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Intracytoplasmic sperm injection in the rat

Published online by Cambridge University Press:  05 April 2001

D. Dozortsev
Affiliation:
Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Hutzel Hospital/Wayne State University, Detroit, MI 48201, USA
T. Wakaiama
Affiliation:
Department of Anatomy and Reproductive Biology, University of Hawaii Medical School, Honolulu, HI 96822, USA
A. Ermilov
Affiliation:
Division of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, Hutzel Hospital/Wayne State University, Detroit, MI 48201, USA
R. Yanagimachi
Affiliation:
Department of Anatomy and Reproductive Biology, University of Hawaii Medical School, Honolulu, HI 96822, USA

Abstract

We applied intracytoplasmic sperm injection (ICSI) to the rat comparing three different sperm injection techniques: conventional setup with a sharp needle bearing a spike (method 1), combination of partial zona dissection (PZD) needle and blunt pipette (method 2) and piezo-injection using a blunt pipette (method 3). We also investigated the timing of sperm pronuclear formation after injection. Survival rates after injection were 8%, 24% and 71% for the methods 1, 2 and 3, respectively. All surviving oocytes formed pronuclei by about 6 h after injection. Although the survival and activation rates following sperm injection using piezo-injection were high, the incidence of normal fertilisation, as evidenced by second polar body extrusion and formation of two pronuclei, was only 10%. The vast majority of the zygotes were multinucleated, although most of them subsequently underwent cleavage. Fixation and staining of injected oocytes at different times after injection revealed that replacement of sperm nuclear protamines by histones takes place by 15 min after injection, sperm head swelling occurs within 0.5–1 h after injection and pronuclei become fully developed by 7 h after injection. Although the rate of normal fertilisation in the rat following ICSI was low under the present experimental conditions, the results indicated that direct ICSI using a piezo-driven pipette would be a potentially valuable method of producing rat offspring.

Type
Research Article
Copyright
1998 Cambridge University Press

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