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1.
Reproduction ; 161(5): 523-537, 2021 05.
Article En | MEDLINE | ID: mdl-33730690

Long ncRNAs regulate a complex array of fundamental biological processes, while its molecular regulatory mechanism in Leydig cells (LCs) remains unclear. In the present study, we established the lncRNA LOC102176306/miR-1197-3p/peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PPARGC1A) regulatory network by bioinformatic prediction, and investigated its roles in goat LCs. We found that lncRNA LOC102176306 could efficiently bind to miR-1197-3p and regulate PPARGC1A expression in goat LCs. Downregulation of lncRNA LOC102176306 significantly supressed testosterone (T) synthesis and ATP production, decreased the activities of antioxidant enzymes and mitochondrial complex I and complex III, caused the loss of mitochondrial membrane potential, and inhibited the proliferation of goat LCs by decreasing PPARGC1A expression, while these effects could be restored by miR-1197-3p inhibitor treatment. In addition, miR-1197-3p mimics treatment significantly alleviated the positive effects of lncRNA LOC102176306 overexpression on T and ATP production, antioxidant capacity and proliferation of goat LCs. Taken together, lncRNA LOC102176306 functioned as a sponge for miR-1197-3p to maintain PPARGC1A expression, thereby affecting the steroidogenesis, cell proliferation and oxidative stress of goat LCs. These findings extend our understanding of the molecular mechanisms of T synthesis, cell proliferation and oxidative stress of LCs.


Leydig Cells/cytology , MicroRNAs/genetics , Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha/metabolism , RNA, Long Noncoding/genetics , Testis/cytology , Animals , Apoptosis , Cell Proliferation , Goats , Leydig Cells/metabolism , Male , Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha/genetics , Testis/metabolism , Testosterone/metabolism
2.
Theriogenology ; 138: 137-144, 2019 Oct 15.
Article En | MEDLINE | ID: mdl-31352175

This study aimed to determine the effects of l-arginine (L-Arg) supplementation on steroid hormone receptors in non-pregnant ovine endometrium. All experimental ewes were randomly assigned to either a control group (n = 6), a nutrient-restricted group (n = 6), or an L-Arg supplemented nutrient-restricted group (n = 6). The effects of L-Arg on estrogen receptor α/ß (ERα/ß) and progesterone receptor (PGR) expression in the ovine endometrium were assessed. Our results showed that levels of ERß and PGR expression were significantly increased by nutrient restriction, but L-Arg counteracted the effect of nutrient restriction on ERß and PGR expression (p < 0.05). Also, expression of endometrial ERα was substantially increased (p < 0.05) by L-Arg supplementation. Furthermore, ERα/ß and PGR were mainly detected in the endometrial luminal epithelium and glandular epithelium. Therefore, we isolated and identified endometrial epithelial cells (EECs) from sheep. Different concentrations of L-Arg were added to investigate the effects on ERα/ß and PGR in EECs. The expression levels of endothelial nitric oxide synthase, ERß, and PGR were significantly increased in response to low-concentration (200 µmol) L-Arg supplementation, which subsequently decreased with a high concentration (800 µmol) (p < 0.05). Otherwise, ERα expression was remarkably increased at both L-Arg concentrations in EECs (p < 0.05). Overall, the results indicated that L-Arg performed crucial roles in the regulation of ovine steroid hormone receptor expression in the endometrium. The results of this study provide a theoretical basis and technical means for the normal function of endometrium in response to low nutrient levels.


