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In vitro development of non-enucleated rat oocytes following microinjection of a cumulus nucleus and chemical activation

Published online by Cambridge University Press:  01 May 2008

Wataru Fujii  and
Hiroaki Funahashi
Wataru Fujii
Affiliation:
Department of Animal Science, Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka, Okayama, 700–8530, Japan.
Hiroaki Funahashi*
Affiliation:
Department of Animal Science, Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka, Okayama, 700–8530, Japan. Department of Animal Science, Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka, Okayama, 700–8530, Japan.
*
All correspondence to: H. Funahashi. Department of Animal Science, Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka, Okayama, 700–8530, Japan. Tel: +81 86 251 8329. Fax: +81 86 251 8388. e-mail: [email protected]
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Summary

The present study examined in vitro development and the cytological status of non-enucleated rat oocytes after microinjection of cumulus nuclei and chemical activation. Oocyte–cumulus complexes were collected from gonadotropin-treated prepubertal female Wistar rats 14 h after human chorionic gonadotropin (hCG) injection. Cumulus nuclei were injected into ovulated oocytes and then stimulated in the presence of 5 mM SrCl2 for 20 min at various time points (0–3.5 h) after injection. Some of the reconstituted eggs were cultured to observe the pronuclear formation, cleavage, and blastocyst formation. The incidences of eggs forming at least one pronucleus or containing two pronuclei were not significantly different among the periods (82.4–83.5% and 43.4–51.9%, respectively). Nor did the incidences of eggs cleaving (86.7–97.7%) and developing to the blastocyst stage (0–3.5%) differ depending on when, after injection, stimulation began. When some of the reconstituted eggs were observed for cytological morphology 1–1.5 h after injection, 71.7% of the eggs caused premature chromatin condensation, but only 46.2% of them formed two spindles around each of maternal and somatic chromatins. However, the morphology of the somatic spindles differed from that of the spindles, which formed around the oocyte chromatins. Only 7.5% of the eggs contained the normal chromosomal number. In many reconstituted oocytes, before activation, an abnormal spindle formation was observed in the somatic chromatins. In conclusion, these results show that non-enucleated rat oocytes injected with cumulus nuclei can form pronuclei and cleave following chemical activation, whereas blastocyst formation is very limited, probably caused by abnormalities in the spindle formation and distribution of somatic chromatids.

Keywords

Cumulus cellIn vitro developmentMicroinjectionOocytesRat
Type
Research Article
Information
Zygote , Volume 16 , Issue 2 , May 2008 , pp. 117 - 125
Copyright
Copyright © Cambridge University Press 2008

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