From 9fb4e460968cd216aec3aaf1c969732e30457f40 Mon Sep 17 00:00:00 2001 From: latoshadyson10 Date: Thu, 2 Apr 2026 17:10:03 +0800 Subject: [PATCH] Add m6A mRNA methylation regulates testosterone synthesis through modulating autophagy in Leydig cells --- ...sis-through-modulating-autophagy-in-Leydig-cells.md | 10 ++++++++++ 1 file changed, 10 insertions(+) create mode 100644 m6A-mRNA-methylation-regulates-testosterone-synthesis-through-modulating-autophagy-in-Leydig-cells.md diff --git a/m6A-mRNA-methylation-regulates-testosterone-synthesis-through-modulating-autophagy-in-Leydig-cells.md b/m6A-mRNA-methylation-regulates-testosterone-synthesis-through-modulating-autophagy-in-Leydig-cells.md new file mode 100644 index 0000000..be9e877 --- /dev/null +++ b/m6A-mRNA-methylation-regulates-testosterone-synthesis-through-modulating-autophagy-in-Leydig-cells.md @@ -0,0 +1,10 @@ +
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CHX chase assay of NHERF2 in Atg7Flox/Flox and SF1-Atg7−/− (E) as well as Atg5Flox/Flox and SF1-Atg5−/− (G) Leydig cells. CHX chase assay of NHERF2 in HEK293T cells in the absence or presence of 3-MA. (C) NHERF2 was stabilized in autophagic flux–disrupted cells. +The firefly luciferase and Renilla luciferase activity in each cell were detected by dual-luciferase reporter assay system (Promega, E1910). Magna MeRIPTM m6A kit was chosen to assess m6A modification levels in target mRNA according to the manufacturer’s instructions (Millipore Sigma, 17–10,499). Camkk2 and Ppm1a mRNA levels were detected by qRT-PCR, and primer sequences were listed in Table S1. +Serum levels of mouse [testosterone store](https://prpack.ru/user/beggarknot4/), LH, FSH, and corticosterone were specifically measured using a RIA kit. These purified Leydig cells were cultured in DMEM/F12 supplemented with 15% FBS, 100 U/ml penicillin, and 100 µg/ml streptomycin at 34°C with 5% CO2. Subsequently, the Leydig cells in the 50% to 60% fraction were carefully collected, washed with PBS, and centrifuged at 500 g for 5 min. +(C and E) The cholesterol uptake defects could be partially rescued by Nherf2 knockdown in autophagy-deficient Leydig cells. The in vitro cholesterol absorption experiment was subsequently performed in the Leydig cells using the fluorescence-labeled cholesterol-rich lipoprotein DiI-HDL (red) and examined using live-cell imaging. (B and D) The cholesterol uptake defects could be partially rescued by Nherf2 knockdown in autophagy-deficient and control Leydig cells. Thus, NHERF2 appears to be the linker between autophagy and cholesterol uptake in Leydig cells. In Nherf2-knockdown autophagy-deficient or WT Leydig cells, cholesterol uptake, both rates and amounts, was significantly increased compared with Nherf2-expressing cells, as indicated by DiI-HDL absorption (Fig. 7, A–E). If yes, then knockdown of Nherf2 in autophagy-deficient Leydig cells might at least partially rescue the impaired cholesterol uptake phenotype of the cells. +[buy testosterone steroids](https://tvoyaskala.com/user/startevent6/) regulates the autophagic clearance of androgen binding protein in rat Sertoli cells. Forty-eight hours after plating, cells were treated with 20 mM Tris (pH 7.4) for 2.5 min to remove residual germ cells. Of note, autophagy degrades ABP by engulfing the protein and the process seems to be selectively regulated by testosterone, which can prolong ABP biological half-life by inhibiting autophagy. In summary, we determined that in rat Sertoli cells, ABP can be cleared by autophagy. +Importantly, within the last few years, compelling evidence has begun to emerge and more detailed classifications of autophagy have been reported. In microautophagy, the lysosome directly engulfs the cytoplasm via the inward invagination of the lysosomal membrane. Every protein has a distinct in vivo half-life time that ranges broadly from a few minutes to more than 100 days 14,15. The essential difference between humans and machines is that our cellular homeostasis requires a dynamic balance between biosynthetic and catabolic processes . In addition, we also determine the occurrence and influence of autophagy on a series of events that take place during testicular development, with the aim of providing a more systematic reference for the study of testicular autophagy. +S3 shows that the steroidogenic cell–specific disruption of autophagy did not affect the concentrations of TGs and TC in testes and serum. S2 shows that the lipids in the adrenal cortex and corticosterone in the serum decreased in the steroidogenic cell–specific autophagy disruption mice. S1 shows that the disruption of autophagy did not affect the fetal–adult Leydig cell transition, proliferation, and [https://lovebookmark.win/](https://lovebookmark.win/story.php?title=best-online-trt-clinics-7-most-trusted-options-2025) Gn secretion. 24 h after transfection, the expression of these proteins was confirmed by immunoblotting. HEK293T cells were maintained at 37°C and 5% CO2 in DMEM supplemented with penicillin, streptomycin, and 10% FBS. The protein lysates (25 µg) were separated by SDS-PAGE and electrotransferred onto a nitrocellulose membrane. +Recent research also suggests that FSH, regardless of the traditional steroidogenic pathway, enhances autophagy by upregulating Beclin1 through the PI3K/JNK/c-Jun pathway, promoting LDs breakdown in pig GCs (Liu et al., 2021). This mouse model showed a nearly 75% reduction in Becn1 levels, with p62 accumulation observed in GCs. These observations imply that autophagy is a vital player in the proper sexual development. Mutually, BFF HD-sEVs enhance macroautophagy and mitophagy in bGCs, inhibit apoptosis in bGCs, and elevate 17β-estradiol release via the PI3K/Akt/mTOR signaling pathways as shown in Figure 4D (Wang et al., 2023). +
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