Biomedical subjects

P Licht

Publications and source records attributed to P Licht.

At least 91 records · Page 5Linked to original sources

PubMed1990

Modulation of neuropeptide-stimulated pituitary hormone secretion in hatchling turtles.

Neuropeptides that have relatively narrow actions on mammalian pituitary secretion may have divergent effects on pituitary hormone secretion in ectothermal vertebrates. In turtles, secretion of both thyrotropin (TSH) and growth hormone (GH) can be stimulated in vitro by thyrotropin-releasing hormone (TRH) and by members of corticotropin-releasing hormone (CRH) and growth hormone-releasing hormone (GHRH) peptide families. To determine if these neuropeptides share common modes of action, and to study other potential regulators of the turtle pituitary, somatostatin-14 (SRIH) and monoamines were tested for direct effects on in vitro basal and neuropeptide-stimulated TSH and GH secretion. Pituitary glands from young turtles (Pseudemys scripta) were cultured in the presence of 25 nM TRH, ovine CRH, or rat GHRH with or without SRIH. Glands were incubated for several 2-hr periods in medium alone or in medium containing peptides. Preincubation for 4 hr with SRIH (6 or 60 nM) significantly reduced basal and TRH-stimulated TSH and GH output (SRIH present during entire incubation). In another experiment, basal hormone secretion was reduced when SRIH (60 nM) was present only during the 2-hr basal period; however, reduction of TSH and GH responses to TRH required the presence of SRIH (60 nM) during the basal period and the period of stimulation. TSH responses to 25 nM oCRH and rGHRH and GH responses to rGHRH were significantly reduced by preincubation with 60 nM SRIH. The biogenic amines, dopamine (DA), serotonin (5HT), and norepinephrine (NE) (50 or 500 nM) were tested for possible direct actions on basal and neuropeptide-stimulated pituitary TSH and GH secretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1990

Effect of gonadotropin-releasing hormone on gonadotropin biosynthesis in pituitaries of the frog, Rana pipiens.

Previous studies indicated that the chronic elevation in LH and FSH secretion from frog pituitaries stimulated with GnRH is dependent upon continued protein synthesis. To determine if GnRH is able to stimulate the de novo synthesis of gonadotropins (Gth: FSH and LH), anterior pituitaries from frogs were incubated in vitro with 35S-methionine in the presence or absence of 100 ng/ml GnRH. Glands stimulated with GnRH secreted elevated levels of FSH and LH, including higher amounts of newly synthesized Gth. Continual exposure to GnRH for 8 h did not affect the pituitary content of either immunoassayable Gth (labeled + unlabeled) or newly synthesized Gth. However, GnRH stimulation did increase the total amount (secreted + pituitary) of newly synthesized Gth. GnRH did not stimulate Gth synthesis in females. The temporal pattern of GnRH-stimulated secretion of newly synthesized Gth from male pituitaries was also determined. A lag period of 60-90 min occurred prior to appreciable secretion of radiolabeled Gth. Secretion of radiolabeled hormone then increased and reached a plateau before decreasing. After 6 h of stimulation, approximately 30% of the radiolabeled Gth still remained within the pituitary. These results are the first direct evidence that GnRH can stimulate the biosynthesis of Gth in a nonmammalian species.

Animals↗

PubMed1990

Testosterone influences hibernation in golden-mantled ground squirrels.

At different phases of the hibernation season, castrated male golden-mantled ground squirrels were implanted with capsules that either were filled with testosterone (T) or left empty (blank). Blank-treated animals hibernated normally when housed at 5 degrees C. Entry into hibernation was prevented in the majority of squirrels treated with T several days before the initial cold challenge. T concentrations that inhibited torpor (greater than 1.2 ng/ml) were comparable with those of intact males at the end of the hibernation season. In some squirrels, moderate T concentrations were compatible with hibernation, but torpor bout duration was shorter than normal. The inhibitory effect of T on hibernation did not appear to require aromatization of T to estradiol. We suggest that a steroid-independent mechanism triggers arousal from hibernation and that T-dependent processes determine whether hibernation is resumed at the end of an arousal period.

Animals↗

PubMed1990

Regulation of pituitary thyrotropin secretion.

