Abstract
Porcine growth hormone (pGH) has many important biological functions and roles, and the biological activity of pGH is closely related with its cell behavior and characteristics. However, so far, the behavior of pGH in swine testicular cell remains unclear. For this, in the current work, the swine testicular cell line (ST) was used as an in vitro model, and CLSM (Confocal laser scanning microscope), IFA (Indirect immunofluorescence assay), FCM (Flow cytometry) and WB (Western-blotting) were used to explore the pGH’s cell behivior and function, and the results showed that pGH and GHR could internalize into ST cell and transported to the nucleus. Furthermore, we studied the internalization kinetics of pGH and GHR on ST cell, and found that pGH and GHR internalizes into ST cell in a time-dependent manner. More importantly, we also investigated the potential molecular functions of pGH-GHR after it entered into the cell nuclei. The results indicated that nuclear-localized GHR could participate in cell proliferation by regulating the signal intensity of STAT5. In summary, our current research shows that the nuclear-localized pGH-GHR participates in the cell proliferation of ST cell, which lays a solid foundation for further research on the regulatory effect of pGH on testicular tissue.
Similar content being viewed by others
Data availability
All data and materials are available for publication. Data related to the paper can be obtained from the corresponding author, based on reasonable requirements.
References
Brooks, A. J., & Waters, M. J. (2010). The growth hormone receptor: Mechanism of activation and clinical implications. Nature Reviews Endocrinology, 6(9), 515–525
Li, W., Lan, H., Liu, H., Fu, Z., Yang, Y., Han, W., Guo, F., Liu, Y., Zhang, H., Liu, J., & Zheng, X. (2013). The activation and differential signalling of the growth hormone receptor induced by pGH or anti-idiotypic monoclonal antibodies in primary rat hepatocytes. Molecular and Cellular Endocrinology, 376(1–2), 51–59
Waxman, D. J., & O’Connor, C. (2006). Growth hormone regulation of sex-dependent liver gene expression. Molecular Endocrinology, 20(11), 2613–2629
Waters, M. J., Hoang, H. N., Fairlie, D. P., Pelekanos, R. A., & Brown, R. J. (2006). New insights into growth hormone action. Journal of Molecular Endocrinology, 36(1), 1–7
Lan, H., Zheng, X., Khan, M. A., & Li, S. (2015). Anti-idiotypic antibody: A new strategy for the development of a growth hormone receptor antagonist. The International Journal of Biochemistry and Cell Biology, 68, 101–108
Zhu, T., Goh, E. L., Graichen, R., Ling, L., & Lobie, P. E. (2001). Signal transduction via the growth hormone receptor. Cell Signaling, 13(9), 599–616
Lan, H.-N., Jiang, H.-L., Li, W., Wu, T.-C., Hong, P., Li, Y. M., Zhang, H., Cui, H.-Z., & Zheng, X. (2015). Development and characterization of a novel anti-idiotypic monoclonal antibody to growth hormone, which can mimic physiological functions of growth hormone in primary porcine hepatocytes. Asian-Australasian Journal of Animal Sciences, 28(4), 573–583
Figueiredo, M. A., Boyle, R. T., Sandrini, J. Z., Junior, A., & Marins, L. F. (2016). High level of GHR nuclear translocation in skeletal muscle of a hyperplasic transgenic zebrafish. Journal of Molecular Endocrinology, 56(1), 47
Mertani, H. C., Raccurt, M., Abbate, A., Kindblom, J., Törnell, J., Billestrup, N., Usson, Y., Morel, G., & Lobie, P. E. (2003). Nuclear translocation and retention of growth hormone. Endocrinology, 144(7), 3182–3195
Harvey, S., Baudet, M. L., Murphy, A., Luna, M., Hull, K. L., & Aramburo, C. (2004). Testicular growth hormone (GH): GH expression in spermatogonia and primary spermatocytes. General and Comparative Endocrinology, 139(2), 158–167
Martínez-Moreno, C., López-Marín, L., Carranza, M., Giterman, D., Harvey, S., Arámburo, C., & Luna, M. (2014). Growth hormone (GH) and GH-releasing hormone (GHRH): Co-localization and action in the chicken testis. General and Comparative Endocrinology, 199, 38–45
Bingol-Kologlu, M., Bahadir, G. B., Vargun, R., Ilkay, H., Bagriacik, E. U., Yolbakan, S., Guven, C., Endogan, T., Hasirci, N., & Dindar, H. (2010). Effects of local and sustained release of FGF, IGF, and GH on germ cells in unilateral undescended testis in rats. Urology, 75(1), 223–228
Swanlund, D. J., N’Diaye, M. R., Loseth, K. J., Pryor, J. L., & Crabo, B. G. (1995). Diverse testicular responses to exogenous growth hormone and follicle-stimulating hormone in prepubertal boars. Biology of Reproduction, 53(4), 749–757
Ihle, J. N., Nosaka, T., Thierfelder, W., Quelle, F. W., & Shimoda, K. (1997). Jaks and Stats in cytokine signaling. Stem Cells, 15(Suppl), 1
