Association of GH gene polymorphism with growth and semen traits in rabbits

[EN] Although growth hormone (GH) gene mutations are described in several species, the studies concerning their variabilities and associations with economic traits in rabbits are scarce, particularly associations with semen traits. A total of 149 rabbit bucks from five populations (V-line=36, Moshtohor line=28, APRI line=42, cross ½A½M=23, and Gabali=20) were used in the present study to identify polymorphism of c.-78 C>T single nucleotide polymorphism (SNP) of GH gene among these populations and to investigate the association of GH gene polymorphism with body weight (BW), daily weight gain (DG) and semen traits. DNA was extracted from blood samples for genotyping of c.-78 C>T SNP of GH gene based on polymerase chain reaction with the restriction fragment length polymorphism (PCR-RFLP) technique. The genetic diversity of SNP C>T of GH gene was assessed in terms of genotypic and allelic frequencies, effective number of alleles (Ne), observed (Ho) and expected (He) heterozygosity, Hardy-Weinberg equilibrium (HWE), reduction in heterozygosity due to inbreeding (FIS) and polymorphism information content (PIC). Three genotypes of TT, CC and TC of PCR product of 231 bp of GH gene were detected and all the populations were in HWE in terms of GH gene. The highest Ne was obtained for the Moshtohor line (1.978), while the lowest allelic numbers were obtained for V-line (1.715) and Gabali breed (1.800). The highest genotype frequency of GH gene was 0.48 in TT genotype of V-line, 0.21 in CC genotype of Moshtohor line, 0.67 and 0.56 in TC genotype of ½A½M and Gabali rabbits (P<0.05). The highest frequency for C allele was recorded by Moshtohor line (0.45) and the lowest frequency by Gabali (0.32). The genetic diversity scores for GH gene were intermediate (Ho=0.551, He=0.471, PIC=0.358). The values of Ho ranged from 0.444 in V-line to 0.667 in ½A½M cross, while the values of He were 0.425 in V-line and 0.508 in Moshtohor line. The values of PIC were moderate and ranged from 0.332 in V-line to 0.375 in M-line. The highest FIS was observed in Moshtohor line (0.042) and the lowest value was observed in ½A½M cross (–0.413). The CT genotype of GH gene showed the highest and significant values for body weights at 4, 8, 10 and 12 wk (542, 1131, 1465 and 1861 g) and daily gains at intervals of 4-6 and 8-10 wk (23.1 and 26.5 g). Additionally, the CT genotype recorded the highest and significant values for volume of ejaculate (1.1 mL), sperm motility (57.6%), live sperm (85.6%), normal sperm (93.1%) and sperm concentration in semen (611×106/mL), along with the lowest and significant values for dead sperms (14.4%) and abnormal sperms (6.9%).The authors are very grateful to the Central Laboratory of the Faculty of Veterinary Medicine, Benha University, Egypt for the help and support in molecular genetic analyses. This work was financially supported by the research project entitled “Genetic improvement of local rabbit breeds by using molecular genetic techniques” from the Scientific Research Fund (SRF), Benha University, Egypt.Khalil, MHE.; Zaghloul, AR.; Iraqi, MM.; El Nagar, AG.; Ramadan, SI. (2021). Association of GH gene polymorphism with growth and semen traits in rabbits. World Rabbit Science. 29(1):31-40. https://doi.org/10.4995/wrs.2021.13001OJS3140291Abdel-Kafy E., Hussein B., Abdel-Ghany S., El-Din A., Badawi Y. 2015. Single nucleotide polymorphisms in growth hormone gene are associated with some performance traits in rabbit. Int. J. Biol. Pharm. Allied Sci., 4: 490-504.Afshari K.P., Javanmard A., Asadzadeh N., Sadeghipanah H., Masomi H., Sabrivand A. 2011. Association between GH encoding gene polymorphism and semen characteristics in Iranian Holstein bulls. Afr. J. Biotechnol., 10: 882-886.Alvariño J. 2000. Reproductive performance of male rabbits. In Proc.: 7th World Rabbit Congress, 4-7 July, Valencia, Spain.Amalianingsih T., Brahmantiyo B. 2014. The variability of growth hormone gene associated with ultrasound imaging of longissimus dorsi muscle and