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B Breton

Publications and source records attributed to B Breton.

At least 55 records · Page 3Linked to original sources

Gonadotropin hormone (GtH) receptors in the ovary of the brown trout Salmo trutta L. in vitro studies.

A method has been developed suitable for the study of the binding of salmon (Oncorhynchus tsawytscha) gonadotropic hormone (GtH) to brown trout (Salmo trutta) ovarian particulate fractions. Several proteolytic enzyme inhibitors were tested to preserve the binding capacity of particulate fractions (1000, 20,000, and 110,000g) from yolky oocytes. The best preservation was obtained with trypsin inhibitor. The effects on binding of the pH, time, and temperature of the incubation were investigated. Binding increased linearly with amount of particulate fraction, and is saturable. In both the 20,000 and 110,000g fractions, there are two classes of receptors: one with a high-affinity constant (3.14 and 0.95 10(10) M-1 for the 20,000 and 110,000g fractions, respectively) and a low capacity (1.76 and 0.63 fmol/g of ovary), the other with a higher-affinity constant (3.44 and 1.78 10(9) M-1) but a higher capacity (7.37 and 7.42 fmol/g of ovary). GtH binding was not affected by ovine luteinizing hormone (oHL), ovine follice-stimulating hormone (oFSH), and human chorionic gonadotropin (hCG), but was partially inhibited by 1 micrograms of bovine thyroid-stimulating hormone (bTSH) and salmon prolactin (sPRL). The results are discussed in relation to the biological specificity of fish GtH and to the unsolved problem concerning the number of gonadotropins in fish.

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Short-term profiles of plasma gonadotropin and estradiol-17 beta levels in the female rainbow trout, from early ovarian recrudescence and throughout vitellogenesis.

At various stages throughout the annual reproductive cycle, female rainbow trout (Salmo gairdneri) were fitted with a catheter in the dorsal aorta. They were bled via the catheter at frequencies of once every 30, 60, or 240 min over periods of 5 to 24 hr. Gonadotropin (GtH), estradiol-17 beta (E(2)17 beta). and estrone levels were measured in the plasma samples. At early ovarian recrudescence (March), short-term (1-2 hr), high-amplitude (delta GtH = up to 100 ng/ml), episodic pulses of GtH were recorded in the plasma of 12 of the 26 studied fish. In the others, GtH levels remained low and constant. No synchronization was found among the individual GtH profiles. E(2)17 beta levels in the same fish were low and constant while estrone was not detectable. At early stages of exogenous vitellogenesis (June), plasma GtH (1-3 ng/ml) and E(2)17 beta (0.5-1.5 ng/ml) levels were low and constant. At advanced stages of exogenous vitellogenesis (September-October), fluctuating GtH levels were found again in most of the females; basal GtH concentrations were only slightly higher than those recorded in June. The fluctuations consisted of short-term (1-3 hr) random GtH pulses of moderate amplitude (delta GtH = up to 5 ng/ml), occurring at a relatively high frequency (up to 5 per 12 hr). Although no regular synchronous daily pattern of GtH was noted, most of the GtH pulses were observed during the photophase and early scotophase. The appearance of GtH pulsatility during exogenous vitellogenesis was accompanied by a large increase in plasma E(2)17 beta up to levels ranging from 6 to 30 ng/ml. In contrast to the GtH profiles, the E(2)17 beta profiles showed continuous and progressive variations, superimposed the abrupt GtH pulses, and a high degree of synchronization. E(2)17 beta levels increased during the photophase and reached maxima toward and during early scotophase.

Animals↗

Short-term profiles of plasma gonadotropin and 17 alpha-hydroxy, 20 beta-dihydroprogesterone levels in the female rainbow trout at the periovulatory period.

Individual free-swimming female rainbow trout in which oocytes underwent final stages of germinal vesicle migration, maturation, or ovulation were bled via a dorsal-aortic catheter at frequencies of once every 1, 3, or 4 hr over periods of 9 to 36 hr. Gonadotropin (GtH) and 17 alpha-hydroxy,20 beta-dihydroprogesterone (17 alpha,20 beta-OHP) levels were measured in the plasma samples. GtH levels were elevated and showed wide and progressive daily variations. A high degree of synchronization appeared among the GtH profiles of individual fish. Two distinct daily GtH surges were observed, one at early photophase and the other during the mid-scotophase. The onset of the GtH increases was closely related to the beginning of the photophase and the scotophase, respectively. In females undergoing oocyte maturation or in ovulated females, 17 alpha,20 beta-OHP levels were increasing or high, showing progressive daily fluctuations that were either synchronized with the GtH changes or somewhat phase-shifted in relation to them. These data are discussed in relation to the seasonal changes in the short-term profiles of reproductive hormones in the trout.

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Effects of [D-Ala6, Pro9-NEt]-LHRH and catecholaminergic drugs on gonadotropin secretion and ovulation in the Chinese loach (Paramisgurnus dabryanus).

The effects of [D-Ala6,Pro9-NEt]-LHRH (LHRH-A) alone and in combination with drugs which influence the actions of dopamine or the synthesis of catecholamines on gonadotropin (GtH) secretion and ovulation in the loach (Paramisgurnus dabryanus) were investigated. LHRH-A alone stimulated an increase in serum GtH levels in the loach, but was a relatively ineffective treatment for the induction of ovulation. Injection of the dopamine receptor antagonist pimozide caused a marked potentiation of the GtH-release response to LHRH-A, and combined injections of pimozide and LHRH-A were an effective treatment for the induction of ovulation. Reserpine, a drug which causes depletion of catecholamines from presynaptic terminals, also caused a marked potentiation of the GtH-release response to LHRH-A and combined treatment induced ovulation. Similarly, administration of alpha-methyl-para-tyrosine to block conversion of tyrosine to L-dopa, or carbidopa to block conversion of L-dopa to dopamine, potentiated the GtH-release response to LHRH-A and induced ovulation. In contrast, the use of diethyldithiocarbamate, to block conversion of dopamine to norepinephrine, failed to augment the action of LHRH-A on GtH release and ovulation. The present results provide further evidence to suggest that dopamine functions as a gonadotropin release-inhibitory factor in teleosts, and demonstrate that the use of drugs which block either the synthesis or the actions of dopamine potentiates the action of LHRH-A in teleosts.

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Endocrine changes associated with spawning behavior and social stimuli in a wild population of rainbow trout (Salmo gairdneri). I. Males.

Rainbow trout (Salmo gairdneri) were collected from a natural spawning population at Pennask Lake, B.C. Blood samples were taken from male trout at different stages of spawning and assayed by radioimmunoassay for gonadotropin (GtH), testosterone (T). 11-ketotestosterone (11-KT), and 17 alpha-hydroxy-20 beta-dihydroprogesterone (17,20-P). Testosterone and 11-KT were detected in the plasma of males in spawning condition: 11-KT was at a higher concentration than T and declined to undetectable levels in postspawning fish. There were striking increases in plasma GtH and 17,20-P in males actively courting nest building females. The increase in 17,20-P was evidently not dependent upon an elevation of the pituitary hormone: 17,20-P remained low in males placed with sexually inactive, unovulated females even though GtH levels rose above levels measured in males isolated from females. The significance of the social control of hormone levels is not clear. There is some evidence that 17,20-P and/or GtH may cause an increase in quantity of the milt, and thus the endocrine response to a sexually active female may serve to synchronize maximum male fertility with oviposition readiness in the female.

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Endocrine changes associated with spawning behavior and social stimuli in a wild population of rainbow trout (Salmo gairdneri). II. Females.

Rainbow trout (Salmo gairdneri) were collected from a natural spawning population at Pennask Lake, B.C. Blood samples taken from female trout at different stages of spawning were assayed by radioimmunoassay for gonadotropin (GtH), estradiol-17 beta (E2), androgens, including testosterone (T), and 17 alpha-hydroxy-20 beta-dihydroprogesterone (17,20-P). Plasma levels of androgen and estradiol were highest in females sampled shortly before ovulation ("green" females) and declined in ovulated and sexually active females, reaching lowest levels in postspawning fish. Concentrations of 17,20-P rose markedly in ovulated females allowed to dig nests and interact with sexually active males. Plasma GtH levels were similar in green unovulated females and ovulated fish prevented from spawning, but showed a marked increase in actively nest building ovulated fish. The results demonstrate that social stimuli affect plasma levels of 17,20-P and perhaps GtH. The functional significance of the endocrine responses to social factors is not clear, but it is suggested that increased hormone levels may contribute to an acceleration or synchronization of breeding, or be responsible for causing and maintaining more vigorous sexual activity.

Androgens↗

Use of pituitary cells in primary culture to study the regulation of gonadotropin hormone (GtH) secretion in rainbow trout: setting up and validating the system as assessed by its responsiveness to mammalian and salmon gonadotropin releasing hormone.

To study the regulation of gonadotropin secretion in rainbow trout in vitro, a method for preparing primary cultures of dispersed pituitary cells is described. Cells were dispersed by collagenase 0.1% in Hank's saline solution for 20 hr at 12 degrees and a high yield of viable cells was obtained. Attempts to improve cell functioning were made by varying culture conditions (density of cells initially plated, age of the culture). Cell functioning was assessed by their ability to respond to increasing doses of mammalian and salmon GnRH. Pituitaries were collected from spermiating males whose pituitaries are known to be sensitive to mammalian GnRH in vivo. Using 96-well plates, optimal conditions for good biological activity, are initial plating with 6.2 X 10(4) cells, incubation with GnRH for 24 hr on the third day after plating. In these conditions mammalian analog and salmon GnRH induced an increase in GtH release for doses ranging from 10(-9) to 10(-6) M. The GtH released during the GnRH incubation period does not decrease the sensitivity of the system since addition of 20 ng of GtH at the beginning of incubation does not modify the response profile.

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Immunoreactive gonadotropin-releasing hormone-like material in the brain and the pituitary gland during the periovulatory period in the brown trout (Salmo trutta L.): relationships with the plasma and pituitary gonadotropin.

In fish there are few data on the gonadotropin-releasing hormone (Gn-RH) neurosecretory activity, which could explain long- and short-term variations of the gonadotropin secretion. There is no biological species specificity between mammal and fish Gn-RH; although there is a structural difference, they are, on the contrary, characterized by a high immunological specificity which does not allow measurement of fish Gn-RH using radioimmunoassay for LH-RH. We have synthesized salmon Gn-RH according to the formula recently proposed by Sherwood (N. Sherwood, L. Eiden, M. Brownstein, J. Spies, J. Rivier, and W. Vale, 1983. Proc. Natl. Acad. Sci. USA 80, 2794-2798). Its activity has been tested by its ability to stimulate the gonadotropin hormone (GtH) secretion in vivo in testosterone-implanted juvenile rainbow trout, and for the recognition of synthesized Gn-RH (s-Gn-RH) perykaria by a specific antibody raised against the s-Gn-RH in regions of the brain described as containing LH-RH immunoreactive-like material. A radioimmunoassay has been developed for the salmon Gn-RH, and its specificity to measure trout Gn-RH has been tested. Using this assay, the brain and pituitary Gn-RH contents have been measured throughout the final phases of maturation and ovulation. Brain Gn-RH increases from the end of vitellogenesis (8.9 +/- 0.76 ng/brain) to ovulation (more than 15 ng/brain). Pituitary Gn-RH is lower (1.58 +/- 0.69 ng/pituitary) at the end of vitellogenesis and follows a similar profile as in the brain, except for a significant decrease just prior the beginning of oocyte maturation. The correlations between Gn-RH levels and GtH pituitary and plasma levels show that total brain Gn-RH is never correlated to the GtH, suggesting that the increase in the brain Gn-RH content is related to a Gn-RH system closely related to maturation and ovulation, which remains to be investigated. On the contrary, pituitary Gn-RH levels are well correlated with pituitary and plasma GtH levels, indicating that pituitary Gn-RH levels might represent a good index of the Gn-RH neurosecretory activity in the fish hypothalamohypophysial complex, given the absence of a portal system in teleost.

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Daily changes in the gonadotropin levels and response of carp oocytes to hypophysial homogenate.

Daily changes in carp gonadotropin levels in adult female carp and daily changes in carp oocyte sensitivity to carp hypophysial homogenate, in vitro and in vivo, were investigated. A total of three series of experiments were carried out. Gonadotropin levels were radioimmunologically determined. The results of series 1 and 2 experiments were subjected to statistical analysis with the use of cosinors circle and ellipse of errors. It has been found that in the mature female carp in the pre-spawning period with the light periods being long (L:D = 16:8) the apogee for gonadotropin occurs 10 hr after the onset of the light period. The sensitivity of the oocytes, in terms of the percentage of mature oocytes (after GVBD) following a 24-hr incubation of ovarian fragments with the hypophysial homogenate, reached the highest value at 1300, i.e. 9 hr after the onset of the light period. It was also found that the injections of carp hypophysial homogenate made at 0900 were much more efficient in inducing ovulation than those at 2100.

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Plasma 11-oxotestosterone and gonadotrophin in relation to the arrest of spermiation in rainbow trout (Salmo gairdneri).

Plasma gonadotrophin (GTH) and 11-oxotestosterone levels have been studied in relation to sperm production during the last weeks of the spermiation period. The level of 11-oxotestosterone decreased in parallel to sperm production (coefficient of correlation: p = 0.59; P less than 0.01) and reached its lowest values (mean +/- SE = 0.33 +/- 0.19 ng/ml; n = 11) when spermiation was over. No significant variations were found in GTH levels; however, there was a slight positive correlation between sperm production and GtH levels (p = 0.39; P less than 0.05).

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[Homologous radioimmunoassay of a hypothalamic factor stimulating salmon pituitary gonodatropic function (sGnRH)].

A radioimmunoassay for Salmon Gn-RH p-gly-His-Trp-Ser-Tyr-Gly-Trp- Leu-Pro-Gly) (NH2) has been developed with a sensitivity of 7 pg/assay tube. The system allows the specific detection of an immunological GnRH related substance in the brain and pituitary of three teleost species but not in an elasmobranch the Dogfish. these results are discussed and some Gn-RH contents of the organs are proposed.

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[Potentiation by pimozide of the effects of LHRH-A on hypophyseal gonadotropin secretion, ovulation and spermiation in the common carp (Cyprinus carpio)].

During the reproductive season in August, we administered to adult male and female Carp an LHRH analog (LHRH-A = D-trp6EA-LHRH) (50 micrograms/kg) alone or in association with pimozide, a dopamine inhibitor (10 mg/kg). LHRH-A caused the circulating portal level of gonadotropin to rise significantly in males and less in females. Spermiation was stimulated in the males and oocyte maturation in females but there was no ovulation. An infection of pimozide and then of LHRH-A 6 h later significantly increased c-GTH secretion and spermiation more than LHRH-A given alone, and induced ovulation in the females.

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[Maturational glycoprotein gonadotropin and estradiol-17-beta during the reproductive cycle of the female brown trout (Salmo trutta)].

Female brown trout were sacrificed every month or twice a month during the reproductive season. Plasma gonadotropin (GtH) and estradiol-17 beta (E2-17 beta) were measured using radioimmunological techniques, in relation with the state of gametogenesis. From ovulation to July egg's diameters remained less than 1 mm, and ovogenesis was characterized from the histological appearance of three types of vitellus: glycoproteique (type I), lipidique (type II), lipidoprotéique (type III), and the immunological plasma detection of the vitellogenin. During this period GtH levels remained lower than 1 ng/ml except in March, when they increased around 5 ng/ml, both with pituitary GtH and plasma E2 17 beta. This rise occurred just before the appearance of the type III vitellus within the oocyte, and might correspond to a critical phase of the reproductive cycle during which vitellogenin could not be detected in 70% of the animals. During rapid growth of the oocyte from 1 to 5 mm, mean GtH levels increased from 0.75 to 2.5 ng/ml when those of the E2-17 beta increased more rapidly but began to drop before the end of vitellogenesis. Maturation and ovulation were accompanied by a rise of the GtH level, in correlation with the lower levels of E2-17 beta measured during the cycle, GtH remained high even after ovulation, and more in fish which had kept their eggs within the body cavity. Correlation among GtH, E2-17 beta, and the diameters of eggs had been calculated. There was a positive correlation among GtH, E2-17 beta levels, and the diameters of eggs during exogenous vitellogenesis, and a negative between GtH and E2-17 beta at the end of the reproductive cycle. These results were discussed, in relation to the existence of a pulsatile mode of GtH secretion.

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[Effect of pulsatile gonadotropic stimulation on the in vitro incorporation of vitellogenin by trout follicles incubated in an open perifusion system].

Rainbow Trout follicles incubated in vitro in an open perifusion system, incorporated statistically more vitellogenin after a pulsatile gonadotropic stimulation than after a perifusion without GtH. Most of the newly incorporated vitellogenin is found within the oocyte cytosol. These results demonstrate the intrinsic stimulatory effect of the "maturing" GtH on the vitellogenin incorporation.

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[Evolution of plasma levels of glycoproteic gonadotropins and of 17 alpha hydroxy-20 beta dihydroprogesterone during maturation and ovulation of rainbow trout, Salmo gairdnerii].

The plasma concentrations of the glycoproteic gonadotropin (GTH) and of the 17 alpha hydroxy-20 beta dihydroprogesterone (17 alpha 20 beta OHP) have been followed in the course of ovulation in 6 rainbow trout, using radioimmunoassays on samples taken every two days. A first slow and limited GTH increase (10 to 20 ng/ml) is detected before and during the ovulation, while the 17 alpha 20 beta OHP rises sharply (up to three to five hundreds nanogrammes per millilitre). Then the progestagen levels drop back to their previous values, for about three weeks after ovulation. At the same time, a second higher GTH increase (25 to 20 ng/ml) is taking place. The relations between these two hormones, maturation and ovulation are discussed.

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Glycoprotein gonadotropin in the plasma and its cellular origin in the adenohypophysis of sham-operated and ovariectomized rainbow trout, Salmo gairdneri.

Among the cells of the pituitary generally believed to produce glycoprotein gonadotropin (GTH) five forms were distinguished, based on the amount and the diameter of granules and globules and the appearance of the rough endoplasmic reticulum. In sham-operated trout so-called "globular" cells predominated, whereas after ovariectomy these were replaced by so-called "cisternal" cells, suggesting that both belong to one GTH-cell type. In addition, ovariectomy caused a strong increase in plasma GTH-levels. This indicates that the transition from globular to cisternal cells is accompanied by extrusion of GTH, and thus points to a storage of GTH in the granules and globules. It is argued that one of the five forms has the morphological characteristics of thyrotropic cells and may not produce glycoprotein GTH.

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