Home LiteratureArticle Details
PMID: 36715674 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Proper phosphorylation of septin 12 regulates septin 4 and soluble adenylyl cyclase expression to induce sperm capacitation.

Journal of cellular physiology ·Vol. 238 ·No. 3 ·2023-00-00 ·Pages 597-609

Wang HY, Shen YR, Tsai YC, Wu SR, Wang CY, Kuo PL

Abstract

Septin-based ring complexes maintain the sperm annulus. Defective annular structures are observed in the sperm of Sept12- and Sept4-null mice. In addition, sperm capacitation, a process required for proper fertilization, is inhibited in Sept4-null mice, implying that the sperm annulus might play a role in controlling sperm capacitation. Hence, we analyzed sperm capacitation of sperm obtained from SEPT12 Ser196 phosphomimetic (S196E), phosphorylation-deficient (S196A), and SEPT4-depleted mutant mice. Capacitation was reduced in the sperm of both the Sept12 S196E- and Sept12 S196A-knock-in mice. The protein levels of septins, namely, SEPT4 and SEPT12, were upregulated, and these proteins were concentrated in the sperm annulus during capacitation. Importantly, the expression of soluble adenylyl cyclase (sAC), a key enzyme that initiates capacitation, was upregulated, and sAC was recruited to the sperm annulus following capacitation stimulation. We further found that SEPT12, SEPT4, and sAC formed a complex and colocalized to the sperm annulus. Additionally, sAC expression was reduced and disappeared in the annulus of the SEPT12 S196E- and S196A-mutant mouse sperm. In the sperm of the SEPT4-knockout mice, sAC did not localize to the annulus. Thus, our data demonstrate that SEPT12 phosphorylation status and SEPT4 activity jointly regulate sAC protein levels and annular localization to induce sperm capacitation.

Keywords
SEPTIN male fertility sperm capacitation
MeSH Terms
Animals Male Mice Adenylyl Cyclases/metabolism Mice, Knockout Phosphorylation Septins/chemistry,deficiency,genetics,metabolism Sperm Capacitation Spermatozoa/metabolism Gene Knock-In Techniques
Chemicals
Adenylyl Cyclases SEPT12 protein, mouse Septins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Wang Han-Yu
Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan. | Department of Obstetrics and Gynecology, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
Shen Yi-Ru
Department of Obstetrics and Gynecology, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
Tsai Yung-Chieh
Department of Obstetrics and Gynecology, Sport Management, and Biotechnology, Chi-Mei Medical Center, Chia Nan University of Pharmacy and Science, Tainan, Taiwan.
Wu Shang-Rung
Institute of Oral Medicine, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
Wang Chia-Yih ORCID
Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan. | Department of Cell Biology and Anatomy, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
Kuo Pao-Lin ORCID
Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan. | Department of Obstetrics and Gynecology, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
References (33)
33 references, click to expand
  1. Chen, S. R., Batool, A., Wang, Y. Q., Hao, X. X., Chang, C. S., Cheng, C. Y., & Liu, Y. X. (2016). The control of male fertility by spermatid-specific factors: Searching for contraceptive targets from spermatozoon's head to tail. Cell Death & Disease, 7(11), e2472. https://doi.org/10.1038/cddis.2016.344
  2. Ded, L., Hwang, J. Y., Miki, K., Shi, H. F., & Chung, J. J. (2020). 3D in situ imaging of the female reproductive tract reveals molecular signatures of fertilizing spermatozoa in mice. eLife, 9. https://doi.org/10.7554/eLife.62043
  3. Finkelstein, M., Etkovitz, N., & Breitbart, H. (2020). Ca(2+) signaling in mammalian spermatozoa. Molecular and Cellular Endocrinology, 516, 110953. https://doi.org/10.1016/j.mce.2020.110953
  4. Freitas, M. J., Vijayaraghavan, S., & Fardilha, M. (2017). Signaling mechanisms in mammalian sperm motility. Biology of Reproduction, 96(1), 2-12. https://doi.org/10.1095/biolreprod.116.144337
  5. Gur, Y., & Breitbart, H. (2006). Mammalian sperm translate nuclear-encoded proteins by mitochondrial-type ribosomes. Genes & Development, 20(4), 411-416. https://doi.org/10.1101/gad.367606
  6. Gur, Y., & Breitbart, H. (2008). Protein synthesis in sperm: Dialog between mitochondria and cytoplasm. Molecular and Cellular Endocrinology, 282(1-2), 45-55. https://doi.org/10.1016/j.mce.2007.11.015
  7. Hereng, T. H., Elgstøen, K. B. P., Cederkvist, F. H., Eide, L., Jahnsen, T., Skålhegg, B. S., & Rosendal, K. R. (2011). Exogenous pyruvate accelerates glycolysis and promotes capacitation in human spermatozoa. Human Reproduction, 26(12), 3249-3263. https://doi.org/10.1093/humrep/der317
  8. Hess, K. C., Jones, B. H., Marquez, B., Chen, Y., Ord, T. S., Kamenetsky, M., Miyamoto, C., Zippin, J. H., Kopf, G. S., Suarez, S. S., Levin, L. R., Williams, C. J., Buck, J., & Moss, S. B. (2005). The “soluble” adenylyl cyclase in sperm mediates multiple signaling events required for fertilization. Developmental Cell, 9(2), 249-259. https://doi.org/10.1016/j.devcel.2005.06.007
  9. Ickowicz, D., Finkelstein, M., & Breitbart, H. (2012). Mechanism of sperm capacitation and the acrosome reaction: Role of protein kinases. Asian Journal of Andrology, 14(6), 816-821. https://doi.org/10.1038/aja.2012.81
  10. Ihara, M., Kinoshita, A., Yamada, S., Tanaka, H., Tanigaki, A., Kitano, A., Goto, M., Okubo, K., Nishiyama, H., Ogawa, O., Takahashi, C., Itohara, S., Nishimune, Y., Noda, M., & Kinoshita, M. (2005). Cortical organization by the septin cytoskeleton is essential for structural and mechanical integrity of mammalian spermatozoa. Developmental Cell, 8(3), 343-352. https://doi.org/10.1016/j.devcel.2004.12.005
  11. Kissel, H., Georgescu, M. M., Larisch, S., Manova, K., Hunnicutt, G. R., & Steller, H. (2005). The Sept4 septin locus is required for sperm terminal differentiation in mice. Developmental Cell, 8(3), 353-364. https://doi.org/10.1016/j.devcel.2005.01.021
  12. Komondor, K. M., & Carlson, A. E. (2020). The secrets of success. eLife, 9, e64379. https://doi.org/10.7554/eLife.64379
  13. Kuo, Y. C., Shen, Y. R., Chen, H. I., Lin, Y. H., Wang, Y. Y., Chen, Y. R., Wang, C. Y., & Kuo, P. L. (2015). SEPT12 orchestrates the formation of mammalian sperm annulus by organizing SEPT12-7-6-2/-4 core complexes. Journal of Cell Science, 128(5), 923-934. https://doi.org/10.1242/jcs.158998
  14. Lhuillier, P., Rode, B., Escalier, D., Lorès, P., Dirami, T., Bienvenu, T., Gacon, G., Dulioust, E., & Touré, A. (2009). Absence of annulus in human asthenozoospermia: Case report. Human Reproduction, 24(6), 1296-1303. https://doi.org/10.1093/humrep/dep020
