Production of transgenic rats by ooplasmic injection of spermatogenic cells exposed to exogenous DNA: A preliminary study

Megumi Kato, Ayako Ishikawa, Ryosuke Kaneko, Takeshi Yagi, Shinichi Hochi, Masumi Hirabayashi

Research output: Contribution to journalArticlepeer-review

33 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)153-158
Number of pages6
JournalMolecular reproduction and development
Volume69
Issue number2
DOIs
Publication statusPublished - 2004 Oct
Externally publishedYes

Keywords

  • DNA microinjection
  • EGFP
  • ELSI
  • ICSI
  • Transgenic rats

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology
  • Cell Biology

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