Interstitial Gland Research Articles

Ovarian development in control and decapitated pig fetuses.

Ovarian development was studied in control and decapitated pig fetuses. Fetuses were decapitated at 42 days postcoitum. At 51, 61, 74, 90 and 112 days postcoitum decapitated and control females were collected. Ovarian weight gradually increased during development in control animals. Deprivation of pituitary hormones as a result of fetal decapitation did not cause a decline in ovarian weight increase. Germ cell maturation in control and decapitated fetuses proceeded in a similar fashion, with secondary follicles being the most advanced stage. Enzyme histochemical activity was present in the primary interstitial gland cells and in granulosa cells and was similar in normal and decapitated fetuses. Both NADH diaphorase activity and 3 beta-hydroxysteroid dehydrogenase activity increased from 51 to 74 days and remained relatively constant thereafter. Since fetal decapitation in the pig hardly influences ovarian development, pituitary dependency of the fetal ovary in the pig is unlikely.

Morphological and morphometric changes in the ovaries of white-footed mice (Peromyscus leucopus) following exposure to long or short photoperiod.

Two experiments were carried out with a total of 36 white-footed mice (Peromyscus leucopus) exposed to either long (LP) or short (SP) photoperiod for six weeks. Ovaries and uteri were weighed. Serial sections of the ovaries were processed for light microscopy. The total number of the interstitial gland cells, preantral and antral follicles with early and advanced stages of atresia as well as corpora lutea, was determined. Uteri from LP mice weighed significantly more than those from SP mice in both experiments. In contrast, the weight of ovaries from LP and SP animals only differed significantly in one experiment; this experiment also produced no obvious changes in the total number of corpora lutea. While interstitial gland cells of LP ovaries were hypertrophic and appeared mature, the interstitial gland cells of SP ovaries were of the degenerating cell type. The total number of follicles as well as their mean follicular diameters were higher in LP animals in comparison with SP animals. In addition, the percentage of antral follicles were increased and there were fewer signs of advanced stages of atresia in the LP group. It is suggested that the changes of the interstitial gland cell morphology combined with alterations of follicular growth during LP or SP appear to support intensified follicular growth.

Effects of aqueous and lipoidal extracts of the wall of preovulatory follicles on the ovary of growing chicks

Effects of continuous intramuscular administration of the lipoidal and aqueous extracts of the wall of the largest preovulatory follicle (3·8–4·0 cm), to chicks from 11 to 29 weeks on alternate days have been studied on the ovarian growth, follicular growth and atresia, and the onset of ovulation. The lipoidal and aqueous extracts appear to have antagonistic effects on the follicular growth and atresia. Chicks injected with lipid extract show increased ovarian weight, enhanced follicular growth and egg laying started at 22nd week which is earlier to 25th week in control birds. The onset of egg laying was delayed in chicks injected with aqueous extract of largest follicle wall by 4 weeks as compared to control birds. Their ovaries showed decreased rate of follicular growth and increased follicular atresia as a result of which abundant amount of lipid-rich interstitial gland tissue of thecal origin accumulates in them.

Gonadal steroid levels in the turtle Emys orbicularis L.: A preliminary study in embryos, hatchlings, and young as a function of the incubation temperature of eggs

Eggs of Emys orbicularis L. were incubated at either 25° (100% male-producing temperature) or at 30° (100% female-producing temperature). Gonadal levels of seven steroids were subsequently measured in embryos (stage 24), hatchlings, and young (3 and 9 months). According to the stage, 22–80 gonads were first removed and cultured in synthetic medium for 2 hr. They were then pooled and homogenized in this medium for steroid assay (RIA). The total steroid levels were weak (10–30 pg/gonad) in embryos and hatchlings and increased in the young turtles. In embryos, total steroid level was higher in males than in females, while at 9 months it was higher in females. As for the different steroids, the main following results were obtained. In embryos, progesterone (Prog), dehydroepiandrosterone (DHA), and testosterone (Testo) were not detected in either testes at 25° or ovaries at 30°, while Δ 4-androstenedione (Δ4A), dihydrotestosterone (DHT), and estrone (E1) were found in higher levels in testes (chiefly E1). Low levels in testes or traces in ovaries of estradiol (E2) were present. At hatching, E2 was no longer detectable, while DHA in both sexes and Testo in males were measurable. At this time, the other hormones displayed either minor variations in females or were decreased in males. At 3 months, Prog and E2 were still missing and only traces of E1 were detected. DHA decreased in males, while it increased in females. Levels of the other steroids, mainly Testo (in both sexes) and DHT (in males) increased. At 9 months, all steroids except E2 were detected. The total steroid content increased markedly, with the highest levels being found for Prog and Δ4A, chiefly in ovaries. This latter increase could be related to the posthatching differentiation of interstitial gland (Leydig cells) in testes and to follicle growth in ovaries.

