Subscribe to RSS
DOI: 10.1055/s-2002-33073
© Johann Ambrosius Barth
Gonadotropin release in periovulatory heifers after GnRH analogs measured by two types of immunoassays
Publication History
received 27 August 2001
first decision 31 December 2001
accepted 27 March 2002
Publication Date:
30 July 2002 (online)
![](https://www.thieme-connect.de/media/eced/200205/lookinside/thumbnails/10.1055-s-2002-33073-1.jpg)
Summary
This study on heifers (n = 27) compared the effects of a GnRH antagonist (Antarelix) and those of an agonist (Triptorelin) on gonadotropin release during the periovulatory period of the oestrous cycle. In three experiments (EXP I-III), an initial injection of GnRH analogs was given 48 h after a single PGF2α pretreatment during the luteal phase, with a further five at 12 h intervals. A challenge by a GnRH agonist (Gonavet®) was performed six hours after the last application of analogs. In EXP I (n = 9), heifers received six times 1.5 mg of Antarelix, 0.5 mg of Triptorelin, and mannitol (5%; control), respectively. In EXP II (n = 12), identical Antarelix and Triptorelin treatments were followed by a single injection of estradiol-17β valerate (6 h after Gonavet®). The dosage of Antarelix was increased to 5 mg for each injection in EXP III (n = 6). Measurement of LH in blood plasma frequently sampled was performed parallely by a competitive RIA method (EXP I + II) and by a sandwich-type electro-chemiluminescence-immunoassay (ECLIA) in EXP III. This non-isotopic technique was also used to additionally analyse FSH levels. Results of EXP I showed that the GnRH antagonist equally suppressed LH surges and ovulation. On the contrary, prior to suppression of LH levels due to down-regulation of pituitary GnRH receptors the agonist Triptorelin induced an initial increase in LH concentration which was followed by ovulation. In the control animals we observed endogenous LH surges as well as smaller elevations after the agonist (Gonavet®) challenge. An increase was also observed in antagonist, but not in Triptorelin treated heifers. Pituitary GnRH receptors were not detectable in animals previously treated by the analogs, whereas concentrations between 2.2-21.0 fmol/mg protein were measured in controls. Results of EXP II confirmed the described effects of GnRH analogs. Additionally, it was shown that exogenous estradiol is able to release LH from the pituitary, although a previous treatment by a GnRH agonist had dropped the pulsatile gonadotropin secretion. Contrary to the LH pattern and despite elevated amounts of the antagonist, the mean concentration and pulse number of FSH were not influenced by the antagonist treatment (EXP III). These data confirmed that (a) the reversibly blocked pituitary function induced by a potent GnRH antagonist may be a useful tool to study gonadotropin-dependent final follicular growth as well as ovulation in cyclic heifers and (b) the novel non-isotopic ECLIA methods for the determination of FSH and LH provided practical alternatives to other immunoassay types.
Key words:
Cattle endocrinology - Gonadotropins - GnRH analog - Methods
References
- 1 Adams G P, Matteri J P, Ko J CH, Ginther O J. Association between surges of follicle-stimulating hormone and the emergence of follicular waves in heifers. J Reprod Fertil. 1992; 94 177-188
- 2 Bellmann A, Schneider F, Kanitz W, Nürnberg G, Tiemann U. Effect of GnRH and its antagonist (Antarelix) on LH release from cultured bovine anterior pituitary cells. Acta Vet Hungar. 2002; 50 79-92
- 3 Bergfelt D R, Smith C A, Adams G P, Ginther O J. Surges of FSH during the follicular and early luteal phases of the estrous cycle in heifers. Theriogenology. 1997; 48 757-768
