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1.
Isolated gonadotrops of the African catfish,Clarias gariepinus, were incubated with dopamine (DA) and/or catecholestrone and the activity of the enzyme catechol-O-methyltransferase (COMT) was determined by measuring the methylated products. From the apparent Km values for DA and catecholestrone of 0.4–1.3 M and 17.9–25.2 M respectively, it was concluded that catecholestrone is a better substrate for the enzyme COMT, compared to DA. Moreover, the methylation of DA is inhibited by comparatively low concentrations of catecholestrone.  相似文献   

2.
PKC activity was detected in spleen extracts from the turbot, Scophthalmus maximus, a teleost flatfish that is farmed commercially in several countries, in assays with the substrate EGF- R651–658 as phosphate acceptor. The activity was purified about 700-fold by a three-step chromatographic procedure (DEAE-cellulose, phenyl-Sepharose and threonine-Sepharose). Maximal activity was obtained in the presence of the typical PKC cofactors Ca2+ (0.1 mM) PtdS (20 g ml–1) and either DAG (2 g ml–1) or PMA (2 g ml–1). Activity was dose-dependently inhibited by H7 and by the PKC-specific inhibitors PKC19–36 and N-myristoylated PKC19–31. The rate of phosphorylation was highest with the PKC-specific substrate MARCKS161–175. In immunoblotting, MC5 (a mouse monoclonal antibody raised against bovine PKC) recognized bands of 80 and 100 kDa. Immunoblotting with antibodies raised against mouse PKC isozymes (, , , , , , and ) indicated the presence of all these isozymes in turbot spleen.  相似文献   

3.
In order to specify the timing of some changes in ovarian steroid production during the transition from vitellogenesis to ovulation, plasma hormones levels andin vivo andin vitro responses of the ovary to salmon gonadotropin (s-GtH) or dibutyryl-cyclic adenosine mono-phosphate (db-cAMP) were recorded in relationship with the state of germinal vesicle migration in the oocyte.In vivo, a small, but significant, increase of plasma 17-hydroxy-20-dihydroprogesterone (17, 20-OH-P) level was detected earlier (at the subperipheral germinal vesicle stage) than the increase of GtH level (detectable at the peripheral germinal vesicle stage) and the decline of oestradiol-17 (E2–17) (also detectable at the peripheral germinal vesicle stage). Negative correlations were established between E2–17 levels and GtH (=–0.53) or 17,20-OH-P (=–0,43) levels while a positive correlation occurred between 17,20-OH-P and GtH levels (=+0,54).In vivo no action of GtH on the decline of E2–17 levels was detected GtH did not stimulate 17,20-OH-P production, within 72h, in females at the end of vitellogenesis stage. It had significant effect in females at other stages closer to ovulation, but the pattern of responses changed according to the stage.In vitro db-cAMP like GtH was able to stimulate 17,20-OH-P output from ovarian follicles. The greatest response was observed at the later stage. (GVBD). Testosterone output was also increased by GtH, but the lowest response was observed at the later stage (GVBD). Androstenedione output was lower than testosterone output.In vitro, a small but significant decline of E2–17 output was induced by GtH. We conclude that substantial changes occur during the very last stages prior to ovulation, both in the steroidogenic potential of the ovary and in the ovarian sensitivity to GtH. 20-oxydoreductase is probably progressively induced during GV migration when GtH basal levels are increasing but still relatively low. Without minimizing the role of discrete pulses of GtH on this induction, we could expect synergic actions of other hormones. Thus a high testosterone/oestradiol ratio in the follicle environment favours 17,20-OH-P secretion.  相似文献   

