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Biomedical subjects

H R Lindner

Publications and source records attributed to H R Lindner.

At least 37 records · Page 2Linked to original sources

Suppression of 20 alpha-hydroxysteroid dehydrogenase activity in cultured rat luteal cells by prolactin.

20alpha-Hydroxysteroid dehydrogenase (20 alpha-SDH) activity increases in the cycling corpus luteum of the rat, beginning at 14.00 h on the day of diestrus, but remains low in corpora lutea of pregnancy throughout the first 19 days of gestation. When cells derived from 7-day-old corpora lutea of pregnant rats were cultured for 7 or 12 days, there was a spontaneous rise in 20 alpha-SDH activity from an initial value of 0.44 +/- 0.27 to 4.1 +/- 0.7 units/mg supernatant protein. Addition of LH (NIH-S-18; 2.0 mug/ml) or prostaglandin F2alpha (2.8 X 10(-5) M) to the medium from day 4 to the end of incubation period caused a slight but significant reduction in 20 alpha-SDH activity (20%, P less than 0.05). Supplementation of the medium with ovine prolactin (HIH-P-S11; 10.0 mug/ml) from the time of seeding or from the 2nd to 4th day of culture reduced the activity of 20 alpha-SDH measured on day 12 by 61% (P less than 0.001). This finding suggests that the suppression of 20 alpha-SDH by prolactin, hitherto demonstrated only in vivo results from a direct action of the hormone on the luteal cell.

Animals

Acute effect of delta1-tetrahydrocannabinol on the hypothalamo-pituitary-ovarian axis in the rat.

Administration of delta1-tetrahydrocannabinol (delta1-THC), the principal psychoactive ingredient of cannabis, to proestrous rats (2 mg/rat, i.p., between 12.00 and 16.00 h) suppressed the proestrous rise in the plasma levels of LH, FSH and prolactin (Prl) and caused a 24 h delay in ovulation. Furthermore, the increased accumulation of prostaglandins of the E-type (PGE) in the ovaries, normally seen on the evening of proestrus, was prevented. Earlier (08.00-10.30 h) or later (18.00 h) administration of the drug on the day of proestrus was only partially effective in inhibiting ovulation. The suppressive effects of delta1-THC on ovulation and gonadotropin secretion were prevented by administration of gonadetropin releasing hormone (GnRH, 0.2 microgram/rat) 1 h after the drug, indicating that the central action of delta1-THC was exerted on the hypothalamus and not on the pituitary gland. Administration of ovine luteinizing hormore (oLH, 2.5 microgram/rat at 16.30 h on the day of proestrus restored ovulation and ovarian PGE accumulation in Nembutal-treated rats, but not in delta1-THC-treated rats; higher doses of oLH (5-10 microgram/rat) reversed the action of delta1-THC on these two parameters.

Animals

Endocrine and reproductive repercussions of immunization against progesterone and oestradiol in female rats.

Mature female rats were immunized with BSA-conjugates of oestradiol-17beta-6-carboxymethyloxime or progesterone-11-hymisuccinate. Sera tested two months later at 1/2000 dilution bound 63.3% +/- 3.1 SEM and 21.2% +/- 2.3 of the homologous tritiated hormone (10 pg),--oestradiol-17beta(Oe2) and progesterone (P), respectively. Rats immunized against Oe2 had a significantly longer oestrous cycle (8.2 days vs. 4.5 days in BSA-immunized rats), with a prolonged leukocytic phase. The cycle of rats immunized against P was also prolonged (10.5 days), but in this group the cornified smear phase was abnormally extended. The luteinizing hormone (LH) normally observed on the afternoon of pro-oestrus failed to occur in the rats immunized against Oe2 and in 70% of rats immunized against P. The latter animals, however, showed sporadic LH discharges during periods of persistent cornification. Immunization against P resulted in elevated total (free plus bound) plasma P levels; immunization against Oe2 prevented the pro-oestrous rise in plasma P. Mating of rats immunized against Oe2 resulated in normal pregnanices, while none of the rats immunized against P delivered.

