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Hum. Reprod. Advance Access originally published online on January 26, 2006
Human Reproduction 2006 21(3):624-631; doi:10.1093/humrep/dei394
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© The Author 2006. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org

An intravital microscopy method permitting continuous long-term observations of ovulation in vivo in the rabbit

Pernilla Dahm-Kähler1,4, Carl Löfman1, Ryota Fujii2, Michael Axelsson3, Per Olof Janson1 and Mats Brännström1

1 Department of Obstetrics and Gynecology, Sahlgrenska Academy, Göteborg University, Sahlgrenska University Hospital, S-413 45 Göteborg, Sweden, 2 Department of Obstetrics and Gynecology, Kanazawa University Medical School, Kanazawa, Japan and 3 Department of Zoology, Göteborg University, Göteborg Sweden

4 To whom correspondence should be addressed. E-mail: pernilla.dahm-kahler{at}vgregion.se

BACKGROUND: A method for intravital microscopy of the rabbit ovary was developed to enable observations of real-time changes during ovulation in vivo. The aim was to correlate these events to biochemical events at specific stages of ovulation. METHODS: Virgin, female rabbits were primed with equine chorionic gonadotrophin (CG) (30–100 IU) then HCG (100 IU) 2 days later to induce ovulation. During anaesthesia, the right ovary was surgically exteriorized and submerged in an organ chamber with a microscopy lens positioned close to the ovary. Continuous video recordings were performed. RESULTS: Initial equine CG priming experiments revealed the highest ovulation rate, without premature luteinization, after 30 IU equine CG. This priming protocol subsequently demonstrated follicular ruptures 11.5–14 h after HCG. Numbers of ovulations from the exteriorized and contralateral non-exteriorized ovary were similar. The sequence of typical features of ovulation was: shutdown of microcirculation in the follicular apex, formation of petechiae in the follicular wall and a cone-shaped structure over the future rupture site, marked bleeding in connection with follicular rupture and a fairly steady extrusion velocity of granulosa cells and the oocyte. CONCLUSION: This method captured a sequence of structural changes during ovulation. It could be combined with blood and follicular fluid sampling for biochemical analysis and could be used in studies on biochemical reactions in relation to specific changes in the follicular structure during ovulation.

Key words: follicle/intravital microscopy/ovary/ovulation/rabbit


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