the 5th day, both granular cells and FS cells increased in number. The above observations suggest that FS cells may play an important role in the restoration of degenerated pituitary glandular tissues during the early stage of the transplantation.

Animals↗
PubMed1985

Fine structural and immunohistochemical studies of goat adenohypophysial cells.

The fine structure of each type of anterior pituitary cell in the male goat was studied through the application of a superimposition technique in which adjacent thick sections were used to identify individual cells beforehand by light-microscopic immunohistochemistry. A cone of the pars intermedia protrudes into the pars anterior, being surrounded by the narrow pituitary cleft; the immunohistochemical appearances of the cells forming the cone resemble those of the pars anterior. Several follicles appear in the pars anterior. Ultrastructurally GH cells resemble prolactin cells. The secretory granules of both types are spherical; the diameter of the former is about 340 nm, whereas that of the latter is about 440 nm. ACTH cells are polygonal in shape with secretory granules, about 180 nm in diameter, scattered throughout the cytoplasm. TSH cells, which are spherical in shape, contain the smallest secretory granules, 150 nm in diameter. The highly electron-dense LH cells contain numerous secretory granules about 210 nm in diameter. Their nuclei are irregular with incisures. Thus, the anterior pituitary cells of the goat are ultrastructurally characteristics and species-specific.

Adrenocorticotropic Hormone↗
PubMed1984

Granulated folliculo-stellate cells and growth hormone cells immunostained with anti-S 100 protein serum in the pituitary glands of the goat.

Goat pituitary glands were immunohistochemically studied with antisera for bovine S-100 protein, rat LH beta, FSH, TSH beta, prolactin, ovine GH, and porcine ACTH1-39 by use of the superimposition technique on adjacent sections. Folliculo-stellate (F-S) cells were divided into two categories on the basis of ultrastructural properties: One consisted of a mass of agranular cells in which the pseudolumina were equipped with microvilli and cilia. Elongate gap junctions were often observed among these cells. The other was a group of granulated cells with or without pseudolumina. In this group the gap junctions were shown to be disintegrated. The dense granules 150-250 nm in diameter began to accumulate in the cells. However, neither type of these F-S cells was immunostained for S-100 protein. On the other hand, numerous polygonal, elongate, irregular or stellate cells containing S-100 protein were distributed throughout the gland. Most of them were immunohistochemically identical with the GH cells laden with the secretory granules 250-450 nm in diameter, but some of them were identical to TSH and prolactin cells which immunostained faintly for S-100 protein. This appears to be the first demonstration of GH cells intensely immunostained for S-100 protein.

Animals↗
PubMed1984

A histometrical study of immunohistochemically identified mitotic adenohypophysial cells in immature and mature castrated rats.

Two weeks after operation the effects of castration on all types of mitotic adenohypophysial cells in 30-day-old (immature) and 75-day-old (mature) male rats were studied immunohistochemically by light microscopy. The total number of mitotic cells in gland specimens increased 1.4 times in immature rats, but remained unchanged in mature animals; mitotic activity (mitotic cell number/mm2) was not affected in either age group. The population of mitotic gonadotrophs increased five to six times in comparison with intact controls. In addition, the percentage of mitotic gonadotrophs and their mitotic activity were raised in both age groups. Castration affected the mitosis of prolactin cells. The percentage of mitotic prolactin cells was reduced by one-third in castrated immature rats, but the total number and percentage of mitotic prolactin cells, as well as their mitotic activity, was reduced by about two-thirds in mature castrated rats. No significant changes in mitotic ACTH, TSH, GH and immunonegative cells were found after castration in either group of animals. Mitotic cells were more numerous in the anterior than in the posterior region of the gland in normal (uncastrated) immature rats, but were uniformly scattered in both regions in normal mature rats. On the other hand, castration induced a high population of mitotic cells in the anterior region regardless of age. This high population was the result of the enhanced mitosis of gonadotrophs. It is concluded that 2 weeks after castration the increment of mitotic divisions of all types of pituitary cells is age-dependent, and that mitosis takes place frequently in gonadotrophs and less frequently in prolactin cells.

Adrenocorticotropic Hormone↗
PubMed1983

Pituitary folliculo-stellate cells immunostained with S-100 protein antiserum in postnatal, castrated and thyroidectomized rats.

Pituitary folliculo-stellate cells and associated cells were studied immunohistochemically throughout the life of male and female rats. The marginal layer cells of the pars distalis and intermedia as well as the folliculo-stellate cells were immunostained with anti-S 100 protein serum. The immunostained folliculo-stellate cells in one section were identified as themselves by their ultrastructural properties on the adjacent section. Corticotrophs, thyrotrophs and prolactin cells, characterized by the stellate shape, were not immunostained with anti-S 100 protein serum. Reactivity for S-100 protein appeared on postnatal Day 6 in the marginal layer cells of the pars intermedia, and appeared on Day 10 in those cells of the pars distalis. No immunostained folliculo-stellate cells appeared before Day 6, but a few of them appeared on Day 10. Thereafter they increased in number, having more intense immunostaining with advancing age. In castrated rats, the immunostained folliculo-stellate cells appeared quite numerously, with branched cytoplasmic processes surrounding the gonadotrophs. In the thyroidectomized rats, however, folliculo-stellate cells lacked ramified cytoplasmic processes, and their topographic affinity for thyrotrophs was negligible.

Age Factors↗
PubMed1982

Immunohistochemical and electron microscopical studies of mitotic adenohypophysial cells in different ages of rats.