- 4 Blödow G, Götze M, Kitzig M, Brüssow K, Duschinski U. Radioimmunologische Steroidhormonbestimmungen in der Follikelflüssigkeit bei Rind und Schwein. Isotopenpraxis. 1988; 24 151-155
- 5 Brooks J, McNeilly A S. Regulation of gonadotrophin-releasing hormone receptor expression in the ewe. Anim Reprod Sci. 1996; 42 89-98
- 6 Brooks J, Struthers W J, McNeilly A S. GnRH-dependent and -independent components of FSH secretion after acute treatment of anoestrous ewes with ovine follicular fluid and a GnRH antagonist. J Reprod Fertil. 1993; 98 591-595
- 7 Brüssow K P, Schneider F, Kanitz W. Die Applikation von GnRH-Agonisten beeinflusst bei Jungsauen die zyklische Gonadotropinausschüttung. Tierärztliche Praxis. 1996; 24 248-255
- 8 Brüssow K P, Schneider F, Nürnberg G. Alteration of gonadotrophin and steroid hormone release, and of ovarian function by a GnRH antagonist in gilts. Anim Reprod Sci. 2001; 66 117-128
- 9 Campbell B K, Dobson H, Scaramuzzi R J. Ovarian function in ewes made hypogonadal with GnRH antagonist and stimulated with FSH in the presence or absence of low amplitude LH pulses. J Endocrinol. 1998; 156 213-222
- 10 Cooke D J, Crowe M A, Roche J F, Headon D R. Gonadotrophin heterogeneity and its role in farm animal reproduction. Anim Reprod Sci. 1996; 41 77-99
- 11 Cooke D J, Crowe M A, Roche J F. Circulating FSH isoform pattern during recurrent increases in FSH throughout the oestrous cycle of heifers. J Reprod Fertil. 1997; 110 339-345
- 12 Crowe M A, Padmanabhan V, Hynes N, Sunderlandr S J, Enright W J, Beitins I Z, Roche J F. Validation of a sensitive radioimmunoassay to measure serum follicle-stimulating hormone in cattle: correlation with biological activity. Anim Reprod Sci. 1997; 48 123-136
- 13 Deaver D R. A new non-isotopic detection system for immunoassays. Nature. 1995; 377 758-760
- 14 Dees W L, Dearth R K, Hooper R N, Brinsko S P, Romano J E, Rahe H, Yu W H, McCann S M. Lamprey gonadotropin-releasing hormone-III selectively releases follicle stimulating hormone in the bovine. Domestic Anim Endocrinol. 2001; 20 279-288
- 15 Deghenghi R, Boutignon R, Wuethrich P. Antarelix (EP 24332) a novel water soluble LHRH antagonist. Biomed & Pharmacother. 1993; 47 107-110
- 16 Dobson H, Campbell B K, Scaramuzzi R J. Use of a GnRH antagonist in conjunction with low amplitude, high frequency LH pulses to induce follicular growth without an LH surge and ovulation. Anim Reprod Sci. 1997; 46 213-222
- 17 D‘Occhio M J, Gifford D R, Earl C R, Weatherly T, v. Rechenberg W. Pituitary and ovarian responses of post-partum acyclic beef cows to continous long-term GnRH and GnRH agonist treatment. J Reprod Fertil. 1989; 85 495-502
- 18 Driancourt M A, Locatelli A, Prunier A. Effects of gonadotrophin deprivation on follicular growth in gilts. Reprod Nutr Dev. 1995; 35 663-673
- 19 Duijkers I JM, Klipping C, Willemsen W N, Krone D, Schneider E, Niebch G, Hermann R. Single and multiple dose pharmacokinetics and pharmacodynamics of the gonadotrophin-releasing hormone antagonist Cetrorelix in healthy female volunteers. Hum Reprod. 1998; 13 2392-2398
- 20 Evans N P, Dahl G E, Mauger D T, Padmanabhan V, Thrun L A, Karsch F J. Does estradiol induce the preovulatory gonadotropin-releasing hormone (GnRH) surge in the ewe by inducing a progressive change in the mode of operation of the GnRH neurosecretory system?. Endocrinology. 1995; 136 5511-5519
- 21 Fike K E, Bergfeld E G, Cupp A S, Kojima F N, Mariscal V, Sanchez T, Wehrmann M E, Grotjan H E, Hamernik D L, Kittok R J, Kinder J E. Gonadotropin secretion and ovarian follicles during oestrous cycles in heifers treated with luteinizing hormone releasing hormone antagonist. Anim Reprod Sci. 1997; 49 83-100
