Mol. Hum. Reprod. Advance Access originally published online on December 10, 2004
Molecular Human Reproduction 2005 11(2):133-140; doi:10.1093/molehr/gah137
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Identification of transcripts by macroarrays, RT–PCR and in situ hybridization in human ejaculate spermatozoa
1Laboratoire de Cytologie et Histologie, Centre Universitaire des Saints-Pères, 75270, Paris, 2EA 1533 Génétique de la Reproduction Humaine, Hôpital Tenon (AP-HP), 75970, Paris and 3Unité INSERM 581, Hôpital Henri-Mondor, 94010 Créteil, Paris, France
4 To whom correspondence should be addressed at: Laboratoire de Cytologie et Histologie, Centre Universitaire des Saints-Pères, 45 rue des Saints-Pères, 75270 Paris Cedex 06, France. Email: jean-pierre.dadoune{at}univ-paris5.fr
Round spermatids contain high levels of extremely varied mRNAs that are synthesized either throughout early spermatogenesis or during spermiogenesis from the haploid genome. Concomitantly, with major changes in the chromatin organization, arrest of transcription occurs at midspermiogenesis. However, previous investigations using RT–PCR have revealed the persistence of numerous and different transcripts in ejaculated spermatozoa. In the present study, a step-by-step analysis by means of macroarray hybridization, RT–PCR and in situ hybridization was performed to identify more accurately the different mRNA species found in the human ejaculated spermatozoa. The data showed an extended pattern of various transcripts encoding a diverse range of proteins involved in signal transduction and cell proliferation. For the first time, they demonstrated that mRNAs coding for the transcription factors NF
B, HOX2A, ICSBP, protein kinase JNK2, growth factor HBEGF and receptors RXRß and ErbB3 accumulate within the sperm nucleus. The origin and fate of the sperm transcripts remain subject to discussion.
Key words: haploid genome/messenger RNAs/spermatozoa
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