Advanced SearchSearch Tips
Intracellular Mechanisms of Growth Hormone Action on Apoptosis in Cultured Porcine Ovarian Granulosa Cells
facebook(new window)  Pirnt(new window) E-mail(new window) Excel Download
 Title & Authors
Intracellular Mechanisms of Growth Hormone Action on Apoptosis in Cultured Porcine Ovarian Granulosa Cells
Sirotkin, A.V.; Makarevich, A.V.; Pivko, J.; Genieser, H.G.;
  PDF(new window)
The aims of this study were to detect spontaneously occurring apoptosis in cultured porcine ovarian cells, to examine the role of growth hormone (GH), tyrosine kinase (TK), protein kinase G (PKG) and cyclin-dependent kinase (CDK) in the control of this process, and to determine whether the effect of GH on apoptosis is mediated by TK-, PKG- and cdc2-dependent intracellular mechanisms. We studied the action of pGH (10 ng/ml), blockers of TK (genistein, lavendustin, both 100 ng/ml), PKG (Rp-Br-PET-cGMPS, 50 nM; KT5823, 100 ng/ml) and CDK (olomoucine, ), as well as combinations of GH with these blockers, on the onset of apoptosis in cultured granulosa cells isolated from antral (3-6 mm) porcine follicles. The functional characteristics of an early apoptotic event, DNA fragmentation, were determined using terminal deoxynucleotidyltransferase (TdT)-mediated dUTP nick end labelling (TUNEL), whilst morphological signs of advanced apoptosis such as pyknosis, chromatin marginalization, shrinkage and fragmentation of nucleus, were detected using routine light microscopy. After culture, some ovarian granulosa cells exhibited DNA fragmentation, which in some cases was associated with morphological apoptosis-related changes (pyknosis, shrinkage and fragmentation of the nucleus). GH significantly reduced the proportion of TUNEL-positive cells. Neither TK nor CDK blockers when given alone, significantly affected the percentage of TUNEL-positive cells although both PKG blockers significantly increased this index. Furthermore, TK and PKG blockers given together with GH, prevented or reversed the inhibitory effect of GH on apoptosis, whilst the CDK blocker olomoucine promoted it. These observations demonstrate apoptosis in porcine ovaries and suggest the involvement of GH, TK, PKG and CDK in the control of this process. They also suggest that the effect of GH on ovarian apoptosis is mediated or regulated by multiple signalling pathways including TK-, PKG- and CDK-dependent intracellular mechanisms.
Growth Hormone (GH);Protein Kinase G;Tyrosine Kinase;CDC Kinase;Apoptosis;Ovary;
 Cited by
Brison, D. R. and R. M. Schultz. 1997. Apoptosis during mouse blastocyst formation: evidence for a role for survival factors including TGF-alpha. Biol. Reprod. 56:1088-1096. crossref(new window)

Byrne, A. T., J. Southgate, D. R. Brison and H. J. Leese. 1999. Analysis of apoptosis in the preimplantation bovine embryo using TUNEL. J. Reprod. Fertil. 117:97-105. crossref(new window)

Campbell, G. S. 1997. Growth hormone signal transduction. J. Pediatry, 131:542-544.

Chun, S. Y., K. M. Eisenhauer, M. Kubo and A. J. Hsueh. 1995. Interleukin-1$\beta$ suppresses apoptosis in rat ovarian follicles by increasing nitric oxide production. Endocrinol. 136:3120-3127. crossref(new window)

Chun, S. Y., K. M. Eisenhauer, S. Minami, H. Billig, E. Perlas and A. J. Hsueh. 1996. Hormonal regulation of apoptosis in early antral follicles: follicle-stimulating hormone as a major survival factor. Endocrinol. 137:1447-1456. crossref(new window)

Costoya, J. A., J. Finidori, S. Moutoussamy, R. Searis, J. Devesa and V. M. Arce. 1999. Activation of growth hormone receptor delivers an antiapoptotic signal: evidence for a role of Akt in this pathway. Endocrinol. 140:5937-5943. crossref(new window)

Danilovich, N. A., A. Bartke and T. A. Winters. 2000. Ovarian follicle apoptosis in bovine growth hormone transgenic mice. Biol. Reprod. 62:103-107. crossref(new window)

Eisenhauer, K. M., S. -Y. Chun, H. Billig and A. J. W. Hsueh. 1995. Growth hormone suppression of apoptosis in preovulatory rat follicles and partial neutralization by insulin-like growth factor binding protein. Biol. Reprod. 53:13-20. crossref(new window)

Guthrie, H. D., W. M. Garrett and B. S. Cooper. 1998. Folliclestimulating hormone and insulin-like growth factor-I attenuate apoptosis in cultured porcine granulosa cells. Biol. Reprod. 58:390-396. crossref(new window)

Guthrie, H. D., W. M. Garrett and B. S. Cooper. 2000. Inhibition of apoptosis in cultured porcine granulosa cells by inhibitors of caspase and serine protease activity. Theriogenol. 54:731-740. crossref(new window)

Hirshfield, A. N. 1991. Development of follicles in the mammalian ovary. Int. Rev. Cytol. 124:43-101. crossref(new window)

Hsueh, A. J. W., H. Billig and A. Tsafriri. 1994. Ovarian follicle atresia: a hormonally controlled apoptotic process. Endocrinol. Rev. 15:1-18. crossref(new window)

Jeay, S., G. E. Sonenshein, M. C. Postel-Vinay and E. Baixeras. 2000. Growth hormone prevents apoptosis through activation of nuclear factor-kappaB in interleukin-3-dependent Ba/F3 cell line. Mol. Endocrinol. 14:650-661. crossref(new window)

Jones, S. M. and A. Kazlauskas. 2001. Growth factor-dependent signalling and cell cycle progression. FEBS Letters 490:110-116. crossref(new window)

Kaipia, A. and A. J. W. Hsueh. 1996. Regulation of ovarian follicular atresia. Annu. Rev. Physiol. 59:349-363. crossref(new window)

Kiya, T., T. Endo, H. Henmi, T. Goto, Y. Kitajima, K. Manase, S. Takahashi and R. Kudo. 1999. The effects of growth hormone on corpus luteum of superovulated rats. Endocrinol. Res. 25:179-193. crossref(new window)

Makarevich, A., A. Sirotkin, T. Taradajnik and P. Chrenek. 1997. Effects of genistein and lavendustin on reproductive processes in domestic animals in vitro. J. Steroid Biochem. Molec. Biol. 63:329-337. crossref(new window)

Petley, T., K. Graff, W. Jiang, H. Yang and J. Florini. 1999. Variation among cell types in the signalling pathways by which IGF-I stimulates specific cellular responses. Horm. Metab. Res. 31:70-76. crossref(new window)

Sirotkin, A. V. and A. V. Makarevich. 1999. Growth hormone regulates secretory activity and apoptosis in cultured bovine granulosa cells through the activation of the cAMP/protein kinase A system. J. Endocrinol. 163:317-327. crossref(new window)

Sirotkin, A. V., A. V. Makarevich, J. Pivko, J. Kotwica, H. -G. Genieser and J. Bulla. 2000. Effect of cGMP analogues and protein kinase G blocker on secretory activity, apoptosis and the cAMP/protein kinase A system in porcine ovarian granulosa cells in vitro. J. Steroid Biochem. Molec. Biol. 74:1-9. crossref(new window)