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The identification and characterization of a testis-specific cDNA during spermatogenesis

Cellular & Molecular Biology Lettersvolume 11, pages80–89 (2006) | Download Citation

Abstract

Using bioinformatics and experimental validation, we obtained a cDNA (named srsf) which was exclusively expressed in the mouse testes. RT-PCR analysis showed that srsf mRNA was not expressed in the gonad during the sex determination period or during embryogenesis. In developing mouse tests, srsf expression was first detected on post-natal day 10, reached its highest level on day 23, and then reduced to and remained at a moderate level throughout adulthood. In situ hybridization analysis demonstrated that srsf mRNA was expressed in pachytene spermatocytes and round spermatids in the testes. The predicted protein contains one RNA-binding domain (RBD) and a serine-arginine rich domain (RS), which are characterized by some splicing factors of SR family members. These findings indicate that srsf may play a role during spermatogenesis.

Abbreviations

A:

type Aspermatogonia

In:

intermediate spermatogonia

B:

type B spermatogonia

PL:

preleptotene spermatocytes

L:

leptotene spermatocytes

Zs:

zygotene spermatocytes

EPs:

early pachytene spermatocytes

LPs:

late pachytene spermatocytes

Ds:

diplotene spermatocytes

MI:

first meiosis

MII:

second meiosis

Rs:

round spermatids

Es:

elongated spermatids

S16:

step 16 spermatids

E:

early

L:

late

SG:

spermatogonia

SC:

spermatocytes

ST:

round spermatid.

References

  1. 1.

    Kleen, K.C. Patterns of transcriptional regulation in the mammalian testes. Mol. Report Dev. 423 (1996) 268–281.

  2. 2.

    Venables, J.P. Alternative splicing in the tests. Curr. Opin. Genet. Dev. 12 (2002) 615–619.

  3. 3.

    Fu, X.D. Specific commitment of different pre-mRNA to splicing by single SR proteins. Nature 365 (1993) 82–85.

  4. 4.

    Portal, D. and Joaqu’ın, M. An early ancestor in the evolution of splicing: a Trypanosoma cruzi serine-arginin-rich protein (TcSR) is functional in cis-splicing. Mol. Biochem. Parasitol. 127 (2003) 37–46.

  5. 5.

    Roland, T. and James, L. Determinants of SR protein specificity. Curr. Opin. Cell Biol. 11 (1999) 358–362.

  6. 6.

    Krämer, A. The structure and function of proteins involved in mammalian pre-mRNA splicing. Annu. Rev. Biochem. 65 (1996) 367–409.

  7. 7.

    Fu, X.D. The superfamily of arginine/serine-rich splicing factors. RNA 1 (1995) 663–680.

  8. 8.

    Manley, J.L. and Tacke, R. SR proteins and splicing control. Genes Dev. 9 (1996) 284–293.

  9. 9.

    Valcárcel, J. and Green, M.R. The SR protein family: pleiotropic functions in pre-mRNA splicing. Trends Biochem. Sci. 21 (1996) 296–301.

  10. 10.

    Hastings, M.L. and Krainer, A.R. Splicing in the new millennium. Curr. Opin. Cell Biol. 13 (2001) 302–308.

  11. 11.

    Meissner, M., Lopato, S., Gotzmann, J., Sauermann, G. and Barta, A. Proto-oncoprotein TLS/FUS is associated to the nuclear mitrix and complexed with splicing factors PTB, SRm160, and SR proteins. Exp. Cell Res. 283 (2003) 184–195.

  12. 12.

    Zhu, J. and Krainer, A.R. Pre-mRNA splicing in the absence of a SR protein SR domain. Genes 14 (2000) 3166–3178.

  13. 13.

    Wang, J., Takegaki, Y. and Manliy, J.L. Targeted disruption of an essential vertebrate gene ASF/SF2 is required for cell viability. Genes Dev. 10 (1996) 2588–2599.

  14. 14.

