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.
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20 March 2000
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March 20 2000
Reverse Transcriptase Activity in Mature Spermatozoa of Mouse
Roberto Giordano,
Roberto Giordano
aCNR Center for the Study of Germinal Cells, University of Siena, 53100 Italy
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Anna Rosa Magnano,
Anna Rosa Magnano
aCNR Center for the Study of Germinal Cells, University of Siena, 53100 Italy
bInstitute of General Biology, University of Siena, 53100 Italy
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Germana Zaccagnini,
Germana Zaccagnini
cCNR Institute of Biomedical Technology, 00161 Rome, Italy
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Carmine Pittoggi,
Carmine Pittoggi
bInstitute of General Biology, University of Siena, 53100 Italy
cCNR Institute of Biomedical Technology, 00161 Rome, Italy
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Nicola Moscufo,
Nicola Moscufo
dIstituto Superiore di Sanità, 00161 Rome, Italy
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Rodolfo Lorenzini,
Rodolfo Lorenzini
dIstituto Superiore di Sanità, 00161 Rome, Italy
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Corrado Spadafora
Corrado Spadafora
cCNR Institute of Biomedical Technology, 00161 Rome, Italy
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Roberto Giordano
aCNR Center for the Study of Germinal Cells, University of Siena, 53100 Italy
Anna Rosa Magnano
aCNR Center for the Study of Germinal Cells, University of Siena, 53100 Italy
bInstitute of General Biology, University of Siena, 53100 Italy
Germana Zaccagnini
cCNR Institute of Biomedical Technology, 00161 Rome, Italy
Carmine Pittoggi
bInstitute of General Biology, University of Siena, 53100 Italy
cCNR Institute of Biomedical Technology, 00161 Rome, Italy
Nicola Moscufo
dIstituto Superiore di Sanità, 00161 Rome, Italy
Rodolfo Lorenzini
dIstituto Superiore di Sanità, 00161 Rome, Italy
Corrado Spadafora
cCNR Institute of Biomedical Technology, 00161 Rome, Italy
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.
Received:
October 13 1999
Revision Requested:
February 08 2000
Accepted:
February 10 2000
Online ISSN: 1540-8140
Print ISSN: 0021-9525
© 2000 The Rockefeller University Press
2000
The Rockefeller University Press
J Cell Biol (2000) 148 (6): 1107–1114.
Article history
Received:
October 13 1999
Revision Requested:
February 08 2000
Accepted:
February 10 2000
Connected Content
Citation
Roberto Giordano, Anna Rosa Magnano, Germana Zaccagnini, Carmine Pittoggi, Nicola Moscufo, Rodolfo Lorenzini, Corrado Spadafora; Reverse Transcriptase Activity in Mature Spermatozoa of Mouse. J Cell Biol 20 March 2000; 148 (6): 1107–1114. doi: https://doi.org/10.1083/jcb.148.6.1107
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