Animals

Differentiation of decidual cells in cultures of rat endometrium.

Endometrial scrapings were collected from rat uteri at various times (0-4 days) after induction of a decidual reaction by i.p. injection of pyrathiazine hydrochloride (20 mg/animal) on the 5th day after a sterile mating. The tissue was dissociated by treatment with trypsin and the cells were cultured as monolayers. The differentiation of decidual cells was followed in these cultures. Two morphologically distinct cell populations were recognized: (i) dispersed spindle-shaped or stellate cells, and (ii) colonies of closely packed polygonal cells, first apparent after 48 h in culture. During culture, [3H] thymidine was incorporated into the nuclei of both cell types, as indicated by autoradiography. There was an increase in the number of cells in the colonies as culture progressed; most mitotic figures and the highest % of labelled nuclei were located within the colonies. Bi- and multinucleated cells, which are a characteristic feature of decidual tissue in vivo, appeared in the colonies on the 3rd day of culture and constituted about 60% of the colony population after 4-5 days. The dispersed cells showed a doubling in nuclear area during 4 dyas in culture, suggesting the formation of polyploid cells; such cells are prominent in fully differentiated decidual cells in vivo. The content of prostaglandin E in the cultured cell, determined by radioimmunoassay, was about 15-fold higher than that in rat embryo fibroblast cultures grown under similar conditions, and was comparable to the level of prostaglandins found in decidual cells in vivo.

Animals

Changes in steroid concentration in the ovaries of immature rats treated with pregnant mare serum gonadotrophin and human chorionic gonadotrophin.

The concentrations of testosterone, progesterone and 20alpha-hydroxypregn-4-en-3-one (20alpha-OHP) were measured in the ovaries of immature rats in which ovulation was induced by treatment with pregnant mare serum gonadotrophin (PMSG) and, 48 h later, with human chorionic gonadotrophin (HCG). The concentration of testosterone in the tissue increased significantly 48 h after treatment with PMSG, reached a peak 4 h after the administration of HCG and declined to the basal level 4 h later. Increases in the levels of progesterone and 20alpha-OHP were observed 4 h after the administration of HCG. Whereas the level of 20alpha-OHP continued to rise during the subsequent 30 h, progesterone levels declined near the presumed time of ovulation (12 h after administration of HCG). It is concluded that 20alpha-hydroxysteroid dehydrogenase activity is present in the immature rat ovary before ovulation and that an increase in the production of testosterone in the ovaries of rats treated with PMSG and HCG precedes increased production of progesterone and 20alpha-OHP in these ovaries.

20-alpha-Dihydroprogesterone

Synchronous generation of ovarian hCG binding sites and LH-sensitive adenylate cyclase in immature rats following treatment with pregnant mare serum gonadotropin.

A concomitant increase in the activity of LH-senstive adenylate cyclase and in the number of LH/hCG binding sites was induced in ovaries of immature rats upon administration of pregnant mare serum gonadotropin (PMSG), a hormone preparation known to have predominantly follicle stimulation (FSH-like) activity. When an optimal dose of PMSG (15 i.u./rat) was administered to 25-day-old rats, specific activity of LH-dependent adenylate cyclase and the number of binding sites for LH/hCG per mg protein remained unchanged during the first 24h, but 48h after injection a 2-to 4-fold increase in both parameters was observed. By contrast, there was no change in basal adenylate cyclase activity or in the response of the enzyme to the stimulatory action of guanosine-5'-(beta gamma-imino) triphosphate (Gpp (NH)p), GTP, or NaF. Specific activity of succinate cytochrome c reductase, glucose-6-phosphatase and 5'-nucleotidase were found to be unaffected by the hormonal pretreatment, although total protein determined in these homogenates increased 3-fold in the course of this treatment. It is inferred that during follicular maturation, FSH enhances the responsiveness of ovarian adenylate cyclase to LH by stimulating the insertion of LH/hCG-receptors into the cell membrane.

