Molecular Human Reproduction, Vol. 6, No. 3, 219-225,
March 2000
© 2000 European Society of Human Reproduction and Embryology
Testis and spermatozoa |
Expression of protamine-1 and -2 mRNA during human spermiogenesis
1 Institut für Veterinäranatomie, Frankfurter Strasse 98, D-35392 Giessen, 2 Institut für Pathologie, Langhansstrasse 10, D-35392 Giessen, and 3 Institut für Anatomie und Zellbiologie, Grosse Steinstrasse 52, D-06097 Halle, Germany
Abstract
During spermiogenesis, the histone-to-protamine replacement causes the compaction of the spermatid chromatin. The genes for protamines, PRM-1 and PRM-2, are transcribed in round and elongating spermatids. The transcripts are stored in a translationally-repressed state by the binding of protein repressors before being translated in elongating and elongated spermatids. RNA extracts from homogenized whole testis samples supply only average data, and cell-specific and stage-specific expression cannot be addressed. Therefore, we used UV-laser-assisted cell-picking (UV-LACP) to select spermatids of defined differentiation steps. Subsequent reverse transcription–polymerase chain reaction (RT–PCR) with intron-spanning primer pairs allowed the detection of DNA-free and pseudogene-free PRM-1 and PRM-2 cDNA. Additional in-situ hybridization with digoxygenin-labelled cRNA probes exhibited PRM-1 and PRM-2 mRNA from step 1/2 spermatids to step 4 spermatids, but not in elongated spermatids. RT–PCR revealed amplicons for PRM-1 and PRM-2 in all spermatids except step 3 round spermatids. Applying proteinase K digestion, PRM-1 and PRM-2 transcripts were also detected in step 3 spermatids indicating that protein repressors may bind to both PRM-1 and PRM-2 mRNA in step 3 round spermatids. These data demonstrate that the combination of UV-LACP and non-radioactive in-situ hybridization appear to be a suitable approach for the study of cell-specific and stage-specific gene expression during spermiogenesis.
human testis/in-situ hybridization/laser-assisted cell-picking/protamines/spermiogenesis
Notes
4 To whom correspondence should be addressed
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  K. Steger, J. Wilhelm, L. Konrad, T. Stalf, R. Greb, T. Diemer, S. Kliesch, M. Bergmann, and W. Weidner Both protamine-1 to protamine-2 mRNA ratio and Bcl2 mRNA content in testicular spermatids and ejaculated spermatozoa discriminate between fertile and infertile men Hum. Reprod., January 1, 2008; 23(1): 11 - 16. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. T. Carrell, B. R. Emery, and S. Hammoud Altered protamine expression and diminished spermatogenesis: what is the link? Hum. Reprod. Update, May 1, 2007; 13(3): 313 - 327. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Zini and J. Libman Sperm DNA damage: clinical significance in the era of assisted reproduction. Can. Med. Assoc. J., August 29, 2006; 175(5): 495 - 500. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. J. Walton, R. A. L. Bayne, I. Wallace, D. T. Baird, and R. A. Anderson Direct Effect of Progestogen on Gene Expression in the Testis during Gonadotropin Withdrawal and Early Suppression of Spermatogenesis J. Clin. Endocrinol. Metab., July 1, 2006; 91(7): 2526 - 2533. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Oliva Protamines and male infertility Hum. Reprod. Update, July 1, 2006; 12(4): 417 - 435. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V.W. Aoki, L. Liu, and D.T. Carrell A novel mechanism of protamine expression deregulation highlighted by abnormal protamine transcript retention in infertile human males with sperm protamine deficiency Mol. Hum. Reprod., January 1, 2006; 12(1): 41 - 50. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y.-T. Chen, M. J. Scanlan, C. A. Venditti, R. Chua, G. Theiler, B. J. Stevenson, C. Iseli, A. O. Gure, T. Vasicek, R. L. Strausberg, et al. Identification of cancer/testis-antigen genes by massively parallel signature sequencing PNAS, May 31, 2005; 102(22): 7940 - 7945. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Mitchell, K. Steger, C. Marchetti, J.-C. Herbaut, P. Devos, and J.-M. Rigot Cellular expression of protamine 1 and 2 transcripts in testicular spermatids from azoospermic men submitted to TESE-ICSI Mol. Hum. Reprod., May 1, 2005; 11(5): 373 - 379. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Steger, M. Slavov, K. Failing, W. Weidner, and M. Bergmann Effect of Vasectomy on Sperm Nuclear Chromatin Condensation in the Rabbit J Androl, March 1, 2005; 26(2): 289 - 295. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Steger, L. Fink, K. Failing, R. M. Bohle, S. Kliesch, W. Weidner, and M. Bergmann Decreased protamine-1 transcript levels in testes from infertile men Mol. Hum. Reprod., June 1, 2003; 9(6): 331 - 336. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Iuchi, T. Kobayashi, T. Kaneko, M. Takahara, T. Ogino, and J. Fujii Expression of a Y-box protein, YB2/RYB-a, precedes protamine 2 expression during spermatogenesis in rodents Mol. Hum. Reprod., November 1, 2001; 7(11): 1023 - 1031. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. E. Franke, S. Kraus, C. Eiermann, K. Pauls, E.-N. Lalani, and M. Bergmann MUC1 in normal and impaired spermatogenesis Mol. Hum. Reprod., June 1, 2001; 7(6): 505 - 512. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Steger, K. Failing, T. Klonisch, H. M. Behre, M. Manning, W. Weidner, L. Hertle, M. Bergmann, and S. Kliesch Round spermatids from infertile men exhibit decreased protamine-1 and -2 mRNA Hum. Reprod., April 1, 2001; 16(4): 709 - 716. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. P. Siffroi, A. Pawlak, M. F. Alfonsi, F. Troalen, G. Guellaen, and J. P. Dadoune Expression of the TAR RNA binding protein in human testis Mol. Hum. Reprod., March 1, 2001; 7(3): 219 - 225. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Banfi, G. Molnar, A. Maturana, K. Steger, B. Hegedus, N. Demaurex, and K.-H. Krause A Ca2+-activated NADPH Oxidase in Testis, Spleen, and Lymph Nodes J. Biol. Chem., September 28, 2001; 276(40): 37594 - 37601. [Abstract] [Full Text] [PDF]
|
|