Lawrence C. Zoller

A quantitative histochemical study of lactate dehydrogenase and succinate dehydrogenase activities in the membrana granulosa of the ovulatory follicle of the rat

SummaryUsing a microdensitometer, lactate dehydrogenase and succinate dehydrogenase activities were measured in the membrana granulosa of the rat ovulatory follicle. Ovaries were removed on each day of the oestrous cycle; oestrus, dioestrus-1, dioestrus-2, and proestrus; and enzyme activities measured in the membrana granulosa as a whole and in four regions within it: peripheral (PR), antral (AR), cumulus oophorus (CO) and corona radiata (CR). Throughout the cycle, lactate dehydrogenase activity was greatest in PR. On oestrus, lactate dehydrogenase activity was progressively less in AR, CO and CR. On dioestrus-1, activity was identical in AR and CO and less in CR. On dioestrus-2, activity was greater in AR than in CO or CR. By proestrus, activity was equal in AR, CO and CR. In the membrana granulosa as a whole, and in each region, lactate dehydrogenase activity declined as ovulation approached. In contrast, succinate dehydrogenase activity in the membrana granulosa as a whole and in PR was constant throughout the cycle. Activity fluctuated in the other regions. Succinate dehydrogenase activity on oestrus was greatest in PR, less in AR and CO and least in CR. On the remaining days, succinate dehydrogenase activity was greatest in PR and less but equal in the remainder of the membrana granulosa.

A quantitative electron microscopic analysis of the membrana granulosa of rat preovulatory follicles

The ultrastructure of the membrana granulosa (MG) of rat preovulatory follicles was examined using stereological techniques. Organelles studied were nuclei, mitochondria, lipid droplets (LD), lysosomes, and smooth and rough endoplasmic reticulum (SER, RER). The peripheral region of the MG contained the greatest volume of mitochondria, LD and SER, organelles associated with steroidogenesis. The volume of RER, an organelle associated with protein production, was greatest in the cumulus oophorus. These results corroborate previous analyses and demonstrate that the rat MG is composed of discrete subregions.

Effects of Silastic progesterone implants on activity cycles and steroid levels in ovariectomized and intact female rats.

We have previously shown that although Silastic implants of progesterone reduce the amount of running of animals living in activity wheels, progesterone-treated animals continue to show periodic fluctuations or peaks in activity. We hypothesized that although progesterone treatment inhibited estrous cycles, ovaries of animals treated with Silastic implants of progesterone continued to secrete estradiol in amounts adequate to stimulate moderate levels of running. In the present study we tested this hypothesis by removing ovaries from progesterone-treated animals and comparing their running behavior and steroid levels to progesterone-treated animals who received sham ovariectomies. Although progesterone treatment significantly inhibited running activity, removal of ovaries in progesterone-treated animals further suppressed running activity. In addition, both estradiol and progesterone levels were significantly reduced following removal of ovaries in progesterone-treated animals. We conclude that although Silastic progesterone implants inhibit normal ovarian and estrous activity cycles, ovaries produce sufficient estradiol to stimulate running behavior.

Quantitative Analysis of the Membrana Granulosa in Developing and Ovulatory Follicles

Ever since Regnier de Graf in 1672 [1] mistakenly identified the entire preovulatory follicle as the ovum, our concept of the structure and function of the follicle has been changing. It is now recognized that the ovarian follicle is a complex structure, composed of subregions that cooperate in the nutrition and expulsion of a healthy oocyte and in the production of numerous steroid hormones. The two major regions of the mature follicle are the membrana granulosa and the theca, separated from each other by a thick and distinct basement membrane. The morphology and ultrastructure of the membrana granulosa dramatically changes as the follicle matures. In this review these structural modifications will be documented and related to the physiology and function of the membrana granulosa both in situ and in vitro.

Effects of tetrahydrocannabinol on rat preovulatory follicles: a quantitative cytochemical analysis

SummaryUsing a microdensitometric histochemical assay, Δ5-3β-hydroxysteroid dehydrogenase activity and glucose-6-phosphate dehydrogenase Types I and II hydrogen generation were measured in preovulatory follicles from normal rats, and in follicles from rats given tetrahydrocannabinol for three days prior to sacrifice. Hydroxysteroid dehydrogenase and Type I hydrogen generation are involved in steroidogenesis, whereas Type II hydrogen generation is involved with general cellular metabolism. All ovaries were removed on pro-oestrus, frozen, sectioned and the sections reacted with the appropriate media. Enzyme activity was measured in the theca and in three regions of the membrana granulosa: peripheral antral and corona radiata. Compared to control animals, the hydroxysteroid dehydrogenase activity was significantly reduced in all follicular regions in rats exposed to tetrahydrocannabinol. Type I hydrogen generation was significantly less in the theca and peripheral region of preovulatory follicles from rats given tetrahydrocannabinol, but the same in the antral region and corona radiata. In all follicular regions examined, Type II hydrogen generation was unchanged following tetrahydrocannabinol administration. Thus, only the enzymes specifically associated with follicular steroidogenesis were affected by administration of the drug.

Effects of daily administration of tetrahydrocannabinol on rat preovulatory follicles. A quantitative electron-microscopic analysis

Using morphometric techniques, we analyzed the effects of delta 9-tetrahydrocannabinol (THC) on the ultrastructure of rat preovulatory follicles. Rats were injected intraperitonally once daily with vehicle alone, or with vehicle plus THC on diestrus (D), metestrus (M) and diestrus (D), or on estrus (E), M and D. All animals were killed on proestrus. Nuclear, mitochondrial, lipid droplet and lysosomal volume density in the membrana granulosa were analyzed. There was no change in nuclear volume, but there were significant THC-induced decreases in mitochondrial and lipid droplet volumes in the peripheral region of the granulosa. There were increases in dense-body volume in all regions of the granulosa from rats given THC either on D alone, or M and D. Dense-body volume returned to control levels in rats injected on E, M and D. It appears that THC can affect follicular ultrastructure and the effects change with the number of injections.

Thein vivo effects of tetrahydrocannabinol on the activity of lactate and succinate dehydrogenases in rat preovulatory follicles

SummaryThe effects of Δ9-tetrahydrocannabinol on the activities of lactate and succinate dehydrogenases in the theca interna and membrana granulosa of rat preovulatory follicles have been analysed microdensitometrically using the same injection regime employed in a previous study on steroidogenic enzymes. A small but statistically significant (18%) decrease in succinate dehydrogenase activity was observed in the theca interna, but none in any region of the membrana granulosa. Lactate dehydrogenase activity was unaffected by THC administration. Thus, a dosage and regimen sufficient to cause significant decreases in the activities of steroidogenic enzymes had little effect on succinate and lactate dehydrogenases in rat preovulatory follicles.

Quantitative Electron-Microscopic Analysis of the in vivo Effects of Tetrahydrocannabinol on Rat Preovulatory Follicles

The ultrastructural composition of 4 regions of rat preovulatory follicles was examined using stereological techniques following an intraperitoneal injection of tetrahydrocannabinol (THC), once each day, for 3 days prior to sacrifice on proestrus. Control rats were injected with solvent alone. Changes in nuclear, mitochondrial, lipid droplet and dense-body volume density were measured. No differences in volume density were noted in the theca or antral region of the membrana granulosa. Compared to controls, the mitochondrial volume density decreased in the peripheral region and the corona radiata. The volume density of lipid droplets decreased in the peripheral region. These changes, and their relationship to the roles of these regions in follicular steroidogenesis, are discussed.