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Fine structure of the lutein cell in the house musk shrew, Suncus murinus.

The ultrastructure of lutein cells during pregnancy and the post partum period was examined by transmission electron microscopy in the house musk shrew, Suncus murinus. Lutein cells on day 13 of pregnancy contained an extensive system of anastomosing tubules or cisternae of smooth ER and many enlarged mitochondria with numerous tubulovesicular cristae. From day 13 on, the number of small granules, possibly lysosomes, increased gradually. Between day 20 and 25, the loss of smooth ER began and elongated, or flattened mitochondria increased. Regressing lutein cells observed after parturition were characterized by abundant large dense bodies, bizarre mitochondria and a decrease in the amount of smooth ER. Unusual forms of mitochondria were always observed after day 5 of pregnancy. Two types could be distinguished; one, found frequently in the second third of pregnancy, was ring-, disc-, cup- or dumb-bell-shaped with tubulovesicular cristae, and the other, found exclusively in the last third of pregnancy and after parturition, was elongated, flattened and sometimes twisted. The paucity of lipid droplets was a characteristic feature of the lutein cells of this species. The significance of these ultrastructural changes of cellular organelles is discussed in relation to the ovarian and plasma levels of progesterone.

Animals↗

The expression of sf-1/Ad4BP is related to the process of luteinization in the marmoset (Callithrix jacchus) ovary.

Expression of the nuclear receptor, steroidogenic factor-1 (SF-1/Ad4BP), was studied in a primate (marmoset) ovary using immunohistochemical, RT-PCR, and immunoblot techniques. The periovulatory phase was compared with the luteal phase. With PCR we found a marmoset homolog of SF-1/Ad4BP to be expressed in ovarian and other steroidogenic tissues. Characteristically, the periovulatory ovaries consisted of growing (non-luteinized) small follicles together with large luteinizing follicles and many corpora lutea accessoria (Clas), which had developed from atretic large follicles. During the luteal phase, true corpora lutea (Cls) were additionally found. In general, we found that small follicles were devoid of any immunoreactivity of SF-1/Ad4BP. In large follicles, the luteinizing theca and granulosa cells express SF-1/Ad4BP. All luteal cells of Clas showed a nuclear staining in both ovary types. In Cls, only a few luteal cells were positive. Large follicles of different sizes showed no differences in expression level, as evidenced by immunoblot analysis. Our results indicate that SF-1/ Ad4BP participates in the activation of genc transcription during the onset of luteinization and that Clas are essential for ovarian luteal function.

Animals↗

Exaggerated gonadotropin response to luteinizing hormone-releasing hormone in amenorrheic runners.

Most studies of exercise-induced amenorrhea have compared amenorrheic athletes (usually runners) with sedentary control subjects. Such comparisons will identify hormonal changes that develop as a result of exercise training but cannot determine which of these changes play a role in causing amenorrhea. To obviate this problem, we assessed reproductive hormone status in a group of five amenorrheic runners and compared them to a group of six eumenorrheic runners matched for body fatness, training intensity, and exercise performance. Compared to the eumenorrheic runners, the amenorrheic runners had lower serum estradiol concentrations, similar basal serum luteinizing hormone and follicle-stimulating hormone concentrations, and exaggerated responses of serum gonadotropins after administration of luteinizing hormone-releasing hormone (100 micrograms intravenous bolus). Serum prolactin levels, both basally and after thyrotropin-releasing hormone administration (500 micrograms intravenous bolus) or treadmill exercise, was similar in the two groups, as were serum thyroid function tests (including thyrotropin response to thyrotropin-releasing hormone). Changes in serum cortisol levels after short-term treadmill exercise were similar in both groups, and serum testosterone levels increased after exercise only in the eumenorrheic group. In neither group did such exercise change serum luteinizing hormone, follicle-stimulating hormone, or thyrotropin levels. We concluded that exercise-induced amenorrhea is not solely related to the development of increased prolactin output after exercise training. The exaggerated gonadotropin response to luteinizing hormone-releasing hormone seen in amenorrheic runners in comparison with matched eumenorrheic runners is consistent with a hypothalamic etiology for the menstrual dysfunction, analogous to that previously described in "stress-induced" or "psychogenic" amenorrhea.