Immunohistochemical localization of ornithine decarboxylase in the rat ovary.

The present report describes the immunocytochemical localization of ornithine decarboxylase in the prepubertal rat ovary after administration of human chorionic gonadotropin (HCG). Numerous ornithine decarboxylase immunoreactive cells appeared in the thecal layer as well as in the interstitial gland tissue after treatment with HCG. The granulosa cells, the ovum and the ovarian stroma were devoid of immunoreactive ornithine decarboxylase. In contrast to the ovary of HCG-treated rats, the ovary of prepubertal rats given the vehicle alone contained only a few weakly immunoreactive cells.

The effect of alkylating agents on the reproductive and hormonal testicular function in patients with rheumatoid arthritis.

An evaluation of testicular function was undertaken in 11 boys and young men aged 14-26 years suffering from rheumatoid arthritis complicated by amyloidosis and treated with cyclophosphamide and chlorambucil. 7 cases were studied during life and 4 at autopsy. Semen analysis showed azoospermia in 4 cases; in one, cryptozoospermia; in one, oligoasthenotheratozoospermia; and in one, asthenoteratozoospermia. In 4 patients with azoospermia and in one with cryptozoospermia, testicular biopsy was done for histo-pathological examination. In patients with azoospermia, only Sertoli cells were present in the seminiferous tubules. In one patient with cryptozoospermia, normal spermato- and spermiogenesis was found only in occasional seminiferous tubules. Microscopic examination of the testis failed to show any changes in the structure of the interstitial gland in any of the patients. Amyloid deposits were found in the vascular walls of the interstitial gland in one patient only. The serum testosterone level was depressed in 3 patients of the 9 examined. The LH level was elevated in 2 patients. The mean value of testosterone-LH ratio was significantly lower in patients treated with alkylating agents as compared with patients not so treated. Serum FSH level was elevated in 6 patients. Post-mortem examination of the testis in 4 patients showed complete absence of the germinal epithelium and only the presence of Sertoli cells. In 2 cases, amyloid deposits were found in the vascular walls in the rete testis. It was concluded that alkylating agents, besides damaging the testicular germinal epithelium, also affect the function of Leydig cells.

Short- and long-term response of the adult dog ovary after 1200 R whole-body X-irradiation and transfusion of mononuclear leukocytes.

The effect of 1200 R whole-body X-irradiation after transfusion of mononuclear blood cells was investigated in 34 dog ovaries (beagle) for 6-1460 days after irradiation. Morphological observations suggest that gonadal and extragonadal function regenerated after irradiation. Follicular growth started 60 days after irradiation. The time required for follicular maturation was inferred to be about 110 days and the time of first ovulation about 170 days. Graafian follicles and corpora lutea were found 2, 3 and 4 years later. A pregnancy occurred 2 years after irradiation. Larger, growing and vesicular follicles were primarily affected by irradiation-induced atresia with damage of the granulosa. Oocytes appeared to be normal, even without the supporting granulosa cells ('naked oocytes'). The interstitial cortical tissue of the ovaries reacted in two ways. Either, it remained inactive (radiation effect I) or it showed the endocrine structure of mature interstitial gland cells (radiation effect II). Microadenomata were observed 195 days after irradiation.

Luteal blood flow and progesterone secretion in pseudopregnant rabbits.

Ovarian and luteal blood flow rates were measured, by means of radioactive microspheres, in anaesthetized rabbits on Day 8 of pseudopregnancy before and after lowering the ovarian perfusion pressure with a sling placed around the aorta. When blood pressure was lowered by 42% luteal flow decreased 9-fold whilst flow to the remaining part of the ovary remained unchanged, indicating the presence of an autoregulatory mechanism in the ovarian interstitial gland. Ovarian progesterone secretion, assessed from progesterone concentrations in ovarian venous blood, was positively correlated to the blood flow per unit of weight of luteal tissue. These data indicate that a high rate of luteal blood flow may be necessary for an optimal steroid production by the corpus luteum.

