Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Jan 5;14(2):183.
doi: 10.3390/ani14020183.

Caveolin 1 Regulates the Tight Junctions between Sertoli Cells and Promotes the Integrity of Blood-Testis Barrier in Yak via the FAK/ERK Signaling Pathway

Qiu Yan  1   2 , Tianan Li  1   2 , Yong Zhang  1   2   3 , Xingxu Zhao  1   2   3 , Qi Wang  1   2 , Ligang Yuan  1   2   3
Affiliations

Caveolin 1 Regulates the Tight Junctions between Sertoli Cells and Promotes the Integrity of Blood-Testis Barrier in Yak via the FAK/ERK Signaling Pathway

Qiu Yan et al. Animals (Basel). .

Abstract

Yaks, a valuable livestock species endemic to China's Tibetan plateau, have a low reproductive rate. Cryptorchidism is believed to be one of the leading causes of infertility in male yaks. In this study, we compared the morphology of the normal testis of the yak with that of the cryptorchidism, and found dysplasia of the seminiferous tubules, impaired tightness of the Sertoli cells, and a disruption of the integrity of the blood-testis barrier (BTB) in the cryptorchidism. Previous studies have shown that CAV1 significantly contributes to the regulation of cell tight junctions and spermatogenesis. Therefore, we hypothesize that CAV1 may play a regulatory role in tight junctions and BTB in Yaks Sertoli cells, thereby influencing the development of cryptorchidism. Additional analysis using immunofluorescence, qRT-PCR, and Western blotting confirmed that CAV1 expression is up-regulated in yak cryptorchidism. CAV1 over-expression plasmids and small RNA interference sequences were then transfected in vitro into yak Sertoli cells. It was furthermore found that CAV1 has a positive regulatory effect on tight junctions and BTB integrity, and that this regulatory effect is achieved through the FAK/ERK signaling pathway. Taken together, our findings, the first application of CAV1 to yak cryptorchidism, provide new insights into the molecular mechanisms of cell tight junctions and BTB. This paper suggests that CAV1 could be used as a potential therapeutic target for yak cryptorchidism and may provide insight for future investigations into the occurrence of cryptorchidism, the maintenance of a normal physiological environment for spermatogenesis and male reproductive physiology in the yak.

