Sperm storage in the oviduct of the Chinese pond turtle Mauremys reevesii depends on oestrogen-based suppression of the TLR2/4 immune pathway
Wenlong Cai A , Wei Chen A , Yajie Wang A , Xingjiang Bu A B , Xingquan Xia A and Liuwang Nie AA Anhui Provincial Key Laboratory of the Conservation and Exploitation of Biological Resources, College of Life Sciences, Anhui Normal University, Wuhu 241000, Anhui, China.
B Corresponding author. Email: buxingjiang@163.com
Reproduction, Fertility and Development 33(12) 736-745 https://doi.org/10.1071/RD20341
Submitted: 28 December 2020 Accepted: 8 July 2021 Published: 4 October 2021
Abstract
The long-term storage of spermatozoa in the female reproductive tract is limited by the innate immune system. Oestrogen plays a role in regulating the innate immune system. Thus, exploring the expression of genes in the Toll-like receptor (TLR) 2/4 pathway and oestrogen receptors in the oviduct of Mauremys reevesii could contribute to our understanding of the mechanism of sperm storage. In this study, three parts of the oviduct (isthmus, uterus and vagina) in three mated and unmated female turtles were used to perform immunohistochemistry and real-time quantitative polymerase chain reaction (qPCR). Immunohistochemistry revealed that the TLR2/4 protein was mainly distributed in epithelial tissues and glandular cell membranes, and that TLR2/4 levels in the oviduct were significantly decreased in mated compared with unmated turtles. Real-time qPCR indicated that TLR2/4, myeloid differentiation factor 88 (MyD88), interleukin 1 receptor associated kinase 4 (IRAK4), TNF receptor associated factor 6 (TRAF6), interferon regulatory factor 3 (IRF3) and interleukin 6 (IL6) mRNA expression was significantly higher in the oviduct of unmated than mated turtles, whereas the opposite was true for the expression of oestrogen receptor 1 (ESR1) and progesterone receptor (PGR). These results indicate that when spermatozoa are stored in the oviduct, an increase in oestrogen suppresses the immune response induced by the TLR2/4 pathway so that spermatozoa are not removed as a foreign substance, but stored until fertilisation. The findings of this study are relevant to our understanding of the relationship between sperm storage and the innate immune system in the oviduct of reptiles.
Keywords: innate immune system, Mauremys reevesii, oestrogen receptor, oviduct, reproductive tract, reptiles, sperm storage, TLR2/4.
References
Ananjeva, N. B., Uteshev, V., Orlov, N., Ryabov, S., Gakhova, E., Kaurova, S., and Browne, R. (2017). Comparison of the modern reproductive technologies for amphibians and reptiles. Russ. J. Herpetol. 24, 275–290.| Comparison of the modern reproductive technologies for amphibians and reptiles.Crossref | GoogleScholarGoogle Scholar |
Barratt, C. L., and Pockley, A. G. (1998). Sperm survival in the female reproductive tract: presence of immunosuppression or absence of recognition? Mol. Hum. Reprod. 4, 309–313.
| Sperm survival in the female reproductive tract: presence of immunosuppression or absence of recognition?Crossref | GoogleScholarGoogle Scholar | 9620828PubMed |
Birkhead, T., and Møller, A. (1993). Sexual selection and the temporal separation of reproductive events: sperm storage data from reptiles, birds and mammals. Biol. J. Linn. Soc. Lond. 50, 295–311.
| Sexual selection and the temporal separation of reproductive events: sperm storage data from reptiles, birds and mammals.Crossref | GoogleScholarGoogle Scholar |
Chen, S., Zhang, L., Le, Y., Waqas, Y., Chen, W., Zhang, Q., and Li, Q. (2015). Sperm storage and spermatozoa interaction with epithelial cells in oviduct of Chinese soft-shelled turtle, Pelodiscus sinensis. Ecol. Evol. 5, 3023–3030.
| Sperm storage and spermatozoa interaction with epithelial cells in oviduct of Chinese soft-shelled turtle, Pelodiscus sinensis.Crossref | GoogleScholarGoogle Scholar | 26357535PubMed |
Chen, H., Liu, T., Holt, W. V., Yang, P., Zhang, L., Han, X., and Chen, Q. (2020). Advances in understanding mechanisms of long-term sperm storage-the soft-shelled turtle model. Histol. Histopathol. 35, 1–23.
