阴囊热应激对山羊血睾屏障结构和功能的影响

doi:10.11843/j.issn.0366-6964.2024.07.017

Abstract

Abstract:

The aim of this study was to investigate the effects of heat stress on the structure and function of blood-testis barrier (BTB) in goats, and to provide a theoretical basis for the prevention and treatment of heat stress in goats. Six healthy male Guanzhong goats aged 1 to 1.5 years old were selected and randomly divided into two groups with three goats in each group. Goats in the control group were fed normally, and the heat stress model was established in the experimental group by the modified scrotal heat stimulation method. The semen quality was detected by CASA, the testicular structure was observed by light microscopy and transmission electron microscopy, the permeability of BTB was detected by biotin tracer, and the expression and localization of key proteins of BTB were detected by Western blot and immunohistochemistry. In heat stress group, the testis index, spermatozoa count, spermatozoa viability, spermatozoa total motility and progressive motility were significantly decreased (P < 0.05). Heat stress caused obvious morphological damage of the testis, and the number of spermatogenic cells decreased and the arrangement was disordered. Endoplasmic reticulum of sertoli cells was expanded and a large number of vacuoles appeared. The tight junction between sertoli cells was loose and discontinuous. The permeability of BTB increased, and the expression level of key junction proteins decreased significantly (P < 0.05), but the distribution and location of BTB were not affected in heat stress group. Heat stress damages the morphology of goat testis, destroys the structure of sertoli cells and reduces the expression of key BTB proteins, thereby impairing the structural and functional integrity of BTB, resulting in the decline of spermatozoa quality and affect the reproductive capacity of male goats.

Key words: goat, heat stress method, blood-testis barrier, ultrastructure

CLC Number:

S827.3

Yongjie WU, Yinghuan XU, Tengfei LIU, Lin MA, Hong CHEN, Yongping XU. Effect of Scrotal Hyperthermia on Structure and Function of Blood-testis Barrier in Goats[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(7): 2973-2982.

