OPN5-TSH-DIO2/DIO3和VIP-PRL在短光照抑制公山麻鸭睾丸活性中的表达规律

doi:10.11843/j.issn.0366-6964.2023.05.021

摘要/Abstract

摘要： 旨在探究短光照抑制公山麻鸭繁殖机能的过程中,OPN5-TSH-DIO2/DIO3通路和VIP-PRL通路主要相关因子表达规律及可能存在的相互关系。本试验选取80只正常繁殖期(220日龄)、体重相近且健康的山麻鸭公鸭作为研究对象,试验前15 d维持正常繁殖期的长光照(18L∶6D),从d16开始每天缩短光照2 h,到每天光照时间为8 h(8L∶16D)后持续至试验结束(d45);于d15、d21、d30、d45采集血液样本和下丘脑、垂体、睾丸等组织样,通过HE染色、ELISA、qRT-PCR、Western blot技术检测睾丸发育情况及下丘脑、垂体、睾丸中相关基因和蛋白表达水平。结果显示,与长光照条件相比,短光照导致公鸭睾丸萎缩,生精细胞和精子数量显著减少(P<0.05);血液LH、PRL、睾酮水平均显著降低(P<0.05);下丘脑GnRH表达下降,GnIH表达上升,但差异均不显著(P>0.05);垂体GnIHR呈上升趋势,FSH呈下降趋势,但均无显著差异(P>0.05),GnRHR试验后期明显上升(P<0.05),LH无明显变化趋势;睾丸中FSHR、LHR的基因表达均下降,但无显著差异(P>0.05)。在OPN5-TSH-DIO2/DIO3通路中,OPN5基因和蛋白表达水平显著降低(P<0.05),DIO2、TSH-β基因表达水平显著降低(P<0.05),DIO2蛋白水平降低(P>0.05),DIO3基因表达趋势则与DIO2相反,缩短光照导致基因表达水平上升(P>0.05),DIO3蛋白水平显著上升(P<0.05);VIP-PRL通路中,下丘脑VIP和垂体PRL的基因表达均显著下降(P<0.05),垂体PRLR的基因表达显著上升(P<0.05),睾丸PRLR的基因表达显著下降(P<0.05)。结果表明,在短光照抑制公山麻鸭繁殖性能的过程中,OPN5-TSH-DIO2/DIO3通路和VIP-PRL通路均参与其中,介导了短光照对生殖轴的抑制作用;与正常繁殖期所需光照条件相比,短光照均会抑制两通路的效率,从而实现对生殖轴活性的下调,但其中两通路间的相互调控关系仍有待进一步研究。

关键词: 山麻鸭, 光照, 睾丸, OPN5, PRL

Abstract: The purpose of this study was to explore the expression and possible relationship of main related factors of OPN5-TSH-DIO2/DIO3 pathway and VIP-PRL pathway in the process of short light inhibiting on reproductive function of male Shanma duck. In this experiment, 80 healthy male Shanma ducks with similar body weight and normal breeding period (220 days old) were selected as subjects. The long light (18L:6D) of the normal breeding period was maintained 15 d before the experiment, from d16, the illumination was shortened by 2 h per day, and the illumination time was 8 h per day (8L:16D) until the end of the experiment (d45). Blood samples and tissue samples of hypothalamus, pituitary and testis were collected on d15, d21, d30 and d45, and testicular development and expression of related genes and proteins in hypothalamus, pituitary and testis were detected by HE staining, ELISA, qRT-PCR and Western blot techniques. The results showed that, compared with long light exposure, short light exposure resulted in testicular atrophy, a significant decrease in the number of spermatogenic cells and sperms(P<0.05), a significant decrease in blood LH, PRL and testosterone levels(P<0.05), a decrease in GnRH expression and an increase in GnIH expression in hypothalamus, but the difference was not significant(P>0.05). GnIHR showed an upward trend and FSH showed a downward trend in pituitary, but the difference was not significant(P>0.05). GnRHR increased significantly in the later stage of the experiment(P<0.05). The gene expression levels of LH in pituitary and FSHR, LHR in testis was decreased, but there was no significant difference(P>0.05). In OPN5-TSH-DIO2/DIO3 pathway, the expression gene levels of OPN5, DIO2, TSH-β and the expression protein levels of OPN5 was decreased significantly(P<0.05), the expression protein levels of DIO2 was decreased, but there was no significant difference(P>0.05). While the expression trend of DIO3 gene was opposite to that of DIO2. Shortening light exposure time led to the increase of gene(P>0.05) and protein(P<0.05) expression level of DIO3. In VIP-PRL pathway, the gene expression of VIP in hypothalamus and PRL in pituitary decreased significantly(P<0.05), while the gene expression of PRLR in pituitary increased significantly(P<0.05), and the gene expression of PRLR in testis decreased significantly(P<0.05). The results showed that both OPN5-TSH-DIO2/DIO3 pathway and VIP-PRL pathway were involved in the process of short light inhibiting on reproductive performance of male Shanma duck, which mediated the inhibitory effect of short light on reproductive axis. Compared with the light conditions needed in normal breeding period, short light could inhibiting the efficiency of two pathways and down-regulate the activity of reproductive axis, but the mutual regulation between the two pathways remains to be further studied.

Key words: Shanma duck, illumination, testis, OPN5, PRL

中图分类号:

S834.3

孙俊峰, 潘建秋, 张焯深, 江丹莉, 沈栩, 陈蓉, 许丹宁, 田允波, 黄运茂. OPN5-TSH-DIO2/DIO3和VIP-PRL在短光照抑制公山麻鸭睾丸活性中的表达规律[J]. 畜牧兽医学报, 2023, 54(5): 2001-2012.

SUN Junfeng, PAN Jianqiu, ZHANG Zhuoshen, JIANG Danli, SHENG Xu, CHEN Rong, XU Danning, TIAN Yunbo, HUANG Yunmao. The Expression Patterns of OPN5-TSH-DIO2/DIO3 and VIP-PRL in the Process of Repressing the Testicular Function of Male Shanma Duck under Short-day Photoperiod[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2001-2012.

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