  15. Luño, V., López-Úbeda, R., García-Vázquez, F. A., Gil, L., & Matás, C. (2013). Boar sperm tyrosine phosphorylation patterns in the presence of oviductal epithelial cells: In vitro, ex vivo, and in vivo models. Reproduction, 146(4), 315-324. https://doi.org/10.1530/REP-13-0159
  16. Miller, D., & Ostermeier, G. C. (2006). Spermatozoal RNA: Why is it there and what does it do? Gynécologie, Obstétrique & Fertilité, 34(9), 840-846. https://doi.org/10.1016/j.gyobfe.2006.07.013
  17. Miller, M. R., Mansell, S. A., Meyers, S. A., & Lishko, P. V. (2015). Flagellar ion channels of sperm: Similarities and differences between species. Cell Calcium, 58(1), 105-113. https://doi.org/10.1016/j.ceca.2014.10.009
  18. Naz, R. K., & Rajesh, P. B. (2004). Role of tyrosine phosphorylation in sperm capacitation/acrosome reaction. Reproductive Biology and Endocrinology, 2(1), 75. https://doi.org/10.1186/1477-7827-2-75
  19. Nolan, M. A., Babcock, D. F., Wennemuth, G., Brown, W., Burton, K. A., & McKnight, G. S. (2004). Sperm-specific protein kinase A catalytic subunit Cα2orchestrates cAMP signaling for male fertility. Proceedings of the National Academy of Sciences, 101(37), 13483-13488. https://doi.org/10.1073/pnas.0405580101
  20. Pozdniakova, S., & Ladilov, Y. (2018). Functional significance of the Adcy10-dependent intracellular cAMP compartments. Journal of Cardiovascular Development and Disease, 5(2), 29. https://doi.org/10.3390/jcdd5020029
  21. Publicover, S. J. (2008). Ca2+ signalling in the control of motility and guidance in mammalian sperm. Frontiers in Bioscience, 13, 5623-5637. https://doi.org/10.2741/3105
  22. Puga Molina, L. C., Luque, G. M., Balestrini, P. A., Marín-Briggiler, C. I., Romarowski, A., & Buffone, M. G. (2018). Molecular basis of human sperm capacitation. Frontiers in Cell and Developmental Biology, 6, 72. https://doi.org/10.3389/fcell.2018.00072
  23. Shen, Y. R., Wang, H. Y., Kuo, Y. C., Shih, S. C., Hsu, C. H., Chen, Y. R., Wu, S. R., Wang, C. Y., & Kuo, P. L. (2017). SEPT12 phosphorylation results in loss of the septin ring/sperm annulus, defective sperm motility and poor male fertility. PLoS Genetics, 13(3), e1006631. https://doi.org/10.1371/journal.pgen.1006631
  24. Shen, Y. R., Wang, H. Y., Tsai, Y. C., Kuo, Y. C., Wu, S. R., Wang, C. Y., & Kuo, P. L. (2020). The SEPT12 complex is required for the establishment of a functional sperm head-tail junction. Molecular Human Reproduction, 26(6), 402-412. https://doi.org/10.1093/molehr/gaaa031
  25. Sugino, Y., Ichioka, K., Soda, T., Ihara, M., Kinoshita, M., Ogawa, O., & Nishiyama, H. (2008). Septins as diagnostic markers for a subset of human asthenozoospermia. Journal of Urology, 180(6), 2706-2709. https://doi.org/10.1016/j.juro.2008.08.005
  26. Sun, X.-H., Zhu, Y.-Y., Wang, L., Liu, H.-L., Ling, Y., Li, Z.-L., & Sun, L.-B. (2017). The Catsper channel and its roles in male fertility: A systematic review. Reproductive Biology and Endocrinology: RB&E, 15(1), 65. https://doi.org/10.1186/s12958-017-0281-2
  27. Touré, A., Lhuillier, P., Gossen, J. A., Kuil, C. W., Lhôte, D., Jégou, B., Escalier, D., & Gacon, G. (2007). The testis anion transporter 1 (Slc26a8) is required for sperm terminal differentiation and male fertility in the mouse. Human Molecular Genetics, 16(15), 1783-1793. https://doi.org/10.1093/hmg/ddm117
  28. Toure, A., Rode, B., Hunnicutt, G. R., Escalier, D., & Gacon, G. (2011). Septins at the annulus of mammalian sperm. Biological Chemistry, 392(8-9), 799-803. https://doi.org/10.1515/bc.2011.074
  29. Toyoda, Y., & Yokoyama, M. (2016). The early history of the TYH medium for in vitro fertilization of mouse ova. Journal of Mammalian Ova Research, 33(1), 3-10.
  30. Tresguerres, M., Levin, L. R., & Buck, J. (2011). Intracellular cAMP signaling by soluble adenylyl cyclase. Kidney International, 79(12), 1277-1288. https://doi.org/10.1038/ki.2011.95
  31. Visconti, P. E., Moore, G. D., Bailey, J. L., Leclerc, P., Connors, S. A., Pan, D., Olds-Clarke, P., & Kopf, G. S. (1995). Capacitation of mouse spermatozoa. II. Protein tyrosine phosphorylation and capacitation are regulated by a cAMP-dependent pathway. Development, 121(4), 1139-1150. https://doi.org/10.1242/dev.121.4.1139
  32. Wang, D., Hu, J., Bobulescu, I. A., Quill, T. A., McLeroy, P., Moe, O. W., & Garbers, D. L. (2007). A sperm-specific Na+/H+ exchanger (sNHE) is critical for expression and in vivo bicarbonate regulation of the soluble adenylyl cyclase (sAC). Proceedings of the National Academy of Sciences, 104(22), 9325-9330. https://doi.org/10.1073/pnas.0611296104
  33. Widmaier, E. P., Raff, H., Strang, K. T., & Vander, A. J. (2016). Vander's human physiology: the mechanisms of body function (Fourteenth edition.ed.). McGraw-Hill.
Article Info
Journal
Journal of cellular physiology
Abbr.
J Cell Physiol
ISSN
1097-4652
DOI
10.1002/jcp.30951
Published
2023-00-00
Epub
2023-00-30
Pages
597-609
Language
English
Region
United States
NLM ID
0050222
Subset
IM
External Links
PubMed source DOI: 10.1002/jcp.30951 View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2