4.
In two separate spawning seasons, spermiating male Atlantic halibut were implanted with pellets containing gonadotrophin-releasing hormone agonist (GnRHa). Males were bled repeatedly, and milt samples were collected. Blood samples were assayed for free and conjugated steroids: testosterone, 11-ketotestosterone, 17,20-dihydroxy-4-pregnen-3-one (17,20-P), 17,20-dihydroxy-4-pregnen-3-one (17,20-P), 17,20,21-trihydroxy-4-pregnen-3-one and steroids with a 17,20 configuration. Towards the end of the first season, pellets were implanted into three wild-caught and three hatchery-reared males. No control fish were available. The major progestogen in plasma was identified as sulphated 5-pregnane-3,17,20-triol (3,17,20-P-5-S). Concentrations of this steroid were stimulated by the GnRHa. Sulphated 17,20-P was also identified in the plasma, but at 10-fold lower concentrations than 3,17,20-P-5-S. In the middle of the second season, pellets were implanted into five hatchery-reared males; five unimplanted males were used as controls. Levels of androgens fell following GnRHa treatment, levels of progestogens rose briefly, and there was a significant increase in the fluidity of the milt. Of all the measured steroids, free and sulphated 17,20-P showed the best correlation with milt fluidity.  相似文献   

5.
Colisa lalia (Ham.) has been used as a model for the development of techniques for induced spawning that are applicable to small teleosts where ovulation requires prolonged exposure to suitable breeding conditions.Human chorionic gonadotrophin (hCG; 4 IU per fish) induced ovulation within 24 hours, whereas homologous pituitary extracts were relatively ineffective. When administered in saline ( 20 g per fish per injection), des-Gly10,[D-Ala6]- LHRH N-ethylamide (LHRHa) was ineffective, but it stimulated ovulation in a proportion of fish when administered ( 1.6 g per fish) as an emulsion in Freund's incomplete adjuvant (FIA). Together, these results suggest that ovulation requires the synthesis as well as the secretion of gonadotrophin in C. lalia.Long-term treatment with thyroid hormones appeared to enhance the ovulatory response to LHRHa in FIA, possibly by effects on the ovary; whereas the various steroids tested were ineffective at the dosages used  相似文献   

6.
The present study investigated the relationships between egg viability and ovarian fluid composition, egg physiology and egg metabolism in lake trout, Salmo trutta lacustris, to obtain biomarkers for egg quality determination. The ovarian fluid pH, protein levels and activities of aspartate aminotransferase and -d-glucuronidase were significantly correlated with egg viability expressed as the number of eyed stage embryos. Regression models demonstrated that an ovarian fluid pH between 8.44 and 8.57, protein levels below 235.56 mg 100 ml–1ovarian fluid, aspartate aminotransferase activity below 31.65 m min–1 l–1ovarian fluid and -d-glucuronidase activity below 8.62 m min–1 l–1 ovarian fluid characterized egg batches with high viability (80%).The increase in the egg wet weight during water hardening was also significantly correlated with the number of eyed stage embryos, and egg batches with high egg viability (80%) increased in wet weight by 13% during water hardening.From the investigated metabolic parameters the number of eyed stage embryos was significantly correlated with activities of NADP-dependent isocitrate dehydrogenase (egg viability 80% at 2.07 nM min–1 mg–1 protein) and NAD-dependent malate dehydrogenase (egg viability 80% at 47.25 nM min–1 mg–1 protein), with the respiration rate (egg viability 80% at 8.71 nM min–1 mg–1 protein), with the ratio of NADH to NAD levels (egg viability 80% 0.872), with the levels of free, non-esterified fatty acids (egg viability 80% 72.34 g mg–1 protein), and the ratio of non esterified to esterified fatty acids (egg viability 80% at 0.749). Also, subjective and visual control methods were described to distinguish between batches with viable and non viable eggs.  相似文献   