Adenylyl Cyclases

Refractoriness of ovarian adenylate cyclase to continued hormonal stimulation.

Culture of preovulatory rat follicles with luteinizing hormone, follicle-stimulating hormone or prostaglandin E2 for 24 h reduced the subsequent response of adenylate cyclase to the homologous by 80, 50 and 90%, respectively; yet follicles refractory to luteinizing hormone fully responded to follicle-stimulating hormone responded to luteinizing hormone and prostaglandin E2, and those refractory to prostaglandin E2 could be stimulated by either gonadotropin. Desensitization of the adenylate cyclase system by luteinizing hormone was achieved by hormone concentrations of 0.8--2.0 mug/ml in the medium; a lower dose of luteinizing hormone (0.4 mug/ml), though effective in stimulating adenylate cyclase, did not induce refractoriness. Prostaglandin E2 caused partial refractoriness at dose levels of 0.1--0.25 mug/ml; higher dose levels were more effective. These findings suggest that continued exposure to the preovulatory follicle to elevated levels of hormones may cause perturbations in either the interaction between the hormone and its specific receptor or in a subsequent step essential for activation of adenylate cyclase.

Adenylyl Cyclases

Demonstration of 8 S-cytoplasmic oestrogen receptor in rat mullerian duct.

1. High affinity macromolecular binding of the non-steroidal synthetic oestrogen [3H]diethylstilboestrol and of [3H] oestradiol-17beta in cytosol of Müllerian duct and uterus, and in blood plasma of perinatal rats, was investigated by sucrose density gradient sedimentation. 2. While [3H] oestradiol was bound to both the characteristic 8 S uterine cytoplasmic receptor and a 4 S component of uterine cytosol and plasma of 11-day-old rats, [3H] diethylstilboestrol was bound almost exclusively by the 8 S cytoplasmic receptor. 3. The greatly reduced binding of [3H] diethylstilboestrol to the 4 S plasma plasmic receptor in the Müllerian duct (precursor of the uterus) of 20-day-old foetuses.

Animals

Capacity of immunologically purified FSH to stimulate cyclic AMP accumulation and steroidogenesis in Graafian follicles and to induce ovum maturation and ovulation in the rat.

Reference preparations of ovine follicle-stimulating hormone (NIH-FSH-S8 and S9; 10-50 mug/ml) induced ovum maturation and stimulated cyclic AMP formation, as well as progesterone and 17beta-estradiol secretion, by rat Graafian follicles in vitro. These actions of NIH-FSH were retained after immunoabsorption of any contaminating luteinizing hormone (LH) present in the preparations, by treatment with an antiserum to the beta-subunit of purified ovine LH (anti-betaLH). In contrast, the corresponding biological actions of NIH-LH-S18 (0.5-10 mug/ml) were abolished by treatment with this anti-betaLH serum. A highly purified FSH preparation (64-96 CD, 0.25 mug/ml) also triggered oocytic meiosis and increased follicular progesterone secretion in vitro. Intraperitoneal (ip) administration of anti-betaLH-treated NIH-FSH-S9 (50 mug/rat at 1430 h) consistently induced ovulation in proestrous rats in which the endogenous gonadotropin surge had been blocked by ip injection of either Nembutal (1345 h) or antiserum to the LH-releasing hormone (1200 h). Injection (ip) of anti-betaLH serum on its own into proestrous rats at 1200 h prevented ovum maturation and follicular rupture. We conclude that currently available reference preparations of ovine FSH possess the capacity to stimulate follicular adenylate cyclase, steroidogenesis, and ovum maturation in vitro, as well as ovulation in vivo, in the rat, and that this capacity cannot be attributed to contamination with material immunochemically identical with LH. However, it is inferred that the physiological triggering of ovulation and related events in this species depends principally on LH.

