Physiology and modulation factors of ovulation in rabbit reproduction management


  • Simona Mattioli University of Perugia
  • Margherita Maranesi University of Perugia
  • Cesare Castellini University of Perugia
  • Alessandro Dal Bosco University of Perugia
  • Maria Arias-Álvarez Complutense University of Madrid
  • Pedro Luis Lorenzo Complutense University of Madrid
  • Pilar García Rebollar Polytechnic University of Madrid
  • Rosa María García-García Complutense University of Madrid



rabbit, ovulation, seminal plasma, β-NGF, GnRH, reproduction


Rabbit is an induced ovulatory species, so ovulation takes place after mating. Traditionally, exogenous and synthetic hormonal factors (administered by intramuscular and intravaginal route) such as GnRH and analogues, or different physical procedures (i.e. stimulation by intravaginal cannula) have been used to induce ovulation in females when artificial insemination is applied in rabbit farms. Restriction and public rejection of the use of hormones is leading to the study of the seminal plasma components with potential action on ovulation induction. The aim of the present review is to collect and summarise the strategies used in recent years to trigger ovulation and improve rabbit fertility management with respect to more animal-friendly manipulation methods. Furthermore, special attention has been paid to the use of a semen component (as endogen molecule) such as beta nerve growth factor (β-NGF) in male and female rabbit reproductive physiology. This neurotrophin and its receptors (TrKA and p75NTR) are abundantly distributed in both male and female rabbit reproductive tracts, and it seems to have an important physiological role in sperm maturation and behaviour (velocity, apoptosis and capacitation), as well as a modulatory factor of ovulation. Endogen β-NGF is diluted in the seminal doses with the extenders; hence it could be considered an innovative and alternative strategy to avoid the current exogenous (by intramuscular route) and stressful hormonal treatments used in ovulation induction. Their addition in seminal dose could be more physiological and improve animal welfare in rabbit farms.


Download data is not yet available.

Author Biographies

Simona Mattioli, University of Perugia

Dept. of Agricultural, Food and Environmental Science

Margherita Maranesi, University of Perugia

Dept. Veterinary Medicine

Cesare Castellini, University of Perugia

Dept. of Agricultural, Food and Environmental Science

Alessandro Dal Bosco, University of Perugia

Dept. of Agricultural, Food and Environmental Science

Maria Arias-Álvarez, Complutense University of Madrid

Dept. of Animal Production, Veterinary Faculty

Pedro Luis Lorenzo, Complutense University of Madrid

Dept. of Animal Production, Veterinary Faculty

Pilar García Rebollar, Polytechnic University of Madrid

Dept. of Agrarian Production. Agronomic, Alimentary and Biosystems

Rosa María García-García, Complutense University of Madrid

Dept. of Physiology, Veterinary Faculty


Adams G.P., Ratto M.H., Huanca W., Singh J. 2005. Ovulationinducing factor in the seminal plasma of alpacas and llamas. Biol. Reprod., 73: 452-457.

Adams G.P., Ratto M.H., Silva M.E., Carrasco R.A. 2016. Ovulation-inducing factor (OIF/NGF) in seminal plasma: a review and update. Reprod. Dom. Anim., 51: 4-17.

Adams, C. E. 1961. Artificial insemination in the rabbit. J. Reprod. Fertil., 2: 251-254.

Bakker J., Baum M.J. 2000. Neuroendocrine regulation of GnRH release in induced ovulators. Frontiers in neuroendocrinology, 21: 220-262.

Benziger D.P., Edelson, J. 1983. Absorption from the vagina. Drug Metab. Rev., 14: 137-168.

Berland M.A., Ulloa-Leal C., Barría M., Wright H., Dissen G.A., Silva M.E., Ojeda S.R., Ratto M.H. 2016. Seminal plasma induces ovulation in llamas in the absence of a copulatory stimulus: role of nerve growth factor as an ovulationinducing factor 1. Endocrinology, 29, en20161310.

Bogle O.A., Carrasco R.A., Ratto M.H., Singh J., Adams G.P. 2018. Source and localization of ovulation-inducing factor/nerve growth factor in male reproductive tissues among mammalian species. Biol. Reprod., 99: 1194-1204.