The pituitary thyrotropin secreting cells of tetrapods share a common responsiveness to TRH, but the specificity of this response may differ among species. TSH responses in amphibians and reptiles appear to be somewhat less specific than in mammals, but each group differs in responsiveness to particular neuropeptides, and it is not clear that TRH is the main regulator of TSH in nonmammalian species. It is not yet known whether these diverse peptides are acting directly on the thyrotrope or possibly through a paracrine mechanism. A CRH-like peptide may be a common neuroregulator of the anuran thyroid and interrenal in metamorphosis. Inhibitory regulation including effects of neuropeptides and thyroid feedback at the pituitary level is evident in reptiles, but is not known for other nonmammalian groups. In ectotherms, the potential actions of temperature on TSH secretion must also be considered. In this regard, it is important to recognize that the thermal relations of the thyrotropin response may be distinct from other pituitary hormones, as well as other components of the hypothalamo-pituitary-thyroid axis (Licht et al., 1989).

Animals↗

PubMed1989

Immunocytochemical and ultrastructural study of the frog (Rana pipiens) pars distalis with special reference to folliculo-stellate cell function during in vitro superfusion.

The colloidal gold immunocytochemical technique was used to determine the ultrastructural features of the glandular cells in the pituitaries of male frogs, Rana pipiens, both in vivo and after superfusion in vitro. Specific reactions to antisera against bullfrog gonadotropins, human prolactin, and synthetic 1-39 corticotropin allowed identification of the 3 corresponding types of glandular cells. No immunoreaction was obtained with antisera against human or ovine-growth hormone, human beta-thyrotropin hormone, and bovine S-100 protein. General morphological features of these immunocytochemically identified glandular cells were similar to those of equivalent cells previously described in other amphibian species. Non-glandular folliculo-stellate cells were distinctive. In freshly removed pituitaries, these folliculo-stellate cells contained lysosome-like structures, but did not show phagocytic vacuoles in the cytoplasm; they contained many mitochondria, and the Golgi complex and endoplasmic reticulum were relatively undeveloped. After 4 or 18 h of superfusion, some immunoreactive gonadotropic, prolactin, and corticotropic cells showed degeneration and destruction. In the same gland, folliculo-stellate cells retained a viable appearance, but showed phagocytic vacuoles containing secretory granule-like structures which were immunoreactive to gonadotropic, prolactin, and corticotropic antibodies. Some folliculo-stellate cells showed phagocytic vacuoles containing complete glandular cells. These results suggest that superfusion causes a destruction of some of the glandular cells, and that folliculo-stellate cells act as phagocytes when cellular debris or moribund cells are present in the intercellular space in the pituitary parenchyma.

Adrenocorticotropic Hormone↗

PubMed1989

Increased levels of arginine vasotocin and neurophysin during nesting in sea turtles.

Arginine vasotocin (AVT) and neurophysin (NP) levels were measured by radioimmunoassay in two species of sea turtle, the olive ridley, Lepidochelys olivacea, and the loggerhead, Caretta caretta, during the brief period of nesting and oviposition. In both species, AVT was low in animals which were not reproductively active. AVT was also low at the time animals emerged from the surf to nest, but increased significantly during oviposition and then declined as the animals returned to the water. NP increased in concert with AVT, also reaching highest levels during oviposition. In both species, however, NP levels remained elevated over prenesting levels at the time of return to the water. These findings are consistent with the hypothesis that an AVT-neurophysin complex is released from the neurohypophysis during nesting, and that AVT is a physiological regulator of oviducal contractions in sea turtles.

Animals↗

PubMed1989

Temperature dependence of in vitro pituitary, testis, and thyroid secretion in a turtle, Pseudemys scripta.

