| Kisspeptin is a kind of neuropeptide of the retinoic acid family, which was found in2001and encoded by the non-metastatic gene of melanoma (kiss-1), many results showedthat kisspeptin played an important role in the regulation of reproductive function in2003.Neurokinin B (NKB) is a polypeptide which contains ten amino acids and belongs to thetachykinin family, the role of NKB in the regulation of reproductive endocrine has beenpaid more and more attention in recent years, and it is considered another important factorin the regulation of reproductive endocrine in addition to kisspeptin. Nowadays, althoughthe regulation of reproductive function of kisspeptin and NKB has become a hot researchtopic at home and abroad, no report is found about the research of kisspeptin/NKB inpoultry especially geese. In the present study, the double immunofluorescence combinedwith laser confocal microscope technology were used to detect the location ofkisspeptin/NKB in the hypothalamus, pituitaries and ovaries of female Wanxi white geeseduring different physiology stages, including adolescence, laying, brooding andrest-circumnatal stages, the software of FV10-ASW3.0was also used to determine thesemi-quantitative fluorescence intensity of kisspeptin/NKB and then analyzed theexpression change of kisspeptin/NKB in the time and space of reproductive axis. Inaddition, the analysis methods of ELISA, RIA and correlation were used to illuminate thepotential connection between the concentration of kisspeptin/NKB and gonadal hormone inserum of Wanxi white geese during the different physiology stages. The results willprovide some scientific theory evidences for the further research to clarify the roles andmechamisms of kisspeptin/NKB in the reproduction of Wanxi white geese.The results showed that (1) During the different physiology stages of Wanxi whitegeese, kisspeptin and NKB were both have different fluorescence intensity expression inthe reproductive axis, but no significant difference was found in the mean fluorescenceintensity between kisspeptin and NKB(P＞0.05).①In the arcuate nucleus andposteromedial nucleus of hypothalamus: the mean fluorescence intensities of adolescenceand brooding stages were significantly higher than that in laying stage (P＜0.05) andrest-circumnated stage (P＜0.05), there was no significant difference between adolescenceand brooding stage, laying and rest-circumnated stage (P＞0.05).②In the paraventricularnucleus: there was no significant difference between adolescence and brooding stage (P＞0.05), but the mean fluorescence intensities of them were both significant higher than those of laying stage (P＜0.05) and rest-circumnatal stage (P＜0.05), while the laying stage wasalso significant higher than that of rest-circumnatal stage (P＜0.05).③In the pituitaries:the immunopositive expressions of kisspeptin and NKB were mainly observed in theanterior pituitary, no immunopositive expression was found in the neurohypophysis. Themean fluorescence intensities of adolescence, laying and brooding stages were bothsignificant higher than that of rest-circumnatal stage (P＜0.05), although there was nosignificant difference between adolescence and brooding stage (P＞0.05), all of them weresignificant higher than rest-circumnatal stage (P＜0.05).④In the ovaries: differentco-expressions were found in the ovaries during the different physiology stages. Theimmunopositive expression of kisspeptin and NKB were only found in the basilarissubstantia, interstitial substance and germinal epithelium of the adolescent Wanxi whitegeese, while in other stages, kisspeptin and NKB were all found in the granular layer,interstitial substance and membrana follicularis of the ovaries. The mean fluorescenceintensities of each part of laying stage were all significant higher than those of broodingstages (P＜0.05) and rest-circumnatal stage (P＜0.05), meanwhile, the mean fluorescenceintensities of brooding stage was significant higher than that of rest-circumnatal stage (P＜0.05). Kisspeptin and NKB in the interstitial substance of adolescent Wanxi white geesewere significant higher than those of brooding stage (P＜0.05) and rest-circumnatal stage(P＜0.05), but no significant difference was found at laying stage (P＞0.05).(2)Theconcentration of kisspeptin and NKB in serum of femal Wanxi white geese during therest-circumnatal stage was significant higher than those of brooding stage (P＜0.05),adolescent age (P＜0.05) and laying stage (P＜0.05), no significant difference was foundbetween adolescence and brooding stage (P＞0.05), but the concentration of kisspeptin andNKB of them were all significant higher than that of laying stage (P＜0.05); Theconcentrations of progesterone and estradiol in serum during the laying stage were thehighest compared to the other stages (P＜0.05), and the level of progesterone and estradiolduring brooding stage was significant higher than rest-circumnatal stage (P＜0.05) andadolescent age (P＜0.05), while rest-circumnatal stage was significant higher thanadolescent age (P＜0.05). The results of correlation analysis showed that the r (correlationcoefficient) value between estradiol and kisspeptin was-0.835, while the r value was-0.780between estradiol and NKB, both of them were highly negatively correlated; Inaddition, the r value between progesterone and kisspeptin or NKB were-0.520and-0.531, which showed that the concentration between progesterone and kisspeptin/NKB weremoderate negatively correlated.The present study demonstrates that:(1)The highest expression of kisspeptin andNKB were found in the hypothalamus of femal Wanxi white geese at adolescence andbrooding stages, the anterior pituitaries and ovaries at adolescence and laying stages werealso have the highest expression, while the lowest expression was observed in reproductiveaxis of rest-circumnatal stage. It suggested that kisspeptin and NKB may play an importantrole of regulation in the reproductive axis of Wanxi white geese during the differentphysiology stages.(2)The average concentration of kisspeption and NKB in serum offemal Wanxi white geese during the different physiology stages were present a regularity,that is, the maximum concentration was at rest-circumnatal stage, then decreased theadolescence and brooding stages, and the minimum level was at laying stage; The highestaverage concentration of progesterone and estradiol was at laying stage, brooding andrest-circumnatal stages were lower than laying stage, while the adolescent age was thelowest.(3) The concentration between progesterone/estrogen and kisspeptin/NKB were allnegatively correlated. |
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