Animals

Migraine attacks. Alleviation by an inhibitor of prostaglandin synthesis and action.

Twenty-six patients with migraine attacks were treated for 3 to 16 months with flufenamic acid (125 mg four to six times per attack), an inhibitor of prostaglandin synthesis and action. In 25 patients the drug afforded symptomatic relief in 195 of 200 treated attacks. Side effects observed were mild dyspepsia (eight patients) and severe upper gastrointestinal symptoms (two patients). None of the eight patients treated with placebo reported any relief (20 attacks). The "common" antimigraine drugs afforded symptomatic relief in 12 of the patients, partial relief in seven, and no relief in seven. Treatment with flufenamic acid was based on the hypothesis that prostaglandins are involved in migraine attack and that the drug relieves migraine by inhibition of the vasoactivity of prostaglandins.

Adolescent

Organization of intramembrane particles in freeze-cleaved gap junctions of rat graafian rollicles: optical-diffraction analysis.

Gap junctions were identified in the membrana granulosa and cumulus oophorus, and between cells of the internal theca, of the preovulatory rat follicle. In replicas of freeze fractured follicles, the A face presented clusters of closely packed intramembrane particles, 7--9 nm in diameter, forming a mosaic pattern, while the B face showed a similar pattern of small pits. Optical diffraction analysis of these electron micrographs revealed that both the intramembrane particles and the corresponding pits were organized in hexagonal lattices with centre-to-centre spacing of 9-10 nm. In small junctions (up to 0.4 mum in diameter), both A and B faces generally consisted of a single lattice, while large junctions (0.5-2 mum) contained multiple lattices differing in orientation. Occasionally intramembrane particles and pits were more loosely arranged throughout the junctional area and failed to show a hexagonal pattern. Both granulosa and thecal cells often contained intracellular vesicles resembling annular junctions. These contained intramembrane particles whose assembly resembled that of the gap junctions with regard to periodicity and lattice organization. Examination of thin section of thin sections suggested that small gap junctions occur also between cytoplasmic processes of coronal cells and the oolemma. No tight junctions were detected between granulosa cells and between thecal cells.

Animals

Dissociation between LH-induced aerobic glycolysis and oocyte maturation in cultured Graafian follicles of the rat.

Luteinizing hormone (NIH-LH-S18; 5 mug/ml) stimulated aerobic glycolysis in cultured Graafian follicles explanted from pro-oestrous rats before the preovulatory gonadotrophin surge: lactate accumulation in the medium was 70% above control levels during 6 h incubations. Iodoacetate (2.5 x 10(-5) M) prevented this effect, without impairing the ability of LH to induce resumption of oocytic meiosis. Enrichment of the medium with pyruvate (3.3 x 10(-4) M) or lactate (2.5 x 10(-2) M) did not in itself cause ovum maturation. The results do not support the hypothesis that termination of meiotic arrest by LH is due to stimulation of glycolytic activity in the follicle cells, resulting in increased availability of an energy source readily urilizable by the oocyte.

Animals

Oocytic meiosis in cultured rat follicles during inhibition of steroidogenesis.

Steroid release by cultured Graafian follicles explanted from rat ovaries on the morning of pro-oestrus was measured by radioimmunoaso the medium reduced the basal level of steroid secretion (progesterone, androstenedione and oestradiol-17beta) and abolished the steroidogenic effect of luteinizing hormone (LH; 5mug/ml) and prostaglandin E2 (PGE2; 10 mug/ml). However, the induction of oocyte maturation by either LH or PGE2 was not impaired by total suppression of the steroidogenic response of the follicles to these hormones by cyanoketone or aminoglutethimide. It is concluded that the meiosis-inducing action of LH on the mammalian egg is not mediated by the effect of the hormone on the rate and pattern of follicular steroidogenesis.

Aminoglutethimide