Amenorrhea↗

Proliferative index of human luteinized granulosa cells varies as a function of ovarian reserve.

OBJECTIVE: We examined whether the proliferative index of luteinized granulosa cells, as determined by flow cytometry, varied as a function of a woman's ovarian reserve, as reserve, as reflected by follicular-phase day 3 serum follicle-stimulating hormone. STUDY DESIGN: This prospective cohort study consisted of 19 women of similar chronologic age preparing for in vitro fertilization-embryo who met specific day 3 serum follicle-stimulating hormone criteria. The "low follicle-stimulating hormone" group consisted of 11 women with day 3 serum follicle-stimulating hormone levels < or = 6 IU/L. The "high follicle-stimulating hormone" group consisted of eight women with day 3 serum follicle-stimulating hormone levels > or = 18 IU/L. A total of 56 preovulatory follicles containing > or = 10(4) luteinized granulosa cells were examined by flow cytometry. The low follicle-stimulating hormone group was compared with the high follicle-stimulating hormone group to examine proliferative index as a function of serum day 3 follicle-stimulating hormone levels. RESULTS: The low follicle-stimulating hormone group had a greater proliferative index (11.1% +/- 0.4%) than did the high follicle-stimulating hormone group (8.3% +/- 0.6%), p < 0.001). This study demonstrates that in spite of the same chronologic age, luteinized granulosa cells from preovulatory follicles of women with day 3 serum follicle-stimulating hormone levels > or = 18 IU/L have a 25% decreased proliferative index compared with luteinized granulosa cells from women with day 3 serum follicle-stimulating hormone levels < or = 6. CONCLUSIONS: This suggests that granulosa cell proliferation is influenced by ovarian reserve and may explain in part the more favorable response to ovulation induction protocols that younger women demonstrate compared with women of more advanced reproductive age.

Adult↗

The presence of luteinized unruptured follicle syndrome and altered folliculogenesis in rats with surgically induced endometriosis.

OBJECTIVE: Our purpose was to determine in the rat model whether endometriosis could influence ovarian function by altering oocyte release or folliculogenesis. STUDY DESIGN: We histologically examined the ovaries of reproductively cycling rats with (n = 16) and without (n = 10) surgically induced endometriosis. The rats in these two groups were further subdivided into unilaterally ovariectomized or ovarian-intact groups. Serial sections of ovaries were examined, and follicular development and frequency of luteinized unruptured follicles were determined. RESULTS: A significant tenfold increase in the number of luteinized unruptured follicles was observed in the ovaries from rats with endometriosis (2.7 per rat) compared with unoperated and sham-operated control groups (overall mean 0.26 per rat, p < 0.05). Additionally, ovaries from unilateral ovariectomized animals with endometriosis contained four times as many luteinized unruptured follicles (four per rat) as did the ovaries from bilaterally ovarian-intact rats with endometriosis (1.40 per rat, p < 0.01). Fewer follicles were present in rats with endometriosis (180 follicles per ovary) than in control rats (231 follicles per ovary, p < 0.05). CONCLUSION: In the rat model the presence of ectopic endometrium is associated with an increased frequency of luteinized unruptured follicles and altered follicular development.

Animals↗

Active immunization to luteinizing hormone releasing hormone to inhibit the induction of mammary tumors in the rat.

Immunization of female rats with a bovine serum albumin-luteinizing hormone releasing hormone conjugate results in suppression of dimethylbenzanthracene mammary tumor incidence. Tumor incidence was 1.3, and 1.29 tumors per rat in bovine serum albumin alone (n = 10) and unimmunized (n = 18) control groups, but no tumors were found in the bovine serum albumin-luteinizing hormone releasing hormone conjugate immunized animals (n = 10). In a second experiment immunization with bovine serum albumin-luteinizing hormone releasing hormone conjugates reduced tumor incidence to 0.3 tumors per rat (n = 10) from the 1.2 tumors per animal seen in the control animals (n = 10) immunized with bovine serum albumin alone. Bovine serum albumin-luteinizing hormone immunization caused the production of anti-LHRH antibodies, an interruption of estrous cycles, lowered serum estradiol and progesterone levels, and atrophy of the ovaries and uteri. Immunization BSA-hormone conjugates is a novel anti-tumor strategy.

