Skip to main content

Advertisement

SpringerLink
Log in

Liprin α3: a putative estrogen regulated acrosomal protein

  • Original Paper
  • Published:
Histochemistry and Cell Biology Aims and scope Submit manuscript

Abstract

Liprin α3 was reported for the first time using sperm proteomics. Present study reports its localization on sperm and immunochemical characterization. Liprin α3 is identified as a 133 kDa protein in testis and epididymal protein extracts. In testis, immunohistochemical localization was seen in pachytenes, diplotenes, round spermatids whereas it was localized in the epithelial cells and luminal sperm in all the three regions of epididymis. Protein was localized in acrosome of rat sperm, which was further confirmed by sequential treatment of sperm with hypertonic solution. In the spermatogenic cells the protein was found to be located in developing acrosome as evident by its co-localization with Golgi marker. Protein was found to be developmentally regulated. In silico analysis of Liprin α3 revealed presence of the estrogen responsive elements upstream to initiation site and its regulation by estrogen was experimentally validated using a tamoxifen treated rat model. Western blot analysis of epididymosomes showed the presence of Liprin α3, indicating its involvement in trafficking of vesicle. The protein expression was seen in both mouse and human sperm indicating conserved nature and a probable role in acrosome reaction.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10

Similar content being viewed by others

References

  • Andrews-Zwilling YS, Kawabe H, Reim K, Varoqueaux F, Brose N (2006) Binding to Rab3A-interacting molecule RIM regulates the presynaptic recruitment of Munc13-1 and ubMunc13-2. J biol chem 281:19720–19731. doi:10.1074/jbc.M601421200

    Article  PubMed  CAS  Google Scholar 

  • Bello OD, Zanetti MN, Mayorga LS, Michaut Ma (2012) RIM, Munc13, and Rab3A interplay in acrosomal exocytosis. Exp Cell Res 318:478–488. doi:org/10.1016/j.yexcr.2012.01.002

    Article  PubMed  CAS  Google Scholar 

  • Bérubé B, Lefièvre L, Coutu L, Sullivan R (1996) Regulation of the epididymal synthesis of P26h, a hamster sperm protein. J Androl 17:104–110

    PubMed  Google Scholar 

  • D’Souza R, Pathak S, Upadhyay R, Gaonkar R, D’Souza S, Sonawane S, Gill-Sharma M, Balasinor NH (2009) Disruption of tubulobulbar complex by high intratesticular estrogens leading to failed spermiation. Endocrinology 150:1861–1869. doi:10.1210/en.2008-1232

    Article  PubMed  Google Scholar 

  • Escalier D, Gallo JM, Schrével J (1997) Immunochemical characterization of a human sperm fibrous sheath protein, its developmental expression pattern, and morphogenetic relationships with actin. J histochem cytochem 45(7):909–922. doi:10.1177/002215549704500701

    Article  PubMed  CAS  Google Scholar 

  • Girouard J, Frenette G, Sullivan R (2011) Comparative proteome and lipid profiles of bovine epididymosomes collected in the intraluminal compartment of the caput and cauda epididymidis. Int J Androl 34:e475–e486. doi:10.1111/j.1365-2605.2011.01203.x

    Article  PubMed  CAS  Google Scholar 

  • Hecht NB (1995) The making of a spermatozoon: a molecular perspective. Dev Genet 16:95–103

    Article  PubMed  CAS  Google Scholar 

  • Iida H, Yoshinaga Y, Tanaka S, Toshimori K, Mori T (1999) Identification of Rab3A GTPase as an acrosome-associated small GTP-binding protein in rat sperm. Dev biol 211:144–155

    Article  PubMed  CAS  Google Scholar 

  • Katafuchi K, Mori T, Toshimori K, Iida H (2000) Localization of a syntaxin isoform, syntaxin 2, to the acrosomal region of rodent spermatozoa. Mol Reprod Dev 57:375–383. doi:10.1002/1098-2795(200012)57:4<375:AID-MRD10>3.0.CO;2-Z

    Article  PubMed  CAS  Google Scholar 

  • Khan SA, Jadhav SV, Suryawanshi AR, Bhonde GS, Gajbhiye RK, Khole VV (2011) Evaluation of contraceptive potential of a novel epididymal sperm protein SFP2 in a mouse model. Am J Reprod Immunol 66:185–198. doi:10.1111/j.1600-0897.2011.01030.x

