Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1978 Apr 15;172(1):147–153. doi: 10.1042/bj1720147

The effect of lutropin on specific protein synthesis in tumour Leydig cells and in Leydig cells from immature rats

Felix H A Janszen 1, Brian A Cooke 1, Maria J A Van Driel 1, Henk J Van Der Molen 1
PMCID: PMC1185673  PMID: 566102

Abstract

The amount of 35S incorporated into the various proteins after separation by electrophoresis on sodium dodecyl sulphate/polyacrylamide gels was used as an estimate of their synthesis in the Leydig cells. Increased synthesis of proteins with apparent mol.wts. 27000 and 29000 was observed 3h after addition of lutropin to tumour Leydig cells. Incubation of Leydig cells from immature rats with lutropin (100ng/ml) for 2h or longer resulted in increased synthesis of proteins with apparent mol.wts. 11000, 21000, 27000 and 29000. At higher concentrations (≥100ng/ml) of lutropin there was a decrease in the synthesis of a protein with apparent mol.wt. 13000. The amount of lutropin required for the stimulation of protein synthesis in both types of Leydig cells was similar to that needed for stimulation of steroidogenesis. Lutropin-stimulated specific protein synthesis was not due to increased concentrations of testosterone, however, because (1) addition of testosterone to the cells had no effect on the synthesis of the proteins, and (2) inhibition of steroidogenesis with elipten phosphate (an inhibitor of the cholesterol side-chain-cleavage enzyme complex) did not abolish the effect of lutropin. The stimulation of specific protein synthesis was also not due to contaminating follitropin in the lutropin preparation. Addition of actinomycin D to the cells at the start of the incubation prevented the effect of lutropin on specific protein synthesis, indicating that mRNA synthesis may be needed for this effect of lutropin. Incubation of the cells with cycloheximide for 30min after labelling of the proteins did not result in a detectable decrease in the amounts of the lutropin-induced proteins, indicating that their half-life is longer than 30min.

Full text

PDF
147

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Cooke B. A., Janszen F. H., Clotscher W. F., van der Molen H. J. Effect of protein-synthesis inhibitors on testosterone production in rat testis interstitial tissue and Leydig-cell preparations. Biochem J. 1975 Sep;150(3):413–418. doi: 10.1042/bj1500413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Garren L. D., Ney R. L., Davis W. W. Studies on the role of protein synthesis in the regulation of corticosterone production by adrenocorticotropic hormone in vivo. Proc Natl Acad Sci U S A. 1965 Jun;53(6):1443–1450. doi: 10.1073/pnas.53.6.1443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Grower M. F., Bransome E. D., Jr Adenosine 3',5'-monophosphate, adrenocorticotropic hormone, and adrenocortical cytosol protein synthesis. Science. 1970 Apr 24;168(3930):483–485. doi: 10.1126/science.168.3930.483. [DOI] [PubMed] [Google Scholar]
  4. Janszen F. H., Cooke B. A., van Driel J. A., van der Molen H. J. Regulation of the synthesis of lutropin-induced protein in rat testis Leydig cells. Biochem J. 1978 Jan 15;170(1):9–15. doi: 10.1042/bj1700009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Janszen F. H., Cooke B. A., van Driel M. J., van der Molen H. J. LH induction of a specific protein (LH-IP) in rat testis Leydig cells. FEBS Lett. 1976 Dec 1;71(2):269–272. doi: 10.1016/0014-5793(76)80948-9. [DOI] [PubMed] [Google Scholar]
  6. Janszen F. H., Cooke B. A., van Driel M. J., van der Molen H. J. Purification and characterization of Leydig cells from rat testes. J Endocrinol. 1976 Sep;70(3):345–359. doi: 10.1677/joe.0.0700345. [DOI] [PubMed] [Google Scholar]
  7. Janszen F. H., Cooke B. A., van der Molen H. J. Specific protein synthesis in isolated rat testis leydig cells. Influence of luteinizing hormone and cycloheximide. Biochem J. 1977 Feb 15;162(2):341–346. doi: 10.1042/bj1620341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Laychock S. G., Rubin R. P. Isolation of ACTH-induced protein from adrenal perfusate. Steroids. 1974 Aug;24(2):177–184. doi: 10.1016/0039-128x(74)90100-7. [DOI] [PubMed] [Google Scholar]
  9. Mendelson C., Dufau M., Catt K. Dependence of gonadotropin-induced steroidogenesis upon RNA and protein synthesis in the interstitial cells of the rat testis. Biochim Biophys Acta. 1975 Dec 5;411(2):222–230. doi: 10.1016/0304-4165(75)90302-5. [DOI] [PubMed] [Google Scholar]
  10. Rubin R. P., Sheid B., McCauley R., Laychock S. G. ACTH-induced protein release from the perfused cat adrenal gland: evidence for exocytosis? Endocrinology. 1974 Aug;95(2):370–378. doi: 10.1210/endo-95-2-370. [DOI] [PubMed] [Google Scholar]
  11. Sowell J. G., Folman Y., Eik-Nes K. B. Androgen metabolism in rat testicular tissue. Endocrinology. 1974 Feb;94(2):346–354. doi: 10.1210/endo-94-2-346. [DOI] [PubMed] [Google Scholar]
  12. Verjans H. L., Cooke B. A., de Jong F. H., de Jong C. M., van der Molen H. J. Evaluation of a radioimmunoassay for testosterone estimation. J Steroid Biochem. 1973 Nov;4(6):665–676. doi: 10.1016/0022-4731(73)90042-3. [DOI] [PubMed] [Google Scholar]
  13. Wiebe J. P. Steroidogenesis in rat leydig cells: changes in activity of 5-ane and 5-ene 3beta-hydroxysteroid dehydrogenases during sexual maturation. Endocrinology. 1976 Feb;98(2):505–513. doi: 10.1210/endo-98-2-505. [DOI] [PubMed] [Google Scholar]
  14. el-Safoury S., Bartke A. Aminoglutethimide inhibits steroidogenesis in the rat testis. Steroids. 1974 Feb;23(2):165–172. doi: 10.1016/0039-128x(74)90149-4. [DOI] [PubMed] [Google Scholar]
  15. van Beurden W. M., Roodnat B., de Jong F. H., Mulder E., van der Molen H. J. Hormonal regulation of LH stimulation of testosterone production in isolated Leydig cells of immature rats: the effect of hypophysectomy, FSH, and estradiol-17beta. Steroids. 1976 Dec;28(6):847–866. doi: 10.1016/0039-128x(76)90035-0. [DOI] [PubMed] [Google Scholar]
  16. van der Molen H. J., Grootegoed J. A., de Greef-Bijleveld M. J., Rommerts F. F., van der Vusse C. J. Distribution of steroids, steroid production and steroid metabolizing enzymes in rat testis. Curr Top Mol Endocrinol. 1975;2:3–23. doi: 10.1007/978-1-4613-4440-7_1. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES