Dataset Information

Inhibition of the fructose transporter Glut5 boosts androgen production by dysregulating the metabolism of murine Leydig cells

ABSTRACT: Over the past few decades, a significant change in global sugar intake has coincided with a rising incidence of male infertility, which is now a major public health concern. Diets rich in fructose have been implicated in both male infertility and an increased susceptibility to metabolic disorders like obesity, diabetes, and related heart problems. While fructose is known to be present in seminal fluid and crucial for sperm motility, the precise role of fructose in testicular function remains largely unknown. While GLUT5 is recognized as the exclusive fructose transporter for dietary uptake, it is known to be expressed in germ and Leydig cells. Our recent study has shown that disrupting the GLUT5 gene in male mice impairs both spermatogenesis and steroidogenesis. However, its specific role within Leydig cells remained unexplored. We investigated this by inhibiting a murine Leydig cell line (mLTC-1) with MSNBA, a specific inhibitor of GLUT5. To comprehensively understand the molecular mechanisms of GLUT5 in these cells, we employed a multi-omics approach. Exposition to MNSBA reduced intracellular fructose content, limited cell proliferation and enhanced androgen production. The increased production of testosterone was linked to the upregulation of steroidogenesis-related genes such as Hsd3b, and steroidogenic acute regulatory protein (StAR). In Leydig cell, inhibition of GLUT5 modify lipid and carbohydrate metabolism. Lipidomic analysis showed a decrease in cholesterol esters and a shift in the ratio of polyunsaturated fatty acids (PUFAs) to monounsaturated fatty acids (MUFAs). These lipid changes were correlated with alterations in the expression of mRNA encoding enzymes involved in lipogenesis, such as Elovl6. Metabolomics analysis showed a reduction of different metabolite of the glycolysis except for pyruvate and lactate. However, pyruvate molecules could be produced from the higher branched-chain amino acids content. BRB-seq approach has revealed an upregulation of several androgen-sensitive genes such as lactate dehydrogenase A (Ldha) producing lactate and a down regulation of several genes associated to insulin pathway. In summary, Glut5 is involved in Leydig cells for the fructose intake. Perturbation of the Glut5 activity led to an alteration of the energy metabolism but stimulated the androgen production. Because androgens are known to influence spermatogenesis, these findings raise the importance of the fructose intake in the function of male fertility.

ORGANISM(S): Mus musculus

PROVIDER: GSE294305 | GEO | 2026/04/10

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Steroid hormones regulate essential physiological processes and inadequate levels are associated with various pathological conditions. In testosterone-producing Leydig cells, steroidogenesis is strongly stimulated by LH via its receptor leading to increased cAMP production and expression of the steroidogenic acute regulatory (STAR) protein, which is essential for the initiation of steroidogenesis. Leydig cell steroidogenesis then passively decreases following the rapid degradation of cAMP into AMP by phosphodiesterases. In this study, we show that AMP-activated protein kinase (AMPK) is activated following cAMP breakdown in MA-10 and MLTC-1 Leydig cells. Activated AMPK then actively inhibits cAMP-induced steroidogenesis by repressing the expression of key regulators of steroidogenesis including Star and Nr4a1. Similar results were obtained in Y-1 adrenal cells and in the constitutive steroidogenic cell line R2C. Our data identify AMPK as an active repressor of steroid hormone biosynthesis in steroidogenic cells that is essential to preserve cellular energy and prevent excess steroid production. Steroid hormones regulate essential physiological processes and inadequate levels are associated with various pathological conditions. In testosterone-producing Leydig cells, steroidogenesis is strongly stimulated by LH via its receptor leading to increased cAMP production and expression of the steroidogenic acute regulatory (STAR) protein, which is essential for the initiation of steroidogenesis. Leydig cell steroidogenesis then passively decreases following the rapid degradation of cAMP into AMP by phosphodiesterases. In this study, we show that AMP-activated protein kinase (AMPK) is activated following cAMP breakdown in MA-10 and MLTC-1 Leydig cells. Activated AMPK then actively inhibits cAMP-induced steroidogenesis by repressing the expression of key regulators of steroidogenesis including Star and Nr4a1. Similar results were obtained in Y-1 adrenal cells and in the constitutive steroidogenic cell line R2C. Our data identify AMPK as an active repressor of steroid hormone biosynthesis in steroidogenic cells that is essential to preserve cellular energy and prevent excess steroid production. MA-10 Leydig cells were treated with either DMSO (control), 10 uM forskolin or forskolin+Aicar (1 mM) for 1.5 h before total RNA extraction

Project description:Evidence that persistent environmental pollutants may target the male reproductive system is increasing. The male reproductive system is regulated by secretion of testosterone by testicular Leydig cells, and perturbation of Leydig cells function may have ultimate consequences. 3-methylsulfonyl-DDE (3-MeSO2-DDE) is a potent adrenal toxicants formed from the persistent insecticide DDT. Although studies have revealed endocrine disruptive effect of 3-MeSO2-DDE, the underlying mechanisms at cellular level in steroidogenic Leydig cells remains to be established. The current study addresses the effect of 3-MeSO2-DDE viability, hormone production and proteome response of primary neonatal porcine Leydig cells. The AlamarBlue™ assay was used to evaluate cell viability. Solid phase radioimmunoassay was used to measure concentration of hormones produced by both unstimulated and luteinizing hormone (LH)-stimulated Leydig cells following 48 h exposure. Protein samples from Leydig cells exposed to a non-cytotoxic concentration of 3-MeSO2-DDE (10µM) were subjected to nano-LC-MS/MS and analyzed on a Q Exactive mass spectrometer and quantified using label-free quantitative algorithm. Gene Ontology (GO) and Ingenuity Pathway Analysis (IPA) were carried out for functional annotation and identification of protein interaction networks. 3-MeSO2-DDE regulated Leydig cell steroidogenesis differentially depending on cell culture condition. Whereas its effect on testosterone secretion at basal condition was stimulatory, the effect on LH-stimulated cells was inhibitory. From triplicate experiments, a total of 7540 proteins were identified in which the abundance of 87 proteins in unstimulated Leydig cells and 146 proteins in LH-stimulated Leydig cells were found to be significantly regulated in response to 3-MeSO2-DDE exposure. These proteins not only are the first reported in relation to 3-MeSO2-DDE exposure, but also display small number of proteins shared between culture conditions, suggesting the action of 3-MeSO2-DDE on several targeted pathways, including mitochondrial dysfunction, oxidative phosphorylation, EIF2-signaling, and glutathion-mediated detoxification. Further identification and characterization of these proteins and pathways may build our understanding to the molecular basis of 3-MeSO2-DDE induced endocrine disruption in Leydig cells.

Dataset Information

Inhibition of the fructose transporter Glut5 boosts androgen production by dysregulating the metabolism of murine Leydig cells

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets