Cytoplasmic granules in luteal cells of pregnant and non-pregnant guinea pigs. A cytochemical study.

Secretory granules, which are released by exocytosis and are speculated to contain progesterone, have been described in luteal cells of sheep and other large domestic animals. These granules are small and densely staining. Gemmell and Stacy ('79) suggested that luteal cells of guinea pigs also contain secretory granules, although they could not document exocytosis of granule content at the fine structural level. For the present study, quantitative methods were used to reexamine the possibility that luteal cells of guinea pigs possess secretory granules. Ovaries of guinea pigs were fixed in situ by vascular perfusion at the time of maximum progesterone secretion, when such granules would be most abundant, as well as other stages. Two types of granules that might be confused with secretory granules are microperoxisomes and lysosomes. Therefore, slices of perfusion-fixed corpora lutea were incubated for the fine structural localization of a peroxisomal enzyme, catalase, or for the lysosomal enzymes, acid phosphatase (ACPase) and arylsulfatase. Other tissue was prepared for conventional electron microscopy. Granule types were classified on the basis of size, morphology, and enzyme content. Quantitation of granule types was carried out on both cytochemically reacted and conventionally prepared luteal tissue. More than 5500 microperoxisomes, 2800 lysosomes, and 1100 multivesicular bodies (MVBs) were tabulated. The results indicate that luteal cells of guinea pigs have three main types of granules: 1) Microperoxisomes, about 0.2 micrometer in diameter and containing catalase; 2) lysosomes, about 0.5 micrometer in diameter and positive for ACPase and arylsulfatase; and 3) MVBs, about 0.4 micrometer in diameter and containing small vesicles. At the time of peak steroid secretion during pregnancy and the estrous cycle, the granule population in luteal cells of guinea pigs consists of 73-80% microperoxisomes, 13-17% lysosomes, and 7-9% MVBs. These proportions are similar in tissue reacted for cytochemistry and tissue prepared by conventional means. Greater than 99% of the small 0.2-0.3 micrometer diameter granules in guinea pig luteal cells are catalase reactive. This finding eliminates from further consideration most of the prime candidates for secretory granules in these cells. Finally, neither a sequential appearance of granules nor exocytosis of secretory product was detected. Our data thus argue against the suggestion that luteal cells of guinea pig have secretory granules of the type observed in corpora lutea of large domestic animals.