Gonadotropin-induced ovulation in a reptile (Anolis carolinensis): histological observations

Although much is known about morphological changes in the apex of the mammalian ovarian follicle prior to its rupture (ovulation), information about this process in nonmammalian vertebrates is limited to only a few species. We describe here the histological changes in the stigma of the lizard (Anolis carolinensis) ovarian follicle preceding FSH-induced ovulation. Females with a large vitellogenic follicle received two injections, separated by 7 hr, of either porcine FSH (25 micrograms) or saline, and then were sampled at 6, 9, 12, 15, 18, and 24 hr after the first injection. Thus, all but the 6-hr group received two injections. By 12 hr, about half of the FSH-treated females had ovulated, and most had ovulated by 15 hr. In contrast, only 1 of 40 control females ovulated. Large unovulated follicles were fixed and bisected through the circular stigma; one hemisphere was embedded in paraffin and stained with Mallory's trichrome, and the other was embedded in plastic and stained with toluidine blue. FSH treatment produced marked histological changes in the stigma region, as well as alterations in nonstigmal areas of the follicle. The membrana granulosa of control follicles consisted of a single layer of squamous, darkly staining granulosa cells. After FSH treatment, nonstigmal granulosa cells separated slightly, and their nuclei became more clear and assumed an oval shape; in the stigma, these cells became widely separated, with round, light-staining nuclei containing one or two prominent nucleoli. The nonstigmal theca of FSH-treated follicles was similar to that of control follicles except that collagen fibers were more dissociated. In the stigma, collagen fibers were widely dissociated, and the theca swelled, presumably due to accumulation of extracellular fluid. Abundant fluid accumulated in the stigma, especially between the granulosa cells and their basement membrane and between the tunica albuginea and the theca externa. These changes in the lizard stigma are similar to those reported in mammals except that no marked inflammatory response occurs in the lizard stigma. We hypothesize that the Anolis follicle undergoes preovulatory luteinization, and that the stigma exhibits ischemic necrosis before rupture.