缺氧诱导基因1C在牦牛隐睾中的表达及其调控机制研究

doi:10.11843/j.issn.0366-6964.2024.07.018

Abstract

Abstract:

The study aimed to investigate the effect of HIG1 hypoxia inducible domain family member 1C (HIGD1C) on apoptosis of sertoli cells in yak testis. In this study, 3 healthy male yaks aged 3-6 years and male yaks with cryptorchidism were selected, respectively as research materials, and their testis was collected. The regulation mechanism of HIGD1C on apoptosis of yak testicular sertoli cells was explored by HE staining, isolation and culture of yak testicular sertoli cells, indirect immunofluorescence, RT-qPCR, Western blot, HIGD1C overexpression and interference vector construction and in vitro cell transfection. The results showed that the basement membrane and interstitial tissue of normal testis were intact, the seminiferous tubules, peritubular muscle cells and sertoli cells were closely arranged, the seminiferous tubules in cryptorchidism were broken and atrophied, and the transcription and protein expression level of HIGD1C in yak cryptorchidism was significantly higher than that in normal testis (P < 0.01); HIGD1C protein was mainly distributed in leydig cells and sertoli cells; yak sertoli cells were successfully isolated and cultured; and HIGD1C protein was located in the nucleus of primary sertoli cells; after overexpression of HIGD1C, the mRNA expression level of Bcl-2 was up-regulated (P < 0.05), the protein expression level was significantly up-regulated (P < 0.01), and the mRNA expression levels and protein expression levels of Bax and Caspase-3 were significantly up-regulated (P < 0.01), the results of flow cytometry showed that the apoptosis rate of sertoli cells in HIGD1C overexpression group was significantly increased (P < 0.01). On the contrary, after HIGD1C knockout, the mRNA and protein expression levels of Bax, Bcl-2 and Caspase-3 were significantly down-regulated (P < 0.01), the results of flow cytometry showed that the apoptosis rate of sertoli cells in HIGD1C knockout group was significantly down-regulated (P < 0.01). The results of this study show that the normal testicular tissue structure is complete, the cryptorchidism stroma is loose, and HIGD1C is highly expressed in the cryptorchidism of yak. In vitro cell experiments showe that up-regulation of HIGD1C could activate pro-apoptotic proteins and promote apoptosis of yak sertoli cells. On the contrary, down-regulation of HIGD1C expression level will inhibit the apoptosis of sertoli cells. It is suggested that HIGD1C is involved in the regulation of apoptosis of yak testicular sertoli cells, which provides a reference for revealing the mechanism of cryptorchidism in yak.

Key words: HIGD1C, cryptorchidism, sertoli cells, apoptosis, yak

CLC Number:

S823.85

Milan MA, Qi WANG, Qiu YAN, Tianan LI, Xingxu ZHAO, Yong ZHANG. Expression of HIG1 Hypoxia Inducible Domain Family Member 1C in Cryptorchidism of Yak and Its Regulatory Mechanism[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(7): 2983-2994.

Figures/Tables 7

Table 1

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引物名称Primer name	引物序列(5′→3′)Primer sequence	片段长度/bp Product length	温度/℃ Tm
HIGD1C	F：TCAGCAGATGAAGACGAAGG R：GGACACCACAGTCACAAAGC	102	56
Caspase-3	F：GGTGGGGTCAGAGCCATAGA R：CATCTTCCACACACACCCGTAG	84	57
Bax	F：CCGAGTGGCGGCTGAAA R：TCCAGATGGTGAGCGAGGC	287	60
Bcl-2	F：CTGCACCTGACGCCCTTCAC R：GCGTCCCAGCCTCCGTTGT	236	60
GAPDH	F：GCTGGTGCTGAGTATGTGGTG R：GCTGACAATCTTGAGGGTGTTG	177	60

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Expression of HIG1 Hypoxia Inducible Domain Family Member 1C in Cryptorchidism of Yak and Its Regulatory Mechanism

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