- 22 Flanagan C A, Millar R P, Illing N. Advances in understanding gonadotrophin-releasing hormone receptor structure and ligand interactions. Rev Reprod. 1997; 2 113-120
- 23 Gibbson J R, Wiltbank M C, Ginther O J. Relationship between follicular development and the decline in the follicle-stimulating hormone surge in heifers. Biol Reprod. 1999; 60 72-77
- 24 Ginther O J, Bergfelt D R, Kulick L J, Kot K. Pulsatility of systemic FSH and LH concentrations during follicular wave development in cattle. Theriogenology. 1998; 50 507-519
- 25 Ginther O J, Bergfelt D R, Beg M A, Kot K. Follicle selection in cattle: role of luteinizing hormone. Biol Reprod. 2001; 64 197-205
- 26 Gong J G, Bramley T A, Gutierrez C G, Peters A R, Webb R. Effects of chronic treatment with a gonadotrophin-releasing hormone agonist on peripheral concentrations of FSH and LH, and ovarian function in heifers. J Reprod Fertil. 1995; 105 263-270
- 27 Gong J G, Campbell B K, Bramley T A, Gutierrez C G, Peters A R, Webb R. Suppression in the secretion of follicle-stimulating hormone and luteinizing hormone, and ovarian follicle development in heifers continuously infused with a gonadotropin-releasing hormone agonist. Biol Reprod. 1996; 55 68-74
- 28 Grasselli F, Baratta M, Tamanini C. Effects of a GnRH analogue (buserelin) infused via osmotic minipumps on pituitary and ovarian acticity of prepubertal heifers. Animal Reprod Sci. 1993; 32 153-161
- 29 Hall J E, Brodie T D, Badger T M, Rivier J, Vale W, Conn P M, Schoenfeld D, Crowley W F. Evidence of differential control of FSH and LH secretion by gonadotropin-releasing hormone (GnRH) from the use of a GnRH antagonist. J Clin Endocrinol Metab. 1988; 67 524-531
- 30 Halmos G, Schally A V, Pinski J, Vadillo-Buenfil M, Groot K. Down-regulation of pituitary receptors for luteinizing hormone-releasing hormone (LH-RH) in rats by LH-RH antagonist Cetrorelix. Proc Natl Acad Sci USA. 1996; 93 2398-2402
- 31 Hennies M, Voglmayr J K, Dietrich E, Stollmann M, Moeller R, Holtz W. Hormonal response of female goats to active immunization against a recombinant human inhibin-subunit, and establishment of an enzyme-linked immunosorbent assay for caprine follicle-stimulating hormone. Reprod Domestic Anim. 2001; 36 65-71
- 32 Illera J C, Munro C J, BonDurant R, Silvan G, Illera M. A sandwich enzyme immunoassay for the measurement of swine plasma LH concentrations. Theriogenology. 1997; 47 144
- 33 Janovick J A, Conn P M. Progesterone diminishes the sensitivity of gonadotropin-releasing hormone-stimulated luteinizing hormone (LH) release and protects an LH pool from desensitization: actions opposed by cholera toxin. Endocrinology. 1996; 137 1823-1827
- 34 Kakar S S, Rahe C H, Neill C D. Molecular cloning, sequencing, and characterizing the bovine receptor for gonadotropin releasing hormone (GnRH). Domestic Anim Endocrinol. 1993; 10 335-342
- 35 Kanitz E, Kanitz W, Delzer W, Schmidt H E, Kitzig M, Schneider F. Die Nutzung der LH-Analytik zur Bestimmung des optimalen Besamungszeitraumes nach Superovulationsbehandlungen. Mh VetMed . 1990; 45 123-125
- 36 Kanitz W, Brüssow K P, Becker F, Torner H, Schneider F, Kubelka M, Tomek W. Comparative aspects of follicular development, follicular and oocyte maturation and ovulation in cattle and pigs. Arch Anim Breeding. 2001; 44 9-23
- 37 Kovacs M, Schally A V, Csernus B, Rekasi Z. Luteinizing hormone-releasing hormone (LH-RH) antagonist Cetrorelix down-regulates the mRNA expression of pituitary rceptors for LH-RH by counteracting the stimulatory effect of endogenous LH-RH. Proc Nat Acad Sci USA. 2001; 98 1829-1834
