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Biomedical subjects

Y Nishimune

Publications and source records attributed to Y Nishimune.

At least 145 records · Page 8Linked to original sources
PubMed1986

Dibutyryl adenosine cyclic monophosphate regulates differentiation of type A spermatogonia with vitamin A in adult mouse cryptorchid testis in vitro.

Surgically prepared cryptorchid mouse testes containing only type A spermatogonia were cultured with (Bu)2cAMP in combination with vitamin A (retinol). Treatment with (Bu)2cAMP and retinol for 12-24 h and with basal medium for an additional 8 days stimulated mitotic activity in type A spermatogonia and induced differentiation of germ cells. However, (Bu)2cAMP alone did not induce differentiation of type A spermatogonia. Moreover, when cryptorchid testes were treated with (Bu)2cAMP for longer than 3 days in the presence or absence of retinol, differentiation of type A spermatogonia did not take place; disintegration of the seminiferous tubules occurred instead. When the cryptorchid testes were cultured for 24 h in a medium containing a fixed concentration of retinol and varying concentrations of (Bu)2cAMP from 0.001-0.4 mM, there was a dose-dependent increase in the number of differentiated and mitotic germ cells and type A spermatogonia. Likewise, at a fixed dose of (Bu)2cAMP and increasing concentrations of retinol, a dose-dependent increase in the number of differentiated and mitotic germ cells occurred. However, the number of type A spermatogonia was decreased. The addition of puromycin, cycloheximide, and actinomycin D to the medium completely blocked retinol-(Bu)2cAMP-induced differentiation of the germ cells. The present results suggest that cAMP and retinol trigger biochemical events promoting the synthesis of specific macromolecules involved in the proliferation and differentiation of type A spermatogonia.

Animals↗
PubMed1986

Effect of duration of cryptorchidism on ability of mouse germ cells to regenerate and differentiate.

When 2-month-old mice were made cryptorchid for periods of up to 19 months, testicular weight fell between 0 and 10 days, remained steady until 7 months, fell rapidly until 12 months, and then remained low. The recovery of weight gain of the testes after surgical reversal of the cryptorchidism was related to the duration of cryptorchidism. After 8 months very little change in testicular weight or specific activity of lactate dehydrogenase X was obtained.

Animals↗
PubMed1985

Separation of germ cells from somatic cells in mouse testis by affinity for a lectin, peanut agglutinin.

A new method for the separation of germ cells from somatic cells in the mouse testis was accomplished by making use of the differences in cell surface affinity for a lectin, peanut agglutinin (PNA). The separation procedure was based on the specific presence of PNA receptor on testicular germ cells and its absence on somatic cells, such as Leydig, Sertoli and peritubular cells. As a result, more than 99% of cells in PNA receptor-positive (PNA+) fractions were identified as germ cells by immunoperoxidase reaction with specific antiserum to mouse testicular germ cells. In contrast, Leydig cells were enriched in PNA receptor-negative (PNA-) fractions, i.e., 65% of cells in these fractions were cytochemically stained for 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activity.

Animals↗
PubMed1985

Effects of steel mutation on spermatogenesis stimulated by fetuin.

The effect of the steel mutation on spermatogenesis was investigated using organ culture. Cultured cryptorchid testes from WB-+/+ mice showed an effective differentiation of type A spermatogonia in response to Pedersen type III fetuin. In contrast, the cryptorchid testes from WB-S1/+ showed neither differentiation nor division of type A spermatogonia. The findings reported here are the first demonstration of retarded response of steel mutation on a well-known agent, fetuin.

Alleles↗
PubMed1984

In vitro demonstration of deleterious effect of steel mutation on spermatogenesis in mice.

The effect of the steel mutation on spermatogenesis was investigated by using organ culture. Cultured cryptorchid testes from C57BL/6- +/+ mice showed an effective differentiation of type A spermatogonia in response to an increased concentration of bovine serum. In contrast, the cryptorchid testes from C57BL/6- Sld/+ mice showed only limited differentiation of type A spermatogonia even in the highest concentration of serum tested. The type A spermatogonia of mutant mice showed the normal mitotic response but did not differentiate further. The findings reported here are the first in vitro demonstration of the deleterious effects of the steel mutation on testicular germ cell differentiation and suggest that the effect of the steel mutation is expressed in Sertoli cells, the only supporting element in seminiferous tubules, or germ cells themselves.

Alleles↗
PubMed1984

Vitamin A and follicle-stimulating hormone synergistically induce differentiation of type A spermatogonia in adult mouse cryptorchid testes in vitro.

To study the mechanism of vitamin A and FSH action on testicular germ cell differentiation, artificially induced cryptorchid testes of adult mice were cultured in vitro, because such testes contain only type A spermatogonia and Sertoli cells in their seminiferous tubules. A synergistic effect of vitamin A compounds with FSH, but not with LH, on testicular germ cell differentiation was observed. Our data suggested that FSH stimulated the proliferation of type A spermatogonia, whereas retinoids induce spermatogenesis from type A spermatogonia into intermediate or type B spermatogonia. Possible mechanisms of action of retinoids and FSH on testicular germ cell differentiation are discussed.

Animals↗
PubMed1984

Production, purification and characterization of the plasminogen activator in teratocarcinoma stem cells induced with sodium butyrate.