7.
Male goldeye were treated with (D-Ala6, Pro9-N ethylamide) — LHRH (LHRH-A) in saline or a silastic pellet (100 µg.kg–1 body weight) and changes in plasma gonadotropin (GtH), 17,20-dihydroxy-4-pregnen-3-one (17,20P) and testosterone (T) levels were determined by radioimmunoassay. LHRH-A increased plasma GtH levels, with the response to LHRH-A in saline being of much greater magnitude and duration than the response to silastic pellet implants. Seasonal differences were found in the response to LHRH-A. Spermiated fish were the most responsive, recrudescing fish the least, and regressed fish displayed an intermediate response. Plasma 17,20P levels were elevated in response to LHRHA in fish of all sexual stages although the magnitude of the increase was not related to the magnitude of the increase in GtH levels. Treatment with LHRH-A also resulted in a transitory increase in plasma T levels. The endocrine control of GtH and steroid secretion in goldeye is discussed in relation to studies in cyprinids and salmonids. Address for reprint requests: Department of Zoology, University of Alberta, Edmonton, Alberta T6G 2E9 Canada.  相似文献   

8.
Plasma levels of 17,20-dihydroxy-4-pregnen-3-one (17,20OHP), which is involved in the regulation of spermiation in male salmonid fish, increase dramatically at the time of spermiation. To advance the understanding of the regulation of 17,20OHP production during the spermatogenetic cycle in trout, we have studied the in vitro effect of gonadotropin type II (GtH II) and the precursor 17-hydroxy-4-pregnene-3,20-dione (17OHP) on the production of 17,20OHP. The sensitivity with which testes secreted 17,20OHP following stimulation with GtH II was maximum during spermatogenesis. The addition of 17OHP (10 to 1600 ng ml-1) to the culture medium of testes fragments induced a significant and dose-related increase in 17,20OHP secretion. Although the capacity to produce 17,20OHP was not saturated by the 17OHP concentrations used, the conversion rate was highest for tested at an immature stage. As to the regulation of 17OHP, in vivo, a single injection of partially purified salmon gonadotropin (50 ng g-1 body weight) induced a significant increase in the circulating levels of 17OHP of immature males. In conclusion, the maximum sensitivity to GtH II stimulation and the highest conversion rate of 17OHP to 17,20OHP in vitro, occurred before the dramatic increase in the 17,20OHP secretion observed in rainbow trout at the time of spermiation.  相似文献   

9.
Administration of Methallibure, a non-steroidal gonadotropin (GTH) inhibitor 20 g g–1 body weight; i.p., daily for 10 days, to prespawning phase female Heteropneustes fossilis inhibited the brain-pituitary-ovarian axis as indicated by significant reductions in plasma and pituitary levels of GTH-II, and plasma levels of 17-estradiol (E2) and testosterone. Concurrently, the treatment resulted in significant reductions in the hypothalamic content of serotonin, noradrenaline (and adrenaline) that stimulate, and a significant elevation of dopamine that inhibits GTH-II release in this species. Activities of the monoamine degrading enzymes, monoamine oxidase and catechol-O-methyltransferase were significantly increased, while that of the synthesizing enzymes, dopamine--hydroxylase and phenylethanolamine-N-methyltransferase were significantly decreased. These results suggest that the mechanism of inhibition of GTH-II secretion includes, among others, differential actions of the drug on hypothalamic monoamine metabolism.  相似文献   

10.
Three experimental approaches were chosen to study the question if the progestin 17-hydroxy-20-dihydroprogesterone (1720OHP) is synthesised in testes of young Oncorhynchus mykiss, in which the absence of spermatozoa was verified histologically: first, in order to detect 20-hydroxysteroid dehydrogenase activity (20HSD), testes homogenates were incubated with 3H-labeled 17OHP.Metabolites were analysed by TLC, HPLC, and repeated crystallization to constant isotope ratios. One of the metabolites was identified as 1720OHP-3H, indicating that already immature testes contain 20HSD activity and are able to produce 20-reduced steroids. Second, 1720OHP was quantified by radioimmunoassay in incubates of testes fragments. The sensitivity of the gonads to gonadotropin II (GtH II) became evident when comparing incubations in the absence and presence of GtH II. Third, plasma levels of 1720OHP were significantly higher in animals injected with partially purified salmon gonadotropin, compared to controls. Thus, for the first time, it could be shown that 20HSD is present in testicular cells other than spermatozoa. Furthermore, 1720OHP is indeed secreted at a very early stage of testicular development; 1720OHP secretion is also responsive to GtH II. Future studies will have to show if the functions of this progestin include the stimulation of spermatogenesis.  相似文献   