Boiti C., Besenfelder U., Brecchia G., Theau Clement M., Zerani M. 2006. Reproductive physiopathology of the rabbit doe. In: Recent Advices in rabbit science. Ed. Maertens L. and Coudert P. Published ILVO. Merelbeke, Belgium.

Boiti C., Castellini C., Canali C., Zampini D., Monaci, M. 1995. Long term effect of PMSG on rabbit does reproductive performance. World Rabbit Sci., 3: 51-56.

Browning J., Reyes L. Wolf R. 1980. Comparison Evidence of Serum Progesterone, in Pregnant Rabbits: Recognition of Pregnancy and Estradiol-1713 and Pseudopregnant for Postimplantation rate. Biol. Reprod., 23: 1014-1019.

Cardinali R., Dal Bosco A., Bonanno A., Di Grigoli A., Rebollar P. G., Lorenzo P. L., Castellini C. 2008. Connection between body condition score, chemical characteristics of body and reproductive traits of rabbit does. Livestock Sci., 116: 209-215.

Carrasco R.A., Ratto, M. H., Adams G. P. 2021. Differential Effects of Estradiol on Reproductive Function in Camelids. Front. Vet. Sci., 8: 124.

Casares-Crespo L., Vicente J. S., Talaván A. M., Viudesde-Castro M.P. 2015. Does the inclusion of protease inhibitors in the insemination extender affect rabbit reproductive performance?. Theriogenology, 85: 928-932.

Casares-Crespo L., Fernandez-Serrano P., Vicente J.S., Marco-Marco-Jimenez F., Viudes-de-Castro M.P. 2018. Rabbit seminal plasma proteome: The importance of the genetic origin. Anim. Reprod. Sci., 189: 30e42.

Castellini C. 1996. Recent advances in rabbit artificial insemination. In Proc.: 6th World Rabbit Congress, 9-12 July 1996, Toulouse, France, 2: 13-26.

Castellini C., Dal Bosco A., Arias-Álvarez M., Lorenzo P. L., Cardinali R., Rebollar P.G. 2010. The main factors affecting the reproductive performance of rabbit does: a review. Animal Reprod. Sci., 122: 174-182.

Castellini C., Mattioli S., Dal Bosco A., Collodel G., Pistilli A., Stabile A.M., Macchioni L., Mancuso F., Luca G., Rende M. 2019. In vitro effect of nerve growth factor on the main traits of rabbit sperm. Reprod. Biol. Endocrinol., 17: 93.

Castellini C., Mattioli S., Dal Bosco A., Cartoni Mancinelli A., Rende M., Stabile A.M., Pistilli A. 2020a. NGF and sperm traits: a review. ICAR 2020, Theriogenology ICAR special issue.

Castellini C., Mattioli S., Dal Bosco A., Cotozzolo E., Mancinelli A. C., Rende M., Pistilli A. 2020b. Nerve growth factor receptor role on rabbit sperm storage. Theriogenology, 153: 54-61.

Cervantes M.P., Palomino J.M., Adams G.P. 2015. In vivo imaging in the rabbit as a model for the study of ovulation-inducing factors. Lab. Anim., 49: 1-9.

Chen B.X., Yuen Z.X., Pan G. 1985. Semen-induced ovulation in the bactrian camel (Camelus bactrianus). Reproduction, 73: 335-339.

Dal Bosco A., Cardinali R., Brecchia G., Rebollar P.G., Fatnassi M., Millán P., Castellini, C. 2014. Induction of ovulation in rabbits by adding Lecirelin to the seminal dose: In vitro and in vivo effects of different excipients. Anim. Rep. Sci., 150: 44-49.

Dal Bosco A., Rebollar P.G., Boiti C., Zerani M., Castellini C. 2011. Ovulation induction in rabbit does: current knowledge and perspectives. Anim. Reprod. Sci.; 129: 106-117.

El Allali K., El Bousmaki N., Ainani H., Simonneaux V. 2017. Effect of the camelid’s seminal plasma ovulation-inducingfactor/β-NGF: a kisspeptin target hypothesis. Front. Vet. Sci., 4: 99.

European Parliament. 2010. Directive 2010/63/EU. European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes. Text with EEA relevance. Available at Accesed September 2017.

