NPFFR1基因对鹅卵泡颗粒细胞激素分泌和细胞凋亡的影响研究

doi:10.11843/j.issn.0366-6964.2021.010.013

Abstract

Abstract: RFamide-related peptide receptor(NPFFR1) is a primary affinity receptor of the gonadotropin-inhibitory hormone (GnIH), which plays an important role in controlling animal reproduction. In order to explore the effect of the NPFFR1 on follicle development of geese, nine healthy laying Sichuan white geese of 42-week-old were used as experimental animals. The RT-qPCR technology was used to determine the expression pattern of NPFFR1 in the granulosa cells (GCs) of pre-hierarchical and hierarchical follicles. The NPFFR1 gene expression was upregulated using the overexpression plasmids technique in GCs, and the concentration of reproduction-related hormones (E2, P4 and AMH) was determined by ELISA in the cellular supernatant, the TUNEL assay was employed to detect apoptosis of the remaining anchorage-dependent cell. The RNA sequencing was performed to screen the differentially expressed genes (DEGs) before and after overexpressing NPFFR1 in GCs of 8-10 mm follicles, respectively. The functional cluster analysis of differentially expressed genes was carried out. The results showed that the gene expression of NPFFR1 was extremely significantly different between the pre-hierarchal and hierarchal periods follicular GCs in geese (P<0.01), except F1 follicles. After overexpressed the NPFFR1 gene (72 h), the concentration of E2 in the supernatant of GCs isolated from hierarchal follicles and AMH in the supernatant of GCs isolated from pre-hierarchal follicles were significantly decreased (P<0.05); There were 267 DEGs (119 down-regulated and 148 up-regulated) were selected. The DEGs were mainly enriched in the process of biological rhythm, follicles rhythm. The qPCR results showed that, compared to the control group, the AMH gene was significantly down-regulated (P<0.05), the Clock, FOS, Per and ANTXR2 were extremely significantly (P<0.01) or significantly (P<0.05) up-regulated, respectively. Therefore, it was specu-lated that the NPFFR1 gene might be involved in follicular development by regulating the expression levels of the rhythmic genes, affecting the steroid hormone secretion and apoptosis of follicular granulosa cells of goose.

Key words: NPFFR1 gene, geese, follicles hierarchical development, sexual hormones, cell apoptosis, circadian genes

CLC Number:

S835.2

ZHANG Keshan, GAO Guangliang, LI Qin, ZHAO Xianzhi, LI Jing, WANG Qigui. The Function of NPFFR1 on Hormone Secretion and Apoptosis of Follicle Granulosa Cell in Geese[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(10): 2822-2831.

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The Function of NPFFR1 on Hormone Secretion and Apoptosis of Follicle Granulosa Cell in Geese

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