In vitro culture was used to examine the direct actions of temperature at the level of pituitary hormone [luteinizing hormone (LH), thyrotropin (TSH), growth hormone (GH), prolactin (PRL)] responses to neuropeptides and two related peripheral endocrine responses [thyroid hormone (T4) and testicular androgen secretion] to pituitary hormones (TSH and gonadotropins) in a turtle, Pseudemys scripta. All these responses were fully suppressed at very low temperatures (5-6 degrees) and maximal near the species' preferred body temperature (28 degrees), but sensitivities differed markedly in intermediate ranges. At the pituitary level, the response of TSH, GH, and PRL to thyrotropin-releasing hormone (TRH) was considerably more temperature sensitive than the response of LH to gonadotropin-releasing hormone (GnRH) stimulation. TSH, GH, and PRL were unresponsive at 20 degrees or below, whereas LH secretion was stimulated almost equally between 12 and 28 degrees; the main effect of cooling on LH secretion was to reduce the duration of the response to GnRH. There was no clear effect of previous thermal history on temperature sensitivity of pituitary neuropeptide responsiveness although the general responsiveness of the gland was altered; however, these latter effects may also be related to variations in other factors such as photoperiod, season, and nutrition. Temperature sensitivities of the thyroid and testes also differed, but in the opposite way from the related pituitary cell types. Thyroid glands were relatively insensitive to temperature and responded to TSH between 12 and 32 degrees, with no difference between 20 and 28 degrees. In contrast, testicular androgen secretion showed an abrupt decline in gonadotropin responsiveness below 28 degrees; dose sensitivity, response rate, and maximal output were affected. Results were similar for sea turtle LH, snapping turtle LH, and ovine follicle-stimulating hormone. Thus, the temperature dependence of the two endocrine systems may have a different rate-limiting component.

Androgens↗

PubMed1989

Effects of gonadectomy and steroids on pituitary gonadotropin secretion in a frog, Rana pipiens.

Secretory dynamics of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were measured at various times following gonadectomy in adult male grass frogs, Rana pipiens. Plasma levels and in vitro initial secretory rates of both LH and FSH increased significantly within 1 wk and remained elevated for 3-4 wk of castration. Pituitary FSH and LH content were unchanged. However, dissociation between the two gonadotropins (Gth) occurred thereafter: Secretion of FSH remained elevated for 70 days, but those of LH declined to control levels after 30 days. In vitro secretion of Gth from gonadectomized (gonadx) frogs declined progressively over time reaching control levels after 24 h incubation. The results indicate that elevated pituitary secretion contributes to the observed circulating LH and FSH levels in gonadx frogs, and that FSH and LH may be controlled independently. Replacement therapy with 17 beta-estradiol (E2) suppressed post-gonadectomy increases in plasma Gth and in vitro responses to GnRH, whereas 5 alpha-dihydrotestosterone (DHT) had little effect in vivo and augmented GnRH responses in long-term castrates. In vitro, E2 also inhibited, while 48 h of DHT treatment had no effect on GnRH responsiveness of pituitaries from gonadx frogs. The actions of these steroids were opposite to those typically observed in mammals (and birds), and support the hypothesis that E2 may contribute to seasonal testicular regression in ranid frogs.

Androgens↗

PubMed1988

Effects of TRH on hormone release from pituitaries of the lizard, Anolis carolinensis.

The ability of thyrotropin-releasing hormone (TRH) to stimulate thyrotropin (TSH) from pituitaries of adult male lizards, Anolis carolinensis, was tested in vivo and in vitro. TSH output by pituitaries in vitro was determined by bioassay of the incubation medium using in vitro T4 output by thyroids from the same lizards. Pituitaries incubated without TRH had no detectable TSH secretion during two consecutive (2 or 3 hr) periods of incubation. Incubation in 10 or 100 ng/ml TRH for 2 or 3 hr significantly augmented release of TSH bioactivity in a dose-dependent manner. Pituitaries taken from goitrogen (100 micrograms methimazole/day for 10 days)-treated lizards had elevated basal TSH secretion but did not respond to TRH. TRH injection in vivo (5 micrograms/hr for 10 hr) appeared to stimulate acute release of MSH activity as judged by darkening of skin color after each injection, and plasma T4 was significantly elevated at the end of treatment. These results provide additional evidence that the reptilian thyrotrope has functional TRH receptors and the TSH-stimulating activity of the tripeptide along with its effects on other pituitary cells was present at an early stage of reptilian evolution.

Animals↗

PubMed1988

Plasma steroid concentrations change in response to sexual behavior in Bufo marinus.