Suspension cultures of pluripotent teratocarcinoma cells were induced with sodium butyrate to produce plasminogen activator (PA), generally regarded as a marker enzyme of differentiation of the teratocarcinoma. The induction of plasminogen activator was very efficient, resulting in production of sufficient enzyme to allow its purification. The activator was inactivated by rabbit anti-human melanoma plasminogen activator antiserum, indicating that it was a tissue-type activator (t-PA). The enzyme was purified by column chromatograph on phosphocellulose, zinc-chelate agarose, Con-A Sepharose and Sephadex G-150. The preparation at the final step of purification gave a single peak of enzyme activity at pH 7.3 +/- 0.1 on isoelectric focusing, and showed a molecular weight of approximately 77,000 on SDS PAGE.

Animals↗
PubMed1983

Mode of cell surface marker changes during retinoic acid-induced differentiation in pluripotent embryonal carcinoma cells.

Pluripotent teratocarcinoma cell line, 311, was cultured in the presence of retinoic acid (RA) and studied for the processes of early marker changes associated with cell differentiation. The cell populations that have lost peanut agglutinin (PNA), Lotus tetragonolobus agglutinin (LTA) or wheat germ agglutinin (WGA) receptor increased in proportion to the period since the start of RA treatment. The kinetics of the appearance of these receptor-negative cell populations suggests that the differentiating cells lose lectin receptors in the order of PNA, LTA and WGA. However, the changes in F9 antigen(s) and LTA receptor occurred at an equal frequency in PNA+ and PNA- cells, indicating that, although the loss of lectin receptors takes place in a distinct order, the change in each receptor itself proceeds independently of the state of other lectin receptors.

Animals↗
PubMed1983

Induction of teratocarcinoma cell differentiation. Effect of the inhibitors of DNA synthesis.

Induction of teratocarcinoma cell (F9) differentiation was studied by using inhibitors of DNA synthesis and several agents known to be differentiation inducers. Inhibition of DNA synthesis induced changes in cell surface marker F9 and stimulated the production of plasminogen activator (PA) in a manner that is dependent upon de novo synthesis of RNA and protein. The results thus indicate close association between inhibition of DNA synthesis and induction of cell differentiation. This approach will be useful in investigating the mechanism of teratocarcinoma cell differentiation.

Animals↗
PubMed1983

Retinoids induce differentiation of type A spermatogonia in vitro: organ culture of mouse cryptorchid testes.

To study the effects of retinoids on testicular germ cell differentiation, especially on stem cells, artificially induced cryptorchid testes of adult mice were cultured in vitro inasmuch as they contain only stem cells, type A spermatogonia. We report here that retinol, retinal, retinol acetate, even retinoic acid, activated cell division in type A spermatogonia and stimulated them to differentiate. These findings are the first demonstration of the in vitro induction of early stage of spermatogenesis by retinoids and also retinoic acid, which is considered to be a biochemically inactive compound for mammalian testicular function.

Animals↗
PubMed1983

In vitro differentiation of Type A spermatogonia from mouse cryptorchid testes in serum-free media.

Artificially induced cryptorchid testes of adult mice were cultured in serum-free media. Pedersen Type III fetuin activated cell division in Type A spermatogonia and stimulated them to differentiate; it also had a synergistic effect with follicle-stimulating hormone (FSH) on testicular germ cell differentiation. However, Pedersen Type IV, Deutsch, and Spiro fetuin did not show any stimulatory effect on the differentiation of germ cells. It was suggested that some serum factor(s) which copurifies with Pedersen Type III fetuin may be the active principle rather than fetuin itself in the differentiation of testicular germ cells in vitro.

Animals↗
PubMed1982

Effects of beta-mercaptoethanol-containing medium and feeder cells on the differentiation of embryonal carcinoma cells.

Pluripotent, feeder-dependent teratocarcinoma cell lines were cultured without a feeder layer in a medium containing 10(-4) M beta-mercaptoethanol (beta-medium) and compared for the development of early markers with cells cultured with a feeder layer. The cells cultured in beta-medium lost the PNA (peanut agglutinin) receptor typically at low density. This change was accompanied with enhanced secretion of plasminogen activator and the loss of sensitivity to anti-F9 serum, indicating the stem cell differentiation. In contrast, the cells cultured on a feeder layer did not show any marker changes, thus indicating the lack of differentiation. These results indicate that while the presence of feeder cells inhibits cell differentiation, cultivation in beta-medium permits the differentiation of pluripotent teratocarcinoma cells.

Animals↗
PubMed1982

Hormones and the differentiation of type A spermatogonia in mouse cryptorchid testes incubated in vitro.

In order to study hormonal effects on testicular germ cell differentiation, especially on type A spermatogonia, artificially induced cryptorchid testes of adult mice were cultured in a medium containing testosterone, dihydrotestosterone, tri-iodothyronine, dibutyryl 3':5' cyclic adenosine monophosphate, human chorionic gonadotrophin, LH, FSH, insulin and transferrin. These substances, with the exception of FSH, showed no stimulatory effect on the differentiation of type A spermatogonia. However, FSH activated cell division in type A spermatogonia and stimulated them to differentiate, while LH showed neither the promotion of differentiation nor a synergistic effect on FSH-mediated germ cell differentiation.

Androgens↗
PubMed1981

Testicular DNA synthesis in vivo: changes in DNA synthetic activity following artificial cryptorchidism and its surgical reversal.

The kinetics of DNA synthesis of whole testis in mice was studied in artificial unilateral cryptorchidism and in its surgical reversal. Cryptorchidism impaired total testicular DNA synthesis. Approximately 1 month after the operation, activity of testicular DNA synthesis decreased to about 30% that of the contralateral intact testis. A complete recovery of DNA synthesis was observed within 1 month after the surgical reversal of cryptorchidism.

Animals↗