miR-122-5p regulates the tight junction of the blood-testis barrier of mice via occludin
Open Access
- 8 April 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Basic and Clinical Andrology
- Vol. 31 (1), 1-10
- https://doi.org/10.1186/s12610-021-00126-8
Abstract
Occludin protein is the primary assembling protein of TJs and the structural basis for tight junction formation between Sertoli cells in the spermatogenic epithelium. The expression of miR-122-5p and occludin are negatively correlated. In order to investigate the regulation mechanism of miR-122-5p on occludin and TJ, the present study isolated primary Sertoli cells from C57BL/6 mice, identified a transcription factor of miR-122-5p in testicle, studied the modulating loci of miR-122-5p on occludin using a dual-luciferase reporter assay, analyzed the regulate of miR-122-5p on the expression of occludin with real-time RT-PCR and Western blot, and studied the effect of miR-122-5p on the tight junction using a Millicell Electrical Resistance System. The relative luciferase activity in the pcDNA-Sp1 + pGL3-miR-122-5p promoter group was significantly higher than that in the pcDNA-Sp1 + pGL3-basic group, which suggests that transcript factor Sp1 promotes the transcription of miR-122-5p. The relative luciferase activity in the occludin 3′-UTR (wt) + miR-122-5p mimic group was significantly lower than that in the other groups (p < 0.01), which indicates that miR-122-5p modulates the expression of occludin via the ACACTCCA sequence of the occludin-3’UTR. The levels of occludin mRNA and protein in the miR-122-5p mimic group were significantly lower than that in the other groups (p < 0.05), which indicates that miR-122-5p reduces the expression of occludin. The trans-epithelial resistance of the miR-122-5p mimic group was significantly lower than that of the blank control group after day 4 (p < 0.05), which indicates that miR-122-5p inhibited the assembly of the inter-Sertoli TJ permeability barrier in vitro. These results displayed that miR-122-5p could regulate tight junctions via the Sp1-miR-122-5p-occludin-TJ axis.Keywords
Funding Information
- National Natural Science Fund (81771567)
This publication has 42 references indexed in Scilit:
- Metastasis to Bone in Human Cancer Is Associated with Loss of Occludin Expression.2016
- Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitroAsian Journal of Andrology, 2016
- MicroRNA-encoded behavior in DrosophilaScience, 2015
- Auto-Regulation of the Sohlh1 Gene by the SOHLH2/SOHLH1/SP1 Complex: Implications for Early Spermatogenesis and OogenesisPLOS ONE, 2014
- Regulation of spermatogenesis by small non-coding RNAs: Role of the germ granuleSeminars in Cell & Developmental Biology, 2014
- An In Vitro System to Study Sertoli Cell Blood-Testis Barrier DynamicsMethods in molecular biology (Clifton, N.J.), 2011
- Occludin regulates macromolecule flux across the intestinal epithelial tight junction barrierAmerican Journal of Physiology-Gastrointestinal and Liver Physiology, 2011
- Expression of CLMP, a novel tight junction protein, is mediated via the interaction of GATA with the kruppel family proteins, KLF4 and Sp1, in mouse TM4 sertoli cellsJournal of Cellular Physiology, 2007
- SP1 transcription factors in male germ cell development and differentiationMolecular and Cellular Endocrinology, 2007
- Transcription factor Sp1 is expressed by three different developmentally regulated messenger ribonucleic acids in mouse spermatogenic cells.Endocrinology, 1996