    Article  PubMed  CAS  Google Scholar 

  • Ko J, Na M, Kim S, Lee J-R, Kim E (2003) Interaction of the ERC family of RIM-binding proteins with the Liprin-α family of multidomain proteins. J biol chem 278:42377–42385. doi:10.1074/jbc.M307561200

    Article  PubMed  CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  PubMed  CAS  Google Scholar 

  • Légaré C, Bérubé B, Boué F, Lefièvre L, Morales CR, El-Alfy M, Sullivan R (1999) Hamster sperm antigen P26h is a phosphatidylinositol-anchored protein. Mol Reprod Dev 52:225–233

    Article  PubMed  Google Scholar 

  • Lowry Oh, Nj Rosebrough, Al Farr, Rj Randall (1951) Protein measurement with the Folin phenol reagent. J biol chem 193:265–275

    PubMed  CAS  Google Scholar 

  • MacGregor JI, Jordan VC (1998) Basic guide to the mechanisms of antiestrogen action. Pharmacol Rev 50:151–196

    PubMed  CAS  Google Scholar 

  • Maier B (2002) Developmental association of the synaptic activity-regulated protein arc with the mouse acrosomal organelle and the sperm tail. Biol Reprod 68:67–76. doi:10.1095/biolreprod.102.004143

    Article  Google Scholar 

  • Majó G, Lorenzo MJ, Blasi J, Aguado F (1999) Exocytotic protein components in rat pituitary gland after long-term estrogen administration. J Endocrinol 161:323–331. doi:10.1677/joe.0.1610323

    Article  PubMed  Google Scholar 

  • Moreno RD, Palomino J, Schatten G (2006) Assembly of spermatid acrosome depends on microtubule organization during mammalian spermiogenesis. Dev Biol 293:218–227. doi:10.1016/j.ydbio.2006.02.001

    Article  PubMed  CAS  Google Scholar 

  • Pathak S, D’Souza R, Ankolkar M, Gaonkar R, Balasinor NH (2010) Potential role of estrogen in regulation of the insulin-like growth factor2-H19 locus in the rat testis. Mol Cell Endocrinol 314:110–117. doi:10.1016/j.mce.2009.08.005

    Article  PubMed  CAS  Google Scholar 

  • Ramalho-Santos J, Moreno RD, Sutovsky P, Chana W, Hewitson L, Wessel GM, Simerly CR, Schatten G (2000) SNAREs in mammalian sperm: possible implications for fertilization. Dev biol 223:54–69. doi:org/10.1006/dbio.2000.9745

    Article  PubMed  CAS  Google Scholar 

  • Rezaul K, Thumar JK, Lundgren DH, Eng JK, Claffey KP, Wilson L, Han DK (2010) Differential protein expression profiles in estrogen receptor-positive and -negative breast cancer tissues using label-free quantitative proteomics. Genes cancer 1:251–271. doi:10.1177/1947601910365896

    Article  PubMed  CAS  Google Scholar 

  • Schoch S, Castillo PE, Jo T, Mukherjee K, Geppert M, Wang Y, Schmitz F, Malenka RC, Südhof TC (2002) RIM1α forms a protein scaffold for regulating neurotransmitter release at the active zone. Nature 415:321–326. doi:10.1038/415321a

    Article  PubMed  CAS  Google Scholar 

  • Schuel H, Burkman LJ (2005) A tale of two cells: endocannabinoid-signaling regulates functions of neurons and sperm. Biol Reprod 73:1078–1086. doi:org/10.1095/biolreprod.105.049106

    PubMed  CAS  Google Scholar 

  • Spangler SA, Hoogenraad CC (2007) Liprin-α proteins: scaffold molecules for synapse maturation. Biochem Soc Trans 35:1278–1282. doi:10.1042/BST0351278

    Article  PubMed  CAS  Google Scholar 

  • Suryawanshi AR, Khan SA, Gajbhiye RK, Gurav MY, Khole VV (2011) Differential proteomics leads to identification of domain-specific epididymal sperm proteins. J Androl 32:240–259. doi:10.2164/jandrol.110.010967

