عنوان مقاله [English]
نویسنده [English]چکیده [English]
Many studies have shown spermatozoa can be used as vectors for transfer of exogenous DNA to
oocyte to produce transgenic animals. However its efficiency is still questionable. The results of
DNA concentration and incubation time on DNA uptake and sperm functionality has been
different or sometimes opposite. In this study the effect of DNA concentration and incubation
time of sheep spermatozoa with foreign DNA on DNA uptake, its dynamic and sperm
functionality and their difference between ejaculated and epididimal sperm was investigated.
For this the ejaculated(N=4) and epididimal(N=6)spermatozoa was incubated with different
concentration (5-500ng/106 sperm/25μl) of rhodamine labaled pEGFP-IRES-hLys in various
time (15-90min) and DNA uptake, dynamic of uptake and sperm functionality was assayed.
Results showed to an average of 65% of incubated spermatozoa uptake the foreign DNA,
depend on DNA concentration and incubation time. Although the amount of transfected
spermatozoa could increase significantly with prolonged incubation time up to 60 min but the
amount of DNA uptake was higher in the first 15min of incubation. Increase of foreign DNA
concentration and incubation time increased the transfected spermatozoa and uptake intensity. It
seems higher DNA concentration, more rapid DNA uptake and more rapid gain of transfection
peak. Slight decrease of sperm total and progressive motility was seen in treatment group in
comparison with Control groups (except in 5min incubation). No spermatozoa with post
acrosomal absorption (true uptake) were motile. So incubation for more than 30 and 60 min in
moderate (20-200ng) and low (5ng) plasmid concentration respectively is not useful.