Activation of endogenous nucleases in mature sperm cells upon interaction with exogenous DNA.
Mature sperm cells, either of epididymal origin or ejaculated and depleted of seminal fluid, are spontaneously able to bind exogenous DNA molecules which are subsequently internalized into sperm nuclei. Southern blot analysis showed that the internalized DNA was specifically cleaved by sperm endonucleases and showed typical fragmentation patterns of localized hypersensitivity. Nucleases were activated in response to the internalization of exogenous DNA by sperm cells and their activity increased with the DNA concentration. Nuclease activation was efficient in epididymal sperm cells, while being drastically reduced in ejaculated washed spermatozoa. Nucleases were Ca++ dependent, and were, respectively, inhibited and activated by preincubating sperm cells with Aurintricarboxylic Acid (ATA) and Ca++ Ionophore A23187, which are known to, respectively, inhibit and activate apoptosis in somatic cells. Moreover, nuclease activation also caused a partial degradation of the sperm endogenous chromosomal DNA; cleaved DNA fragments were released from the sperm cells to the medium. Taken together, these results suggest that a metabolically active process similar to apoptosis is triggered in the nuclei of mature sperm cells upon interaction with exogenous DNA.
Maione
B
B
Istituto di Tecnologie Biomediche, CNR, Rome, Italy.
Pittoggi
C
C
Achene
L
L
Lorenzini
R
R
Spadafora
C
C
D000818:Animals; D017209:Apoptosis; D001312:Aurintricarboxylic Acid / Q000494:pharmacology; D001483:Base Sequence; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000502:physiology; D002417:Cattle; D004247:DNA / Q000378:metabolism*; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D004789:Enzyme Activation; D007476:Ionophores / Q000494:pharmacology; D007782:Lactoglobulins / Q000235:genetics; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D013094:Spermatozoa / Q000201:enzymology*; D013552:Swine
D007476:Ionophores; D007782:Lactoglobulins; D000001:Calcimycin; D001312:Aurintricarboxylic Acid; D004247:DNA; D004706:Endodeoxyribonucleases; D002118:Calcium