9,10-Dimethyl-1,2-benzanthracene↗

Lymphoma of ovary with stromal luteinization, presenting as secondary amenorrhea.

A case of primary lymphoma of the ovary with extensive stromal luteinization is presented. Immunohistochemical markers that work satisfactorily in fixed tissues allowed a diagnosis of B cell lymphoma to be made on formalin-fixed tissue sections. Immunohistochemical analysis of frozen tissue and genotypic analysis confirmed the diagnosis of B cell lymphoma. By electron microscopy two populations of cells, including large atypical lymphoid cells and luteinized stromal cells, were identified. Presentation with amenorrhea and resumption of menstrual periods after removal of the tumor suggested the possibility of functional activity in the luteinized stromal cells. Stromal luteinization has not been previously described in primary ovarian lymphoma.

Adult↗

Immunocytochemical demonstration of neuropeptide Y and luteinizing hormone-releasing hormone-immunoreactive structures in the organum vasculosum laminae terminalis of juvenile gilts.

Immunocytochemical investigations on the immature gilt organum vasculosum laminae terminalis showed extensive neuropeptide Y- and luteinizing hormone-releasing hormone-immunoreactive innervation of the organ. The luteinizing hormone-releasing hormone-containing varicose fibers ran along the organum vasculosum laminae terminalis in close association with blood vessels. The nerve processes originating from well-stained luteinizing hormone-releasing hormone-immunoreactive perikarya were distributed around the organum vasculosum laminae terminalis. The matrix of the organum vasculosum laminae terminalis was abundantly supplied by neuropeptide Y-immunoreactive varicose fibers. Numerous neuropeptide Y-immunoreactive terminals seemed to penetrate the ependymal lining of the organ. From these observations, it is concluded that there are favorable morphological conditions for secretion of neuropeptide Y into the cerebrospinal fluid of the third ventricle and release of luteinizing hormone-releasing hormone into fenestrated capillaries of the organ.

Animals↗

Application of the silver-gold intensified 3,3'-diaminobenzidine chromogen to the light and electron microscopic detection of the luteinizing hormone-releasing hormone system of the rat brain.

A highly sensitive, recently developed immunohistological method is introduced in the present study with special emphasis on the luteinizing hormone-releasing hormone system of the rat brain. The method utilizes the specific capability of the diaminobenzidine endproduct, a frequently used chromogen in immunocytochemistry, to produce and bind silver grains from a special physical developer, following suppression of the argyrophilia of the nervous tissue by thioglycolic acid. Metal deposition into the immunolabelled structures results in a real Golgi-like appearance of immunoreactive profiles. Specificity of this silver method was confirmed by ultrastructural analysis, which showed that unlabelled elements did not bind silver. Using this method, more immunoreactive neurons and fibres were visualized than compared with the results of the traditional peroxidase-antiperoxidase method. The luteinizing hormone-releasing hormone-synthesizing neurons proved to be fusiform, exhibiting either smooth or rough surfaced contour. Unlabelled terminals established axo-somatic synapses on labelled perikarya. The juxtaposition of immunoreactive luteinizing hormone-releasing hormone profiles suggest the possibility of self-regulation within the luteinizing hormone-releasing hormone system. The main advantages of the method are the increased sensitivity with preserved selectivity and wide applicability in different fields of neuroscience (peptide and transmitter immunocytochemistry, double labelling, horseradish peroxidase tract tracing, X-ray analysis).

3,3'-Diaminobenzidine↗

Axovascular relationships in developing median eminence of perinatal rats with special reference to luteinizing hormone-releasing hormone projections.