Steroid hormone concentrations change in response to social or environmental stimuli in many vertebrates. To test this phenomenon in an amphibian, we examined plasma androgen (A) and corticosterone (B) concentrations in male marine toads (Bufo marinus), a tropical species exhibiting intermale competition, amplectic clasping of females, and bouts of breeding behavior following rains. When males clasped females for 0, 1, 2, or 3 hr, plasma A concentrations were significantly and positively correlated with hours spent in amplexus. In field-sampled males, plasma A concentrations were higher in amplexing males than in single males. Among single males those found closer to breeding ponds had higher A concentrations than those more distant. These data support the hypothesis that sexual stimuli enhance plasma A concentrations in this amphibian. In 3-hr experimental tests and field-sampled males, B concentrations were higher in amplexing than in single males. Unlike some amphibians, short-term elevations of B apparently are not associated with decreased reproductive function. However, as in other amphibians in which high B concentrations are associated with stress-induced inhibition of reproduction, after 48-72 hr in captivity male toads showed high B concentrations and low plasma androgen concentrations. The bursts of sexual activities exhibited by B. marinus following heavy rains were associated with no changes in A concentration and with increased B concentration.

Animals↗

PubMed1988

Plasma androgen and gonadotropin levels during hibernation and testicular maturation in golden-mantled ground squirrels.

The 4-5-mo hibernation season of golden-mantled ground squirrels consists of extended torpor bouts interspersed with brief, periodic intervals of normothermic arousal. Plasma levels of testosterone (T), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) and degree of scrotal pigmentation were measured in torpid and aroused male ground squirrels throughout a season of hibernation and in active animals after the termination of torpor. T was basal in torpid animals; beginning 3 weeks before torpor ended, T was elevated in normothermic males during the first half of periodic arousals but returned to basal levels before animals reentered torpor. After the last (terminal) arousal from torpor, T levels were moderately elevated for 4 wk and maximal for the next 6 wk before they returned to basal values. LH patterns were similar to those of T; however, levels of T and LH were positively correlated only in aroused or posthibernation males. FSH levels remained constant and low during most of the heterothermic season but increased in several torpid males within 3 days of terminal arousal. FSH levels peaked 2 wk after the end of heterothermy. Scrotal pigmentation developed over the first 4 wk after terminal arousal. Maturation of reproductive function occurs during the 4 wk after termination of heterothermy, but elevated levels of T during arousals and variable levels of FSH in the last days of torpor suggest that activation or increased sensitivity of the hypothalamic-pituitary-gonadal axis is important in the termination of heterothermy in ground squirrels.

Androgens↗

PubMed1987

Effects of thyrotropin-releasing hormone in vitro on thyrotropin and prolactin release from the turtle pituitary.

Effects of synthetic thyrotropin-releasing hormone (TRH) on thyrotropin (TSH) and prolactin (PRL) release by hemipituitaries of adult turtles, Chrysemys picta, were studied in an in vitro superfusion system. Significant increases in the rates of secretion of both immuno-reactive TSH and PRL occurred at doses between 0.01 and 10 ng/ml TRH. TSH secretion increased acutely by two-, to sixfold over nonstimulated secretion levels; responses tended to decline after many hours of continual stimulation, but output remained elevated above baseline in most cases. PRL secretion increased, parallel to TSH secretion during TRH stimulation. No significant difference was found in secretion rates between males and females, and no clear relationship between TRH responsiveness and reproductive stage was evident. These data provide the first direct evidence for the stimulation of TSH secretion by TRH in a reptile and confirm earlier reports that TRH stimulates the release of PRL in the turtle. Although previous in vivo studies indicated that TSH secretion was not affected by TRH in turtles, the present data indicate that the dose sensitivity of the chelonian gland is comparable with that of mammalian and avian pituitaries. Evidence for the role of TRH in endogenous TSH regulation is still lacking in reptiles but the present data provide evidence for functional TRH receptors on the chelonian thyrotrope and, hence, argue against the hypothesis that TSH stimulating activity of TRH evolved relatively recently in association with endothermy.

Animals↗

PubMed1987

Specificity of amphibian and reptilian pituitaries for various forms of gonadotropin-releasing hormones in vitro.

