Register      Login
Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

Reproduction and plasma concentrations of leptin, insulin and insulin-like growth factor 1 in growth-hormone-transgenic female sheep before and after artificial insemination

H. Kadokawa A B C D , J. R. Briegel A , M. A. Blackberry C , D. Blache C , G. B. Martin C and N. R. Adams A
+ Author Affiliations
- Author Affiliations

A Division of Livestock Industries, Private Bag #5, PO Wembley, WA 6913, Australia.

B Department of Animal Production, National Agricultural Research Center for Hokkaido Region, Hitsujigaoka 1, Toyohira, Sapporo, 062-8555, Japan.

C School of Animal Biology, Faculty of Natural and Agricultural Sciences, The University of Western Australia, Crawley, WA 6009, Australia.

D To whom correspondence should be addressed. email: hiroya@affrc.go.jp

Reproduction Fertility and Development 15(1) 47-53 https://doi.org/10.1071/RD02074
Submitted: 10 September 2002  Accepted: 21 January 2003   Published: 21 January 2003

Abstract

The transgenic sheep used in this study expressed an additional copy of the gene for ovine growth hormone (GH), so they had continuously high plasma concentrations of GH. They were used to test whether the GH transgene affected plasma concentrations of the metabolic hormones leptin, insulin-like growth factor 1 (IGF-1) and insulin, and whether these effects were associated with changes in conception, pregnancy or parturition following artificial insemination. Compared with control animals, the GH-transgenic sheep had higher bodyweight, lower body condition score and less subcutaneous fat (Pthinsp;< 0.05). These sheep also had lower plasma concentrations of leptin, higher plasma concentrations of insulin, and higher plasma concentrations of IGF-1 (Pthinsp;< 0.001). A similar proportion of GH-transgenic and control ewes came into oestrus, but the conception rate to artificial insemination was lower in GH-transgenic ewes than in the controls. Only four live lambs were recovered from 12 GH-transgenic ewes (33%) compared with 38 lambs from 43 controls (88%). This outcome was not associated with any difference in plasma progesterone profile in the period leading up to artificial insemination (Day 0). The GH-transgenic ewes had lower concentrations of FSH at all times measured (Day −19, Day −2 and Day 19). These results indicate that appropriate regulation of GH secretion from pituitary or peripheral tissues is necessary for normal reproduction and normal levels of metabolic hormones. Chronically high concentrations of GH were associated with increased levels of IGF-1 and insulin, and decreased levels of leptin.

Extra keywords: body fatness, fertility, FSH.


Acknowledgments

We thank Dr B. Brown (CSIRO) for supplying the frozen semen from GH-transgenic rams and Dr K. Ward (CSIRO) for identifying the sheep that carried the transgene. Antiserum for the insulin assay was kindly provided by Dr P. Wynn (Sydney University). Reagents for the assays of IGF-1 and FSH were kindly donated by National Institute of Diabetes, Digestive & Kidney Disease and Dr A.F. Parlow (Pituitary Hormones and Antisera Center, Harbour-UCLA Medical Center, Torrance, CA, USA). Development of the leptin assay was supported by the National Health and Medical Research Council of Australia (Grant no. 981672). This study was supported in part by an Australian S and T Award 2000–2001 of the Australian Academy of Science.


References

Adams, N. R. , Briegel, J. R. , and Ward, K. A. (2002). The impact of a transgene for ovine growth hormone on the performance of two breeds of sheep. J. Anim. Sci. 80, 2325–33.
PubMed |

Amstalden, M. , Garcia, M. R. , Williams, S. W. , Stanko, R. L. , Nizielski, S. E. , Morrison, C. D. , Keisler, D. H. , and Williams, G. L. (2000). Leptin gene expression, circulating leptin, and luteinizing hormone pulsatility are acutely responsive to short-term fasting in prepubertal heifers: relationships to circulating insulin and insulin-like growth factor I. Biol. Reprod. 63, 127–33.
PubMed |

Bartke, A. , Cecim, M. , Tang, K. , Steger, R. W. , Chandrashekar, V. , and Turyn, D. (1994). Neuroendocrine and reproductive consequences of overexpression of growth hormone in transgenic mice. Proc. Soc. Exp. Biol. Med. 206, 345–59.
PubMed |

