Visualization of the three-dimensional distribution of collagen fibrils over preovulatory follicles in the hamster

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Abstract
Before an oocyte can escape from a preovulatory follicle, the apical wall must thin to the point of rupture. Although numerous layers of cells are present, it is the collagen fibrils in the theca externa that provide most of the strength to the developing follicle. The three-dimensional distribution and integrity of these fibrils over a follicle cannot be appreciated with standard used methods such as examination of thin sections by transmission electron microscopy. In this paper we describe a technique that removes cells superficial to the collagen fibrils so that their distribution may be examined by scanning electron microscopy. On the third day of the hamster's 4-day estrous cycle, bundles of fibrils pass from intrafollicular areas and ascend follicles. Approximately halfway up the follicle wall, the bundles fan out and form a meshwork of fibrils which covers the apex. As the time of ovulation approaches, the number of layers of fibrils decreases over the apex until a tear forms in the weakened matrix. Experimental results demonstrating that the meshwork is composed of collagen fibrils are presented. The usefulness of this technique in visualizing the collagen content in preovulatory follicles is discussed as well as factors that may aid in weakening this layer so that follicle rupture may occur.