ラット体細胞核移植のためのストロンチウムによる卵子の活性化誘起

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The objective of this study was to determine the parthenogenetic activation regimen of ovulated rat oocytes using strontium (Sr2+). Spontaneous activation of Wistar rat oocytes was triggered immediately after dissociation from the animals, but such activation seemed to be incomplete because no cleavage occurred even after 34 h culture in modified KRB medium. The survival rate of oocytes treated with 5 mM Sr2+ for 6 h (28%) was lower than that of those treated with 0, 0.31 and 1.25 mM Sr2+ (91-99%). Overall activation efficiency, as determined by the proportion of cleaved oocytes at 28 h of subsequent culture, was the highest in the oocytes treated with 1.25 mM Sr2+ (57%), followed by those treated with 0.31 mM Sr2+ (45%), 5 mM Sr2+ (20%) and 0 mM Sr2+ (8%). When oocytes from Sprague-Dawley (SD) or Wistar rats were incubated with 1.25 mM Sr2+ for 6 h, the oocytes from the SD strain tended to be more resistant to the Sr2+ treatment than the oocytes from the Wistar strain (survival rate 99 vs 87%), and the overall activation efficiency for SD oocytes was higher than that for Wistar oocytes (74 vs 45%). Therefore, nuclei of cumulus cells were injected into enucleated SD oocytes using piezo-driven micropipettes and the reconstructed oocytes were then activated with 1.25 mM Sr 2+ for 6 h. The cleavage rate of reconstructed oocytes 18 h post-activation was 22%. One implantation site but no live offspring was observed in the 9 surrogate mothers receiving a total of 224 presumptive cloned zygotes. This study contains fundamental data concerning oocyte activation and somatic cell nuclear transplantation in rats.
日本繁殖生物学会の論文

ラット体細胞核移植のためのストロンチウムによる卵子の活性化誘起

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