Japon Bıldırcınlarında Tavuk GnRH'nin Serum Testesteron Konsantrasyonu ve Yumurtalarda Döllülük Üzerine Etkileri

Abstract

Yaşlı erkeklerin fertilitesindeki düşüş çıkış oranında azalmayla sonuçlanır. Erkeğin fertilitesi hipotalamo-hipofisyal ve testis eksenin kontrolü altındadır. Böylece bu projenin amacı tavuk GnRH-I (cGnRH-I)’in serum testosteron konsantrasyonuna ve yumurta döllülük oranına etkisini ölçmektir. Yirmi haftalık erkek ve dişi Japon bıldırcınları kafeslere konuldu ve ad libitum beslendiler. Bıldırcınlar random olarak; fosfatla tamponlanmış tuz çözeltisi (PBS), 5 µg cGnRH-I, 20µg cGnRH-I enjektedilen grup ve enjeksiyon yapılmayan doğal grup (N) olmak 4 gruba ayrıldı. Her grup 8 alt grupa ayrıldı ve her bir alt gruba 8 dişi 2 erkek konuldu. Sadece erkekler haftada bir kez olmak üzere üç hafta boyunca 200 µl PBS, 5 veya 20 µg cGnRH-I içeren 200 µl PBS ile enjekte edildiler. Son enjeksiyondan bir hafta sonra yumurtalar haftalık toplandı, 11 gün boyunca kuluçka makinasında tutuldu ve 12. günde kırılarak döllü ve dölsüz yumurtalar belirlendi. Kan numuneleri erkelerin jugular damarından alındı. Serum; pıhtilaşan kan örneklerinden santrifüjle ayrıldı. Serumdaki toplam testesteron kompetetif ümmünokimya yöntemiyle ölçüldü. 5µg cGnRH enjeksiyonu serum testesteron konsantrasyonunu PBS enjekte dilen gruba kıyasla artırırken diğer faklar istatistiksel olarak önemli bulunmadı. GnRH enjeksiyonu fertilite oranında önemli değişmeye sebep olmadı. Bu çalışmada 20 haftalık genç bıldırcınlara cGnRH enjeksiyonu yapıldı. Hayvanlar genç oldukları için kanlarında yeteri kadar GnRH vardı ve fazladan GnRH enjekte etme bir farka sebep olmadı.

Keywords

• GnRH,Döllülük,Serum,Bıldırcın,Testosteron

The Effects of Chicken GnRH on Serum Testosterone Concentration and Egg Fertility in Japanese Quail

Abstract

The decrease in fertility of aged cocks results with decreased hatching rate. Male fertility is under the control of the hypothalamus-pituitary and testicular axis. Therefore, the objective of this study is to measure the effect of chicken GnRH-I (cGnRH-I) on serum testosterone concentration and egg fertility. Twenty weeks old male and female Japanese quail were kept in cages and fed ad libitum. Quails were randomly divided into 4 groups as phosphate buffered saline group (PBS), 5 µg cGnRH-I group, 20µg cGnRH-I group and group received no injection (Naturel group, N group). Each group included 8 replicates and each replicate made up randomly selected 8 females and 2 males. Only male birds were subcutaneously injected once a week for three weeks with 200 µl as PBS, 200 µl PBS containing 5 or 20µg cGnRH-I. One week after the last injection, eggs from each group were weekly collected and incubated for 11 days and broken on day 12 then the numbers of fertilized and unfertilized eggs were determined. Blood samples from the males were obtained from the jugular vein. Serum was extracted from the clotted blood by centrifuging. Total serum testosterone levels were measured by competitive immunochemistry. Injection of 5µg cGnRH increased serum testosterone concentration over PBS injected group other differences were not statistically different. Injection of cGnRH did not cause significant changes in fertility rates. To conclude, 20 weeks old male quails were injected with cGnRH-I and giving extra GnRH-I did not cause any difference in fertility, since there was plenty of GnRH in their blood.

Keywords

• GnRH,Fertility,Serum,Quail,Testosterone

References

1. Anjum, S., Krishna, A., Sridaran, R., Tsutsui, K. 2012. Localization of gonadotropin-releasing hormone (GnRH), gonadotropin-inhibitory hormone (GnIH), kisspeptin and GnRH receptor and their possible roles in testicular activities from birth to senescence in mice. Journal of Experimental Zoology. Part A, Ecological Genetics and Physiology 317:630-644.
2. Avital-Cohen, N., Heiblum, R., Argov-Argaman, N., Rosenstrauch, A., Chaiseha, Y., Mobarkey, N., Rozenboim, I. 2013. Age-related changes in gonadal and serotonergic axes of broiler breeder roosters. Domestic Animal Endocrinology 44: 145-150.
3. Bahk, J.Y., Hyun, J.S., Chung, S.H., Lee H., Kim, M.O., Lee, B.H., Choi, W.S. 1995. Stage specific identification of the expression of GnRH mRNA and localization of the GnRH receptor in mature rat and adult human testis. The Journal of Urology 154: 1958-1961.
4. Bull, P., Morales, P., Huyser, C., Socías, T., Castellón, E.A. 2000. Expression of GnRH receptor in mouse and rat testicular germ cells. Molecular Human Reproduction 6: 582-586.
5. Cho, R.N., Hahn ,T.P., MacDougall-Shackleton, S., Ball, G.F. 1998. Seasonal variation in brain GnRH in free-living breeding and photorefractory house finches (Carpodacus mexicanus). General and Comparative Endocrinology 109: 244-250.
6. Hahn, T.P., Ball, G.F. 1995. Changes in brain GnRH associated with photo refractoriness in house sparrows (Passer domesticus). General and Comparative Endocrinology 99: 349-363.
7. Kochman, K., Domański, E. 1969. Purification of the hypothalamic substances responsible for the release of gonadotropins from the pituitary gland. Acta Physiologica Polonica 20: 521-536.
8. Latimer, V.S., Kohama, S.G., Garyfallou, V.T., Urbanski, H.F. 2001. A developmental increase in the expression of messenger ribonucleic acid encoding a second form of gonadotropin-releasing hormone in the rhesus macaque hypothalamus. The Journal of Clinical Endocrinology and Metabolism 86: 324-329.

