Changes in luteinizing hormone-releasing hormone content of discrete hypothalamic areas associated with spontaneous and induced preovulatory luteinizing hormone surges in the domestic hen

It is widely assumed that luteinizing hormone-releasing hormone (LHRH) neuronal activation is involved in the preovulatory surge of LH in the hen. In addition, this LH surge may be initiated by ovarian progesterone (P4) release. Thus, spontaneous and P4-induced LH surges should be associated with acute changes in LHRH content of discrete hypothalamic areas associated with LHRH cell bodies and/or LHRH axon terminals. Medial preoptic area (mPOA) and infundibulum (INF) LHRH content was measured by radioimmunoassay at intervals before, at, and following peak LH levels of a spontaneous preovulatory surge of LH, as well as when this surge was advanced by P4 administration in laying hens. Nonlaying birds served as additional controls. Levels of serum LH, P4, 17 beta-estradiol and pituitary LH were also measured. Increased (P less than 0.05) LHRH content in mPOA without changes in the INF are associated with peak serum LH levels of the spontaneous LH surge. By contrast, decreased (P less than 0.05) LHRH content in both mPOA and INF is associated with peak serum LH levels when the spontaneous surge was advanced 8 h by P4 administration to laying hens. Medial preoptic area and INF LHRH contents were significantly lower (P less than 0.05) in nonlaying than in laying hens.(ABSTRACT TRUNCATED AT 250 WORDS)     

Hypothalamic alterations and reproductive aging in female rats: evidence of altered luteinizing hormone-releasing hormone neuronal function

Prior to the age-related loss of regular estrous cycles, female rats exhibit an attenuated preovulatory LH surge, a sign that reproductive decline is imminent. Numerous studies have revealed an important role for the hypothalamus in aging of the reproductive axis in this species. Because LHRH represents the primary hypothalamic signal that regulates gonadotropin release, assessments of LHRH neuronal activity can provide a window into hypothalamic function during reproductive aging. Studies of the dynamic activity of LHRH neurons during times of enhanced secretion have revealed deficits in middle-aged females. Available data are consistent with a decline in LHRH synthesis, transport, and secretion in middle-aged females during times of increased demand for LHRH output. Moreover, the alterations noted in LHRH neuronal function could account, in part, for the attenuation and eventual loss of the preovulatory LH surge with age. Elements extrinsic to LHRH neurons undoubtedly contribute to the decline in the parameters of LHRH neuronal function observed in middle-aged females. Whether alterations intrinsic to LHRH neurons also play a role in the age-associated reduction in LHRH synthesis and secretion remains to be determined. Recent examinations of hormone profiles during the perimenopausal period suggest that a potential hypothalamic contribution to aging of the reproductive axis in women warrants further examination.     

Effect of adrenalectomy and 5-hydroxytryptophan on phasic release of luteinizing hormone

The effect of 5-hydroxytryptophan (5-HTP) on serum progesterone and the possible role of adrenal progesterone in mediating stimulation by 5-HTP of phasic release of luteinizing. hormone (LH) were investigated in estradiol benzoate (EB)-treated ovariectomized rats. LH surges were induced in long-term (at least two weeks) ovariectomized rats by two injections of EB (20 micrograms/rat, s.c.) with an interval of 72 hrs. Administration of 5-HTP (50 mg/kg, i.p.) at 1000 hr in EB-treated ovariectomized rats resulted in a four-fold increase in serum progesterone within 30 mins, and significantly stimulated the LH surge at 1600 hr. This facilitative effect of 5-HTP on serum LH, but not progesterone, was further potentiated in rats pretreated with P-chlorophenylalanine (PCPA) 72 hrs earlier. Adrenalectomy shortly before 5-HTP administration attenuated the LH surge in saline treated controls, and completely blocked the facilitative effect of 5-HTP on the afternoon surge of LH in rats pretreated with PCPA 72 hrs earlier. On the other hand, chronic adrenalectomy (for 6 days) followed by hydrocortisone (0.2 mg/rat/day) replacement not only had no effect on the LH surge in saline treated controls, but also failed to prevent 5-HTP from facilitating the LH surge in PCPA pretreated rats. On the first day of bleeding, the basal LH value at 1000 hr in sham operated controls was significantly suppressed by PCPA pretreatment 48 hrs earlier. The second dose of 5-HTP administered on the next day failed to potentiate LH surges in either sham operated or adrenalectomized rats.(ABSTRACT TRUNCATED AT 250 WORDS)     

A possible role for progesterone in the preovulatory gonadotropin surge through modulation of LHRH degrading activity