Keywords: RAN-seq; blood–testis barrier; cryptorchidism; tight junctions; yak.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
TJs and BTB was detected in yak testis and cryptorchidism. (A) Histomorphology was analyzed by H&E staining in yak testis and cryptorchidism. (B) The mRNA expression level of ZO-1, occludin and claudin-11 were detected by qRT-PCR; values represent mean ± SD, n = 3. * p < 0.05, ** p < 0.01. (C) The protein expression of ZO-1, occludin and claudin-11 were analyzed using Western blotting. (D–F) Localization of ZO-1, occludin and claudin-11 protein in testis and cryptorchidism were analyzed using immunohistochemistry. PMC: peridubular myoid cells; S: spermatogonium; SZ: spermatozoa; ST: Seminiferous tubule; PS: primary spermatocyte; SC: Sertoli cell; LC: Leydig cell; CV: capillary vessel.
Figure 2
Figure 2
GO and KEGG Analysis of Differently Expressed Target Genes. (A) The histogram shows number of mRNA. Green and yellow represent total mRNAs and novel mRNAs; red and blue represent increased and decreased mRNA abundance. (B) GO annotation functional classification of differentially expressed genes. (C) KEGG pathways with significant enrichment of differentially expressed genes. (D–F) GO and KEGG analysis difference expression genes for immunity response to stimulus, biological adhesion, development process and cell junction.
Figure 3
Figure 3
Identifying differentially expressed microRNA of small RNA sequencing. (A) The fragment length distribution of microRNAs in yak testis and cryptorchidism. (B) The number statistics of miRNA in per samples. (C) The total of miRNA in all samples. (D) The heatmap of all microRNAs in the two groups. (E) The statistical graph of the differentially expressed miRNAs (DERs) between groups. (F) TGFB1 and CAV1 were potentially regulated by miRNAs.
Figure 4
Figure 4
Verification of DE miRNAs and DE mRNAs. (A) qPCR analysis for 8 randomly selected miRNAs. Data represent the mean ± SD, n = 3, * p < 0.05, ** p < 0.01. (B) qPCR analysis for 8 randomly selected mRNAs. Data represent the mean ± SD, n = 3, * p < 0.05, ** p < 0.01. (C) Expression patterns of TGFB1 and CAV1 proteins by Western blotting analysis, n = 3. (D) Immunohistochemical stain assay for expression and location of CAV1 in testis and cryptorchidism. (E) Intracellular localization analysis of CAV1 protein in Sertoli cells. PMC: peritubular myoid cells; S: spermatogonium; SZ: spermatozoa; ST: seminiferous tubule; PS: primary spermatocyte; SC: Sertoli cell; LC: Leydig cell.
Figure 5
Figure 5
Expression analysis of CAV1 over-expression efficiency in yak Sertoli cells. (A) Immunofluorescence staining identified the isolated yak SCs using antibodies against WT1. Original magnification: ×200. (B) Fluorescence microscopy of Sertoli cells transfected with an p-EGFP-CAV1 overexpression plasmid or control empty vector (p-EGFP). Cells were examined at 72 h after transfection. Original magnification: ×400. (C) qRT-PCR analysis of CAV1 mRNA in Sertoli cells transfected to pIRES2-EGFP-CAV1; values represent mean ± SD, n = 3, ** p < 0.01. (D) The protein expression of CAV1 was detected using Western blotting, n = 3.
Figure 6
Figure 6
Silencing of CAV1 expression in Sertoli cells. (A) qRT-PCR analysis of CAV1 mRNA levels in Sertoli cells transfected for 72 h with a control siRNA or two CAV1-targeting siRNAs. Values represent mean ± SD, n = 3, * p < 0.05, ** p < 0.01. (B) Western blotting analysis protein level of CAV1 in Sertoli cells after transfection for siRNA-NC, siRNA-CAV1-1 and siRNA-CAV1-2; GAPDH was probed as an internal control, n = 3.
Figure 7
Figure 7
CAV1 promoted TJs and BTB of Sertoli cells. (A–G) The mRNA expression of FAK, SRC, JNK, ERK, ZO-1, occludin and claudin-11 was detected by qRT-PCR; values represent mean ± SD, n = 3. * p < 0.05, ** p < 0.01. (H) The protein expression of FAK, SRC, p-SRC, JNK, ERK1, p-ERK1, ZO-1, occludin and claudin-11 was detected using Western blotting, n = 3.
Figure 8
Figure 8
CAV1 suppression of TJs and BTBs of Sertoli cells. (A–G) The mRNA expression of FAK, SRC, JNK, ERK, ZO-1, occludin and claudin-11 was detected by qRT-PCR after being transfected for siRNA-CAV1; values represent mean ± SD, n = 3. * p < 0.05, ** p < 0.01. (H) The protein expression of FAK, SRC, p-SRC, JNK1, ERK1, p-ERK1, ZO-1, occludin and claudin-11 was detected using Western blotting, n = 3.
Figure 9
Figure 9
CAV1 regulates TJs and the integrity of BTBs of SCs via FAK/ERK signaling pathway. Abbreviations: CAV1: caveolin-1; SRC: SRC proto-oncogene; ERK: mitogen-activated protein kinase 1; JNK: c-Jun NH2-terminal kinase; ZO-1: TJP1 (ZO1) tight junction protein 1.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Goda Y., Mizutani S., Mizutani Y., Kitahara G., Siswandi R., Wakabayashi K., Torisu S., Kaneko Y., Hidaka Y., Osawa T., et al. Usefulness of computed tomography for cryptorchidism in bulls. J. Vet. Med. Sci. 2022;84:1430–1436. doi: 10.1292/jvms.22-0124. - DOI - PMC - PubMed
    1. St Jean G., Gaughan E.M., Constable P.D. Cryptorchidism in North American cattle: Breed predisposition and clinical findings. Theriogenology. 1992;38:951–958. doi: 10.1016/0093-691X(92)90169-R. - DOI - PubMed
    1. Mieusset R., Bujan L.E., Massat G., Mansat A., Pontonnier F. Inconstant ascending testis as a potential risk factor for spermatogenesis in infertile men with no history of cryptorchism. Hum. Reprod. 1997;12:974–979. doi: 10.1093/humrep/12.5.974. - DOI - PubMed
    1. Kellaway R.C., Seamark R.F., Farrant R.K. Sterilisation of cattle by induced cryptorchidism. Aust. Vet. J. 1971;47:547–550. doi: 10.1111/j.1751-0813.1971.tb02052.x. - DOI - PubMed
    1. Goto A., Yoshida N., Nakada K., Inoue Y., Hisaeda K., Inaba T., Domoto N., Ishiguro Y., Itoh M., Takahashi E., et al. Efficiency of immunocastration with an anti-gonadotropin-releasing hormone vaccine on cryptorchid bulls. J. Vet. Med. Sci. 2023;85:551–556. doi: 10.1292/jvms.22-0571. - DOI - PMC - PubMed

LinkOut - more resources