Characteristics of DNA Binding to Sheep Spermatozoa

概要

論文の詳細を見る
Spermatozoa were incubated in PBS with [<SUP>32</SUP>P] labeled DNA fragments at 37 C. After 10 min incubations, DNA fragments were tightly bound to spermatozoa. Pre-treatment of sperm with 3% glutaraldehyde prevented this binding ability. Bound DNA could not be removed from sperm, even after treatment of the sperm with 0.1% Triton X-100 or DNase. After the induction of capacitation and the acrosome reaction of the spermatozoa, which are prerequisites for normal fertilization, more than 40% of the DNA fragments remained bound. The site of binding of DNA on spermatozoa was determined by membrane fractionation on discontinuous sucrose gradient and electronmicroscopic observation and found to be on the plasma membrane of the post-acrosomal region. Artificial insemination was carried out with spermatozoa incubated with DNA fragments, and 20 fetuses at 35 days of pregnancy and 11 lambs were collected. The possibility that DNA integration into the chromosome had occurred was determined by Southern hybridization. Under these experimental conditions, DNA did not integrate into the fetal genome in any sample tested..
日本繁殖生物学会の論文