11.
The effect of [d-Lys6] salmon gonadotropin-releasing hormone alone (sGnRH) and in combination with domperidone, a dopamine antagonist, on the spawning of common carp, Cyprinus carpio, was investigated during the late spawning season (when mature fish stop spawning under natural conditions when males and females are put together). Fish were induced to spawn by [d-Lys6]-sGnRH salmon gonadotropin-releasing hormone analogue (sGnRH-X) only when injected in combination with domperidone at doses of 10 g kg-1 and 20 mg kg-1 body weight respectively. [d-Lys6]-sGnRH-X alone did not induce spawning at a dose as high as 100 g kg-1 body weight. Fish injected with domperidone alone did not spawn either. These results demonstrate that [d-Lys6]-sGnRH-X in combination with a dopamine antagonist is effective in inducing spawning during the late spawning season.  相似文献   

12.
The objective of the present study was to investigate the in vivo effects of different doses of 17-estradiol (E2) and testosterone (T) on the levels of plasma and pituitary gonadotropin II (GTH II) in 2-year-old black porgy, Acanthopagrus schlegeli, during the spawning season. Male fish were distributed among 7 groups (n = 49), control, E2or T (with 3 doses, 2.4 ng, 72 ng and 2.2 g g–1 body weight). Fish were injected with the respective vehicle or different doses of E2 or T on days 1 and 14. Plasma E2 levels were significantly increased in the 72 ng E2 group on days 8 and 14. Plasma vitellogenin levels were significantly higher in the 72 ng E2 group on days 14 and 20, and 2.2 g E2 group on days 8, 14 and 20 than those in the control group. Plasma GTH II concentrations were significantly higher in the 2.2 g E2 group than in the control and other E2groups on days 8, 14 and 20. Pituitary GTH II contents was significantly higher in the 7.2 ng E2 group compared to the control and other E2groups on day 20. Plasma GTH II concentrations were similar in the control and all the T groups on days 8, 14 and 20. None of the doses of T treatment stimulated pituitary GTH II content on day 20, although plasma vitellogenin levels were elevated. It is concluded that GTH II synthesis and secretion in black porgy is stimulated by an estrogen-specific effect.  相似文献   

13.
The accumulation of immunoreactive corticosteroids in the bile of rainbow trout during stress was monitored by radioimmunoassay and GUMS. Although plasma cortisol levels were elevated by confinement for 1 hour, biliary levels of free and conjugated steroids in the bile were unaffected. However, after 24 hours confinement, in addition to elevated plasma cortisol levels, free and conjugated steroids in the bile were also significantly higher than in control, unstressed fish. The time-course of change in plasma and biliary corticosteroid levels was determined in rainbow trout subject to 96 hours confinement stress. Free steroid levels in the bile of stressed fish were elevated within 2 hours of the onset of stress, while levels of conjugated steroids were significantly elevated within 4 hours of the onset of confinement. Analysis of bile from stressed fish, by GC/MS, established the major conjugated steroids present to be tetrahydrocortisone (230 g ml–1 bile), tetrahydrocortisol (75 g ml–1), cortisone (33.5 g ml–1), cortisol (25 g ml–1) and -cortolone (5 g ml–1). The data are discussed with reference to the role of cortisone and conjugating enzymes in the clearance of cortisol, and further data are presented to suggest that the analysis of biliary steroid content may provide a suitable means of identifying stressed fish under conditions in which an additional sampling stress is unavoidable.  相似文献   