Figures/Tables 6

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Fig. 6

References 41

1	RAHMAN M B , SCHELLANDER K , LUCEÑO N L , et al. Heat stress responses in spermatozoa: Mechanisms and consequences for cattle fertility[J]. Theriogenology, 2018, 113, 102- 112. doi: 10.1016/j.theriogenology.2018.02.012
2	CAI H , QIN D Z , PENG S . Responses and coping methods of different testicular cell types to heat stress: overview and perspectives[J]. Biosci Rep, 2021, 41 (6): BSR20210443. doi: 10.1042/BSR20210443
3	WANG X N , MENG Z , MA A J . Genotype by temperature interaction for plasma physiological indexes in turbot (Scophthalmus maximus) under acute heat stress─exploring a method for screening physiological biomarkers of nontoxic stress in aquatic environments[J]. Environ Sci Technol, 2023, 57 (7): 2813- 2825. doi: 10.1021/acs.est.2c07958
4	VAN WETTERE W H E J , KIND K L , GATFORD K L , et al. Review of the impact of heat stress on reproductive performance of sheep[J]. J Anim Sci Biotechnol, 2021, 12 (1): 26. doi: 10.1186/s40104-020-00537-z
5	霍元楠, 邱美佳, 张姣姣, 等. 精氨酸及其代谢物抑制热应激诱导仔猪支持细胞凋亡的机制[J]. 畜牧兽医学报, 2024, 55 (2): 587- 597.
	HUO Y N , QIU M J , ZHANG J J , et al. Arginine and its metabolites attenuate heat stress-induced apoptosis of immature boar sertoli cells[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55 (2): 587- 597.
6	RIZZOTO G , KASTELIC J P . A new paradigm regarding testicular thermoregulation in ruminants?[J]. Theriogenology, 2020, 147, 166- 175. doi: 10.1016/j.theriogenology.2019.11.019
7	SHAHAT A M , THUNDATHIL J C , KASTELIC J P . Melatonin improves post-thaw sperm quality after mild testicular heat stress in rams[J]. Reprod Domest Anim, 2023, 58 (3): 423- 430. doi: 10.1111/rda.14302
8	QIN D Z , CAI H , HE C , et al. Melatonin relieves heat-induced spermatocyte apoptosis in mouse testes by inhibition of ATF6 and PERK signaling pathways[J]. Zool Res, 2021, 42 (4): 514- 524. doi: 10.24272/j.issn.2095-8137.2021.041
9	李延森, 李照见, 贾潇潇, 等. 环境高温诱导巴马香猪睾丸细胞凋亡并激活Nrf2/HO-1抗氧化信号通路[J]. 畜牧兽医学报, 2018, 49 (6): 1154- 1162.
	LI Y S , LI Z J , JIA X X , et al. Ambient high temperature induced cellular apoptosis and Nrf2/HO-1 signal pathway in testes of Bama miniature pigs[J]. Acta Veterinaria et Zootechnica Sinica, 2018, 49 (6): 1154- 1162.
10	LI S D , WANG Q M , YU H , et al. Polystyrene microplastics induce blood-testis barrier disruption regulated by the MAPK-Nrf2 signaling pathway in rats[J]. Environ Sci Pollut Res Int, 2021, 28 (35): 47921- 47931. doi: 10.1007/s11356-021-13911-9
11	ZHANG P F , ZHENG Y , LV Y H , et al. Melatonin protects the mouse testis against heat-induced damage[J]. Mol Hum Reprod, 2020, 26 (2): 65- 79. doi: 10.1093/molehr/gaaa002
12	GOEL A . Heat stress management in poultry[J]. J Anim Physiol Anim Nutr (Berl), 2021, 105 (6): 1136- 1145. doi: 10.1111/jpn.13496
13	张禛. 热应激对猪睾丸转化生长因子-β3和血睾屏障紧密连接分子表达与定位的影响[D]. 晋中: 山西农业大学, 2019.
	ZHANG Z. Effect of heat stress on the expression and localization of transforming growth factor-β3 and blood-testis barrier tight junctions molecules in boar testes[D]. Jinzhong: Shanxi Agricultural University, 2019. (in Chinese)
14	赵雅君, 付红梅, 李全凤, 等. 微波辐射家兔阴囊对血睾屏障的影响[J]. 中国男科学杂志, 2000, 14 (2): 100- 102.
	ZHAO Y J , FU H M , LI Q F , et al. Effect of blood testis microwave irradiation on scrotum in rabbits[J]. Chinese Journal of Andrology, 2000, 14 (2): 100- 102.