Fluorescence Localization of Luteinizing Hormone/Human Chorionic Gonadotropin Uptake in the Primate Ovary: Characterization of the Preovulatory Ovary

A fluorescent derivative of human chorionic gonadotropin (hCG) for binding studies was prepared by conjugation of hightly purified hCG with fluorescein isothiocynate (FITC-hCG) in a molar ratio of 3:1. The resulting FITC-hCG conjugate retained full cross-reactivity with antibodies directed against intact hCG, the hCG beta-subunit, the carboxyl-terminal peptide sequence of beta-hCG. The fluorescent hormone retained binding activity similar to that of native hCG, and the capacity to induce progesterone production from primate corpora luteal cells in vitro. After intravenous administration to female rhesus monkeys, FITC-hCG was localized within the theca cell layer of certain antral follicles, including the dominant follicle, but no fluorescence was noted around preantral follicles. Areas of fluorescence were also localized to discrete groups of interstitial gland cells in the stroma. The degree of fluorescence associated with these cells was indistinguishable from that observed in the thecal cells. The development of a fluorescent derivative that is biologically and immunologically similar to native hCG has permitted differential microscopic localization of luteinizing hormone/hCG uptake in the preovulatory ovary.

Interstitium: site of steroid synthesis in the ovary of the long term hypophysectomized hamster.

The ovaries of hamsters hypophysectomized (H̄) on the afternoon of proestrus synthesize in vitro substantial amounts of progesterone for as long as 20 days post-H̄ (Taya and Greenwald, 1979b). The purpose of the present study was to identify by histochemistry and ultrastructure the ovarian cell type which still possessed the enzymes capable of steroidogenesis despite the long term absence of gonadotropins. After Day 6 post-H̄, the ovaries contained interstitium and viable and atretic small preantral follicles. The viable preantral follicles were histochemically devoid of Δ5-3β-hydroxysteroid dehydrogenase (3β-HSDH). Histochemically, there were no appreciable changes in lipids in the interstitial gland cells (IGC) until Day 20 post-H, followed by a slight decrease by Day 30. The 3β-HSDH activity in the IGC showed no appreciable changes until Day 6 and thereafter decreased gradually until Day 30. Trace amounts of glucose-6-phosphate dehydrogenase were found in the IGC until Day 6 and thereafter the enzyme was undetectable. The IGC contained only trace amounts of acid phosphatase until Day 10, followed by a gradual increase by Day 30. A single i.p. injection of 5 μg ovine LH on Day 10, 20 or 30 resulted 3 h later in dramatic increases in the concentration of serum progesterone and ovarian progesterone and testosterone but not estradiol-17β. Concomitantly, the depletion of lipid droplets in viable interstitial cells was observed by histochemical and ultrastructural criteria. However, lipid accumulations in degenerating interstitial cells and atretic granulosa cells were unaffected by LH treatment. These data indicate that the interstitium is an active steroidogenic tissue for at least 30 days post-H and that LH can rapidly stimulate synthesis and secretion of progesterone and testosterone by the interstitium in long term hypophysectomized hamsters.

The origin of the extrinsic adrenergic innervation to the rat ovary.

Cutting the suspensory ligament reduced the ovarian content of norepinephrine (NE) to less than half that of controls and only a few blood vessels had perivascular fibers and an occasional nerve remained in the interstitial gland. Cutting the ovarian plexus had a less drastic, but similar effect on the ovarian content of NE and on the pattern of ovarian adrenergic nerves. Cutting both the suspensory ligament and ovarian plexus eliminated visualization of ovarian adrenergic nerves, but some ovarian NE was still measurable. Fluorescence and electron microscopic studies of the suspensory ligament revealed a large adrenergic nerve embedded in smooth muscle of the ligament. The nerve was also acetylcholinesterase-positive. Cutting the celiac plexus or incising a small nerve lateral to the plexus and medial to the origin of the suspensory ligament, had the same effect on the ovarian adrenergic nerves as cutting the suspensory ligament. It is concluded that the extrinsic adrenergic nerves to the rat ovary reach the organ by two routes: one via the nerve in the suspensory ligament (superior ovarian nerve), and one via the traditionally described ovarian plexus along the ovarian artery.

Localization of some steroidogenic enzymes in the ovary of the rabbit.

The distribution of delta 5-3 beta-HSD, peroxidase and cytochrome oxidase in immature, sexually mature and pregnant rabbit ovary has been studied histochemically. Corpora lutea are found only in pregnant rabbits. delta 5-3 beta-HSD is present in the theca interna of mature follicles, corpora lutea and interstitial gland cells but is absent in the granulosa cells of both developing and mature follicles. The granulosa cells of mature and developing follicles, hypertrophied theca interna and the luteal cells all show intense cytochrome oxidase activity. Peroxidase is present in the corpora lutea only. It is suggested that delta 5-3 beta-HSD in the theca interna and interstitial gland cells is the enzyme responsible for steroid synthesis in the ovaries of immature as well as sexually mature rabbits, while peroxidase and delta 5-3 beta-HSD present in the corpora lutea together regulate luteal steroidogenesis during pregnancy. The intense cytochrome oxidase activity together with peroxidase and delta 5-3 beta-HSD confirms the observations that this tissue is a site of intense oxidative activity.