| 31290136PubMed |
De Lorenzo, G., Ferrari, S., Cervone, F., and Okun, E. (2018). Extracellular DAMPs in plants and mammals: immunity, tissue damage and repair. Trends Immunol. 39, 937–950.
| Extracellular DAMPs in plants and mammals: immunity, tissue damage and repair.Crossref | GoogleScholarGoogle Scholar | 30293747PubMed |
Ellington, J. E., Jones, A. E., Davitt, C. M., Schneider, C. S., Brisbois, R. S., Hiss, G. A., and Wright, R. W. (1998). Human sperm function in co-culture with human, macaque or bovine oviduct epithelial cell monolayers. Hum. Reprod. 13, 2797–2804.
| Human sperm function in co-culture with human, macaque or bovine oviduct epithelial cell monolayers.Crossref | GoogleScholarGoogle Scholar | 9804233PubMed |
Ezz, M. A., Marey, M. A., Elweza, A. E., Kawai, T., Heppelmann, M., Pfarrer, C., and Zaabel, S. M. (2019). TLR2/4 signaling pathway mediates sperm-induced inflammation in bovine endometrial epithelial cells in vitro. PLoS One 14, e0214516.
| TLR2/4 signaling pathway mediates sperm-induced inflammation in bovine endometrial epithelial cells in vitro.Crossref | GoogleScholarGoogle Scholar | 30995239PubMed |
Fischer, M., and Ehlers, M. (2008). Toll-like receptors in autoimmunity. Ann. N. Y. Acad. Sci. 1143, 21–34.
| Toll-like receptors in autoimmunity.Crossref | GoogleScholarGoogle Scholar | 19076342PubMed |
Gan, F., Liu, Q., Liu, Y., Huang, D., Pan, C., Song, S., and Huang, K. (2018). Lycium barbarum polysaccharides improve CCl4-induced liver fibrosis, inflammatory response and TLRs/NF-kB signaling pathway expression in wistar rats. Life Sci. 192, 205–212.
| Lycium barbarum polysaccharides improve CCl4-induced liver fibrosis, inflammatory response and TLRs/NF-kB signaling pathway expression in wistar rats.Crossref | GoogleScholarGoogle Scholar | 29196051PubMed |
Gist, D. H., and Jones, J. M. (1989). Sperm storage within the oviduct of turtles. J. Morphol. 199, 379–384.
| Sperm storage within the oviduct of turtles.Crossref | GoogleScholarGoogle Scholar | 29865618PubMed |
Hasan, M. M., Viil, J., Lättekivi, F., Ord, J., Reshi, Q. U. A., Jääger, K., and Salumets, A. (2020). Bovine follicular fluid and extracellular vesicles derived from follicular fluid alter the bovine oviductal epithelial cells transcriptome. Int. J. Mol. Sci. 21, 5365.
| Bovine follicular fluid and extracellular vesicles derived from follicular fluid alter the bovine oviductal epithelial cells transcriptome.Crossref | GoogleScholarGoogle Scholar |
Holt, W. V., and Fazeli, A. (2016). Sperm storage in the female reproductive tract. Annu. Rev. Anim. Biosci. 4, 291–310.
| Sperm storage in the female reproductive tract.Crossref | GoogleScholarGoogle Scholar | 26526545PubMed |
Holt, W. V., and Lloyd, R. E. (2010). Sperm storage in the vertebrate female reproductive tract: how does it work so well? Theriogenology 73, 713–722.
| Sperm storage in the vertebrate female reproductive tract: how does it work so well?Crossref | GoogleScholarGoogle Scholar | 19632711PubMed |
Huck, U. W., Tonias, B. A., and Lisk, R. D. (1989). The effectiveness of competitive male inseminations in golden hamsters, Mesocricetus auratus, depends on an interaction of mating order, time delay between males, and the time of mating relative to ovulation. Anim. Behav. 37, 674–680.