Arginine/pharmacology , Caloric Restriction , Endometrium/drug effects , Estrogen Receptor alpha/genetics , Estrogen Receptor beta/genetics , Receptors, Progesterone/genetics , Sheep , Animal Nutritional Physiological Phenomena/drug effects , Animals , Caloric Restriction/veterinary , Cells, Cultured , Endometrium/metabolism , Epithelial Cells/drug effects , Epithelial Cells/metabolism , Estrogen Receptor alpha/metabolism , Estrogen Receptor beta/metabolism , Female , Gene Expression Regulation/drug effects , Nutrients , Pregnancy , Receptors, Progesterone/metabolism , Sheep/genetics , Sheep/metabolism , Uterus/drug effects , Uterus/metabolism
3.
Gene ; 710: 131-139, 2019 Aug 20.
Article En | MEDLINE | ID: mdl-31158446

As a fundamental regulator of mitochondrial function, peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PPARGC1A) acts as a powerful coactivator of many transcriptional factors that relate to steroidogenesis, while the regulatory mechanism remains unclear. In the present study, testosterone secretion of goat Leydig cells (LCs) mediated by miR-1197-3p via PPARGC1A was investigated. We found PPARGC1A protein was diversely localized in testis, and the expression of PPARGC1A in testis of 9-month-old goat was significantly higher than that in 3-month-old goat. In addition, suppression of PPARGC1A significantly decreased the testosterone secretion in goat LCs, as well as reduced the expressions of key steroidogenesis related genes [steroidogenic acute regulatory protein (StAR), cytochrome P450 family 11 subfamily A member 1 (CYP11A1), and 3 beta-hydroxysteroid dehydrogenase (3BHSD)], and overexpression of PPARGC1A showed the opposite effects. Moreover, we observed suppression of miR-1197-3p increased the synthesis of testosterone and promoted the expressions of PPARGC1A, StAR, CYP11A1, and 3BHSD by directly targeting PPARGC1A in the LCs. Furthermore, overexpression of PPARGC1A could alleviate miR-1197-3p induced aberrant steroidogenesis related gene expressions and testosterone synthesis. Taken together, miR-1197-3p could act as an essential regulator of LC testosterone secretion in goat testis by targeting PPARGC1A. These results provide a novel view of the regulatory mechanisms involved in male sexual maturation and help us to understand the molecular role of PPARGC1A in testosterone synthesis.


Leydig Cells/metabolism , MicroRNAs/genetics , Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha/genetics , Testosterone/metabolism , Animals , Cells, Cultured , Cholesterol Side-Chain Cleavage Enzyme/metabolism , Gene Expression Regulation, Developmental , Goats , Male , Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha/metabolism , Real-Time Polymerase Chain Reaction
4.
Guang Pu Xue Yu Guang Pu Fen Xi ; 34(11): 2923-8, 2014 Nov.
Article Zh | MEDLINE | ID: mdl-25752032

The objective of the present study was to reveal different tolerance of peanut plants to Ca deficiency by determining Ca uptake and Fourier transform infrared spectral (FTIR) differences of two peanut cultivars grown in nutrition solution. Peanut cultivars LH11 and YZ9102 were selected. Seedlings at the first leaf stage were cultivated for 28 days in nutrient solution with 0, 0.01 and 2.0 mmol x L(-1) Ca treatments, respectively. The results showed that under 0 and 0.01 mmol x L(-1) Ca supply, YZ9102 did not show Ca deficiency symptoms and the plant biomass did not change, whereas LH11 exhibited shoot-tip necrosis, smaller plant size, more lateral branches, and plant dry matter weights decreased significantly. YZ9102 had higher plant Ca concentration and Ca accumulation than LH11. Besides, for LH11, Ca was mainly accumulated in roots, while for YZ9102 mainly in leaves. As compared with plants cultivated in 2.0 mol x L(-1) Ca nutrition, root, stem and leaf of LH11 plants under Ca deficiency stress showed higher transmittance at peaks 1 060, 1 380, 1 655, 2 922, and 3 420 cm(-1) in FTIR spectra, indicating that the contents of protein, sugar and lipid decreased obviously in LH11 plants in condition that Ca supply was limited. However, the FTIR spectra of YZ9102 were less affected by Ca deficiency. It is suggested that YZ9102 might be more tolerant to Ca deficiency.


Arachis/physiology , Calcium/metabolism , Biomass , Plant Leaves , Plant Roots , Seedlings , Spectroscopy, Fourier Transform Infrared , Stress, Physiological
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