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Mol. Hum. Reprod. Advance Access published online on April 22, 2005
Molecular Human Reproduction, doi:10.1093/molehr/gah169
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Molecular Human Reproduction. Published by Oxford University Press [2005] on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oupjournals.org
Received February 1, 2005
Accepted March 14, 2005

Article

Cellular expression of protamine 1 and 2 transcripts in testicular spermatids from azoospermic men submitted to TESE-ICSI

Valérie Mitchell 1*, Klaus Steger 2, Carole Marchetti 3, Jean-Claude Herbaut 4, Patrick Devos 5, and Jean-Marc Rigot 6

1 Spermiologie-Biologie de la Reproduction, et unité 422 INSERM, hôpital A. Calmette, Boulevard du Professeur Jules Leclercq, CHRU-Faculté de Médecine, F-59037,
2 Klinik für Urologie und Kinderurologie, Rudolf-Buchheim-Strasse 7, D-35385 Giessen, Germany,
3 Biologie de la Reproduction, hôpital Jeanne de Flandre, CHRU, F-59037, Lille cedex, France,
4 Biolille, 17 rue de la Digue, BP 117, F-59016,
5 Unité de Biostatistiques, Faculté de Médecine, Pôle Recherche, CHRU, F-59037 Lille cedex, France and
6 Service d'Andrologie, hôpital A. Calmette, Boulevard du Professeur Jules Leclercq, CHRU, F-59037 Lille cedex, France

* To whom correspondence should be addressed.
Valérie Mitchell, E-mail: mitchell{at}lille.inserm.fr


  Abstract

Testicular sperm extraction (TESE) combined with ICSI is used to treat azoospermia. However, the factors that influence the outcome of ICSI in this situation are ill-defined. We sought to investigate the expression of protamine 1 (PRM1) and protamine 2 (PRM2) transcripts in testicular spermatids from obstructive and non-obstructive azoospermic men with impaired spermatogenesis. The relationship between PRM1 and PRM2 transcript levels and the TESE-ICSI outcome was evaluated. The cellular expression of PRM1 and PRM2 mRNAs in single testicular spermatids from 41 azoospermic patients (in whom testicular spermatozoa were subsequently recovered and submitted for TESE-ICSI) was determined by radioactive in situ hybridization. Group I contained seven men with congenital, obstructive azoospermia and whose testicular biopsies indicated quantitatively normal spermatogenesis. Group II consisted of 18 azoospermic men with moderately impaired spermatogenesis. Sixteen men with non-obstructive azoospermia and severely deranged spermatogenesis (i.e. mixed atrophy with small foci of spermatids and spermatozoa) constituted group III. The spermatids of men with severely deranged spermatogenesis exhibited significant lower PRM1 mRNA expression than in the other patient groups. There were no significant inter-group differences in PRM2 mRNA expression. Spermatid PRM1 expression was lower in non-pregnant couples than in pregnant couples. The low number of spermatids in cases of mixed atrophy with small spermatogenic foci is associated with significantly lower PRM1 expression and a lower pregnancy rate. These results emphasize the role of PRM1 as a potentially critical factor in post-ICSI embryonic development.

Keywords: azoospermia; in situ hybridization; protamine; testis.
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