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© The Rockefeller University Press, 0021-9525/2000//1107 $5.00
The Journal of Cell Biology, Volume 148, Number 6, , 2000 1107-1114

Brief Report

Reverse Transcriptase Activity in Mature Spermatozoa of Mouse



Roberto Giordanoa, Anna Rosa Magnanoa,b, Germana Zaccagninic, Carmine Pittoggib,c, Nicola Moscufod, Rodolfo Lorenzinid, and Corrado Spadaforac

a CNR Center for the Study of Germinal Cells, University of Siena, 53100 Italy
b Institute of General Biology, University of Siena, 53100 Italy
c CNR Institute of Biomedical Technology, 00161 Rome, Italy
d Istituto Superiore di Sanità, 00161 Rome, Italy
CNR Institute of Biomedical Technology, Via G.B. Morgagni 30/E, 00161 Rome, Italy.+39 06 49387120+39 06 49903117/2972

cspadaf{at}tin.it

We show here that a reverse transcriptase (RT) activity is present in murine epididymal spermatozoa. Sperm cells incubated with human poliovirus RNA can take up exogenous RNA molecules and internalize them in nuclei. Direct PCR amplification of DNA extracted from RNA-incubated spermatozoa indicate that poliovirus RNA is reverse-transcribed in cDNA fragments. PCR analysis of two-cell embryos shows that poliovirus RNA-challenged spermatozoa transfer retrotranscribed cDNA molecules into eggs during in vitro fertilization. Finally, RT molecules can be visualized on sperm nuclear scaffolds by immunogold electron microscopy. These results, therefore, reveal a novel metabolic function in spermatozoa, which may play a role during early embryonic development.

Key Words: spermatozoa • reverse transcriptase • retroposon • nuclear scaffold • fertilization

© 2000 The Rockefeller University Press

Abbreviations used in this paper: CTAB, cetyltrimethylammonium bromide; FISH, fluorescent in situ hybridization; FM, fertilization medium; IVF, in vitro fertilization; LCMV, lymphocytic choriomeningitis virus; ORF, open reading frame; RT, reverse transcriptase.


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