[The incorporation of macromolecules into the germ cells of male mice via electroporation and dimethyl sulfoxide].
Electroporation (incorporation of macromolecules into the living cells by means of electric pulses) provides inclusion of plasmid 14C-DNA into immature cells of spermatogenic epithelium. The highest level of foreign DNA incorporation into spermatocytes and spermatids has been induced by 8kV electric pulses applied 3 times with 20 sec intervals. Meanwhile, mature sperms are found to be exclusively resistant to exogenous DNA irrespective of the voltage level, the number of pulses and Ca++ uptake (contents). Incubation of mature sperms for two hours in the medium with Ca++ (10 mM) and dimethylsulfoxide--(DMSO, 33%) provides highly reliable incorporation of plasmid 14C-DNA into sperm heads. The sperm cells with foreign DNA incorporated by means of Ca++ and DMSO treatment still remain alive and mobile. The possibilities of mature sperms loaded with foreign DNA for the creation of transgenic mammals are discussed.
Baranov
V S
VS
Hapala
I
I
Hrijac
P
P
Kovac
L
L
Boda
K
K
D000818:Animals; D001345:Autoradiography; D002250:Carbon Radioisotopes; D004271:DNA, Fungal / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D004560:Electricity; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D046911:Macromolecular Substances; D008297:Male; D008722:Methods; D051379:Mice; D008808:Mice, Inbred CBA; D010957:Plasmids; D013091:Spermatogenesis / Q000187:drug effects / Q000502:physiology; D013094:Spermatozoa / Q000187:drug effects / Q000378:metabolism*
D002250:Carbon Radioisotopes; D004271:DNA, Fungal; D046911:Macromolecular Substances; D004121:Dimethyl Sulfoxide
Vkliuchenie makromolekul v polovye kletki samtsov mysheĭ pri pomoshchi élektroporatsii i dimetilsul'foksida.