Biomedical subjects

N T Phamantu

Publications and source records attributed to N T Phamantu.

4 recordsLinked to original sources

PubMed1999

Activation of protein kinase C increases proteoglycan synthesis in immature rat Sertoli cells.

In order to determine the signal transduction pathways involved in the regulation of proteoglycan (PG) synthesis in immature rat Sertoli cells (SC), we have examined the effect of the tumor promoter phorbol ester PMA (phorbol myristate acetate) on [35S]sulfate and [3H]glucosamine incorporation into PG molecules neosynthesized by cultured rat SC. PMA induced a dose- and time-dependent stimulation of labeled cell-associated PG as determined by quantitative solid phase assay. The overall effect of PMA resulted from enhancement of both glycosylation and catabolism of cell PG, this latter effect leading to a drastic decrease of their residence time in the membrane. Besides these quantitative effects, activation of protein kinase C by PMA induced qualitative changes as reflected by increase in relative proportion of heparan sulfate PG (HSPG) in cell membrane PG. In light of our previous results suggesting an inverse relationship between PG synthesis and FSH responsiveness in immature rat Sertoli cells, the PMA-induced upregulation of cell membrane PG, and particularly HSPG, could constitute one mechanism involved in the repression of FSH-stimulated steroidogenesis induced by PKC activation.

Animals↗

PubMed1999

Inhibition of transmembrane calcium influx induces decrease in proteoglycan synthesis in immature rat Sertoli cells.

Beyond increased cAMP synthesis, calcium influx has been involved in signal transduction triggered by the gonadotropin follicle-stimulating hormone (FSH), the main regulator of Sertoli cells functions. In order to delineate a possible involvement of calcium in the regulation of proteoglycan synthesis, we have examined the effect of low-voltage-activated calcium channel blocker verapamil on both [(35)S]-sulfate and [(3)H]-glucosamine incorporation into proteoglycan molecules neosynthesized by cultured Sertoli cells from 20-day-old rats. Verapamil induced a dose- and time-dependent decrease in labeling of both secreted and cell-associated proteoglycans, as determined by quantitative solid-phase assay. This effect was mimicked by the addition of the calcium chelator EGTA, suggesting that verapamil effect resulted from the inhibition of transmembrane calcium influx. The decrease in apparent proteoglycan synthesis appeared to be attributable primarily to a lowering of the glycanation process, as shown by experiments using an exogenous acceptor for glycosaminoglycan synthesis. Moreover, verapamil induced a decrease in relative proportion of heparan sulfate proteoglycans in the cell layer. Pulse-chase kinetics demonstrated that verapamil also altered proteoglycan catabolism, leading to glycosaminoglycan retention in the cell layer and inhibiting the proteoglycan desulfation step. We conclude that intracellular calcium is essential to maintain Sertoli cell proteoglycan expression and could thus be involved in the repression of Sertoli cell cAMP-dependent syntheses such as estradiol production.

Animals↗

PubMed1995

Inhibition of proteoglycan synthesis induces an increase in follicle stimulating hormone (FSH)-stimulated estradiol production by immature rat Sertoli cells.

In order to define the possible involvement of proteoglycans (PG) in the regulation of Sertoli cell functions, we have examined the effect of para-nitrophenyl-beta-D-xyloside (PNPX), a specific inhibitor of PG synthesis, on follicle stimulating hormone (FSH)-dependent estradiol production by immature rat Sertoli cells. Addition of PNPX to the culture medium induced a dose-dependent inhibition of 35S-labeled PG synthesis in Sertoli cells both in the medium and the cell layer. Simultaneously there was a drastic increase in 35S-labeled secreted glycosaminoglycans. By 1 mM PNPX, syntheses of chondroitin sulfate proteoglycans released into culture medium and of heparan sulfate proteoglycans associated with the cell layer were 35% of values from untreated cells. Simultaneously, PNPX induced a twofold (mean of seven experiments, range 17-250%) enhancement of FSH (100 ng/ml)-stimulated estradiol production. In each individual experiment, there was an inverse relationship between the amplitude of PNPX-induced increase in FSH responsiveness and the FSH capability to stimulate basal estradiol production in cultured rat Sertoli cells. The effect of PNPX on FSH-stimulated aromatase activity was not mimicked by para-nitrophenyl-beta-D-galactoside, a structural analog of PNPX that has no effect on PG synthesis. The (Bu)2cAMP-stimulated estradiol synthesis was not modified in the presence of PNPX. Moreover, PNPX enhancement of FSH-stimulated estradiol synthesis disappeared when Sertoli cells were cultured in the presence of 1-methyl-3-isobutylxanthine, an inhibitor of phosphodiesterase activity. These findings suggest that inhibition of PG synthesis under PNPX conditions did not affect signal transduction steps distal to cAMP but rather decreased the phosphodiesterase activity in Sertoli cells.(ABSTRACT TRUNCATED AT 250 WORDS)

1-Methyl-3-isobutylxanthine↗

PubMedDate not supplied

Sodium chlorate induces undersulfation of cellular proteoglycans and increases in FSH-stimulated estradiol production in immature rat Sertoli cells.

The functional influence of cell proteoglycan (PG) undersulfation on estradiol synthesis by immature rat Sertoli cell cultures was investigated by using sodium chlorate, an inhibitor of the active sulfate donor for sulfotransferases. The addition of sodium chlorate to 20-day-old rat Sertoli cell cultures abolished [35S]-sulfate incorporation into neosynthesized PG and consequently reduced the residence time of undersulfated PG in cell membrane. Simultaneously, follicle-stimulating hormone (FSH)-stimulated estradiol synthesis was increased by 45%. The effects of sodium chlorate upon Sertoli cell PG synthesis and steroidogenesis were not reproduced with the addition of sodium chloride. Addition of phosphodiesterase inhibitors (MIX or Ro20-1724) decreased the magnitude of the chlorate effect on FSH-stimulated steroidogenesis, suggesting that part of chlorate's effect on steroidogenesis resulted from a decrease in adenosine cyclic 3',5'-phosphate (cAMP)-specific phosphodiesterase activity. Additionally, chlorate 1) increased Sertoli cell steroidogenesis at a step located beyond cAMP (restricted to Sertoli cell cultures exhibiting moderate steroidogenic response to (Bu)2cAMP) and 2) abolished the inhibition of steroidogenesis induced by transforming growth factor-beta. These results support our previous data, which showed that alteration in PG synthesis and the consequent decrease in cell membrane PG content induce an increase in FSH-stimulated estradiol synthesis in Sertoli cell cultures. The identification of cAMP-specific phosphodiesterase activity as a signal transduction step modified by PG undersulfation suggests the possible involvement of cell PG in the regulation of phosphodiesterase activity and, therefore, of FSH responsiveness during testicular development.

Animals↗