9. Lescheid, D.W., Terasawa, E., Abler, L.A., Urbanski, H.F., Warby, C.M., Millar, R.P., Sherwood, N.M. 1997. A second form of gonadotropin releasing hormone (GnRH) with characteris-tics of chicken GnRHII is present in the primate brain. Endocrinology, 138: 5618-5629.
10. McCann, S.M., Taleisnik, S., Friedman, H.M. 1960. LH-releasing activity in hypothalamus extracts. Proceedings of the Society for Experimental Biology and Medicine 104: 432-434.
11. Qasimi, M.I., Nagaoka, K., Watanabe, G. 2018. Feeding of phytosterols reduced testosterone production by modulating GnRH and GnIH expression in the brain and testes of male Japanese quail (Coturnix coturnix japonica). Poultry Science 97:1066-1072.
12. Ottinger, MA., Duchala, S., Mason, M. 1983. Age-related reproductive decline in the male japanese quail. Hormones and Behavior 17: 197-207.
13. Ottinger, M., Kubakawa, K., Kikuchi, M., Thompson, N., Ishii, S. 2002. Effects of exogenous testosterone on testicular luteinizing hormone and follicle-stimulating hormone receptors during aging. Experimental Biology and Medicine 227: 830-836.
14. Parry, D.M., Goldsmith, A.R., Millar, R.P., Glennie, L.M. 1997. Immunocytochemical Localization of GnRH Precursor in the Hypothalamus of European Starlings during Sexual Maturation and Photorefractoriness. Journal of Neuroendocrinology 9: 235-243.
15. Ramakrishnappa, N., Rajamahendran, R., Lin, Y-M., Leung, P.C.K. 2005. GnRH in non-hypothalamic reproductive tissues. Animal Reproduction Science 88: 95-113.
16. Rosenstrauch, A., Degen, A.A., Friedländer, M. 1994. Spermatozoa retention by Sertoli cells during the decline in fertility in aging roosters. Biology of Reproduction 50: 129-136.
17. Rosenstrauch, A., Weil, S., Degen, A.A., Friedländer, M. 1998. Leydig cell functional structure and plasma androgen level during the decline in fertility in aging roosters. General and Comparative Endocrinology 109: 251-258.
18. Schally, A.V., Arimura, A., Baba, Y., Nair, R.M., Matsuo, H., Redding, T.W., Debeljuk, L. 1971. Isolation and properties of the FSH and LH-releasing hormone. Biochemical and Biophysical Research Communications 43: 393-399.
19. Sharp, P., Talbot, R., Main, G., Dunn, I., Fraser, H., Huskisson, N. 1990. Physiological roles of chicken LHRH-I and-II in the control of gonadotrophin release in the domestic chicken. Journal of. Endocrinology 124: 291-299.
20. Terasawa, E., Kurian, J.R., Guerriero, K.A., Kenealy, B.P., Hutz, E.R., Keen, K.L. 2010. Recent Discoveries on the control of GnRH neurons in nonhuman primates. Journal of Neuroendocrinology 22: 630-638.
21. Vizcarra, J., Kirby, J. Kreider, D. 2010. Testis development and gonadotropin secretion in broiler breeder males. Poultry Science 89: 328-334.
22. Weil, S., Degen, A.A., Rosenstrauch, A., Friedländer, M. 1996. Intratesticular spermatozoa retention in low fertility ageing roosters is related to malformations of Sertoli cell ectoplasmic specializations. Journal of Experimental Zoology 275; 317-325.
23. Weil, S., Degen, A.A., Friedländer, M., Rosenstrauch, A.1999a. Low fertility in aging roosters is related to a high plasma concentration of insulin and low testicular contents of ACTH and lactate. General and Comparative Endocrinology 115, 110-115.
24. Weil, S., Rozenboim, I., Degen, A.A., Dawson, A., Friedländer, M., Rosenstrauch, A. 1999b. Fertility decline in aging roosters is related to increased testicular and plasma levels of estradiol. General and Comparative Endocrinology 115: 23-28.
25. White, R.B., Eisen, J.A., Kasten, T.L., Fernald, R.D. 1998. Second gene for gonadotropin-releasing hormone in humans. Proceedings of the National Academy of Sciences of the United States of America 95: 305-309.