15	国晓瞳, 张倩, 于枫, 等. 黄芩苷对热应激犊牛睾丸支持细胞胶质细胞源性神经生长因子(GDNF)和干细胞因子(SCF)表达的影响[J]. 农业生物技术学报, 2015, 23 (4): 441- 449. doi: 10.3969/j.issn.1674-7968.2015.04.003
	GUO X T , ZHANG Q , YU F , et al. Baicalin affects glial cell line-derived neurotrophic factor (GDNF) and stem cell factor (SCF) expression in heat-stressed calf (Bos taurus) sertoli cells[J]. Journal of Agricultural Biotechnology, 2015, 23 (4): 441- 449. doi: 10.3969/j.issn.1674-7968.2015.04.003
16	XI H , FAN X , ZHANG Z , et al. Bax and Bcl-2 are involved in the apoptosis induced by local testicular heating in the boar testis[J]. Reprod Domest Anim, 2017, 52 (3): 359- 365. doi: 10.1111/rda.12904
17	ZENG Y X , YANG Q , OUYANG Y J , et al. Nickel induces blood-testis barrier damage through ROS-mediated p38 MAPK pathways in mice[J]. Redox Biol, 2023, 67, 102886. doi: 10.1016/j.redox.2023.102886
18	MAURYA V P , SEJIAN V , KUMAR D , et al. Impact of heat stress, nutritional restriction and combined stresses (heat and nutritional) on growth and reproductive performance of Malpura rams under semi-arid tropical environment[J]. J Anim Physiol Anim Nutr (Berl), 2016, 100 (5): 938- 946. doi: 10.1111/jpn.12443
19	SHAHAT A M , RIZZOTO G , KASTELIC J P . Amelioration of heat stress-induced damage to testes and sperm quality[J]. Theriogenology, 2020, 158, 84- 96. doi: 10.1016/j.theriogenology.2020.08.034
20	HASANI A , KHOSRAVI A , RAHIMI K , et al. Photobiomodulation restores spermatogenesis in the transient scrotal hyperthermia-induced mice[J]. Life Sci, 2020, 254, 117767. doi: 10.1016/j.lfs.2020.117767
21	WEI Y X , ZHOU Y , LONG C L , et al. Polystyrene microplastics disrupt the blood-testis barrier integrity through ROS-Mediated imbalance of mTORC1 and mTORC2[J]. Environ Pollut, 2021, 289, 117904. doi: 10.1016/j.envpol.2021.117904
22	李悦. 五子衍宗丸对睾丸支持细胞及血睾屏障的作用和机制[D]. 北京: 北京中医药大学, 2021.
	LI Y. Mechanism of Wuziyanzong pill on Sertoli cells and blood-testis barrier[D]. Beijing: Beijing University of Chinese Medicine, 2021. (in Chinese)
23	HU S Q , LIU D L , LIU S J , et al. Lycium barbarum polysaccharide ameliorates heat-stress-induced impairment of primary sertoli cells and the blood-testis barrier in rat via androgen receptor and Akt phosphorylation[J]. Evid Based Complement Alternat Med, 2021, 2021, 5574202.
24	LI X X , CHEN S R , SHEN B , et al. The heat-induced reversible change in the blood-testis barrier (BTB) is regulated by the androgen receptor (AR) via the partitioning-defective protein (Par) polarity complex in the mouse[J]. Biol Reprod, 2013, 89 (1): 12.
25	李治利, 刘容序, 麻冰洁, 等. 蜂毒素对热应激鹌鹑睾丸损伤的保护作用[J]. 养殖与饲料, 2023, 22 (10): 24- 29.
	LI Z L , LIU R X , MA B J , et al. Protective effect of melittin on testicular injury in heat-stressed quails[J]. Animals Breeding and Feed, 2023, 22 (10): 24- 29.
26	和玉丹, 刘慧娟, 张佳琦, 等. 日粮添加绿原酸对热应激小鼠睾丸抗氧化功能的影响[J]. 畜牧与兽医, 2022, 54 (4): 40- 46.
	HE Y D , LIU H J , ZHANG J Q , et al. Effects of dietary chlorogenic acid on the antioxidant capacity of testis in mice under heat stress[J]. Animal Husbandry & Veterinary Medicine, 2022, 54 (4): 40- 46.
27	张宁, 张黎, 丁晓萍, 等. 精子染色质结构分析与精液参数的相关性研究[J]. 中华男科学, 2003, 9 (3): 166- 169.
	ZHANG N , ZHANG L , DING X P , et al. Analysis of correlation between parameters of sperm chromatin structure assay and semen routine Analysis[J]. National Journal of Andrology, 2003, 9 (3): 166- 169.