Bartke, A. , Chandrashekar, V. , Turyn, D. , Steger, R. W. , and Debeljuk, L. (1999). Effects of growth hormone overexpression and growth hormone resistance on neuroendocrine and reproductive functions in transgenic and knock-out mice. Proc. Soc. Exp. Biol. Med. 222, 113–23.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Blache, D. , Chagas, L. M. , Blackberry, M. A. , Vercoe, P. E. , and Martin, G. B. (2000a). Metabolic factors affecting the reproductive axis in male sheep. J. Reprod. Fertil. 120, 1–11.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Blache, D. , Tellam, R. L. , Chagas, L. M. , Blackberry, M. A. , Vercoe, P. E. , and Martin, G. B. (2000b). Level of nutrition affects leptin concentrations in plasma and cerebrospinal fluid in sheep. J. Endocrinol. 165, 625–37.
PubMed |

Breier, B. H. , Gallaher, B. W. , and Gluckman, P. D. (1991). Radio-immunoassay for Insulin-like growth factor-I: solutions to some potential problems and pitfalls. J. Endocrinol. 128, 347–57.
PubMed |

Brozos, C. N. , Saratsis, Ph. , Boscos, C. , Kyriakis, S. C. , and Alexopoulos, C. (1999). The effect of bovine somatotropin (bST) administration on reproduction, progesterone concentration during lactation and LH secretion during estrus, in dairy ewes. Anim. Reprod. Sci. 56, 177–87.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Cecim, M. , Kerr, J. , and Bartke, A. (1995). Infertility in transgenic mice overexpressing the bovine growth hormone gene: luteal failure secondary to prolactin deficiency. Biol. Reprod. 52, 1162–6.
PubMed |

Chandrashekar, V. , and Bartke, A. (1998). The role of growth hormone in the control of gonadotropin secretion in adult male rats. Endocrinology 139, 1067–74.
PubMed |

Chandrashekar, V. , Bartke, A. , and Wagner, T. E. (1992). Neuroendocrine function in adult female transgenic mice expressing the human growth hormone gene. Endocrinology 130, 1802–8.
PubMed |

Chandrashekar, V. , Bartke, A. , Awoniyi, C. A. , Tsai-Morris, C. H. , Dufau, M. L. , Russell, L. D. , and Kopchick, J. J. (2001). Testicular endocrine function in GH receptor gene disrupted mice. Endocrinology 142, 3443–50.
PubMed |

Curlewis, J. D. , and McNeilly, A. S. (1991). Prolactin short-loop feedback and prolactin inhibition of luteinizing hormone secretion during the breeding season and seasonal anoestrus in the ewe. Neuroendocrinology 54, 279–85.
PubMed |

Curlewis, J. D. , Sibbald, A. M. , Milne, J. A. , and McNeilly, A. S. (1991). Chronic treatment with long-acting bromocriptine does not affect duration of the breeding season, voluntary food intake, body weight, or wool growth in the Scottish blackface ewe. Reprod. Fertil. Dev. 3, 25–33.
PubMed |

Delavaud, C. , Bocquier, F. , Chilliard, Y. , Keisler, D. H. , Gertler, A. , and Kann, G. (2000). Plasma leptin determination in ruminants: effect of nutritional status and body fatness on plasma leptin concentration assessed by a specific RIA in sheep. J. Endocrinol. 165, 519–26.
PubMed |

Elimam, A. , Lindgren, A. C. , Norgren, S. , Kamel, A. , Skwirut, C. , Bang, P. , and Marcus, C. (1999). Growth hormone treatment downregulates serum leptin levels in children independent of changes in body mass index. Horm. Res. 52, 66–72.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Fowlkes, J. , and Freemark, M. (1992). Binding of placental lactogen and growth hormone to fetal sheep fibroblasts. Pediatr. Res. 32, 200–3.
PubMed |

Fruhbeck, G. (2001). A heliocentric view of leptin. Proc. Nutr. Soc. 60, 301–18.
PubMed |

Furuhata, Y. , Kagaya, R. , Hirabayashi, K. , Ikeda, A. , Chang, K.-T. , Nishihara, M. , and Takhashi, M. (2000). Development of obesity in transgenic rats with low circulating growth hormone levels: involvement of leptin resistance. Eur. J. Endocrinol. 143, 535–41.
PubMed |

Furuhata, Y. , Hirabayashi, K. , Yonezawa, T. , Takahashi, M. , and Nishihara, M. (2002). Effects of pair-feeding and growth hormone treatment on obese transgenic rats. Eur. J. Endocrinol. 146, 245–9.
PubMed |

Gales, N. J. , Williamson, P. , Higgins, L. V. , Blackberry, M. A. , and James, I. (1997). Evidence for a prolonged postimplantation period in the Australian sea lion (Neophoca cinerea). J. Reprod. Fertil. 111, 159–63.