Topographical relationships of neurosecretory axons with the capillaries of the primary portal plexus were studied in the median eminence of rats from the 14th fetal till the 9th postnatal day by means of electron microscopy combined with morphometric analysis. Special attention was given to the light and electron microscopic immunocytochemical examination of luteinizing hormone-releasing hormone projections to the median eminence. Neurosecretory axons possessing secretory granules and clear microvesicles were first observed in the median eminence at the 14th fetal day. However, all of them were situated at a distance from the primary portal plexus. By the 20th fetal day, neurosecretory axons reached the external basal lamina of the primary portal plexus giving rise to so-called axovascular contacts. Some axons even penetrated into the perivascular space, apparently facilitating the neurohormone delivery into the hypophysial portal circulation. From that time on, both the number of the axons abutting on the external basal lamina and the entire area of axovascular contacts increased gradually. As to luteinizing hormone-releasing hormone axons, they grew into the median eminence from the 18th fetal day concentrating in older fetuses and neonates either over the primary portal plexus, or around the infundibular recess of the 3rd ventricle. After birth, the concentration and distribution of luteinizing hormone-releasing hormone axons within the median eminence became similar to those of adults. Luteinizing hormone-releasing hormone axons were found to arise from the neurons of septopreoptic area including the diagonal band of Broca. These data suggest the onset of neurohormone release in the median eminence from the 14th fetal day followed by the establishment of the hypothalamic control over the pituitary functions during the perinatal period in rats.

Animals↗

Acetylcholinesterase and luteinizing hormone-releasing hormone distinguish separate populations of terminal nerve neurons.

The terminal nerve is composed of a morphologically heterogeneous population of unipolar, bipolar and multipolar neurons located in the nasal and intracranial cavities of vertebrates. The question has arisen as to whether these neurons are neurochemically heterogeneous and therefore possibly functionally different as well. Among the substances localized in the terminal nerve are acetylcholinesterase and luteinizing hormone-releasing hormone-like immunoreactive material. We have developed a double-label procedure, combining immunocytochemistry and enzyme histochemistry to determine whether these two substances are localized within different populations of terminal nerve neurons. Compatibility of the two procedures was accomplished by modifications of the fixative and primary antibody solutions. In the immunocytochemical step, the avidin-biotin-peroxidase complex coupled to a new chromogen, Chromo-red, produced a bright red reaction product in neurons containing luteinizing hormone-releasing hormone-like material. This reaction product was easily differentiated from the black silver-intensified acetylcholinesterase label. In both neonatal and adult preparations, a large population of terminal neurons contained the acetylcholinesterase label only, whereas a smaller population contained both acetylcholinesterase and luteinizing hormone-releasing hormone-like material. The acetylcholinesterase-containing population of neurons was concentrated peripherally and included multipolar neurons. In contrast neurons with the two substances co-localized were unipolar or bipolar and were concentrated centrally. The simultaneous visualization of acetylcholinesterase and luteinizing hormone-releasing hormone-like material in the same tissue section enable the differentiation of two separate neurochemically defined populations of terminal neurons. The distribution of these two neuronal types was the same in neonatal and adult animals. These data provide support for a functional diversity of terminal neurons.

Acetylcholinesterase↗

Modelling the luteinizing hormone-releasing hormone pulse generator.

Pituitary hormones are released in pulses as a result of episodic patterns of electrical activity in neuroendocrine neurons. The mechanisms underlying such pulsatility have, however, been difficult to elucidate. For example, the luteinizing hormone-releasing hormone neurons regulating reproductive functioning have a sparse and scattered distribution within the hypothalamus which has made definitive electrophysiological investigation impracticable. Little is known not only of their electrical characteristics, but also of the critical neural components with which they interact to form the so-called "luteinizing hormone-releasing hormone pulse generator". We have used here a neural modelling approach, based on the FitzHugh-Nagumo model of a single neuron, to provide a simple dynamical network model of this neuroendocrine pulse generator. We have found that the minimal components required to generate pulsatile luteinizing hormone secretion arise from combining luteinizing hormone-releasing hormone neurons with reciprocally connected inhibitory interneurons and an external stimulatory input. Local GABA neurons and ascending noradrenergic and/or adrenergic inputs have been used as the biological basis for these respective components. The network displays a wide repertoire of behaviours comparable with experimental observations, including some thought previously to be paradoxical. The capacity of this model network to display complex behavioural features interpretable against experimental evidence suggests that this type of modelling may become a necessary adjunct to empirical studies of pulsatile neuroendocrine systems.