Histological and Histochemical Studies on Follicular Atresia in the Ovary of the Lizard, Calotes versicolor

Histological study of atretic follicles in the ovary of Calotes versicolor shows the presence of yolky, glandular, bursting, haemorrhagic and cystic types of atresia. The granulosa cells of the atretic follicles exhibit A5-3p,-HSDH, 17,/-HSDH, 11/3-HDSH, G-6-PDH, NADH dia phorase, LDH and ICDH activity for a short duration, indicating their transient steroidogenic capacity and then degenerate. Theca interna cells show an increased activity for these enzymes in the atretic follicles and later persist as interstitial gland cells in the ovarian stroma.

Steroid synthesizing cellular sites in the ovary of the domestic pigeonColumba livia (Gmelin): A histochemical study

The ovary of the domestic pigeon, Columba livia, has been assayed histochemically for the localization of delta 5-3 beta-hydroxysteroid dehydrogenase (delta 5-3 beta-HSDH), 17 beta-hydroxysteroid dehydrogenase (17 beta-HSDA), 11 beta-hydroxysteroid dehydrogenase (11 beta-HSDH), glucose-6-phosphate dehydrogenase (G6P-DH) and NADH-diaphorase activities during different periods of the reproductive cycle. delta 5-3 beta-HSDH, 17 beta-HSDH, 11 beta-HSDH, G6P-DH and NADH-diaphorase activity was found in the theca interna of growing, atretic and postovulatory follicles, the granulosa of ovulatory, atretic and postovulatory follicles, and interstitial gland cells during the pre-incubation and the laying periods. During the incubation and squab feeding periods only delta 5-3 beta-HSDH, G6P-DH and NADH-diaphorase activities were observed in the above mentioned cells. The steroidogenic potential of atretic follicles depends upon the type of atresia a follicle undergoes.

Ovarian Interstitial Gland Tissue and Serum Progestin Levels during the First Periovulatory Period in the Mated Rabbit1

In order to determine the morphology of the interstitial gland tissue and possibly correlate these findings with the serum levels of 20α-dihydroprogesterone, mature New Zealand virgin rabbits were each assigned to 1 of 4 groups: estrous controls; 5 h; 10 h; and 2 days after mating. Data indicated that the amount of interstitial gland tissue in the ovaries varied between rabbits regardless of the group to which they were assigned. The virgin estrous rabbits generally had minimal amounts of interstitial gland tissue and low levels of serum 20α-dihydroprogesterone. Ovaries of some rabbits had an abundance of interstitial gland tissue which was composed mainly of large cells and had an output of 20α-dihydroprogesterone greater than those ovaries with a lesser amount of interstitial gland tissue. In those ovaries where an abundance of interstitial gland tissue was present, some morphological changes were seen between the 3 periovulatory stages which can be related to variations in 20α-dihydroprogesterone. At 5 h after mating, the ovaries with large amounts of interstitial gland tissue had cells with an abundance of smooth endoplasmic reticulum and serum 20α-dihydroprogesterone levels were elevated. At 10 h after mating, some follicles had ovulated and the large interstitial gland cells displayed an increase in numbers of lipid droplets which was accompanied by a decrease in 20α-dihydroprogesterone. At 2 days after mating, most large interstitial gland cells had an accumulation of large lipid droplets and progestin levels continued to decrease.

Ultrastructure of the cells of the ovarian interstitial gland in hypophysectomized rats. The effects of stimulation of the ovarian plexus and of denervation.

The fine structure of the interstitial gland of the ovary was studied in hypophysectomized rats and in hypophysectomized rats after denervation of the ovary or stimulation of the ovarian plexus. Hypophysectomized rats were used to eliminate gonadotropic influences on interstitial cells. In hypophysectomized rats, there was a large amount of intercellular space and cells had irregularly shaped nuclei and a large nuclear-cytoplasmic ratio. Prominent cytoplasmic features included small mitochondria with an electron-dense matrix, rough and smooth endoplasmic reticulum, polysomes and large osmiophilic lipid droplets. Interstitial cells from stimulated ovaries had reduced intercellular space and a reduced nuclear-cytoplasmic ratio. Mitochondria had tubular cristae; smooth endoplasmic reticulum-surrounded lipid droplets, and large polysomes were present. After section of the ovarian plexus, intercellular space was increased and filopodia were numerous. Cytoplasmic features included mitochondria with a dense matrix and indistinct cristae, large electronlucent lipid droplets, and variously sized multivesicular structures. These observations suggest that stimulation of the ovarian plexus in hypophysectomized rats causes regressed cells to assume the fine structural features of active steroidogenic cells. In contrast, interruption of the ovarian nerve supply causes a qualitative and quantitative increase in ultrastructural features characteristic of regressed steroidogenic cells. These responses of interstitial gland cells to denervation and stimulation provide morphological evidence for a functional role for the adrenergic nerves to this ovarian compartment.