Hainan, L., Huilin, L., Khan, M. A., Xin, Z., YuJiang, Y., Hui, Z., & Naiquan, Y. (2018). The basic route of the nuclear translocation porcine growth hormone (GH)-growth hormone receptor (GHR) complex (pGH/GHR) in porcine hepatocytes. General and Comparative Endocrinology, 266, 101–109
Chung, C. S., Etherton, T. D., & Wiggins, J. P. (1985). Stimulation of swine growth by porcine growth hormone. Journal of Animal Science, 60(1), 118–130
Vize, P. D., & Wells, J. R. (1987). Isolation and characterization of the porcine growth hormone gene. Gene, 55(2–3), 339–344
Lobie, P. E., Breipohl, W., Aragón, J. G., & Waters, M. J. (1990). Cellular localization of the growth hormone receptor/binding protein in the male and female reproductive systems. Endocrinology, 126(4), 2214–2221
van Kerkhof, P., Govers, R., Alves dos Santos, C. M., & Strous, G. J. (2000). Endocytosis and degradation of the growth hormone receptor are proteasome-dependent. Journal of Biological Chemistry, 275(3), 1575–1580
Conway-Campbell, B. L., Wooh, J. W., Brooks, A. J., Gordon, D., Brown, R. J., Lichanska, A. M., Chin, H. S., Barton, C. L., Boyle, G. M., Parsons, P. G., Jans, D. A., & Waters, M. J. (2007). Nuclear targeting of the growth hormone receptor results in dysregulation of cell proliferation and tumorigenesis. Proceedings of the National Academy Sciences of the United States of America, 104(33), 13331–13336
Conway-Campbell, B. L., Brooks, A. J., Robinson, P. J., Perani, M., & Waters, M. J. (2008). The extracellular domain of the growth hormone receptor interacts with coactivator activator to promote cell proliferation. Molecular Endocrinology, 22(9), 2190–2202
Lobie, P. E., Mertani, H., Morel, G., Morales-Bustos, O., Norstedt, G., & Waters, M. J. (1994). Receptor-mediated nuclear translocation of growth hormone. Journal of Biological Chemistry, 269(33), 21330–21339
Lan, H., Liu, H., Hong, P., Li, R., & Zheng, X. (2018). Porcine growth hormone induces the nuclear localization of porcine growth hormone receptor. Asian-Australasian Journal of Animal Sciences, 31(4), 499–504
Lobie, P. E., Sadir, R., Graichen, R., Mertani, H. C., & Morel, G. (1999). Caveolar internalization of growth hormone. Experimental Cell Research, 246(1), 47–55
van Kerkhof, P., Sachse, M., Klumperman, J., & Strous, G. J. (2001). Growth hormone receptor ubiquitination coincides with recruitment to clathrin-coated membrane domains. Journal of Biological Chemistry, 276(6), 3778–3784
Yang, N., Huang, Y., Jiang, J., & Frank, S. J. (2004). Caveolar and lipid raft localization of the growth hormone receptor and its signaling elements: impact on growth hormone signaling. Journal of Biological Chemistry, 279(20), 20898–20905
Strous, G. J., van Kerkhof, P., Govers, R., Ciechanover, A., & Schwartz, A. L. (1996). The ubiquitin conjugation system is required for ligand-induced endocytosis and degradation of the growth hormone receptor. The EMBO Journal, 15(15), 3806–3812
Strous, G. J., & Govers, R. (1999). The ubiquitin-proteasome system and endocytosis. Journal of Cell Science, 112(Pt 10), 1417–1423
Lobie, P. E., Wood, T. J., Chen, C. M., Waters, M. J., & Norstedt, G. (1994). Nuclear translocation and anchorage of the growth hormone receptor. Journal of Biological Chemistry, 269(50), 31735–31746
Acknowledgements
Y.Z. thanks S.L. (The Third Operating Room, Jilin University First Hospital, Changchun, China, 130118) and all colleagues for assistance with this study.
Funding
This work was supported by the National Natural Science Foundation of China (Grant No. 30162022).
Author information
Authors and Affiliations
Contributions
Y.Z. and H.L. conceived and designed the experiments. Y.Z., Q.Z., and D.W. performed the experiments. Y.Z., M.W., S.L., and X.Z. analyzed the data. H.L. and S.L. (The Third Operating Room, Jilin University First Hospital, Changchun, China, 130118) contributed reagents/materials/analysis tools. Y.Z. and H.L. contributed to the writing of the manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no competing interests.
Ethical approval
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
Consent for publication
All authors read and approved the final manuscript.
Additional information
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Zhang, Y., Zhao, Q., Wu, D. et al. The Cellular Behavior, Intracellular Signaling Profile and Nuclear-Targeted Potential Functions of Porcine Growth Hormone (pGH) in Swine Testicular Cells. Cell Biochem Biophys 80, 403–414 (2022). https://doi.org/10.1007/s12013-022-01068-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12013-022-01068-2