The preovulatory surge of gonadotropins is triggered by estradiol and enhanced to its full magnitude by progesterone. Progesterone may exert this effect through several mechanisms. One of the mechanisms is through the ability of progesterone to induce an increase in the hypothalamic content and release of LHRH. The purpose of this study was to determine if progesterone might not act through yet another mechanism and facilitate LHRH release of the proestrous gonadotropin surge through modulation of luteinizing hormone releasing hormone (LHRH) degrading activity. Sixty-day-old Sprague-Dawley rats were ovariectomized; 14 days later, the estradiol-progesterone milieu of proestrous was mimicked in these animals through the use of estradiol containing silastic implants and subcutaneous progesterone injections. The LHRH degrading activity of the hypothalamus, pituitary and serum were monitored subsequently at preselected time points. In the hypothalamus, estradiol alone was capable of inducing significant increase in degrading activity; progesterone alone had no effect; however, progesterone subsequent to estradiol priming suppressed the increase induced by estradiol alone. In the pituitary, neither estradiol alone nor progesterone alone nor progesterone subsequent to estradiol priming had any significant effect on degrading activity. In the serum, estradiol induced a rapid and significant increase in activity; progesterone alone suppressed activity; progesterone subsequent to estradiol priming induced a similar but more rapid suppression. Therefore, the overall tendency was for estradiol to stimulate and progesterone to suppress LHRH degrading activity in the tissues studied. The results of this study indicate that progesterone has the capacity to suppress LHRH degrading activity and may be one of the mechanisms capable of increasing the availability of LHRH to the anterior pituitary gland thereby facilitating the preovulatory gonadotropin surges.     

Dose-dependent inhibition of the preovulatory surges of gonadotropins and prolactin by the antiestrogen CI-628: possible sites of action

The present series of experiments was conducted in an attempt to correlate previously reported dose-dependent and site-selective inhibitory effects of an antiestrogen, CI-628, on 17 beta-estradiol (E2)-receptor interactions in the anterior pituitary gland (AP) and hypothalamus with its effects on the preovulatory surges of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and prolactin. The effects of CI-628 on the response of the AP to luteinizing hormone-releasing hormone (LHRH) and thyrotropin-releasing hormone (TRH) also were examined. In the first study, rats exhibiting 4-day estrous cycles were injected with various doses (0.02, 0.20, 2.0, and 20 mg/kg) of CI-628 or vehicle at 0900 h on diestrus-2 and proestrus. The preovulatory LH surge and both preovulatory and secondary FSH surges were marginally affected by 0.02 mg/kg CI-628, but were completely abolished by higher doses. In contrast, a dose of 0.20 mg/kg only delayed the prolactin surge; however, higher doses were effective in extinguishing cyclic prolactin release. In a second experiment, CI-628 in rats treated on diestrus-2 and proestrus exerted a dose-dependent suppression of the AP LH response to an initial injection of LHRH on proestrous afternoon in rats whose endogenous LH surges were blocked by phenobarbital. However, AP LH responses to a second LHRH injection to assess the self-priming capacity of LHRH were attenuated only in rats given 0.20, 2.0, and 20 mg/kg CI-628. Contrastingly, the AP prolactin response to TRH was suppressed only in rats given 0.20 mg/kg CI-628.(ABSTRACT TRUNCATED AT 250 WORDS)     

A comparison of human chorionic gonadotropin and luteinizing hormone releasing hormone on the induction of spermiation and amplexus in the American toad (Anaxyrus americanus)

ABSTRACT: BACKGROUND: Captive breeding programs for endangered amphibian species often utilize exogenous hormones for species that are difficult to breed. The purpose of our study was to compare the efficacy of two different hormones at various concentrations on sperm production, quantity and quality over time in order to optimize assisted breeding. METHODS: Male American toads (Anaxyrus americanus) were divided into three separate treatment groups, with animals in each group rotated through different concentrations of luteinizing hormone releasing hormone analog (LHRH; 0.1, 1.0, 4.0 and 32 micrograms/toad), human chorionic gonadotropin (hCG; 50, 100, 200, and 300 IU), or the control over 24 hours. We evaluated the number of males that respond by producing spermic urine, the sperm concentration, percent motility, and quality of forward progression. We also evaluated the effects of hCG and LHRH on reproductive behavior as assessed by amplexus. Data were analyzed using the Generalized Estimating Equations incorporating repeated measures over time and including the main effects of treatment and time, and the treatment by time interaction. RESULTS: The hormone hCG was significantly more effective at stimulating spermiation in male Anaxyrus americanus than LHRH and showed a dose-dependent response in the number of animals producing sperm. At the most effective hCG dose (300 IU), 100 % of the male toads produced sperm, compared to only 35 % for the best LHRH dose tested (4.0 micrograms). In addition to having a greater number of responders (P < 0.05), the 300 IU hCG treatment group had a much higher average sperm concentration (P < 0.05) than the treatment group receiving 4.0 micrograms LHRH. In contrast, these two treatments did not result in significant differences in sperm motility or quality of forward progressive motility. However, more males went into amplexus when treated with LHRH vs. hCG (90 % vs. 75 %) by nine hours post-administration. CONCLUSION: There is a clear dichotomy between the two hormones' physiological responses on gamete production and stimulation of amplexus. Understanding how these two hormones influence physiology and reproductive behaviors in amphibians will have direct bearing on establishing similar breeding protocols for endangered species.     

