In vivo imaging reveals an essential role of vasoconstriction in rupture of the ovarian follicle at ovulation

• Published in: Proceedings of the National Academy of Sciences - PNAS 2016-02, Vol.113 (8), p.2294-2299
• Main Authors: Migone, Fernando F., Cowan, Robert G., Williams, Rebecca M., Gorse, Kiersten J., Zipfel, Warren R., Quirk, Susan M.
• Format: Article
• Language: English
• Subjects: Animals; Biological Sciences; Cells; Endothelin-2 - deficiency; Endothelin-2 - genetics; Endothelin-2 - physiology; Female; Fertility; Intravital Microscopy; Mice; Mice, 129 Strain; Mice, Transgenic; Microscopy; Microscopy, Fluorescence, Multiphoton; Ovarian Follicle - blood supply; Ovarian Follicle - diagnostic imaging; Ovarian Follicle - physiology; Ovulation - genetics; Ovulation - physiology; Protease inhibitors; Receptors, G-Protein-Coupled - deficiency; Receptors, G-Protein-Coupled - genetics; Receptors, G-Protein-Coupled - physiology; Receptors, Peptide - deficiency; Receptors, Peptide - genetics; Receptors, Peptide - physiology; Receptors, Transforming Growth Factor beta - deficiency; Receptors, Transforming Growth Factor beta - genetics; Receptors, Transforming Growth Factor beta - physiology; Reproductive system; Smoothened Receptor; Ultrasonic imaging; Ultrasonography; Vasoconstriction - genetics; Vasoconstriction - physiology
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.1512304113

Rupture of the ovarian follicle releases the oocyte at ovulation, a timed event that is critical for fertilization. It is not understood how the protease activity required for rupture is directed with precise timing and localization to the outer surface, or apex, of the follicle. We hypothesized that vasoconstriction at the apex is essential for rupture. The diameter and blood flow of individual vessels and the thickness of the apical follicle wall were examined over time to expected ovulation using intravital multiphoton microscopy. Vasoconstriction of apical vessels occurred within hours preceding follicle rupture in wild-type mice, but vasoconstriction and rupture were absent in Amhr2cre/+SmoM2 mice in which follicle vessels lack the normal association with vascular smooth muscle. Vasoconstriction is not simply a response to reduced thickness of the follicle wall; vasoconstriction persisted in wild-type mice when thinning of the follicle wall was prevented by infusion of protease inhibitors into the ovarian bursa. Ovulation was inhibited by preventing the periovulatory rise in the expression of the vasoconstrictor endothelin 2 by follicle cells of wild-type mice. In these mice, infusion of vasoconstrictors (either endothelin 2 or angiotensin 2) into the bursa restored the vasoconstriction of apical vessels and ovulation. Additionally, infusion of endothelin receptor antagonists into the bursa of wild-type mice prevented vasoconstriction and follicle rupture. Processing tissue to allow imaging at increased depth through the follicle and transabdominal ultrasonography in vivo showed that decreased blood flow is restricted to the apex. These results demonstrate that vasoconstriction at the apex of the follicle is essential for ovulation.