28	张琛. 热应激对新西兰白兔种公兔精液品质和精浆代谢物的影响[D]. 扬州: 扬州大学, 2022.
	ZHANG C. Effects of heat stress on semen quality and seminal plasma metabolites in male New Zealand white rabbits[D]. Yangzhou: Yangzhou University, 2022. (in Chinese)
29	SHE J J , FENG N N , ZHENG W L , et al. Zearalenone exposure disrupts blood-testis barrier integrity through excessive Ca²⁺-mediated autophagy[J]. Toxins (Basel), 2021, 13 (12): 875.
30	LI Y Y , ZAFAR M I , WANG X T , et al. Heat stress and pulsed unfocused ultrasound: the viability of these physical approaches for drug delivery into testicular seminiferous tubules[J]. Curr Drug Deliv, 2020, 17 (5): 438- 446.
31	范小瑞. 猪隐睾睾丸精子发生障碍的分子机制研究[D]. 晋中: 山西农业大学, 2021.
	FAN X R. Study on the molecular mechanism of spermatogenesis disorder in cryptorchidism boars[D]. Jinzhong: Shanxi Agricultural University, 2021. (in Chinese)
32	魏琳婷, 葛蓬勃, 马小琴, 等. TRPC6参与内质网应激诱导的肾小球系膜细胞凋亡[J]. 西安交通大学学报(医学版), 2023, 44 (6): 898- 903.
	WEI L T , GE P B , MA X Q , et al. TRPC6 involves in endoplasmic reticulum stress induced apoptosis of glomerular mesangial cells[J]. Journal of Xi'an Jiaotong University (Medical Sciences), 2023, 44 (6): 898- 903.
33	赵玉华, 李俊莹, 王春苗, 等. 大黄酸衍生物4B诱导内质网应激触发顺铂耐药卵巢癌细胞凋亡的活性研究[J]. 广西医科大学学报, 2021, 38 (3): 431- 437.
	ZHAO Y H , LI J Y , WANG C M , et al. Apoptosis of cisplatin-resistant ovarian cancer cells induced by Rhein derivative 4B via triggering endoplasmic reticulum stress[J]. Journal of Guangxi Medical University, 2021, 38 (3): 431- 437.
34	刘洪茂, 姬洋, 曹博阳, 等. PERK信号通路在热诱导睾丸生殖细胞凋亡中的作用[J]. 环境与健康杂志, 2016, 33 (1): 19- 22.
	LIU H M , JI Y , CAO B Y , et al. Role of PERK signal pathway in scrotal hyperthermia-induced germ cell apoptosis in testes[J]. Journal of Environment and Health, 2016, 33 (1): 19- 22.
35	胡煜. 热应激通过miR-145-5p-花生四烯酸调节仔猪睾丸支持细胞自噬的分子机制[D]. 重庆: 西南大学, 2021.
	HU Y. Major: animal genetics breeding and reproduction field: animal reproductive physiology and reproduction regulation[D]. Chongqing: Southwest University, 2021. (in Chinese)
36	CHANG L L , LU Z , LI D L , et al. Melamine causes testicular toxicity by destroying blood-testis barrier in piglets[J]. Toxicol Lett, 2018, 296, 114- 124.
37	ȘENTÜRK G E , CANILLIOĜLU Y E , UMAY C , et al. Distribution of Zonula Occludens-1 and Occludin and alterations of testicular morphology after in utero radiation and postnatal hyperthermia in rats[J]. Int J Exp Pathol, 2012, 93 (6): 438- 449.
38	MAZAUD-GUITTOT S , MEUGNIER E , PESENTI S , et al. Claudin 11 deficiency in mice results in loss of the Sertoli cell epithelial phenotype in the testis[J]. Biol Reprod, 2010, 82 (1): 202- 213.
39	张禛, 范小瑞, 席华明, 等. 热应激对性成熟猪睾丸TGF-β3和Claudin-11蛋白表达的影响[J]. 畜牧兽医学报, 2016, 47 (10): 2136- 2142.
	ZHANG Z , FAN X R , XI H M , et al. Effect of heat stress on the expression of TGF-β3 and Claudin-11 protein in mature boar testis[J]. Acta Veterinaria et Zootechnica Sinica, 2016, 47 (10): 2136- 2142.
40	LI M W M , MRUK D D , LEE W M , et al. Connexin 43 is critical to maintain the homeostasis of the blood-testis barrier via its effects on tight junction reassembly[J]. Proc Natl Acad Sci U S A, 2010, 107 (42): 17998- 18003.
41	JIANG X H , MA T L , ZHANG Y W , et al. Specific deletion of Cdh2 in Sertoli cells leads to altered meiotic progression and subfertility of mice[J]. Biol Reprod, 2015, 92 (3): 79.