| The effectiveness of competitive male inseminations in golden hamsters, Mesocricetus auratus, depends on an interaction of mating order, time delay between males, and the time of mating relative to ovulation.Crossref | GoogleScholarGoogle Scholar |
Iwasaki, A., and Medzhitov, R. (2015). Control of adaptive immunity by the innate immune system. Nat. Immunol. 16, 343–353.
| Control of adaptive immunity by the innate immune system.Crossref | GoogleScholarGoogle Scholar | 25789684PubMed |
Jochimsen, D. M., Peterson, C. R., Andrews, K. M., and Whitfield Gibbons, J. (2004). ‘A literature review of the effects of roads on amphibians and reptiles and the measures used to minimize those effects.’ (Idaho Fish and Game Department, USDA Forest Service.)
Klinge, C. M. (2001). Estrogen receptor interaction with estrogen response elements. Nucleic Acids Res. 29, 2905–2919.
| Estrogen receptor interaction with estrogen response elements.Crossref | GoogleScholarGoogle Scholar | 11452016PubMed |
Kumar, H., Kawai, T., and Akira, S. (2011). Pathogen recognition by the innate immune system. Int. Rev. Immunol. 30, 16–34.
| Pathogen recognition by the innate immune system.Crossref | GoogleScholarGoogle Scholar | 21235323PubMed |
Liu, T., Yang, P., Chen, H., Huang, Y., Liu, Y., Waqas, Y., and Chen, Q. (2016). Global analysis of differential gene expression related to long-term sperm storage in oviduct of Chinese Soft-Shelled Turtle Pelodiscus sinensis. Sci. Rep. 6, 33296.
| Global analysis of differential gene expression related to long-term sperm storage in oviduct of Chinese Soft-Shelled Turtle Pelodiscus sinensis.Crossref | GoogleScholarGoogle Scholar | 27628424PubMed |
Marey, M. A., Yousef, M. S., Kowsar, R., Hambruch, N., Shimizu, T., Pfarrer, C., and Miyamoto, A. (2016). Local immune system in oviduct physiology and pathophysiology: attack or tolerance? Domest. Anim. Endocrinol. 56, S204–S211.
| Local immune system in oviduct physiology and pathophysiology: attack or tolerance?Crossref | GoogleScholarGoogle Scholar | 27345318PubMed |
Nagai, Y., Garrett, K. P., Ohta, S., Bahrun, U., Kouro, T., Akira, S., and Kincade, P. W. (2006). Toll-like receptors on hematopoietic progenitor cells stimulate innate immune system replenishment. Immunity 24, 801–812.
| Toll-like receptors on hematopoietic progenitor cells stimulate innate immune system replenishment.Crossref | GoogleScholarGoogle Scholar | 16782035PubMed |
Nechaeva, M. V. (2011). Physiological responses to acute changes in temperature and oxygenation in bird and reptile embryos. Respir. Physiol. Neurobiol. 178, 108–117.
| Physiological responses to acute changes in temperature and oxygenation in bird and reptile embryos.Crossref | GoogleScholarGoogle Scholar | 21513821PubMed |
Orr, T. J., and Zuk, M. (2012). Sperm storage. Curr. Biol. 22, R8–R10.
| Sperm storage.Crossref | GoogleScholarGoogle Scholar | 22240479PubMed |
Rafferty, A. R., and Reina, R. D. (2012). Arrested embryonic development: a review of strategies to delay hatching in egg-laying reptiles. Proc. R. Soc. B. 279, 2299–2308.
| Arrested embryonic development: a review of strategies to delay hatching in egg-laying reptiles.Crossref | GoogleScholarGoogle Scholar | 22438503PubMed |
Rhodin, A. G., Iverson, J. B., Bour, R., Fritz, U., Georges, A., Shaffer, H., and Van Dijk, P. (2017). ‘Turtles of the world: annotated checklist and atlas of taxonomy, synonymy, distrubution, and conservation status.’ 8th edn. Chelonian Research Monographs, no. 7. (Chelonian Research Foundation, Turtle Conservancy.)