Action Potentials↗

Solitary luteinized follicle cyst of pregnancy and puerperium.

We report a huge ovarian solitary luteinized follicle cyst (SLFC) that contained 17 liters of fluid. Computed tomography scan revealed a 32 x 32 x 20 cm right simple cystic ovarian mass without calcification or septation. The fluid was removed, and the collapsed unilocular cyst was examined pathologically. No solid or papillary areas were identified. The cyst wall lining was composed of multiple layers of luteinized cells of variable size and shape with abundant eosinophilic cytoplasm and focally enlarged, pleomorphic, and hyperchromatic nuclei. Nests of luteinized cells were embedded within the fibrous tissue of the cyst wall. These findings are characteristic of solitary luteinized follicle cyst of pregnancy. Even though less than 15 cysts of this kind have been recorded, one should consider this uncommon, benign, non-neoplastic cystic ovarian lesion in the differential diagnosis of other cystic ovarian lesions.

Adolescent↗

Transforming growth factor-beta regulates the expression of luteinizing hormone receptors in ovarian granulosa cells.

In cultured granulosa cells, addition of 1 to 50 ng follicle-stimulating hormone induced a 350-fold rise in luteinizing hormone receptors, while larger amounts of gonadotropin up to 200 ng reduced these receptors to approximately 50% of peak levels. Transforming growth factor-beta (16 pM) enhanced the stimulatory actions of low levels of gonadotropin (2.5-10 ng) by 2 to 3-fold, and inhibited the induction of luteinizing hormone receptors by higher levels of follicle-stimulating hormone (greater than or equal to 50 ng) by 30-50%. The actions of the growth factor were concentration-dependent over the range from 0.8 to 16 pM and included a similar biphasic effect upon gonadotropin-induced cAMP production. Modulation of cAMP formation and luteinizing hormone receptor expression by transforming growth factor-beta could influence the ability of the granulosa cell to respond to luteinizing hormone during ovarian follicular maturation and ovulation.

Animals↗

Depletion of Raf-1 protooncogene by geldanamycin causes apoptosis in human luteinized granulosa cells.

OBJECTIVE: To investigate the hypothesis that epidermal growth factor (EGF) signaling in luteinized granulosa cells works through Raf-1 and mitogen-activated protein (MAP) kinases and that depletion of Raf-1 by geldanamycin will inhibit the signaling pathway and cause apoptosis. DESIGN: Laboratory study. SETTING: University of Minnesota. PATIENT(S): Human luteinized granulosa cells from IVF patients. INTERVENTION(S): The cells were treated with vehicle (DMSO), 0.5 microM of geldanamycin, 10 ng/mL of EGF, and geldanamycin + EGF. MAIN OUTCOME MEASURE(S): Radiochemical MAP kinase assay, Western blotting, confocal microscopy, and flow cytometry. RESULT(S): Geldanamycin treatment depleted Raf-1 and lowered MAP kinase activity in luteinized granulosa cells. EGF treatment increased MAP kinase phosphorylation and translocation of the phosphorylated MAP kinase to the nucleus. Geldanamycin blocked this effect. Cleavage of caspase-3, the executioner protein in apoptosis, into an active 17 kD fragment was observed by Western blotting in geldanamycin-treated cells. Finally, by flow cytometry we observed significantly increased percentages of subdiploid apoptotic nuclei in geldanamycin-treated cells. CONCLUSION(S): In human luteinized granulosa cells, EGF works through Raf-1, and MAP kinase and depletion of Raf-1 by geldanamycin resulted in decreased MAP kinase activity, increased activated caspase-3, and, ultimately, apoptosis.

