Is Autophagy a Prerequisite for Steroidogenesis in Leydig Cells?
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
We investigated the involvement of autophagy with steroidogenesis in testicular Leydig cells. Human chorionic gonadotropin (hCG)-stimulated T production in Leydig cells was not remarkably altered in the presence of an autophagy inhibitor 3-methyladenine (3-MA). Although pretreatment with 3-MA demonstrated a tendency to decrease hCG-induced T production, the differences were significant only at a higher time point of 24 h following hCG. Microtubule associated protein light chain 3 (LC3)-II was detectable in the control cells in all the experiments. The hCG-induced increase in steroidogenic acute regulatory protein (StAR) and cytochrome P450 side chain cleave (P450scc) protein levels were not significantly altered by 3-MA. Leydig cells isolated from immature rat testes 12 h following hCG treatment showed relatively increased levels of LC3-II protein compared to the control group. Furthermore, LC3-II levels shown in these cells reached almost the identical to those from normal adult testes. However, LC3-II protein levels were almost comparable or even slightly lower than the controls at 48 h following hCG. Expression of StAR and P450scc was upregulated at both 12 and 48 h after hCG. We also used MA-10 cells, the mouse Leydig cell line, in this experiment. When dibutyryl cyclic-AMP was treated with MA-10 cells, P4 levels were significantly increased in the cell culture medium. However, P4 levels tended to decrease in the presence of 3-MA, but the difference was not statistically significant. This was consistent with the results of the rat Leydig cell experiments. Together, we believe that although autophagy participates in steroidogenesis and enhances steroidogenic efficacy of Leydig cells, it may not be a decisive cellular process for steroidogenesis, specifically in the mature Leydig cells.
Abstract
We investigated the involvement of autophagy with steroidogenesis in testicular Leydig cells. Human chorionic gonadotropin (hCG)-stimulated T production in Leydig cells was not remarkably altered in the presence of an autophagy inhibitor 3-methyladenine (3-MA). Although pretreatment with 3-MA demonstrated a tendency to decrease hCG-induced T production, the differences were significant only at a higher time point of 24 h following hCG. Microtubule associated protein light chain 3 (LC3)-II was detectable in the control cells in all the experiments. The hCG-induced increase in steroidogenic acute regulatory protein (StAR) and cytochrome P450 side chain cleave (P450scc) protein levels were not significantly altered by 3-MA. Leydig cells isolated from immature rat testes 12 h following hCG treatment showed relatively increased levels of LC3-II protein compared to the control group. Furthermore, LC3-II levels shown in these cells reached almost the identical to those from normal adult testes. However, LC3-II protein levels were almost comparable or even slightly lower than the controls at 48 h following hCG. Expression of StAR and P450scc was upregulated at both 12 and 48 h after hCG. We also used MA-10 cells, the mouse Leydig cell line, in this experiment. When dibutyryl cyclic-AMP was treated with MA-10 cells, P4 levels were significantly increased in the cell culture medium. However, P4 levels tended to decrease in the presence of 3-MA, but the difference was not statistically significant. This was consistent with the results of the rat Leydig cell experiments. Together, we believe that although autophagy participates in steroidogenesis and enhances steroidogenic efficacy of Leydig cells, it may not be a decisive cellular process for steroidogenesis, specifically in the mature Leydig cells.
참고문헌 (29)
- Chen H, "Leydig cells: From stem cells to aging", Mol Cell Endocrinol, 306, pp.9~16 (2009)
- Chen H, "Age-related decreased Leydig cell testoster-one production in the brown Norway rat", J Androl, 15, pp.551~557 (1994)
- Choi AM, "Autophagy in human health and disease", N Engl J Med, 368, pp.651~662 (2013)
- Ewing LL, "Leydig cell structure and steroidogenic function", Recent Prog Horm Res, 39, pp.599~635 (1983)
- Gao F, "Autophagy regulates testosterone synthesis by facilitating cholesterol uptake in Leydig cells", J Cell Biol, 217, pp.2103~2119 (2018)
- Glick D, "Autophagy: Cellular and molecular mechanisms", J Pathol, 221, pp.3~12 (2010)
- Klinefelter GR, "Effect of luteinizing hormone deprivation in situ on steroidogenesis of rat Leydig cells purified by a multistep procedure", Biol Reprod, 36, pp.769~783 (1987)
- Klionsky DJ, "Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes", Autophagy, 4, pp.151~175 (2008)
- Klionsky DJ, "Autophagy as a regulated pathway of cellular degradation", Science, 290, pp.1717~1721 (2000)
- Lamark T, "Mechanisms of Selective Autophagy", Annu Rev Cell Dev Biol, 37, pp.143~169 (2021)
- Levine B, "Autophagy in the pathogenesis of disease", Cell, 132, pp.27~42 (2008)
- Levine B, "Biological functions of autophagy genes: A disease perspective", Cell, 176, pp.11~42 (2019)
- Li WR, "Autophagic deficiency is related to steroidogenic decline in aged rat Leydig cells", Asian J Androl, 13, pp.881~888 (2011)
- Miller WL, "Mitochondrial specificity of the early steps in steroidogenesis", J Steroid Biochem Mol Biol, 55, pp.607~616 (1995)
- Miller WL, "Steroidogenic enzymes", Endocr Dev, 13, pp.1~18 (2008)
- Miller WL, "Role of mitochondria in steroidogenesis", Endocr Dev, 20, pp.1~19 (2011)
- Miller WL, "Early steps in steroidogenesis: Intracellular cholesterol trafficking", J Lipid Res, 52, pp.2111~2135 (2011)
- Mizushima N, "Autophagy: Renovation of cells and tissues", Cell, 147, pp.728~741 (2011)
- Noda T, "Tor, a phosphatidylinositol kinase homologue, controls autophagy in yeast", J Biol Chem, 273, pp.3963~3966 (1998)
- Roberts KP, "Androgen regulation of spermatogenesis in the rat", Ann N Y Acad Sci, 637, pp.90~106 (1991)
- Schnebert S, "Diving into the evolutionary history of HSC70-linked selective autophagy pathways: Endosomal microautophagy and chaperone-mediated autophagy", Cells, 11, pp.1945~ (2022)
- Seglen PO, "3-Methyladenine: Specific inhibitor of autophagic", lysosomal protein degradation in isolated rat hepatocytes, 1982, pp.79, 1889~1892
- Stocco DM, "Regulation of the acute production of steroids in steroidogenic cells", Endocr Rev, 17, pp.221~244 (1996)
- Tang XM, "Origin and fate of autophagosomes in Leydig cells of normal adult rats", J Androl, 9, pp.284~293 (1988)
- Tsujimoto Y, "Another way to die: Autophagic programmed cell death", Cell Death Differ, 12(Suppl 2), pp.1528~1534 (2005)
- Turner TT, "On the androgen microenvironment of maturing spermatozoa", Endocrinology, 115, pp.1925~1932 (1984)
- Vargas JNS, "The mechanisms and roles of selective autophagy in mammals", Nat Rev Mol Cell Biol, 24, pp.167~185 (2023)
- Yi J, "Functional implication of autophagy in steroid-secreting cells of the rat", Anat Rec, 242, pp.137~146 (1995)
- Zirkin BR, "Leydig cells: Formation, function, and regulation", Biol Reprod, 99, pp.101~111 (2018)