perirenal fat in rabbits. Media Peternakan, 37: 1-7. https://doi.org/10.5398/medpet.2014.37.1.1Amiri S., Jemmali B., Ferchichi M. A., Jeljeli H., Boulbaba R., Ben Gara A. 2018. Assessment of growth hormone gene polymorphism effects on reproductive traits in Holstein dairy cattle in Tunisia. Arch. Anim. Breed., 61: 481-489. https://doi.org/10.5194/aab-61-481-2018Bindu, K., Raveendran, A., Antony, S., Raghunandanan, K. 2011. Association of myostatin gene (MSTN ) polymorphism with economic traits in rabbits. In: Fibre production in South American Camelids and other fibre animals. Springer. https://doi.org/10.3920/978-90-8686-727-1_16Darwish S.F., Tarek S.H., Badr M.R. 2016. Association between genotypes of growth hormone gene and semen quality traits in cow bulls and buffalo bulls. Alex. J. Vet. Sci., 49: 147-152. https://doi.org/10.5455/ajvs.221998El-Aksher, S.H., Sherif, H.S., Khalil, M.H., El-Garhy, H.A.S., Ramadan, S. 2017. Molecular analysis of a new synthetic rabbit line and their parental populations using microsatellite and SNP markers. Gene Rep., 8: 17-23.https://doi.org/10.1016/j.genrep.2017.05.001El-Sabrout K., Aggag S. 2017a. Associations between single nucleotide polymorphisms in multiple candidate genes and body weight in rabbits, Vet. World, 10: 136-139. https://doi.org/10.14202/vetworld.2017.136-139El-Sabrout K., Aggag S. 2017b. The gene expression of weaning age and its effect on productive performance of rabbits. World Rabbit Sci., 25: 1-7. https://doi.org/10.4995/wrs.2017.4777El-Tarabany M.S., El-Bayomi K., Abdelhamid T. 2015. Semen characteristics of purebred and crossbred male rabbits. PloS one, 10: e0128435. https://doi.org/10.1371/journal.pone.0128435Estany J., Baselga M., Blasco A., Camacho J. 1989. Mixed model methodology for the estimation of genetic response to selection in litter size of rabbits. Lives. Prod. Sci., 21: 67-75. https://doi.org/10.1016/0301-6226(89)90021-3Fontanesi L., Tazzoli M., Scotti E., Russo V. 2008. Analysis of candidate genes for meat production traits in domestic rabbit breeds. In Proc.: 9th World Rabbit Congress, 10-13 June 2008,.Verona, Italy.Fontanesi L., Dall'Olio S., Spaccapaniccia E., Scotti E., Fornasini D., Frabetti A.. 2012. A single nucleotide polymorphism in the rabbit growth hormone (GH1) gene is associated with market weight in a commercial rabbit population. Livest. Sci., 147: 84-88. https://doi.org/10.1016/j.livsci.2012.04.006Gencheva D., Georgieva S., Velikov K., Koynarski T., Tanchev S. 2017. Single nucleotide polymorphism of the Growth Hormone Receptor (GHR) encoding gene in Oryctolagus cuniculus. J. BioSci. Biotechnol. 6: 197-201.Gidenne T., Garreau H., Drouilhet L., Aubert C., Maertens L. 2017. Improving feed efficiency in rabbit production, A review on nutritional, technico-economical, genetic and environmental aspects. Anim. Feed Sci. Technol., 225: 109-122. https://doi.org/10.1016/j.anifeedsci.2017.01.016Groeneveld E. 2006. PEST user's manual. Institute of Animal Husbandry and Animal Behaviour, Mariensee, Germany, FAL, Germany.Hristova D., Tanchev S., Velikov K., Gonchev P., Georgieva S. 2017. Rabbit growth hormone and myostatin gene polymorphisms. J. Agric. Res., 2: 000133 (Open Access). https://doi.org/10.23880/OAJAR-16000133Hristova D.G., Tanchev S., Velikov K., Gonchev P., Georgieva S. 2018. Single nucleotide polymorphism of the growth hormone (GH) encoding gene in inbred and outbred domestic rabbits. World Rabbit Sci., 26: 49-55. https://doi.org/10.4995/wrs.2018.7211Iraqi M., Afifi E., Baselga M., Khalil M., García M. 2008. Additive and heterotic components for post-weaning growth traits in a crossing project of V-line with Gabali rabbits in Egypt. In Proc.: 9th World Rabbit Congress, 10-13 June 2008, Verona, Italy.Kalinowski, S.T., Taper, M.L., Marshall, T.C. 2007. Revising how the computer program CERVUS accommodates genotyping error increases success in paternity assignment. Molecular Ecology, 16: 1099-1106. https://doi.org/10.1111/j.1365-294X.2007.03089.xKhalil M., Baselga M. 2002. Rabbit genetic resources in Mediterranean countries. In