Hardie, L. J. , Rayner, D. V. , Holmes, S. , and Trayhurn, P. (1996). Circulating leptin levels are modulated by fasting, cold exposure and insulin administration in lean but not Zucker (fa/fa) rats as measured by ELISA. Biochem. Biophys. Res. Commun. 223, 660–5.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Herman, A. , Helman, D. , Livnah, O. , and Gertler, A. (1999). Ruminant placental lactogens act as antagonists to homologous growth hormone receptors and as agonists to human or rabbit growth hormone receptors. J. Biol. Chem. 274, 7631–9.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Houseknecht, K. L. , Portocarrero, C. P. , Ji, S. , Lemenager, R. , and Spurlock, M. E. (2000). Growth hormone regulates leptin gene expression in bovine adipose tissue: correlation with adipose IGF-I expression. J. Endocrinol. 164, 51–7.
PubMed |

Hull, K. L. , and Harvey, S. (2001). Growth hormone: role in female reproduction. J. Endocrinol. 168, 1–23.
PubMed |

Hull, K. L. , and Harvey, S. (2002). GH as a co-gonadotropin: the relevance of correlative changes in GH secretion and reproductive state. J. Endocrinol. 172, 1–19.
PubMed |

Isozaki, O. , Tsushima, T. , Miyakawa, M. , Nozoe, Y. , Demura, H. , and Seki, H. (1999). Growth hormone directly inhibits leptin gene expression in visceral fat tissue in fatty Zucker rats. J. Endocrinol. 161, 511–16.
PubMed |

Jimenez-Krassel, F. , Binelli, M. , Tucker, H. A. , and Ireland, J. J. (1999). Effect of long-term infusion with recombinant growth hormone-releasing factor and recombinant bovine somatotropin on development and function of dominant follicles and corpora lutea in Holstein cows. J. Dairy Sci. 82, 1917–26.
PubMed |

Joyce, I. M. , Khalid, M. , and Haresign, W. (2000). The effect of recombinant GH treatment on ovarian follicle growth and atresia in sheep. Theriogenology 54, 327–38.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Kieffer, T. J. , and Habener, J. F. (2000). The adipoinsular axis: effects of leptin on pancreatic beta-cells. Ame. J. Physiol. Endocrinol. Metab. 278, E1–14.


Kolaczynski, J. W. , Nyce, M. R. , Considine, R. V. , Boden, G. , Nolan, J. J. , Henry, R. , Mudaliar, S. R. , Olefsky, J. , and Caro, J. F. (1996). Acute and chronic effect of insulin on leptin production in humans, studies in vivo and in vitro. Diabetes 45, 699–701.
PubMed |

Lincoln, G. A. (1990). Correlation with changes in horns and pelage, but not reproduction, of seasonal cycles in the secretion of prolactin in rams of wild, feral and domesticated breeds of sheep. J. Reprod. Fertil. 90, 285–96.
PubMed |

Liu, X. , Andoh, K. , Yokota, H. , Kobayashi, J. , Abe, Y. , Yamada, K. , Mizunuma, H. , and Ibuki, Y. (1998). Effects of growth hormone, activin, and follistatin on the development of preantral follicle from immature female mice. Endocrinology 139, 2342–7.
PubMed |

Marie, M. , Findlay, P. A. , Thomas, L. , and Adam, C. L. (2001). Daily patterns of plasma leptin in sheep: effects of photoperiod and food intake. J. Endocrinol. 170, 277–86.
PubMed |

Miller, D. W. , Blache, D. , and Martin, G. B. (1995). The role of intracerebral insulin in the effect of nutrition on gonadotrophin secretion in mature male sheep. J. Endocrinol. 147, 321–9.
PubMed |