European Parliament. 2013. Report from the Commission to the Council and the European Parliament. Sixth Report on the statistics on the number of animals used for experimental and other scientific purposes in the Member States of the European Union SEC 2010, 1107. Available at ad69-61ead7e377e8.0001.03/DOC_1&format=PDF. Accesed September 2017

Fernández-Serrano P., Casares-Crespo L., Viudes-de-Castro M.P. 2017. Chitosan-dextran sulphate nanoparticles for Gn RH release in rabbit insemination extenders. Reprod. Domest. Anim., 52: 72-74.

García-García R.M., Masdeu M.D. M., Sánchez Rodríguez A., Millán P., Arias-Álvarez M., Sakr,O.G., Rebollar P. G. 2018a. β-nerve growth factor identification in male rabbit genital tract and seminal plasma and its role in ovulation induction in rabbit does. It. J. Anim. Sci., 17: 442-453.

García-García R.M., Arias-Álvarez M, Sánchez Rodríguez A., García Rebollar P. Lorenzo P.L. 2018b. NGF systems is differentially expressed in the ovary, oviduct and uterus of rabbit does although independent of serum hormonal levels. In: 22nd Annual Conference of the European Society for Domestic Animal Reproduction (ESDAR). Reprod. Dom. Anim. 53, S2: 88.

García-García R.M., Arias-Álvarez M, Sánchez Rodríguez A., García Rebollar P. Lorenzo P.L. 2020. Role of nerve growth factor in the reproductive physiology of female rabbits: a review. Theriogenology, 150: 321-328.

Goodman S.B., Kugu K., Chen S.H., Preutthipan S, Tilly K.I., Tilly J.L., Dharmarajan A.M. 1998. Estradiol-mediated suppression of apoptosis in the rabbit corpus luteum with a shift in expression of Bcl-2 family members favouring cellular survival. Biol Reprod., 59: 820-827.

Harcout-Brown F. 2002. Textbook of Rabbit Medicine. Elsevier Health Sciences, 3: 356-360. Available at

Harper, M.J.K. 1970. Factors influencing sperm penetration of rabbit eggs in vivo. J. Exp. Zool., 173: 47-62.

Hassanein E.M., Hashem N.M., El-Azrak K.E.M., Gonzalez-Bulnes A., Hassan G.A., Salem M.H. 2021. Efficiency of GnRH-Loaded Chitosan Nanoparticles for Inducing LH Secretion and Fertile Ovulations in Protocols for Artificial Insemination in Rabbit Does. Animals, 11: 440.

Holgado-Madruga M., Moscatello D.K., Emlet D.R., Dieterich R., Wong A.J. 1997. Grb2-associated binder-1 mediates phosphatidylinositol 3-kinase activation and the promotion of cell survival by nerve growth factor. In Proc.: National Academy of Sciences, 94: 12419-24.

Johnston S.D., O’Callaghan P.O., Nilsson K., Tzipori G., Curlewis J.D. 2004. Semen-induced luteal phase and identification of a LH surge in the koala (Phascolarctos cinereus). Reproduction, 128: 629-634.

Lebas F., Theau-Clement M., Remy B., Drion P., Beckers J.F. 1996. Production of anti-PMSG antibodies and its relation to the productivity of rabbit does. World Rabbit Sci., 4: 57-62.

Levi-Montalcini R., Hamburger V. 1951. Selective growth stimulating effects of mouse sarcoma on the sensory and sympathetic nervous system of the chick embryo. J. Exp. Zool., 116: 321-361.

Li C., Sun Y., Yi K., Ma Y., Zhang W., Zhou X. 2010. Detection of nerve growth factor (NGF) and its specific receptor (TrkA) in ejaculated bovine sperm, and the effects of NGF on sperm function. Theriogenology, 74: 1615-1622.

Lin W.W., Ramirez V.D. 1991. Effect of mating behavior on luteinizing hormone-releasing in female rabbits as monitored with push-pull cannulae. Neuroendocrinology, 53: 229-325.

Lytton, F.D.C., Poyser N.L. 1982. Prostaglandin production by the rabbit and placenta in vitro uterus. J. Reprod. Fertil., 66: 591-599.

Maranesi M., Zerani M., Lilli L., Dall’Aglio C., Brecchia G., Gobbetti A., Boiti C. 2010. Expression of luteal estrogen receptor, interleukin-1, and apoptosis-associated genes after PGF2alpha administration in rabbits at different stages of pseudopregnancy. Domest. Anim. Endocrinol., 39: 116-30.