In vitro perifusion was employed to compare the potencies of mammalian, avian, salmon, and lamprey gonadotropin-releasing hormones (GnRHs) on the release of luteinizing hormone (LH) from the pituitaries of an amphibian (Rana pipiens) and a reptile (Chrysemys picta). The chicken-I and salmon GnRH variants were equipotent with mammalian GnRH in both the frog and the turtle glands. By contrast, the lamprey GnRH was inactive (less than 1% as potent as the others). Lamprey GnRH also failed to stimulate LH release or to induce GnRH priming when administered chronically to the frog gland. These results support the hypothesis that the GnRH receptors on nonmammalian pituitary cells are much less specific than those of the mammal with regard to the amino acid at position 8 of the GnRH molecule. These data suggest that the native GnRH variant or the one most like that found in the brain of a species is not necessarily the most potent biologically in that species. However, the nonmammalian pituitary does show some specificity with regard to the structure of natural GnRHs in that none of the tetrapod species studied is responsive to lamprey GnRH.

Animals↗

PubMed1987

Subcellular localization of gonadotrophic hormones LH and FSH in frog adenohypophysis using double-staining immunocytochemistry.

We applied double post-embedding immunocytochemical methods using specific antibodies against bullfrog (Rana catesbeiana) luteinizing hormone (LH) and follicle-stimulating hormone (FSH) with immunogold staining (5- and 20-nm particles) to determine the subcellular localization of both gonadotropins and to observe their immunostaining patterns in anterior pituitary of the frog Rana pipiens. Results showed that individual gonadotrophs may store either one or both gonadotropins in a given secretory granule and in large globules (lysosomes?). Most gonadotrophs (50-88%) contain both hormones; 12-50% contain only FSH, and only a few (0-7%) contain LH alone. Individual secretory granules, even in cells that contain both hormones, may contain only one or both gonadotropin molecules. Evaluation of the percentage of monohormonal and multihormonal secretory granules revealed that multihormonal secretory granules were the most numerous and that LH monohormonal secretory granules were the least numerous. These results indicate that cellular storage of gonadotropin in amphibian pituitary is similar to that described for mammals, where a single cell type containing both gonadotropins predominates. Variability in hormone content both of cells and of granules in all individuals is consistent with the hypothesis that frog pituitary possesses a single multipotential gonadotroph.

Animals↗

PubMed1986

Photothermal effects on the testicular cycle in the musk turtle, Sternotherus odoratus.

A series of photothermal treatments was initiated at different times of year to examine the relative contributions of exogenous and endogenous factors affecting the timing of the testicular cycle of the kinosternid turtle, Sternotherus odoratus. Elevated temperature appeared to be the major factor stimulating testicular recrudescence (spermatocytogenesis, spermiogenesis, and, to some extent, androgen secretion). However, depending on the time of year, short or long daylengths, in conjunction with elevated temperatures, enhanced testicular responses (recrudescence or regression). Also, high temperatures are not requisite for recrudescence. Spermatogenic activity and testis growth differ in their temperature sensitivity and may become dissociated, and an endogenous mechanism may have affected peak androgen secretion. Testicular regression exhibited a dependence on an interaction among temperature, photoperiod, and endogenous regulation. Turtles with testes in advanced stages of spermiogenesis underwent accelerated regression when exposed to even slightly reduced temperatures, especially at short daylengths. However, regression was delayed by a combination of high temperature and short daylength in fall. Results are compared to other experimental studies of photothermal effects on turtles and discussed in terms of the described latitudinal variations in the timing of the testicular cycle of S. odoratus.

Acclimatization↗

PubMed1986

Dependence of GnRH action on Na+, K+, and Ca2+ in the frog, Rana pipiens, pituitary.