Steroidogenic cellular sites in the cat ovary: A histochemical study

The distribution of lipids, NADH, NADPH diaphorase, glucose 6-phospate, the isocitrate dehydrogenases, Δ 5-3 β- and 17β-hydroxysteroid dehydrogenases, and acid phosphatase in immature, cycling, and pregnant cat ovaries has been histochemically studied. Corpora lutea were found only in pregnant cats. Acid phosphatase was present in the degenerating follicles and only in trace amounts in the corpora lutea and was absent in the interstitial gland cells, germinal epithelium, and the normal developing follicles. Lipids and the various enzymes studied were localized in the theca interna of preantral and antral follicles, interstitial gland cells, and in the corpora lutea. Germinal epithelium lacked lipids, diaphorases, and all the enzymes investigated. It is suggested that theca interna and the interstitial gland cells form principal sites of steroid synthesis in the ovaries of immature as well as cycling cats while corpora lutea form additional sites of steroidogenesis during pregnancy.

Effect of abdominal vagotomy of the pregnant rat on LH and progesterone concentrations and fetal resorption.

Abdominal vagotomy on Day 8 of pregnancy in rats decreased the number of live fetuses at Day 16 and increased the number of resorbing fetuses. The activity of delta5-3beta-hydroxysteroid dehydrogenase (3beta-HSD) in the corpus luteum and interstitial gland, LH and progesterone values in plasma and progesterone values in ovarian tissue were all lower in vagotomized rats than in sham-operated controls. Ovarian PGF levels were not affected. We suggest that these effects were caused by a direct effect of vagotomy on LH secretion which in turn lowers 3beta-HSD activity and progesterone levels in ovarian tissue and plasma, leading to fetal resorption.

Sites of production of sex steroids: secretion of steroids from x-irradiated and polycystic ovaries of rats.

Ovaries of immature rats and PMS-induced pregnant rats were unilaterally x-irradiated. Ten days later, the concentrations of pregnane compounds in the ovarian venous plasma were measured. LH (2 μg) was injected iv 30 min before bleeding. A comparison of steroid levels in the ovarian venous effluent of rats with and without destruction of selected tissue components by irradiation of the ovaries suggests that the follicles contribute to the secretion of 5α-pregnane-3,20-dione and 3α-hydroxy-5α-pregnan-20-one in the presence of interstitial gland tissue. Because it is known that follicular tissue is involved in the production of estrogens, we studied the interrelationship between the secretion of the two progesterone metabolites and estrogens in follicular polycystic ovaries of androgen-sterilized rats. Normal ovaries of diestrus-2 rats were used as controls for the polycystic ovaries. The injection of LH greatly increased the secretion of 5a-pregnane-3,20-dione and 3α-hydroxy-5α-pregnan-20-one within 1h in ...

Ultrastructure of rat ovarian interstitial gland cells during pregnancy.

The fine structure of the interstitial gland of the rat ovary was studied at estrus and on Days 4, 6, 10, 14 and 18 of pregnancy. At estrus, ovarian interstitial cells have small nuceli with dense irregular clumps of heterochromatin. Mitochondria are small and rod-shaped and have predominantely lamellar cristae. Numerous osmiophilic lipid droplets are present. At Days 4 and 6, nuclear heterochromatin is reduced, and nucleoli are larger and complex. Mitochondria are enlarged and often bizarre-shaped and have tubular cristae. Golgi and smooth endoplasmic reticulum are more conspicuous. At Day 10, prominent ultrastructural features include nuclei with conspicuous heterochromatin, smaller mitochondria with both lamellar and tubular cristae, numerous ribosomes and lipid droplets with decreased osmiophilia. At Days 14 and 18, nuclei have increased heterochromatin, mitochondria are small and have lamellated cristae and an increase in the size and number of lipid droplets occurs. These observations suggest that steroidogenic activity of interstitial cells is highest during the first half of pregnancy and regresses during the last half. It is suggested that the interstitial gland is an important ovarian source of pregnancy hormone(s) during the first half of gestation and that LH may modulate steroidogenic activity is this ovarian component.