[1]	LI Longping, LI Tuo, CAO Peiwen, ZHU Haijing, ZHANG Xiaoling, ZHANG Chen, XIAO Puhui, DONG Shuwei, FENG Ping, QU Lei, BI Taifei. Effects of Diets with Different Energy Levels on Rumen Fermentation Characteristics and Microbial Composition of Weaned Male Shaanbei White Cashmere Goats [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(7): 3011-3023.
[2]	DONG Shucan, MAO Shuaixiang, WU Cuiying, LI Yaokun, SUN Baoli, GUO Yongqing, DENG Ming, LIU Dewu, LIU Guangbin. The Effect of the Androgen Receptor Inhibitor Enzalutamide on Proliferation and Apoptosis of Goat Ovarian Granulosa Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2022-2031.
[3]	LI Qiuyun, TIAN Xinyuan, LIAO Wensheng, ZHANG Huanrong, REN Yupeng, YANG Falong, ZHU Jiangjiang, XIANG Hua. Effects of SOCS2 on Proliferation, Cycle and Apoptosis of Turbinate Bone Cells in Goats [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2226-2240.
[4]	LI Pengfei, GAO Guiqin, ZHOU Guangqing, WU Jinyan, YAN Xinmin, CAO Xiaoan, HE Jijun, YUAN Ligang, SHANG Youjun. Establishment and Application of TaqMan Fluorescence Quantitative RT-PCR Detection Method for Enzootic Nasal Tumor Virus of Goats [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2259-2266.
[5]	PENG Peiya, CHEN Yuhan, YANG Long, WANG Ming, ZHAO Ruiting, HE Jun, YIN Yulong, LIU Mei. Research Progress of Copy Number Variation in Livestock [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1356-1369.
[6]	KANG Jia, DUAN Xiangru, YIN Xuejiao, YANG Ruochen, LI Taichun, SHAN Xinyu, CHEN Meijing, ZHANG Yingjie, LIU Yueqin. Effects of Cysteine and Methionine on Secondary Hair Follicle Growth and Hair Dermal Papilla Cell Proliferation in vitro in Cashmere Goats [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 515-527.
[7]	SONG Yan, YUAN Yongfeng, QIAN Hongyu, LI Xincan, LUO Hongyan, WANG Zhiying, ZHOU Zuoyong. Identification and Partial Biological Characteristics Analysis of Corynebacterium pseudotuberculosis Isolated from Goats [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 680-687.
[8]	YAN Xiaochun, XI Haijiao, LI Jinquan, WANG Zhiying, SU Rui. Study on Estimates of Genomic Breeding Value of Fleece Traits in Inner Mongolia Cashmere Goats [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 120-128.
[9]	ZHANG Chenjian, LI Yinxia, DING Qiang, LIU Weijia, WANG Huili, HE Nan, WU Jiashun, CAO Shaoxian. Efficient Preparation of CRISPR/Cas9-mediated Goat SOCS2 Gene Edited Embryos [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 129-141.
[10]	ZHANG De'an, YANG Ruozhu, LIU Jie, LIU Dewu, DENG Ming, LIU Guangbin, SUN Baoli, GUO Yongqing, LI Yaokun. Expression Analysis of Transcriptome in the Liver of Chuanzhong Black Goats Fed with Silage Neolamarckia Cadamba Substitute for Silage Corn [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(1): 232-244.
[11]	TANG Yinmei, LI Qi, LI Haiyang, LIN Yaqiu, WANG Yong, XIANG Hua, HUANG Lian, ZHU Jiangjiang. Cloning of the Goat FATP2 Gene and Its Regulatory Effect on Lipid Deposition in Precursor Adipocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3642-3652.
[12]	SHAO Peng, TANG Yinmei, LIN Yaqiu, WANG Yong, XIANG Hua, HUANG Lian, ZHU Jiangjiang. Regulation Effect of PSMD9 on Lipid Deposition in Goat Precursor Adipocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3653-3663.
[13]	LIU Xinhuan, YUN Jialei, MAO Li, LI Jizong, HAO Fei, HE Miaofeng, YANG Leilei, ZHANG Wenwen, CHENG Zilong, SUN Min, LIU Maojun, WANG Shaohui, BAI Juan, LI Wenliang. Isolation, Identification, Virulence Genes and Drug Resistance Analysis of Escherichia coli Isolated from Diarrheal Goat and Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3445-3454.
[14]	ZHANG Renbao, ZHOU Donghui, ZHOU Lisheng, GAO Xiaoxiao, LIU Nan, HE Jianning. Analysis of Genetic Structure of Conservation Population in Jining Gray Goats Based on 70 K SNP Chip [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 2836-2847.
[15]	SHAO Yuexin, ZHANG Xinyu, GE Liyan, SHI Huaiping. Cloning and RNA Interference Analysis of ATF4 Gene in Xinong Saanen Dairy Goat [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2353-2364.

Effect of Scrotal Hyperthermia on Structure and Function of Blood-testis Barrier in Goats

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 6

References 41

Related Articles 15

Recommended Articles

Metrics

Comments