The roles of K+, Ca2+, and Na+ ions in the mechanism of gonadotropin releasing hormone (GnRH) action on frog (Rana pipiens) hemipituitaries were studied using an in vitro superfusion system. The effects of elevated K+ alone or in combination with Ca2+-depleted medium, tetrodotoxin (TTX), or with 100 ng/ml GnRH were examined. The involvement of K+ was also studied indirectly through the use of tetraethyl ammonium chloride (TEA). The importance of Ca2+ was established by the loss of responsiveness to GnRH in Ca2+-depleted medium, or in the presence of the Ca2+ competitor CoCl2. The absence of a major dependence of GnRH on Na+ was revealed by the continued gonadotropin secretion after addition of 1 microM TTX to medium containing GnRH or 36.3 mM KCl, or by replacement of NaCL with choline chloride. High (10 X normal) KCl (36.3 mM) stimulated gonadotropin--both LH and FSH--secretion, but the response was more gradual than for GnRH. The inclusion of TEA (to block K+ efflux) in medium with GnRH accentuated the effect of GnRH, and the effects of elevated (36.3 mM) KCl and 100 ng/ml GnRH (a relatively high dose) were additive. Responses to high K+, like GnRH, were abolished by removal of Ca2+ from the medium. Overall, the roles of K+, Ca2+, and Na+ ions in the mechanism of GnRH action are very similar between mammals and frogs; Ca2+ apparently serves a critical function in the mechanism of GnRH action, while Na+ appears not to be involved. K+ can induce gonadotropin secretion, but it is not clear that it plays a direct role in the mediation of the action of GnRH.

Animals↗

PubMed1986

The cellular basis of the calcium dependence of GnRH-stimulated gonadotropin release from frog, Rana pipiens, pituitaries.

An in vitro superfusion system was used in an attempt to identify the cellular systems involved in the Ca2+ dependence of gonadotropin-releasing hormone (GnRH) actions in the frog, Rana pipiens. Superfusion with 5 microM A23187 (a calcium ionophore) or phorbol myristate acetate (an analog of diacylglycerides) caused marked increases in luteinizing hormone (LH) and follicle-stimulating hormone (FSH) secretion. Exclusion of Ca2+ from the medium prevented the stimulatory effects of PMA. The potent stimulator of adenylate cyclase, forskolin, caused only slight stimulation of LH and FSH secretion, which was also prevented by removal of Ca2+. The cytoskeletal disruptive agents colchicine, nocodazole, and cytochalasin B, and the calmodulin inhibitors, trifluoperazine and pimozide, had no significant effects on the action of GnRH. Overall, these results indicate that the major mechanisms of Ca2+ involvement in the response to GnRH by the frog pituitary are similar to that of mammals, with the possible exceptions of lesser roles for calmodulin and the cytoskeleton in the frog. The study suggests that polyphosphatidyl-inositol-diacylglyceride metabolism may be critical in understanding the mechanism of GnRH action in frogs.

Animals↗

PubMed1986

Seasonal cycles in gonadal activity and plasma gonadotropin in the musk turtle, Sternotherus odoratus.

Plasma gonadotropin cycles were examined in relation to the seasonal gonadal cycle in freshly captured musk turtles, Sternotherus odoratus, from South Carolina. Acute and chronic stress effects of captivity on testicular growth and circulating plasma testosterone (T) and follicle-stimulating hormone (FSH) also were examined. Monthly mean FSH levels in freshly captured males were correlated significantly with plasma T. Both T and FSH were minimal in spring when testes were small, and highest in the fall in males that had begun spermiation and whose testes had begun to regress. Individual plasma T and FSH values were significantly correlated during the fall when both hormones were at elevated levels. In contrast, plasma luteinizing hormone (LH) remained undetectable throughout the year in males. Females had nondetectable levels of both plasma LH and FSH throughout the year. There was a seasonal pattern in plasma 17 beta-estradiol which was correlated with follicular growth. Holding males in captivity (outdoors) reduced plasma T by 35-60% within 24 hr of capture (depending on season) and plasma T remained at these lower levels after 2 weeks. However, FSH was not significantly affected by such short-term "stress." Testicular weight and FSH cycles of captive males were similar to those of males in the field. Monthly plasma T of long-term captives also exhibited a seasonal cycle but at a level significantly lower than in the field and with a delayed peak. Thus, while the data suggest that FSH may be important to both seasonal testicular growth and androgen secretion, fluctuations in FSH alone cannot account for all observed patterns or changes in spermatogenesis or plasma androgen levels.

Animals↗