M. H. Khalil & M. Baselga (Eds.), Options Méditerranéennes (Vol. Série B, pp. 262). Zaragoza, Spain: CIHEAM-IAMZ.Khalil M.H., Al-Sobayil K., Al-Saef A., García M., Baselga M. 2007. Genetic evaluation for semen characteristics in a crossbreeding project involving Saudi and Spanish V-line rabbits. Animal, 1: 923-928. https://doi.org/10.1017/S1751731107000341Kmieć M., Terman A., Wierzbicki H., Zych S. 2007. Association of GH gene polymorphism with semen parameters of boars. Acta Vet. Brno., 76: 41-46. https://doi.org/10.2754/avb200776010041Lavara R., García M., Torres C., Vicente J., Baselga M. 2008. Genetic parameters for semen traits of rabbit males: II. Motility. In Proc.: 9th World Rabbit Congress., 10-13 June 2008, Verona, Italy.Lavara, R., Vicente, J., Baselga, M. 2011. Genetic parameter estimates for semen production traits and growth rate of a paternal rabbit line. J. Anim. Breed. Genet., 128: 44-51. https://doi.org/10.1111/j.1439-0388.2010.00889.xLavara, R., David, I., Moce, E., Baselga, M., Vicente, J. 2013. Environmental and male variation factors of freezability in rabbit semen. Theriogenology, 79: 582-589. https://doi.org/10.1016/j.theriogenology.2012.11.007Maertens L., Gidenne T. 2016. Feed efficiency in rabbit production: nutritional, technico-economical and environmental aspects. In Proc.: 11th World Rabbit Congress, 15-18 June, Qingdao, China.Migdal L., Palka S., Kmiecik M., Derewicka O. 2019. Association of polymorphisms in the GH and GHR genes with growth and carcass traits in rabbits (Oryctolagus cuniculus). Czech J. Anim. Sci., 64: 255-264. https://doi.org/10.17221/27/2019-CJASNikbin S., Panandam J.M., Yaakub H., Murugaiyah M. 2018. Association of novel SNPs in gonadotropin genes with sperm quality traits of Boer goats and Boer crosses. J. Applied Anim. Res., 46: 459-466. https://doi.org/10.1080/09712119.2017.1336441Pal A., Chakravarty A.K., Chatterjee P.N. 2014. Polymorphism of growth hormone gene and its association with seminal and sexual behavioral traits in crossbred cattle. Theriogenology, 81: 474-480. https://doi.org/10.1016/j.theriogenology.2013.11.002Peakall P., Smouse R. 2012. GenAlEx 6.5: Genetic analysis in Excel. Population genetic software for teaching and research. An update. Bioinformatics, 28: 2537-2539. https://doi.org/10.1093/bioinformatics/bts460Raymond M. 1995. GENEPOP (version 1.2): Population Genetics Software for exact tests and ecumenicism. J. Hered., 86: 248-249. https://doi.org/10.1093/oxfordjournals.jhered.a111573Sahwan F.M., El-Sheik A.I., Sharaf M.M., El-Nahas A.F. 2014. Genetic polymorphism in growth hormone receptor gene (GHR) and its relationship with growth trait in pure and hybrid rabbit breeds. Alex. J. Vet. Sci., 43: 45-51. https://doi.org/10.5455/ajvs.165197Tanchev S. 2015. Conservation of genetic resources of autochthonous domestic livestock breeds in Bulgaria. A review. Bulgarian J. Agric. Sci., 21: 1262-1271.Tusell L., Legarra A., García-Tomás M., Rafel O., Ramon J., Piles M. 2012. Genetic basis of semen traits and their relationship with growth rate in rabbits. J. Anim. Sci. 90: 1385-1397. https://doi.org/10.2527/jas.2011-4165Yardibi H., Hosturk G.T., Paya I., Kaygisiz F., Ciftioglu G., Mengi A., Oztabak K. 2009. Associations of growth hormone gene polymorphisms with milk production traits in South Anatolian and East Anatolian red cattle. J. Anim. Vet. Adv. 8: 1040-1044. https://doi.org/10.3923/javaa.2009.1040.1044Youssef Y., Iraqi M., El-Raffa A., Afifi E., Khalil M., García M., et al. 2008. A joint project to synthesize new lines of rabbits in Egypt and Saudi Arabia: emphasis for results and prospects. In Proc.: 9th World Rabbit Congress, 10-13 June, Verona-Italy.Zachmann M. 1992. Interrelations between growth hormone and sex hormones: physiology and therapeutic consequences. Horm. Res. Paediat., 38: 1-8. https://doi.org/10.1159/000182562Zhang W.X., Zhang G.W., Peng J., Lai S.J. 2012. The polymorphism of GHR gene associated with the growth and carcass traits in three rabbit breeds. In Proc.: 10th World Rabbit Congress, Sharm El-Sheikh, Egypt, 75-7