    Article  PubMed  CAS  Google Scholar 

  • Syntin P, Cornwall Ga (1999) Immunolocalization of CRES (cystatin-related epididymal spermatogenic) protein in the acrosomes of mouse spermatozoa. Biol Reprod 60:1542–1552. doi:10.1095/biolreprod60.6.1542

    Article  PubMed  CAS  Google Scholar 

  • Tomes CN, Michaut M, De Blas G, Visconti P, Matti U, Mayorga LS (2002) SNARE complex assembly is required for human sperm acrosome reaction. Dev biol 243:326–338. doi:org/10.1006/dbio.2002.0567

    Article  PubMed  CAS  Google Scholar 

  • Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354

    Article  PubMed  CAS  Google Scholar 

  • Wakle MS, Joshi SA, Khole VV (2005) Monoclonal antibody from vasectomized mouse identifies a conserved testis-specific antigen TSA70. J androl 26(6):761–771. doi:10.2164/jandrol.05045

    Article  PubMed  CAS  Google Scholar 

  • Wang Y, Okamoto M, Schmitz F, Hofmann K, Südhof TC (1997) Rim is a putative Rab3 effector in regulating synaptic-vesicle fusion. Nature 388:593–598

    Article  PubMed  CAS  Google Scholar 

  • WHO laboratory manual for the examination and processing of human semen (2010), 5th edn. ISBN 978 92 4 154778 9

  • Wright PK, May FE, Darby S, Saif R, Lennard TW, Westley BR (2009) Estrogen regulates vesicle trafficking gene expression in EFF-3, EFM-19 and MCF-7 breast cancer cells. Int J Clin Exp Pathol 2:463–475

    PubMed  CAS  Google Scholar 

  • Zhen M, Jin Y (1999) The Liprin protein SYD-2 regulates the differentiation of presynaptic termini in C. elegans. Nature 401:371–375

    PubMed  CAS  Google Scholar 

  • Zhou Y, Zheng M, Shi Q, Zhang L, Zhen W, Chen W, Zhang Y (2008) An epididymis-specific secretory protein hongres1 critically regulates sperm capacitation and male fertility. PLoS ONE 3:12. doi:10.1371/journal.pone.0004106

    Google Scholar 

  • Zitranski N, Borth H, Ackermann F, Meyer D, Viewig L, Breit A, Gudermann T, Boekhoff I (2010) The “acrosomal synapse”: subcellular organization by lipid rafts and scaffolding proteins exhibits high similarities in neurons and mammalian spermatozoa. Comm Integr biol 3:513–521. doi:10.4161/cib.3.6.13137

    Article  Google Scholar 

Download references

Acknowledgments

The authors thank Ms. Shobha and Ms. Reshma at the confocal department of National Institute for Research in Reproductive Health for their assistance in image acquisition and processing. The authors also wish to thank Dr. Rahul Gajabhiye, clinician for providing semen samples from normal individuals. Financial support for implementation of this project and award of junior research fellowship to one of us (CJ) from the Department of Science and Technology, New Delhi is gratefully acknowledged.

Author information

Author notes

  1. Amol R. Suryawanshi

    Present address: Institute of Life Sciences, Bhubaneswar, 751023, India

Authors and Affiliations

  1. Department of Gamete Immunobiology, National Institute for Research in Reproductive Health (ICMR), J. M. Street, Parel, Mumbai, 400012, India

    Chetanchandra S. Joshi, Amol R. Suryawanshi, Shagufta A. Khan & Vrinda V. Khole

  2. Department of Neuroendocrinology, National Institute for Research in Reproductive Health (ICMR), J. M. Street, Parel, Mumbai, 400012, India

    Nafisa H. Balasinor

Authors
  1. Chetanchandra S. Joshi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Amol R. Suryawanshi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Shagufta A. Khan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nafisa H. Balasinor
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Vrinda V. Khole
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Vrinda V. Khole.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Joshi, C.S., Suryawanshi, A.R., Khan, S.A. et al. Liprin α3: a putative estrogen regulated acrosomal protein. Histochem Cell Biol 139, 535–548 (2013). https://doi.org/10.1007/s00418-012-1044-y

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00418-012-1044-y

Keywords

Search

Navigation