- 38 Krsmanovic L Z, Martinez-Fuentes J, Arora K K, Mores N, Tomic M, Stojilkovic S S, Catt K J. Local regulation of gonadotroph function by pituitary gonadotropin-releasing hormone. Endocrinology. 2000; 141 1187-1195
- 39 Lowry O H, Rosebrough N J, Farr A L, Randall R J. Protein measurement with the folin phenol reagent. J Biol Chem. 1951; 193 265-275
- 40 Ludwig M, Felberbaum R E, Küpker W, Diedrich K. The development and applications of LHRH antagonists. Reprod Med Rev. 2000; 8 41-56
- 41 Madill S, Rieger D, Johnson W H, Walton J S, Coy D H, Rawlings N C. Effects of an LHRH antagonist on the time of occurrence and amplitude of the preovulatory LH surge, progesterone, and oestradiol secretion, and ovulation in superovulated Holstein heifers. Theriogenology. 1994; 41 951-960
- 42 Manikkam M, Rajamahendran R. Effects of bovine follicular fluid and LHRH antagonist on the dominant follicle maintained by a norgestomet ear implant in heifers. Theriogenology. 1994; 41 247
- 43 Matteri R L, Roser J F, Baldwin R L, Lipovetsky V, Papkoff H. Characterization of a monoclonal antibody which detects luteinizing hormone from diverse mammalian species. Domestic Anim Endocrinol. 1987; 4 157-165
- 44 McCann S M, Kimura M, Walczewska S, Karanth S, Rettori V, Yu W H. Hypothalamic control of gonadotropin secretion by LHRH, FSHRF, NO, cytokines, and leptin. Domestic Anim Endocrinol. 1998; 15 333-344
- 45 McLeod B J, Dodson S E, Peters A R, Lamming G E. Effects of a GnRH agonist (buserelin) on LH secretion in post-partum beef cows. Anim Reprod Sci. 1991; 24 1-11
- 46 Merriam G R, Wachter K W. Algorithms for the study of episodic hormone secretion. Am J Physiol. 1982; 243 E310-E318
- 47 Mihm M, Good T EM, Ireland J LH, Knight P G, Roche J F. Decline in serum follicle-stimulating hormone concentrations alters key intrafollicular growth factors involved in selection of the dominant follicle in heifers. Biol Reprod. 1997; 57 1328-1337
- 48 Moguilevski J A, Wuttke W. Changes in the control of gonadotrophin secretion by neurotransmitters during sexual development in rats (Review). Exp Clin Endocrinol Diabetes. 2001; 109 188-195
- 49 Mutayoba B M, Meyer H HD, Schams D, Schallenberger E. Development of a sensitive enzyme immunoassay for LH determination in bovine plasma using the streptavidin-biotin technique. Acta Endocrinologica. 1990; 122 227-232
- 50 Nett T M, Cermak D, Braden T, Manns J, Niswender G D. Receptors for GnRH and estradiol and pituitary content of gonadotropins in beef cows. I. Changes during the estrous cycle. Domestic Anim Endocrinol. 1987; 4 123-132
- 51 Oussaid B, Mariana J C, Poulin N, Fontaine J, Lonergan P, Beckers J F, Cognie Y. Reduction of the developmental competence of sheep oocytes by inhibition of LH pulses during the follicular phase with a GnRH antagonist. J Reprod Fertil. 1999; 117 71-77
- 52 Oussaid B, Lonergan P, Khatir H, Guler A, Monniaux D, Touze J L, Beckers J F, Cognie Y, Mermillod P. Effect of GnRH antagonist-induced prolonged follicular phase on follicular atresia and oocyte developmental competence in vitro in superovulated heifers. J Reprod Fertil. 2000; 118 137-144
- 53 Padmanabhan V, McNeilly A S. Is there an FSH-releasing factor?. Reproduction. 2001; 121 21-30
- 54 Pappa A, Seferiais K, Marselos M, Tsolas O, Messinis I E. Development and application of competitive ELISA assays for rat LH and FSH. Theriogenology. 1999; 51 911-926
- 55 Patterson-Day D J, Geisert R D, Hill C M, Minton J E, McCann J P, Morgan G L. GnRH antagonist inhibition of luteinizing hormone secretion and ovulation in the pig. Anim Reprod Sci. 1997; 49 207-214
- 56 Prakash B S, Wallenhorst S, Metten M, Holtz W, Wuttke W. Development of a sensitive enzymeimmunoassay (EIA) for FSH determination in bovine plasma. Anim Reprod Sci. 1999; 55 183-192