Mitotic rates of the six types of immunohistochemically identifiable adenohypophysial cells were histometrically calculated in colchicine-pretreated male rats 5, 17, 30 and 70 days old. Sections were stained with the antisera against rLH, rFSH, rTSH, oGH, rPRL and pACTH1-39. The mitotic growth rate of the anterior pituitary gland at 30 days of age was much higher than at other times. Mitotic growth rates of GH and PRL cells increased with advancing age, while those of ACTH-TSH- and immunonegative cells decreased with advancing age. LH/FSH cells showed no variation in mitotic growth rate with age. Mitotic cells can be classified into six cell types based on their fine structural properties: (1) agranular cells associated with the folliculo-stellate cells; (2) ambiguous cells with scanty minute secretory granules (50-150 nm in diameter); (3) basophils with a number of small secretory granules (130-200 nm); (4) immature acidophils whose large secretory granules (130-300 nm) are sporadically scattered; (5) acidophils with numerous spherical larger secretory granules (200-300 nm); and (6) prolactin cells with large polymorphic granules. At day 5 there was a high mitotic rate of the agranular and ambiguous cells [types (1) and (2)]; at day 70 a high mitotic rate was found in immature and mature acidophils [types (4) and (5)]. The mitotic rate of basophils (type 3) was high only at day 17 and low at all other times. The mitotic rate of prolactin cells (type 6) showed a slight increment with advancing age. It is concluded that the mitotic rates of the six cell types are age-dependent.

Aging↗
PubMed1981

A whole range of fine structural criteria for immunohistochemically identified LH cells in rats.

Pituitaries from normal, young and adult male rats were fixed either in sublimate-formalin or in glutaraldehyde-osmium. In adjacent Paraplast sections, almost all the gonadotrophs were immunostained with both LH and FSH antisera. The rat LH beta and FSH antisera used were shown to be highly specific by the absorption test and by double antibody radioimmunoassay. Thin and thick adjacent Epon sections were prepared for EM and immunohistochemical examination. Cells stained with the rat LH beta antiserum were identified by LM, and the observed in detail by EM. On the basis of these observations we suggest that the LH cells are arranged in a sequence of basophils, i.e., Types II/III, III, III/IV and IV: Type II/III basophils are elongate with a cytoplasmic process and less vesiculated. They have morphological features of Type II (classical thyrotrophs) and also of Type III basophils. Type III basophils are oval in shape and moderately vesiculated. Both Types II/III and III basophils can be divided into two classes of cell characterized mainly by the existence of only small secretory granules (150-220 nm in diameter) (Type A) or by the coexistence of small and large (350-500 nm) (Type B). Type III/IV basophils are cells intermediate between types III and IV basophils, and moderately vesiculated with an abundance of secretory granules (150-300 nm in diameter). Type IV basophils are large, spherical or oval cells whose RER cisternae are conspicuously dilated; they contain less numerous secretory granules (150-300 nm in diameter). It is concluded that LH cells are not a single cell type, but include a wide range of subtypes.

Animals↗
PubMed1979

Suppression of the accumulation of steroid-inducible glutamine synthetase mRNA on embryonic chick retinal polysomes by interferon preparation.

The suppression mechanism of glutamine synthetase [EC 6.3.1.2] induction mediated by chick interferon preparation was investigated in embryonic chick neural retina cell cultures. A translational assay was used to measure the level of mRNA coding for the enzyme. RNA extracted from the retinal polysomes was chromatographed on oligo(dT)-cellulose and translated in a cell-free protein synthesizing system derived from wheat germ. The newly synthesized enzyme was isolated by immunoprecipitation with anti-enzyme gamma-globulin and identified using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Neither polysomal profiles nor total protein synthesis directed by the RNA preparations was affected by interferon treatment. Comparing the amount of glutamine synthetase synthesized in vitro by equal amounts of mRNA fraction from retinas treated with or without interferon indicated that the suppression was associated with reduced level of the enzyme mRNA on polysomes.

Animals↗
PubMed1978

Interferon suppresses steroid-inducible glutamine synthetase biosynthesis in embryonic chick neural retina.

Effect of chick interferon on the biosynthesis of glutamine synthetase (L-glutamate: ammonia ligase (ADP-forming), EC 6.3.1.2) was studied in the embryonic chick neural retina cultures induced for the enzyme activity by hydrocortisone. The retinal enzyme radioactively labelled with [3H]leucine was precipitated by specific antibody against the enzyme isolated from adult chick liver. The immunological determination offered evidence that the suppressive effect of interferon on the hormonal induction of the enzyme was primarily due to reduced rate of its synthesis and accumulation.

Animals↗
PubMed1978

Susceptibility of chick neural retina to viral multiplication in vitro during embryonic development.

Decrease in the susceptibility of embryonic chick neural retina cultures to the multiplication of various viruses was observed with increasing age of the embryo. In contrast the retinal cells supported the multiplication of Sindbis virus irrespective of the age when they were infected with the viral RNA. These results suggest that the restricted multiplication of the viruses observed is due to the modulated inability of the cell to process the adsorbed viruses for subsequent replication.

Aging↗
PubMed1976

Reversible inhibition of interferon-induced antiviral state by deoxyadenosine.

Deoxyadenosine reversibly inhibited the development of antiviral state (AVS) in chick embryo fibroblasts stimulated by interferon. Studies on cellular macromolecular syntheses in deoxyadenosine-treated cells suggested that the inhibition was related to nucleolar RNA synthesis. Reference experiments with actinomycin D and cordycepin (3'deoxyadenosine) also implied causal- or interrelation between AVS development and nucleolar RNA synthesis.

Adenine↗