14.
The effects of (ip, 10 injections over 20 days) of melatonin (75 g 100 g–1 BW), the serotonin (5-HT)-synthesis blocker, para-cholorophenylalanine (p-CPA, 10 mg 100g–1 BW) and the catecholamine-synthesis blocker, -methylparatyrosine (-MPT, 10 mg 100 g–1 BW) on gonadotropin (GTH) secretion and ovarian activity were studied in Heteropneustes fossilis during late preparatory to early prespawning (April–May). The treatments resulted in significant reductions of plasma GTH and estradiol-17 levels, the gonadosomatic index, frequency distribution of vitellogenic and postvitellogenic oocytes, and ovarian and serum 32p-labelled alkali-labile phosphoprotein (a marker of vitellogenic activity). Most of the oocytes were nonvitellogenic or had undergone atretic changes. The hepatic 32-phosphoprotein content increased significantly over the saline control value. The effects were similar and pronounced in the p-CPA and melatonin-treated groups but were moderate in the -MPT-treated group. Hypothalamic 5-HT content and turnover were significantly inhibited in the p-CPA and melatonin-treated groups but the content and turnover of catecholamines were not. The -MPT treatment decreased significantly the content and turnover of dopamine (DA), noradrenaline (NA), and adrenaline (A) but did not influence the 5-HT content or turnover. These results suggest that 5-HT, NA and A are stimulatory to GTH secretion and that melatonin may act on the serotonergic system to inhibit the pituitary-gonadal axis.to whom correspondence should be addressed.A part of the results was presented at the International Workshop on Pineal gland: Its molecular signals and published as an abstract in Neuroendocrinol. Lett. 14: 399 pp., 1992.  相似文献   

15.
16.
The plasma concentrations of fourteen ovarian steroids were measured in postvitellogenic African catfish,Clarias gariepinus, which had been injected with pimozide and LHRHa. Postvitellogenesis persisted for at least four hours after pimozide and LHRHa administration. During this stage a limited rise in the plasma gonadotropin (GTH) level was accompanied by an increase in the testosterone concentration. The estradiol level was high and remained high except for a passing drop during the stage of germinal vesicle migration. At the stage of germinal vesicle migration a strong increase in the plasma GTH level coincided with a maximum in the testosterone concentration and a concomitant increase in the levels of 17,20-dihydroxy-4-pregnen-3-one and of five 5-reduced pregnanes. During germinal vesicle breakdown the GTH concentration remained high, the plasma level of 17-hydroxyprogesterone tended to increase, and the levels of 5-pregnane-3, 17-diol-20-one, 5-pregnane-3,17,20-triol and 5-pregnane-3,17,20-triol reached a maximum. At pre-ovulation the GTH concentration did not change, and peak levels were reached of 17,20-dihydroxy-4-pregnen-3-one and 5-pregnane-3,6,17-triol-20-one. Shortly after ovulation the GTH concentration slightly decreased together with a sharp decline in the concentrations of 17,20-dihydroxy-4-pregnen-3-one and the 5-reduced steroids, with the exception of 5-pregnane-3,17,20-triol, 5-pregnane-3,6,17,20-tetrol and 5-dihydrotestosterone. The plasma concentrations of androstenedione, estrone, etiocholanolone and 5-androstane-3,17-diol showed marginal fluctuations during oocyte maturation and ovulation. Apart from 17,20-dihydroxy-4-pregnen-3-one, the 5-reduced pregnanes might be candidates for the function of oocyte maturation inducing hormone inC. gariepinus.  相似文献   

