Effects of &alpha;-Ketoglutaric Acid/Oxoglutarate Receptor 1 System on Testosterone Secretion and Spermatogenesis of Male Animals

XU Chang; ZHANG Cha; TAN Chengquan; XI Qianyun; WANG Shenghui; JIANG Qingyan; SHU Gang

doi:10.3969/j.issn.1006-267x.2022.05.054

2022 , Vol. 34 >Issue 5: 3286 - 3296

DOI: https://doi.org/10.3969/j.issn.1006-267x.2022.05.054

RESEARCH PAPER

Effects of α-Ketoglutaric Acid/Oxoglutarate Receptor 1 System on Testosterone Secretion and Spermatogenesis of Male Animals

XU Chang ,
ZHANG Cha ,
TAN Chengquan ,
XI Qianyun ,
WANG Shenghui ,
JIANG Qingyan ,
SHU Gang

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1. Guangdong Provincial Key Laboratory of Animal Nutrition Control, Scientific Observing and Experimental Station of Animal Nutrition and Feed Science in South China, Ministry of Agriculture, College of Animal Science, South China Agricultural University, Guangzhou 510642, China;
2. Guangdong Wenshi Breeding Swine Technology Co., Ltd., Yunfu 527400, China

Received date: 2021-10-24

Online published: 2022-05-14

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Abstract

This experiment was conducted to investigate effects of α-ketoglutarate acid (AKG)/oxoglutarate receptor 1 (OXGR1) system on testosterone secretion and spermatogenesis of male animals. Firstly, the localization of OXGR1 in testicular tissue was detected by immunofluorescence. Then, OXGR1 knockout (OXGR1KO) mice were adopted to test the serum testosterone content by enzyme linked immunosorbent assay (ELISA) method, the hematoxylin-eosin (HE) staining was used to analyze the differences of testicular seminiferous tubule lumen area and testicular tissue morphology, and the mRNA expression level of testicular spermatogenesis marker genes were detected by real-time quantitative PCR. Then, in the acute test, sixteen 10-week-old mal mice were randomly divided into 4 groups according to the body weight, and the serum testosterone content and aggressiveness of the mice were detected after injection of 0, 1, 5 and 10 mg/kg AKG for 6 h, respectively; then twelve 10-week-old male mice and OXGR1KO mice were randomly divided into 4 groups according to the body weight, and the serum testosterone content was detected after 6 h injection of 5 mg/kg AKG in each group. In vitro test, the leydig cells (TM3) cells were treated with different concentrations of AKG for 24 h, and the content of testosterone in supernatant of TM3 cells was detected; the TM3 cells were treated with 200 μmol/L AKG to detect the intracellular calcium ion concentration. In long-term test, twelve 8-week-old male mice and OXGR1KO mice were selected and randomly divided into 4 groups according to the body weight:WT group, WT+2%AKG group, OXGR1KO group and OXGR1KO+2%AKG group, after 28 days of experiment, the serum testosterone content and testicular seminiferous tubule lumen area were detected. The results showed as follows:1) the OXGR1 was specifically distributed in leydig cells of testis. 2) OXGR1KO could significantly reduce the serum testosterone content and the mRNA relative expression of class Ⅲ beta-tubulin (Tubb3) and sex-determining region Y box 9 (Sox9) (P<0.05 or P<0.01). 3) After acute intraperitoneal injection of 5 mg/kg AKG for 6 h, the serum testosterone content of mice was significantly increased (P<0.05), and the aggressiveness of mice was increased. 4) The in vitro study showed that AKG could significantly increase the testosterone content in supernatant of TM3 cells in a dose-dependent manner (P<0.01), and 200 μmol/L AKG could significantly promote the release of instantaneous intracellular calcium of TM3 cells (P<0.01). 5) Long-term drinking water supplemented with 2%AKG for 4 weeks could significantly increase the serum testosterone content (P<0.01), significantly decrease the lumen area of testicular seminiferous tubules (P<0.01), while OXGR1KO could significant reversal AKG-induced serum testosterone content and seminiferous tubule morphology change (P<0.05). In conclusion, AKG, as an important bioactive metabolic intermediate, can increase the serum testosterone content of male animals through OXGR1 and affect the development of seminiferous tubules and spermatogenesis.

Key words： AKG; OXGR1; testosterone; lumen area

Cite this article

XU Chang , ZHANG Cha , TAN Chengquan , XI Qianyun , WANG Shenghui , JIANG Qingyan , SHU Gang . Effects of α-Ketoglutaric Acid/Oxoglutarate Receptor 1 System on Testosterone Secretion and Spermatogenesis of Male Animals[J]. Chinese Journal of Animal Nutrition, 2022 , 34(5) : 3286 -3296 . DOI: 10.3969/j.issn.1006-267x.2022.05.054

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