Apoptosis↗

Ryudocan expression by luteinized granulosa cells is associated with the process of follicle atresia.

OBJECTIVE: To evaluate the presence of ryudocan in follicular fluid (FF) and its possible correlation with FF E(2) and P, and to study the levels of ryudocan in granulosa-lutein cells stimulated with hCG. DESIGN: Controlled clinical study and in vitro experiment. SETTING: University teaching hospital. PATIENT(S): One hundred seven patients undergoing IVF. INTERVENTION(S): The FF and granulosa-lutein cells were aspirated from follicles 34 hours after an ovulatory gonadotropin bolus. MAIN OUTCOME MEASURE(S): FF ryudocan, E(2), and P levels as well as hCG-mediated induction of ryudocan. RESULT(S): Ryudocan was abundant in the FF; the concentration of ryudocan in human FF was estimated to be 305.5 +/- 200.8 ng/mL (mean +/- SD). Atretic follicles had higher concentrations of ryudocan (559.1 +/- 156.5 ng/mL). FF ryudocan levels were inversely correlated with FF E(2) (r = -0.5023) and P concentrations (r = -0.4459). A detectable amount of ryudocan was found in pooled granulosa-lutein cells. Ryudocan production was augmented by surge levels of hCG. CONCLUSION(S): Ryudocan is expressed in luteinized granulosa cells in vitro. The higher concentrations of ryudocan in FF of atretic follicles suggest an involvement of ryudocan in the process of atresia.

Adult↗

Detection of premature luteinization with serum progesterone levels at the time of the postcoital test.

Poor cervical mucus (CM) may be caused by a number of factors, including premature luteinization, local cervical effects, and inadequate folliculogenesis. In an attempt to distinguish between these causes of poor CM, we obtained progesterone (P) levels at the time of postcoital tests (PCTs) in infertile women during spontaneous or clomiphene citrate (CC)-stimulated cycles. The amount of CM, viscosity, ferning, spinnbarkeit, and cellularity were each scored from 0 to 3 points on the day after detection of the urinary luteinizing hormone surge (luteal day 1). The charts of 46 such patients were retrospectively reviewed. Eleven control patients with good CM scores (greater than 10) had low P levels (less than 2.5 ng/mL). Of the remaining 35 cycles, 19 were marked by low P levels, and 16 were accompanied by P levels of greater than 2.5 ng/mL. Overall, 94.4% of CC-stimulated cycles versus 64.3% of spontaneous cycles had abnormal CM scores (less than 9). On the basis of these inappropriately elevated P levels, premature luteinization can be cited as the cause of poor mucus quality. In fact, premature luteinization may be responsible for some of the purported antiestrogenic effects of CC. Therefore, it is appropriate to draw a P level at the time of a poor PCT, particularly in cycles stimulated by CC.

Cervix Mucus↗

Results of in vitro fertilization and embryo transfer by combined long-acting gonadotropin-releasing hormone analog D-Trp-6-luteinizing hormone-releasing hormone and gonadotropins.

To avoid cancellation of in vitro fertilization (IVF) because of early luteinization, pituitary suppression by gonadotropin-releasing hormone (GnRH) was carried out in 111 cycles. D-Trp-6-luteinizing hormone-releasing hormone (LH-RH) microcapsules were administered intramuscularly at menstruation and menotropin (hMG) stimulation was started 19 days (mean) later. In 3 cycles (2.7%), only early luteinization occurred. The mean number of oocytes per cycle was 6.7, with a fertilization and cleavage rate of 50 and 95%, respectively. A mean of 3.4 embryos were transferred per cycle. The 111 cycles resulted in 34 clinical pregnancies, 41% per cycle with embryo transfer. The early abortion, multiple pregnancy, and ovarian hyperstimulation rates were 24, 18, and 11%, respectively. It is concluded that D-Trp-6-LH-RH/hMG cycles are associated with a very low occurrence of early luteinization, high number of oocytes and embryos, and a substantial incidence of ovarian hyperstimulation syndrome.

Adult↗