Maranesi M., Zerani M., Leonardi L., Pistilli A., Arruda-Alencar J., Stabile A.M., Rende M., Castellini C., Petrucci L., Parillo F.,

Moura A., Boiti C. 2015. Gene expression and localization of NGF and its cognate receptors NTRK1 and NGFR in the sex organs of male rabbits. Reprod. Dom. Anim., 50: 918-925.

Maranesi M., Parillo F., Leonardi L., Rebollar P.G., Alonso B., Petrucci L., Zerani M. 2016. Expression of nerve growth factor and its receptors in the uterus of rabbits: functional involvement in prostaglandin synthesis. Domest. Anim. Endocrinol., 56: 20-28.

Maranesi M., Petrucci L., Leonardi L., Piro F., Rebollar P.G., Millán P., Cocci P., Vullo C., Parillo F., Moura A., Mariscal G.G., Boiti C., Zerani M. 2018. New insights on a NGF-mediated pathway to induce ovulation in rabbits (Oryctolagus cuniculus). Biol Reprod., 98: 634-643.

Maranesi M., Palermo F.A., Bufalari A., Mercati F., Paoloni D., Cocci P., Moretti G., Crotti S., Zerani M., Dall'Aglio C. 2020. Seasonal Expression of NGF and Its Cognate Receptors in the Ovaries of Grey Squirrels (Sciurus carolinensis). Animals (Basel). 10:1558.

Maranesi M., Cocci P., Tomassoni D., Mercati F., Palermo F.A., Anipchenko P., Boiti C., Bufalari A., Zerani M., Dall’Aglio C. 2020a. Ovulation inducing factors in rabbits (Oryctolagus cuniculus): the role of IL1. ESDAR 2021, Reprod. Dom. Anim., submitted.

Milligan, S., 1982. Induced ovulation in mammals. In: Oxford Reviews of Reproductive Biology, Vol. 4 (Ed. by C.A. Finn). Clarendon Press, Oxford, UK. pp.1-46.

Molina I., Pla M., Vicente J.S., Martin A., Romeu A. 1991. Induction of ovulation in rabbits with pure urinary luteinizing hormone and human chorionic gonadotrophin: comparison of oocyte and embryo quality. Hum. Reprod., 6: 1449-1452.

Pei Y. 2010. Effect of nerve growth factor (NGF) on the development of preimplantation rabbit embryos in vitro. Veter. Res. Commun., 34: 11-18.

Quintela L.A., Peña A.I., Vega M.D., Gullón J., Prieto M.C., Barrio M., Herradón P.G. 2004. Ovulation induction in rabbit does submitted to artificial insemination by adding buserelin to the seminal dose. Reprod. Nutr. Dev., 44: 79-88.

Ratto M.H., Leduc Y.A., Valderrama X.P., van Straaten K.E., Delbaere L.T.J., Pierson R.A., Adams G.P. 2012. The nerve of ovulation- inducing factor in semen. In Proc.: National Academy of Sciences, 109: 15042-15047.

Rebollar P.G., Ubilla E., Alvariño J.M.R 1994. Grouping of rabbit reproduction management by means of artificial insemination. World Rabbit Sci., 2: 87-91.

Rebollar P.G., Dal Bosco A., Millán P., Cardinali R., Brecchia G., Sylla L., Castellini C. 2012. Ovulating induction methods in rabbit does: the pituitary and ovarian responses. Theriogenology, 77: 292-298.

Sánchez-Rodríguez A., Abad P., Arias-Álvarez M., Rebollar P.G., Bautista J.M., Lorenzo P.L. 2018. Recombinant production of rabbit β-Nerve Growth Factor and its biological effect on rabbit sperm. Bio Rxiv., 458612.

Sánchez-Rodríguez A., Arias-Álvarez M., Timón P., Bautista J.M., Rebollar P.G., Lorenzo P.L. 2019a. Characterization of β-Nerve Growth Factor-TrkA system in male reproductive tract of rabbit and the relationship between β-NGF and testosterone levels with seminal quality during sexual maturation. Theriogenology, 126: 206-213.

