Strain Differences in Superovulatory Response, Embryo Development and Efficiency of Transgenic Rat Production |
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Authors: | Elena Popova Michael Bader Alexander Krivokharchenko |
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Institution: | 1. Max-Delbrück-Center for Molecular Medicine (MDC), Robert-R?ssle-Str. 10, D-13092, Berlin-Buch, Germany
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Abstract: | The differences between rat strains in superovulation response, in vitro and in vivo development of preimplantation embryos and overall transgenic efficiency was studied. The protocols for induction of superovulation
using single injections of pregnant mare’s serum gonadotropin (PMSG) or minipumps with follicle stimulating hormone (FSH)
were compared in Lewis (LEW), Wistar-Kyoto (WKY), and stroke-prone spontaneously hypertensive rats (SHRSP) or Sprague–Dawley
(SD) and Wistar rats as representative inbred or outbred strains, respectively. The percentage of mated animals with positive
superovulatory response was similar in all strains (60.0–100%). The mean number of ova per donor was not dependent on the
kind of hormonal treatment used within each rat strain. In general, females from outbred SD and Wistar rats were more responsive
to hormonal treatments than animals from inbred rat strains. In addition, SD female rats produced a significantly higher number
of embryos per female in response to PMSG-treatment compared to all other strains. Between the inbred strains, SHRSP was the
most effective for superovulation. In vitro development of intact zygotes to the blastocyst stage was not different between SD, Wistar and SHRSP rats. In contrast, in vitro development of WKY zygotes was significantly less efficient than in other strains. However, 2-cell stage embryos in vivo produced from SD, SD × Wistar and WKY animals showed no difference in competence to develop to blastocyst stage in vitro. The proportion of offspring developing after oviduct transfer of intact zygotes was similar in all strains (44.0–56.4%)
with the exception of WKY rats (35.9%). We also compared the survival rate after injection, ability of manipulated zygotes
to develop to term and overall transgenic efficiency in various rat strains. SD and SHRSP zygotes survived after microinjection
better than the WKY and Lewis zygotes. No differences were found in the efficiency of transgene integration per newborn in
different strains ranging from 5.7 to 16.7%. The results of this study demonstrate that different rat strains have varying
responses to superovulation, sensitivity to microinjection, capability to develop in vitro until blastocyst stage or in vivo to term after transfer to foster mothers. Despite these differences all studied strains can be used for efficient transgenic
rat production. |
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Keywords: | embryo culture embryo development in vitro rat strain variation superovulation transgenesis |
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