- 57 Quesnel H, Paquier A, Mounier A M, Prunier A. Feed restriction in cyclic gilts: gonadotropin-independent effects on follicular growth. Reprod Nutr Dev. 2000; 40 405-414
- 58 Roelants H, Schneider F, Göritz F, Streich J, Blottner S. Seasonal changes of spermatogonial proliferation in Roe Deer, demonstrated by flow cytometric analysis of c-kit receptor, in relation to follicle-stimulating hormone, luteinizing hormone, and testosterone. Biol Reprod. 2002; 66 305-312
- 59 Roberge S, Schramm R D, Schaaly, Reeves J J. Reduced postpartum anestrus of suckled beef cows treated with microencapsulated luteinizing hormone-releasing hormone. J Anim Sci. 1992; 70 3825-3830
- 60 Rolf C, Gottschalk I, Behre H M, Rauch Ch, Thyroff U, Nieschlag E. Pharmacokinetics of new testosterone transdermal therapeutic systems in gonadotropin-releasing hormone antagonist-suppressed normal men. Exp Clin Endocrinol Diabetes. 1999; 107 63-69
- 61 Rose M W, Gaines-Das R E, Balen A H. Definition and measurement of follicle stimulating hormone. Endocrine Reviews. 2000; 21 5-22
- 62 Roth C, Leonhardt S, Seidel C, Lakomek M, Wuttke W, Jarry H. GnRH antagonist cetrorelix prevents sexual maturation of peripubertal male rats. Exp Clin Endocrinol Diabetes. 2000; 108 358-363
- 63 Savio J D, Thatcher W W, Badinga L, de la Sota R L, Wolfenson D. Regulation of dominant follicle turnover during the oestrous cycle in cows. J Reprod Fertil. 1993; 97 77-84
- 64 Schally A V, Comaru-Schally A M. Rational use of agonists and antagonists of luteinizing hormone-releasing hormone (LH-RH) in the treatment of hormone-sensitive neoplasms and gynaecologic conditions. Adv Drug Delivery Rev. 1997; 28 157-169
- 65 Taft R, Ahmad N, Inskeep E K. Exogenous pulses of luteinizing hormone cause persistence of the largest bovine ovarian follicle. J Anim Sci. 1996; 74 2985-2991
- 66 Turzillo A M, Nett T M. Regulation of GnRH receptor gene expression in sheep and cattle. J Reprod Fertil (Suppl) 54 75-86 1999;
- 67 Turzillo A M, Nolan T E, Nett T M. Regulation of gonadotropin-releasing hormone (GnRH) receptor gene expression in sheep: interaction of GnRH and estradiol. Endocrinology. 1998; 139 4890-4894
- 68 Viczarra J A, Wettemann R P, Braden D T, Turzillo A M, Nett T M. Effect of gonadotropin-releasing hormone (GnRH) pulse frequency on serum and pituitary concentrations of luteinizing hormone and follicle-stimulating hormone, GnRH receptors, and messenger ribonucleic acid for gonadotropin subunits in cows. Endocrinology. 1997; 138 594-601
- 69 Williams B, Brooks A N, Aldridge T C, Pennie W D, Stephenson R, McArdle C A. Oestradiol is a potent mitogen and modulator of GnRH signalling in αT3-1 cells: are these effects causally related?. J Endocrinol. 2000; 164 31-43
- 70 Wu J C, Salfon S C, Miller W L. Gonadal hormones and gonadotropin-releasing hormone (GnRH) alter messenger ribonucleic acid levels for GnRH receptors in sheep. Endocrinology. 1994; 135 1846-1850
- 71 Yu W H, Karanth S, Walczewska A, Sower S A, McCann S M. A hypothalamic follicle-stimulating hormone-releasing decapeptide in the rat. Proc Natl Acad Sci USA. 1997; 94 9499-9503
- 72 Zhou W, Rodic V, Kitanovic S, Flanagan C A, Chi L, Weinstein H, Maayani S, Millar R P, Sealfon S C. A locus of the gonadotropin-releasing hormone receptor that differentiates agonist and antagonist binding sites. J Biol Chem. 1995; 270 18853-18857
Dr. Falk Schneider
Unit of Reproductive Biology, Research Institute for the Biology of Farm Animals
Wilhelm-Stahl-Allee 2
18196 Dummerstorf, Germany
Phone: +49-38208-68757
Fax: +49-38208-68752
Email: falk.schneider@fbn-dummerstorf.de