Related Experiment Video
Updated: May 23, 2026
A Seminiferous Tubule Squash Technique for the Cytological Analysis of Spermatogenesis Using the Mouse Model
Published on: February 6, 2018
MicroRNA-34c enhances murine male germ cell apoptosis through targeting ATF1
Xiaoxuan Liang1, Doudou Zhou, Chao Wei
1State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing, People's Republic of China.
Background:
MicroRNAs (miRNAs) play vital regulatory roles in many cellular processes. The expression of miRNA (miR)-34c is highly enriched in adult mouse testis, but its roles and underlying mechanisms of action are not well understood.
Methodology/Principal Findings:
In the present study, we show that miR-34c is detected in mouse pachytene spermatocytes and continues to be highly expressed in spermatids. To explore the specific functions of miR-34c, we have established an in vivo model by transfecting miR-34c inhibitors into primary spermatocytes to study the loss-of-function of miR-34c. The results show that silencing of miR-34c significantly increases the Bcl-2/Bax ratio and prevents germ cell from apoptosis induced by deprivation of testosterone. Moreover, ectopic expression of the miR-34c in GC-2 cell trigger the cell apoptosis with a decreased Bcl-2/Bax ratio and miR-34c inhibition lead to a low spontaneous apoptotic ratio and an increased Bcl-2/Bax ratio. Furthermore, ectopic expression of miR-34c reduces ATF1 protein expression without affecting ATF1 mRNA level via directly binding to ATF1's 3'UTR, indicating that ATF1 is one of miR-34c's target genes. Meanwhile, the knockdown of ATF1 significantly decreases the Bcl-2/Bax ratio and triggers GC-2 cell apoptosis. Inhibition of miR-34c does not decrease the GC-2 cell apoptosis ratio in ATF1 knockdown cells.
Conclusions/Significance:
Our study shows for the first time that miR-34c functions, at least partially, by targeting the ATF1 gene in germ cell apoptosis, providing a novel mechanism with involvement of miRNA in the regulation of germ cell apoptosis.
Insights
MicroRNA-34c regulates germ cell apoptosis by targeting ATF1. Inhibiting miR-34c prevents testosterone deprivation-induced cell death, revealing a novel mechanism in male reproduction.
Area of Science:
- Molecular Biology
- Reproductive Biology
- Cell Biology
Background:
- MicroRNAs (miRNAs) are crucial regulators of cellular processes.
- MicroRNA (miR)-34c is abundant in adult mouse testes, but its function is unclear.
Purpose of the Study:
- To investigate the role and mechanism of miR-34c in male germ cell apoptosis.
- To identify target genes of miR-34c involved in apoptosis regulation.
Main Methods:
- Established an in vivo loss-of-function model using miR-34c inhibitors in primary spermatocytes.
- Utilized GC-2 cells for ectopic miR-34c expression and ATF1 knockdown studies.
- Analyzed apoptosis via Bcl-2/Bax ratio and direct ATF1 targeting using 3'UTR assays.
Main Results:
- Silencing miR-34c protected germ cells from testosterone deprivation-induced apoptosis.
- Ectopic miR-34c induced apoptosis by decreasing the Bcl-2/Bax ratio.
- miR-34c directly targets ATF1, reducing its protein levels and promoting apoptosis.
- ATF1 knockdown mimicked miR-34c's pro-apoptotic effects, and miR-34c inhibition was ineffective in ATF1-depleted cells.
Conclusions:
- miR-34c plays a significant role in regulating germ cell apoptosis.
- The ATF1 gene is a direct target of miR-34c, mediating its pro-apoptotic function.
- This study uncovers a novel miRNA-dependent mechanism regulating male germ cell apoptosis.
Related Concept Videos
01:22MicroRNAs
01:22MicroRNAs
01:16In-vitro Mutagenesis