双峰驼<i>AGBL5</i>基因在发情期和乏情期睾丸及附睾中的表达及定位分析

doi:10.7668/hbnxb.20196717

摘要/Abstract

摘要：

为探究AGBL5在双峰驼睾丸及附睾组织中的表达及分布规律,及对季节性繁殖的调控作用,基于转录组学筛选的差异基因AGBL5,以发情期、乏情期双峰驼睾丸及附睾组织为材料,通过H&E染色、免疫荧光(IF)、实时荧光定量PCR(qRT-PCR)和蛋白免疫印迹(Western Blot)等方法,检测睾丸及附睾不同发育时期AGBL5基因的表达情况。测序结果显示,AGBL5基因的mRNA水平在乏情期睾丸组织中高表达;H&E结果显示,发情期睾丸组织生精小管管腔大于乏情期,有大量精细胞存在,发情期附睾管壁较厚,乏情期则相反,发情期和乏情期都能观察到精细胞的存在;在附睾组织中,发情期的附睾管壁厚度增加,且周围结缔组织呈现疏松状态;乏情期的附睾管壁厚度变薄,结缔组织则较为致密,同时附睾尾管腔内可见大量精细胞积聚;qRT-PCR结果显示,在睾丸组织中,乏情期AGBL5基因的mRNA表达水平极显著高于发情期;在附睾组织中,乏情期附睾头及附睾尾中AGBL5基因的mRNA表达水平均显著高于发情期,而附睾体AGBL5基因的mRNA表达水平显著低于发情期;Western Blot结果显示,AGBL5蛋白表达量同mRNA表达趋势一致。IF结果显示,AGBL5蛋白在睾丸间质细胞、支持细胞、附睾上皮及平滑肌细胞中均存在不同程度荧光表达信号。综上所述,结果提示,AGBL5的差异表达,可能与睾丸的发育及精子的成熟过程密切相关,为进一步探究AGBL5参与雄性双峰驼季节性繁殖调控奠定数据基础。

关键词: 双峰驼, AGBL5, 差异表达, 睾丸, 附睾

Abstract:

To investigate the expression and distribution patterns of AGBL5 in the testis and epididymis tissues of Bactrian camels,as well as its regulatory role in seasonal reproduction,this study focused on AGBL5,a differentially expressed gene identified through transcriptomic screening,using tissues from the testis and epididymis of Bactrian camels during the breeding season and non-breeding season,using H&E staining,immunofluorescence(IF),Real-time Quantitative PCR(qRT-PCR)and Western Blot to examine the expression of the AGBL5 gene in testicular and epididymal tissues at different developmental stages.Sequencing results indicated that the AGBL5 gene was highly expressed at mRNA level in testicular tissue during the non-breeding season.H&E results showed that the lumen of the seminiferous tubules in testicular tissue was larger during the breeding season than during the non-breeding season,with a large number of spermatozoa present.The walls of the epididymal ducts were thicker during the breeding season,whereas the opposite was true during the non-breeding season.Spermatozoa were observed during both the breeding and non-breeding seasons;in epididymal tissue,the thickness of the epididymal duct walls increased during the breeding season,and the surrounding connective tissue appeared loose;during the non-breeding season,the epididymal duct walls were thinner and the connective tissue was denser,whilst a large accumulation of spermatozoa was visible within the lumen of the epididymal tail.qRT-PCR results showed that in testicular tissue,the mRNA expression level of the AGBL5 gene was significantly higher during the non-breeding season than during the breeding season.In epididymal tissue,the mRNA expression levels of the AGBL5 gene in both the head and tail of the epididymis were significantly higher during the non-breeding season than during the breeding season,while the mRNA expression levels in the body of the epididymis were significantly lower during the non-breeding season than during the breeding season.Western Blot results showed that AGBL5 protein expression levels were consistent with the trends observed for mRNA expression.IF results showed that the AGBL5 protein exhibited varying degrees of fluorescent expression signals in testicular interstitial cells,sertoli cells,epididymal epithelial cells and smooth muscle cells.In summary,the results suggest that the differential expression of AGBL5 may be closely related to testicular development and sperm maturation,laying a data foundation for further investigation into the role of AGBL5 in the regulation of seasonal reproduction in male Bactrian camels.

Key words: Bactrian camel, AGBL5, Differential expression, Testis, Epididymis

中图分类号:

马文瑞, 南京宏, 葛婷婷, 刘媛媛, 张勇, 赵兴绪, 王琪. 双峰驼AGBL5基因在发情期和乏情期睾丸及附睾中的表达及定位分析[J]. 华北农学报, 2026, 41(5): 230-238. doi: 10.7668/hbnxb.20196717.

MA Wenrui, NAN Jinghong, GE Tingting, LIU Yuanyuan, ZHANG Yong, ZHAO Xingxu, WANG Qi. Analysis of AGBL5 Expression in Testis and Epididymis of Bactrian Camels During the Breeding and Non-breeding Seasons[J]. Acta Agriculturae Boreali-Sinica, 2026, 41(5): 230-238. doi: 10.7668/hbnxb.20196717.

http://www.hbnxb.net/CN/Y2026/V41/I5/230

图/表 7

表1 引物序列信息

Tab.1 Primer sequence information

基因 Gene	引物序列(5'—3') Primer sequence (5'—3')	片段长度/bp Product size	扩增效率/% Amplification efficiency	退火温度/℃ Annealing temperature
GAPDH	F: AACATCATCCCTGCTTCTACC	184	93	58
	R: ATGCCTGCTTCACTACCTTCT
AGBL5	F: AACCCCAGGCAGGTCTACTT	173	108	59
	R: CTCTCTCCTGGGTGAACTCG

图1 AGBL5基因的转录组表达量分析 FQ.发情期;NC.乏情期。不同大写字母表示差异极显著(P<0.01)。图3,6同。

Fig.1 Analysis of transcriptional expression levels of the AGBL5 gene FQ.The estrus phase;NC.The anestrus phase.Different uppercase letters indicate extremely significant differences(P<0.01).The same as Fig.3,6.

图2 H&E染色 LC.间质细胞;ST.生精小管;PC.管周肌样细胞;PS.初级精母细胞;S.精原细胞;CV.微血管;CA.纤毛;BC.基细胞;EEC.附睾上皮细胞;SZ.精子;SMC.平滑肌细胞。

Fig.2 H&E staining LC.Interstitial cells;ST.Spermatogenic tubules;PC.Peritubular myoid cells;PS.Primary spermatocytes;S.Spermatogonia;CV.Capillaries;CA.Cilia;BC.Basal cells;EEC.Epididymal epithelial cells;SZ.Spermatozoa;SMC.Smooth muscle cells.

图3 AGBL5在双峰驼睾丸的表达分析不同小写字母表示差异显著(P<0.05)。图4—6同。

Fig.3 Expression analysis of AGBL5 in Bactrian camel testes Different lowercase letters indicate significant differences(P<0.05).The same as Fig.4—6.

图4 AGBL5在双峰驼附睾头的表达分析

Fig.4 Expression analysis of AGBL5 in the head of the epididymis of Bactrian camel

图5 AGBL5在双峰驼附睾体的表达分析

Fig.5 Expression analysis of AGBL5 in the body of the epididymis of Bactrian camels

图6 AGBL5在双峰驼附睾尾的表达分析

Fig.6 Expression analysis of AGBL5 in the tail of the epididymis of Bactrian camels

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双峰驼AGBL5基因在发情期和乏情期睾丸及附睾中的表达及定位分析

Analysis of AGBL5 Expression in Testis and Epididymis of Bactrian Camels During the Breeding and Non-breeding Seasons

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