Sánchez-Rodríguez A., Abad P., Arias-Álvarez M., Rebollar P. G., Bautista J. M., Lorenzo P.L., García-García R.M. 2019b. Recombinant rabbit beta nerve growth factor production and its biological effects on sperm and ovulation in rabbits. PloS one, 14.

Sánchez-Rodríguez A., Arias-Álvarez M., Millan P., Lorenzo P. L., García-García R. M., Rebollar P. G. 2020. Physiological effects on rabbit sperm and reproductive response to recombinant rabbit beta nerve growth factor administered by intravaginal route in rabbit does. Theriogenology, 157: 327-334.

Sari L.M., Zampini R., Arganaraz M.E., Carretero M.I., Fumuso F.G., Barraza D.E. 2018 Expression of beta-NGF and highaffinity NGF receptor (TrKA) in llama (Lama glama) male reproductive tract and spermatozoa. Mol. Reprod. Dev., 85: 934-44.

Sari L. M., Zampini R., Del Pino F. G., Argañaraz M. E., Ratto M. H., Apichela S. A. 2020. Effects of NGF Addition on Llama (Lama glama) Sperm Traits After Cooling. Front. Vet. Sci., 7.

Schneidgenova M., Vašíček J., Cupka P., Chrenek P. 2011. Is it necessary to control seasonal quality of the rabbit ejaculate? Slovak J Anim Sci., 44: 48-51.

Silva M., Niño A., Guerra M., Letelier C., Valderrama X.P., Adams G.P., Ratto M.H. 2011. Is an ovulationinducing factor (OIF) present in the seminal plasma of rabbits? Anim. Reprod. Sci., 127: 213-221.

Silva M., Ulloa-Leal C., Norambuena C., Fernández A., Adams G.P., Ratto M.H. 2014. Ovulation-inducing factor (OIF/NGF) from seminal plasma origin enhances Corpus Luteum function in llamas regardless the preovulatory follicle diameter. Anim. Reprod. Sci., 148: 221-227.

Silva M., Paiva L., Ratto M.H. 2020. Ovulation mechanism in South American Camelids: The active role of β-NGF as the chemical signal eliciting ovulation in llamas and alpacas. Theriogenology, 150: 280-287.

Simmon J.A., Danfbrth D.R., Hutchinson J.S., Hodgen G.D. 1988. Characterization of recombinant DNAderived human luteinizing hormone in vitro and in vivo: efficacy in ovulation induction and corpus luteum support. J. Am. Med. Assoc., 259: 3290-3295.

Theau-Clément M., Maertens L., Castellini C., Besenfelder U., Boiti C. 2005. Recommendations and guidelines for applied reproduction trials with rabbit does. World Rabbit Sci., 13: 147-164.

Theau-Clément M. 2007. Preparation of the rabbit doe to insemination: a review. World Rabbit Sci., 15: 61-80.

Tometten M., Blois S., Arck P.C. 2005. Nerve growth factor in reproductive biology: link between the immune, endocrine and nervous system? Chem. Immunol. Allergy, 89: 135-48.

Ubilla E., Rebollar P.G. 1995. Influence of the postpartum day on plasma estradiol 17 beta levels, sexual behaviour and conception rate, in artificially inseminated rabbit does. Anim. Reprod. Sci., 38: 337-344.

Ulloa-Leal C., Bogle O.A., Adams G.P., Ratto M.H. 2014. Luteotrophic effect of ovulation-inducing factor/nerve growth factor present in the seminal plasma of llamas. Theriogenology, 8: 1101-1107.

Viudes-de-Castro M.P., Moce E., Lavara R., Marco-Jiménez F., Vicente J.S. 2014. Aminopeptidase activity in seminal plasma and effect of dilution rate on rabbit reproductive performance after insemination with an extender supplemented with buserelin acetate. Theriogenology, 81: 1223-1228.

Viudes-de-Castro M.P., Casares-Crespo L., Marco-Jiménez F., Vicente J.S. 2017. Physical effect of insemination cannula on ovulation induction in rabbit doe. XVII Jornadas sobre Producción Animal, Zaragoza, España, 30 y 31 de mayo de 2017: 380-382.

Zerani M., Polisca A., Boiti C., Maranesi M. 2021. Current Knowledge on the Multifactorial Regulation of Corpora Lutea Lifespan: The Rabbit Model. Animals, 1: 296.