    Stefan, W., Simion, C., Ivshina, M. and Nickerson, J.A. In vitro FRAP reveals the ATP-dependent nuclear mobilization of the exon junction complex protein SRm160. J. Cell Biol. 164 (2004) 843–850.

  15. 15.

    Stefan, W., Simion, C. and Nickerson, J.A. The spatial targeting and nuclear matrix binding domains of SR m160. Proc. Natl. Acad. Sci. U.S.A. 100 (2003) 3269–3274.

  16. 16.

    Dijkman, H.B.P.M., Mentzel, S, de Jong, A.S., Assmann, K.J.M. RNA in situ hybridization using digoxigenin-labeled cRNA probes. Biochemica 2(1995) 21–25

  17. 17.

    Thompson, J.D., Higgins, D.G., Gibson, and T.J. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 11 (1994) 4673–4680.

  18. 18.

    Birney, E., Kumar, S. and Krainer, A.R. Analysis of the RNA-recognition motif and RS and RGG domains: conservation in metazoan pre-mRNA splicing factors. Nucleic Acids Res. 21 (1993) 5803–5816.

  19. 19.

    Zhang, D., Penttila, T.L., Morris, P.L. and Roeder, R.G. Cell-and stage-specific high-level expression of TBP-related factor 2 (TRF2) during mouse spermatogenesis. Mech. Dev. 106 (2001) 203–205.

  20. 20.

    Ina, S., Tsunekawa, N., Nakamura, A. and Noce, T. Expression of the mouse Aven gene during spermatogenesis, analyzed by subtraction screening using Mvh-knockout mice. Gene Expr. Patterns 5 (2003) 635–638.

  21. 21.

    Bellve, A.R., Cavicchia, J.C., Millette, C.F., O’Brien, D.A., Bhatnagar, Y.N. and Dym, M. Spermatogenic cells of the prepuberal mouse: isolation and morphological characterization. J. Cell Biol. 74 (1977) 68–85.

  22. 22.

    Taizo, K. and Masaki, F.T. Unique and redundant functions of SR proteins, a conserved family of splicing factors, in Caenorhabditis elegants development. Mech. Dev. 95 (2000) 67–76.

  23. 23.

    Mattox, W., McGuffin, M.E. and Baker, B.S. A Negative feedback mechanism revealed by functional analysis of the alternative isoforms of the Drosophila splicing regulator transformer-2. Genetics 143 (1996) 303–314.

  24. 24.

    Rugh, R. The mouse, its reproduction and development. Burgess Publishing Company, Minneapolis. (1968).

  25. 25.

    Sassone-Corsi, P. Unique chromatin remodeling and transcriptional regulation in spermatogenesis. Science 296 (2002) 2176–2178

  26. 26.

    Kimmins, S., Kotaja, N., Davidson, I. and Sassone-Corsi, P. Testis-specific transcription mechanisms promoting male germ-cell differentiation. Repro. 128 (2004) 5–12

  27. 27.

    Baoan, L., Mahalakshmi, N. and Mackay, D.R. Ovoll regulates meiotic pachytene progression during spermatogenesis by repressing Id2 expression. Development 132 (2005) 1463–1473

  28. 28.

    Tung, K-S. and Bilanchone, V. The pachytene checkpoint prevents accumulation and phosphorylation of the meiosis-specific transcription factor Ndt80. Proc. Natl. Acad. Sci. U.S.A. 97 (2000) 12187–12192.

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Affiliations

  1. Laboratory of Molecular Genetics and Developmental Biology, College of Life Science, Wuhan University, Wuhan, 430072, China

    • Ying Chen
    • , Ping Song
    •  & Wuming Gong

  2. Department of Gynecology and obstetrics, Zhongnan Hospital, Wuhan University, Wuhan, 430071, China

    • Jiarui Hu

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Corresponding author

Correspondence to Ping Song.

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Key Words

  • Expression pattern
  • Spermatogenesis
  • Splicing factor

Cellular & Molecular Biology Letters

ISSN: 1689-1392

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