17.
18.
Doppler flow probes were fitted around the ventral aorta of rainbow trout, which were exposed to combinations of pH and aluminum (pH 5.1–6.2, Al 0–80 g l–1) for 60 h. Fish accumulated Al at the gill and exhibited decreased blood Na+, Cl, and Ca2+ concentrations, and increased K+, hematocrit, hemoglobin, glucose, and lactate, indicating increasing ionoregulatory disturbance with increasing Al concentration. Fish exposed to ambient water (6.2) or low-pH (5.3) water without Al exhibited slight reductions in heart rates, as well as increased stroke volume, resulting in little variation in cardiac output. In the presence of Al (20 to 40 g l–1) at low pH (5.1–5.3), fish increased their heart rate slightly and generally maintained their stroke volume, resulting in increased cardiac output in the first two days of exposure. At the highest Al concentration (80 g l–1, pH 5.1), tachycardia was observed, concomitant with a decrease in stroke volume. The ionoregulatory imbalance and resulting increased blood viscosity explain these increases in heart rate rather than stroke volume in fish exposed to high concentrations of Al.  相似文献   

19.
A particulate fraction obtained from trout testis at the time of spermiation shows saturable binding sites for125I-labeled salmon gonadotropin (125I-GtH). Non-gonadal tissues (liver, muscle and spleen) did not demonstrate specific125I-GtH binding. The tracer's specific activity was determined by the self-displacement method (18 to 30 Ci/g). Maximal specific binding ability of125I-GtH varied from 20 to 30% of the labelled ligand added, depending on the hormone preparation. Specific binding of125I-GtH to 20 mg of the testis membrane varied from 40 to 85% of the total binding depending on the method of membrane prepratation, and was competitively inhibited by concentrations of unlabelled GtH ranging from ca 1 to 1000 ng/ml of incubate. Gonadotropin of mammalian origin, ovine TSH or salmon prolactin competed only weakly, or not at all, for testicular gonadotropin binding sites (relative potencies s-GtH>>FSH=hCG>s-PRL>bTSH). Scatchard analysis of equilibrium binding studies shows that saturable gonadotropin binding was due to a class of high affinity binding sites (sites I Ka3×1010 M–1) and possibly to a second class of lower affinity binding sites (sites II Ka=5 to 14×108 M–1). The binding capacity of sites I, as measured in enriched membrane preparations, was 45±18 fmoles/g of testis during the period of spermiation. The concentration of GtH required to obtain half maximal displacement of125I-GtH in the binding studies was of the same order of magnitude as the apparent ED50 for GtH stimulation of 11-Cetotestosterone (11KT) secretion by trout testesin vitro. Mammalian LH and FSH were 100 to 1000 folds less potent than salmor GtH to increase 11 KT secretion.  相似文献   

20.
The in vitro effects of several steroids on the maturation of intact white sturgeon (Acipenser transmontanus) ovarian follicles were investigated. At the highest concentration (1024 ng ml–1 for the C21 steroids and 1139 ng ml–1 for the C19 steroids), all of the C21 steroids tested, progesterone (P4), 17-hydroxyprogesterone (17OHP), 17,20-dihydroxy-4-pregnen-3-one (17,20-P), 17,(20,21-trihydroxy-4-pregnen-3-one 20-S), 11-deoxycortisol (S) and cortisol (F), as well as testosterone (T) induced germinal vesicle breakdown (GVBD) at 14 and 22 h. At 6 h, only P4 and 17,20-P induced maturation at the highest concentration (1024 ng ml–1). At 14 and 22 h, 11-deoxycortisol was the most potent steroid inducer of GVBD followed by P4, 17OHP, 17,20-P, and 20-S. The steroid 11-hydroxytestosterone (11OHT) was completely ineffective at all concentrations and exposure times. The C21 steroids induced oocyte maturation at concentrations ranging from 4 to 1024 ng ml–1, whereas T induced GVBD at 225 to 1139 ng ml–1. Calculation of the mean effective concentration that induced 50% GVBD (EC50) from the 22 h incubations revealed the following order of potencies: S > P4 > 17OHP > 17,20-P > 20-S >> F > T. These bioassay results, together with previous findings on the endogenous production of steroids by ovarian follicles from gonadotropin-primed females, indicate that more than one steroid has a biological role in the resumption of meiosis in sturgeon oocytes and provides empirical evidence for P4, 17OHP, S, 20-S, and 17,20-P as maturation-inducing steroids in white sturgeon.  相似文献   

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