Mizuno, T. M. , Bergen, H. , Funabashi, T. , Kleopoulos, S. P. , Zhong, Y. G. , Bauman, W. A. , and Mobbs, C. V. (1996). Obese gene expression – reduction by fasting and stimulation by insulin and glucose in lean mice, and persistent elevation in acquired (diet-induced) and genetic (yellow agouti) obesity. Proc. Natl Acad Sci. USA 93, 3434–8.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Monget, P. , and Martin, G. B. (1997). Involvement of insulin-like growth factors in the interactions between nutrition and reproduction in female mammals. Hum. Reprod. 12, 33–52.(Suppl.)
PubMed |

Moschos, S. , Chan, J. L. , and Mantzoros, C. S. (2002). Leptin and reproduction: a review. Fertil. Steril. 77, 433–44.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Naar, E. M. , Bartke, A. , Majumdar, S. S. , Buonomo, F. C. , Yun, J. S. , and Wagner, T. E. (1991). Fertility of transgenic female mice expressing bovine growth hormone or human growth hormone variant genes. Biol. Reprod. 45, 178–87.
PubMed |

Nancarrow, C. D. , Marshall, J. T. A. , Clarkson, J. L. , Murray, J. D. , Millard, R. M. , Shanahan, C. M. , Wynn, P. C. , and Ward, K. A. (1991). Expression and physiology of performance regulating genes in transgenic sheep. J. Reprod. Fertil. Suppl. 43, 277–91.
PubMed |

Picazo, R. A. , Gonzalez, D. B. A. , Gomez, B. A. , del Campo, A. , Granados, B. , Gresguerres, J. , and Lopez, S. A. (2000). Effects of bromocriptine administration during the follicular phase of the oestrous cycle on prolactin and gonadotrophin secretion and follicular dynamics in merino monovular ewes. J. Reprod. Fertil. 120, 177–86.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Pomp, D. , Geisert, R. D. , Durham, C. M. , and Murray, J. D. (1995). Rescue of pregnancy and maintenance of corpora lutea in infertile transgenic mice expressing an ovine metallothionein 1a-ovine growth hormone fusion gene. Biol. Reprod. 51, 170–8.


Russel, A. J. F. , Doeny, J. M. , and Gunn, R. G. (1969). Subjective assessment of body fat in live sheep. J. Agric. Sci. Cambridge 72, 451–4.


Saladin, R. , Devos, P. , Guerremillo, M. , Leturque, A. , Girard, J. , Staels, B. , and Auwerx, J. (1995). Transient increase in obese gene expression after food intake or insulin administration. Nature 377, 527–9.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Silbergeld, A. , Klinger, B. , Schwartz, H. , and Laron, Z. (1992). Serum prolactin in patients with Laron-type dwarfism: effect of insulin-like growth factor I. Horm. Res. 37, 160–4.
PubMed |

Sonnenberg, G. E. , Krakower, G. R. , Hoffman, R. G. , Maas, D. L. , Hennes, M. M. I , and Kissebah, A. H. (2001). Plasma leptin concentrations during extended fasting and graded glucose infusion: relationships with changes in glucose, insulin and FFA. J. Clinic. Endocrinol. Metab. 86, 4895–900.


Spicer, L. J. (2001). Leptin: a possible metabolic signal affecting reproduction. Dom. Anim. Endocrinol. 21, 251–70.
Crossref | GoogleScholarGoogle Scholar |

Thomas, L. , Wallace, J. M. , Aitken, R. P. , Mercer, J. G. , Trayhurn, P. , and Hoggard, N. (2001). Circulating leptin during ovine pregnancy in relation to maternal nutrition, body composition and pregnancy outcome. J. Endocrinol. 169, 465–76.
PubMed |

Tindal, J. S. , Knaggs, G. S. , Hart, I. C. , and Blake, L. A. (1978). Release of growth hormone in lactating and non-lactating goats in relation to behaviour, stages of sleep, electroencephalograms, environmental stimuli and levels of prolactin, insulin, glucose and free fatty acids in the circulation. J. Endocrinol. 76, 333–46.
PubMed |

Ward, K. A. , and Brown, B. W. (1998). The production of transgenic domestic livestock: successes, failures and the need for nuclear transfer. Reprod. Fertil. Dev. 10, 659–65.
Crossref | GoogleScholarGoogle Scholar | PubMed |