Sever, D. M., and Hamlett, W. C. (2002). Female sperm storage in reptiles. J. Exp. Zool. 292, 187–199.
| Female sperm storage in reptiles.Crossref | GoogleScholarGoogle Scholar | 11754034PubMed |
Sever, D. M., and Staub, N. L. (2011). Hormones, sex accessory structures, and secondary sexual characteristics in amphibians. In ‘Hormones and reproduction of vertebrates.’ (Eds D. O. Norris and K. H. Lopez.) pp. 83–98. (Elsevier.)
Smith, T. T., and Nothnick, W. B. (1997). Role of direct contact between spermatozoa and oviductal epithelial cells in maintaining rabbit sperm viability. Biol. Reprod. 56, 83–89.
| Role of direct contact between spermatozoa and oviductal epithelial cells in maintaining rabbit sperm viability.Crossref | GoogleScholarGoogle Scholar | 9002636PubMed |
Stark, G., Tamar, K., Itescu, Y., Feldman, A., and Meiri, S. (2018). Cold and isolated ectotherms: drivers of reptilian longevity. Biol. J. Linn. Soc. Lond. 125, 730–740.
| Cold and isolated ectotherms: drivers of reptilian longevity.Crossref | GoogleScholarGoogle Scholar |
Su, S. B., Silver, P. B., Grajewski, R. S., Agarwal, R. K., Tang, J., Chan, C. C., and Caspi, R. R. (2005). Essential role of the MyD88 pathway, but nonessential roles of TLRs 2, 4, and 9, in the adjuvant effect promoting Th1-mediated autoimmunity. J. Immunol. 175, 6303–6310.
| Essential role of the MyD88 pathway, but nonessential roles of TLRs 2, 4, and 9, in the adjuvant effect promoting Th1-mediated autoimmunity.Crossref | GoogleScholarGoogle Scholar | 16272281PubMed |
Suarez, S. S. (2008). Regulation of sperm storage and movement in the mammalian oviduct. Int. J. Dev. Biol. 52, 455–462.
| 18649258PubMed |
Tu, X.-K., Yang, W.-Z., Shi, S.-S., Chen, Y., Wang, C.-H., Chen, C.-M., and Chen, Z. (2011). Baicalin inhibits TLR2/4 signaling pathway in rat brain following permanent cerebral ischemia. Inflammation 34, 463–470.
| Baicalin inhibits TLR2/4 signaling pathway in rat brain following permanent cerebral ischemia.Crossref | GoogleScholarGoogle Scholar | 20859668PubMed |
Uematsu, S., and Akira, S. (2006). Toll-like receptors and innate immunity. J. Mol. Med. 84, 712–725.
| Toll-like receptors and innate immunity.Crossref | GoogleScholarGoogle Scholar | 16924467PubMed |
Uller, T., and Olsson, M. (2008). Multiple paternity in reptiles: patterns and processes. Mol. Ecol. 17, 2566–2580.
| Multiple paternity in reptiles: patterns and processes.Crossref | GoogleScholarGoogle Scholar | 18452517PubMed |
Verstak, B., Hertzog, P., and Mansell, A. (2007). Toll-like receptor signalling and the clinical benefits that lie within. Inflamm. Res. 56, 1–10.
| Toll-like receptor signalling and the clinical benefits that lie within.Crossref | GoogleScholarGoogle Scholar | 17334664PubMed |
Wira, C. R., Rodriguez-Garcia, M., and Patel, M. V. (2015). The role of sex hormones in immune protection of the female reproductive tract. Nat. Rev. Immunol. 15, 217–230.
| The role of sex hormones in immune protection of the female reproductive tract.Crossref | GoogleScholarGoogle Scholar | 25743222PubMed |
Yousef, M. S., Marey, M. A., Hambruch, N., Hayakawa, H., Shimizu, T., Hussien, H. A., and Miyamoto, A. (2016). Sperm binding to oviduct epithelial cells enhances TGFB1 and IL10 expressions in epithelial cells as well as neutrophils in vitro: prostaglandin E2 as a main regulator of anti-inflammatory response in the bovine oviduct. PLoS One 11, e0162309.
| Sperm binding to oviduct epithelial cells enhances TGFB1 and IL10 expressions in epithelial cells as well as neutrophils in vitro: prostaglandin E2 as a main regulator of anti-inflammatory response in the bovine oviduct.Crossref | GoogleScholarGoogle Scholar | 27662642PubMed |