Production of transgenic rats by ooplasmic injection of spermatogenic cells exposed to exogenous DNA: a preliminary study.
The aim of the present study was to investigate the efficiencies of producing transgenic rats by the ooplasmic injection of sperm heads (intracytoplasmic sperm injection: ICSI) and elongating spermatids (elongating spermatid injection: ELSI) exposed to the EGFP DNA solution. A slightly lower proportion of ICSI oocytes using sperm heads exposed to a concentration of 0.5 microg/ml DNA solution for 1 min developed into offspring (13.3%, 48/361) when compared to that of oocytes injected with nontreated sperm heads (19.4%, 32/165). Eight ICSI offspring were found to be EGFP-carrying transgenic rats (16.7% per offspring; 2.2% per embryo). After a 1-min exposure of the elongating spermatids to 5 microg/ml of DNA solution, 8.8% (45/511) of the ELSI oocytes developed into offspring while 12.7% (22/173) of the ELSI oocytes using nontreated spermatids developed. Six ELSI offspring carried the EGFP DNA (13.3% per offspring; 1.2% per embryo). The conventional pronuclear microinjection of 5 microg/ml of DNA solution resulted in the higher production of offspring (29.7%, 104/350) and the birth of three transgenic rats (2.9% per offspring; 0.9% per embryo). Thus, sperm heads and elongating spermatids were practically useful as the vector of exogenous DNA if the DNA-exposed spermatogenic cells were microinseminated into rat oocytes.
National Institute for Physiological Sciences, Okazaki, Aichi, Japan.
Megumi
M
Kato
Ayako
A
Ishikawa
Ryosuke
R
Kaneko
Takeshi
T
Yagi
Shinichi
S
Hochi
Masumi
M
Hirabayashi
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D004247:DNA / Q000235:genetics*; D046149:Embryo Culture Techniques; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics; D008297:Male; D009865:Oocytes / Q000166:cytology*; D051381:Rats / Q000235:genetics*; D013077:Sperm Head / Q000637:transplantation*; D020554:Sperm Injections, Intracytoplasmic; D013087:Spermatids / Q000637:transplantation*
D049452:Green Fluorescent Proteins; D004247:DNA