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Production of transgenic rats by ooplasmic injection of spermatogenic cells exposed to exogenous DNA: A preliminary study
Author(s) -
Kato Megumi,
Ishikawa Ayako,
Kaneko Ryosuke,
Yagi Takeshi,
Hochi Shinichi,
Hirabayashi Masumi
Publication year - 2004
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.20161
Subject(s) - biology , offspring , spermatid , microinjection , andrology , embryo , intracytoplasmic sperm injection , sperm , transgenesis , spermatogenesis , transgene , exogenous dna , fertilisation , embryogenesis , genetics , dna , reproductive technology , gene , microbiology and biotechnology , endocrinology , pregnancy , in vitro fertilisation , medicine
Abstract 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 μg/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 μg/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 μg/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. Mol. Reprod. Dev. 69: 153–158, 2004. © 2004 Wiley‐Liss, Inc.