Leydig cell protein synthesis and steroidogenesis in response to acute stimulation by luteinizing hormone in rats

Lindi Luo, Haolin Chen, Douglas M. Stocco, Barry R Zirkin

Research output: Contribution to journalArticle

Abstract

We examined the temporal relationship between protein synthesis and testosterone production by rat Leydig cells in primary culture. Leydig cells were isolated from adult control Sprague-Dawley rats and from rats that had received LH-suppressive testosterone and estradiol (TE) implants in vivo for 10 days. The cells were incubated for 1-4 h with [35S]methionine in the presence or absence of maximally stimulating ovine LH, and newly synthesized proteins were examined by two-dimensional PAGE autoradiography. Approximately 800-900 newly synthesized polypeptides were readily visible on all autoradiograms, most of which did not differ in the cells from intact control and TE-treated rats. Incubation of cells from the control and treated rats with maximally stimulating LH for 4 h in both cases resulted in significant increases in testosterone production and in three newly synthesized polypeptides. These polypeptides, along with two others that changed little in response to LH, were similar in apparent molecular mass, 30 kDa, but differed in isoelectric point. Time-course studies revealed a temporal relationship between stimulation of the three 30-kDa proteins and of testosterone production. Western blot analysis identified the 30-kDa proteins as steroidogenic acute regulatory protein (STAR). The results of these studies, for the first time utilizing primary cultures of highly purified, testosterone-producing Leydig cells, provide further correlative evidence of a role for STAR protein in the acute regulation of Leydig cell testosterone biosynthesis by Lh.

Original languageEnglish (US)
Pages (from-to)263-270
Number of pages8
JournalBiology of Reproduction
Volume59
Issue number2
DOIs
StatePublished - Aug 1998

Fingerprint

Leydig Cells
Luteinizing Hormone
Testosterone
Proteins
Peptides
Estradiol
Isoelectric Point
Autoradiography
Methionine
Sprague Dawley Rats
Sheep
Western Blotting

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Embryology

Cite this

Leydig cell protein synthesis and steroidogenesis in response to acute stimulation by luteinizing hormone in rats. / Luo, Lindi; Chen, Haolin; Stocco, Douglas M.; Zirkin, Barry R.

In: Biology of Reproduction, Vol. 59, No. 2, 08.1998, p. 263-270.

Research output: Contribution to journalArticle

@article{a02dcd215e7f4250bca2d15ca8e0c5da,
title = "Leydig cell protein synthesis and steroidogenesis in response to acute stimulation by luteinizing hormone in rats",
abstract = "We examined the temporal relationship between protein synthesis and testosterone production by rat Leydig cells in primary culture. Leydig cells were isolated from adult control Sprague-Dawley rats and from rats that had received LH-suppressive testosterone and estradiol (TE) implants in vivo for 10 days. The cells were incubated for 1-4 h with [35S]methionine in the presence or absence of maximally stimulating ovine LH, and newly synthesized proteins were examined by two-dimensional PAGE autoradiography. Approximately 800-900 newly synthesized polypeptides were readily visible on all autoradiograms, most of which did not differ in the cells from intact control and TE-treated rats. Incubation of cells from the control and treated rats with maximally stimulating LH for 4 h in both cases resulted in significant increases in testosterone production and in three newly synthesized polypeptides. These polypeptides, along with two others that changed little in response to LH, were similar in apparent molecular mass, 30 kDa, but differed in isoelectric point. Time-course studies revealed a temporal relationship between stimulation of the three 30-kDa proteins and of testosterone production. Western blot analysis identified the 30-kDa proteins as steroidogenic acute regulatory protein (STAR). The results of these studies, for the first time utilizing primary cultures of highly purified, testosterone-producing Leydig cells, provide further correlative evidence of a role for STAR protein in the acute regulation of Leydig cell testosterone biosynthesis by Lh.",
author = "Lindi Luo and Haolin Chen and Stocco, {Douglas M.} and Zirkin, {Barry R}",
year = "1998",
month = "8",
doi = "10.1095/biolreprod59.2.263",
language = "English (US)",
volume = "59",
pages = "263--270",
journal = "Biology of Reproduction",
issn = "0006-3363",
publisher = "Society for the Study of Reproduction",
number = "2",

}

TY - JOUR

T1 - Leydig cell protein synthesis and steroidogenesis in response to acute stimulation by luteinizing hormone in rats

AU - Luo, Lindi

AU - Chen, Haolin

AU - Stocco, Douglas M.

AU - Zirkin, Barry R

PY - 1998/8

Y1 - 1998/8

N2 - We examined the temporal relationship between protein synthesis and testosterone production by rat Leydig cells in primary culture. Leydig cells were isolated from adult control Sprague-Dawley rats and from rats that had received LH-suppressive testosterone and estradiol (TE) implants in vivo for 10 days. The cells were incubated for 1-4 h with [35S]methionine in the presence or absence of maximally stimulating ovine LH, and newly synthesized proteins were examined by two-dimensional PAGE autoradiography. Approximately 800-900 newly synthesized polypeptides were readily visible on all autoradiograms, most of which did not differ in the cells from intact control and TE-treated rats. Incubation of cells from the control and treated rats with maximally stimulating LH for 4 h in both cases resulted in significant increases in testosterone production and in three newly synthesized polypeptides. These polypeptides, along with two others that changed little in response to LH, were similar in apparent molecular mass, 30 kDa, but differed in isoelectric point. Time-course studies revealed a temporal relationship between stimulation of the three 30-kDa proteins and of testosterone production. Western blot analysis identified the 30-kDa proteins as steroidogenic acute regulatory protein (STAR). The results of these studies, for the first time utilizing primary cultures of highly purified, testosterone-producing Leydig cells, provide further correlative evidence of a role for STAR protein in the acute regulation of Leydig cell testosterone biosynthesis by Lh.

AB - We examined the temporal relationship between protein synthesis and testosterone production by rat Leydig cells in primary culture. Leydig cells were isolated from adult control Sprague-Dawley rats and from rats that had received LH-suppressive testosterone and estradiol (TE) implants in vivo for 10 days. The cells were incubated for 1-4 h with [35S]methionine in the presence or absence of maximally stimulating ovine LH, and newly synthesized proteins were examined by two-dimensional PAGE autoradiography. Approximately 800-900 newly synthesized polypeptides were readily visible on all autoradiograms, most of which did not differ in the cells from intact control and TE-treated rats. Incubation of cells from the control and treated rats with maximally stimulating LH for 4 h in both cases resulted in significant increases in testosterone production and in three newly synthesized polypeptides. These polypeptides, along with two others that changed little in response to LH, were similar in apparent molecular mass, 30 kDa, but differed in isoelectric point. Time-course studies revealed a temporal relationship between stimulation of the three 30-kDa proteins and of testosterone production. Western blot analysis identified the 30-kDa proteins as steroidogenic acute regulatory protein (STAR). The results of these studies, for the first time utilizing primary cultures of highly purified, testosterone-producing Leydig cells, provide further correlative evidence of a role for STAR protein in the acute regulation of Leydig cell testosterone biosynthesis by Lh.

UR - http://www.scopus.com/inward/record.url?scp=0031904576&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031904576&partnerID=8YFLogxK

U2 - 10.1095/biolreprod59.2.263

DO - 10.1095/biolreprod59.2.263

M3 - Article

C2 - 9687294

AN - SCOPUS:0031904576

VL - 59

SP - 263

EP - 270

JO - Biology of Reproduction

JF - Biology of Reproduction

SN - 0006-3363

IS - 2

ER -