Human chorionic gonadotropin and luteinizing hormone-releasing hormone reverse the blockade of ovulation in pregnant mare's serum gonadotropin-primed immature rats by the anti-androgenic drug, hydroxyflutamide

The present study was designed to examine mechanism(s) of the anti-ovulatory action of the anti-androgen, hydroxyflutamide (OH-F). Prepubertal rats were treated with 4 IU pregnant mare's serum gonadotropin (PMSG) (day -2) to induce first estrus and ovulation. They received OH-F in sesame oil or oil alone at 08:00 and 20:00 h on day 0 (the day of proestrus) and ovulations were assessed on the morning of day 1. Eighty-three percent of control animals ovulated with a mean of 7.7 +/- 1.1 corpora lutea per rat. Hydroxyflutamide blocked ovulation in all but 2 of the 12 rats receiving this drug alone. All of OH-F treated rats that received 5 and 25 IU human chorionic gonadotropin (hCG) ovulated with means +/- SEM of 9.1 +/- 0.1 and 7.3 +/- 1.4 corpora lutea per rat, respectively. The dose of 0.2 IU hCG was essentially ineffective, while the effect of 1.0 IU hCG was intermediate. At the dose of 20 ng and above (100 and 500 ng) luteining hormone-releasing hormone (LHRH) completely overcame the ovulation blockade in the OH-F treated animals, while a 4-ng dose was ineffective. At 18:00 h on the day of proestrus, serum LH levels in control animals were 17.56 +/- 2.60 ng/mL, which were 920% above basal levels (1.90 +/- 0.13) indicating a spontaneous LH surge. This surge was suppressed in OH-F treated rats. Injection of LHRH, at the dose of 20 ng and above, reinstated the LH release in OH-F treated animals. Thus, the anti-androgen, OH-F, inhibits ovulation in PMSG-treated immature rats through its interference with the preovulatory LH surge; the inhibition can be reversed by hCG or LHRH. Hydroxyflutamide does not appear to interfere at the level of the pituitary, but may have direct action at the hypothalamic and (or) extrahypothalamic sites involved in the generation of positive feedback signals that control LH release.     

Induced ovulation and embryonic development of ocellated puffer, Takifugu ocellatus

Acute injections of different hormones to induce ovulation in mature ocellated puffer, Takifugu ocellatus, collected from natural waters during the spawning season, were carried out to develop a reliable protocol for mass production of seed in this species. All experimental fish were divided into seven groups treated with: a saline injection (control), single or two injections of luteinizing hormone‐releasing hormone analog (LHRH‐a; single injection: 50 μg kg^?1, two injections: 10 and 40 μg kg^?1), single or two injections of pituitary (single injection: 6 mg kg^?1, two injections: 1 and 5 mg kg^?1) and single or two injections of human chorionic gonadotropin (hCG; single injection: 2500 IU kg^?1, two injections: 500 and 2000 IU kg^?1), respectively. The percentage of fish that ovulated in six hormonal treatments reached 100%, either with a single injection or with two injections whereas the fish in control group failed to spawn. There were no significant differences among all hormonal treatments in egg production, fertilization rate, or hatch rate (P > 0.05) except time to ovulation between a single injection group and the two‐injection group (P < 0.05). The fertilized eggs of ocellated puffer were spherical, demersal, and adhesive. They had a mean oocyte diameter of 1.487 ± 0.106 mm (range: 1.404–1.560). The egg membrane was transparent and yolk was buff in color, containing a cluster of small oil globules. Thirty‐four successive stages of embryonic development were identified and characterized. Fertilized eggs incubated at 18–20°C generally commenced hatching at 144 h after fertilization. Newly hatched larvae were about 3.26–3.45 mm in length. The induced ovulation technique using acute injections of hormones is an important step in the development of the culture of the ocellated puffer.     

IGF—ImRNA在不同年龄鲤表达的差异和LHRH—A对IGF—ImRNA表达的影响

利用RNA酶保护法对7月龄性未成熟幼鲤和2龄性成熟鲤组织胰岛素样生长因子-I（IGF-I）mRNA的表达水平进行测定,结果表明成鱼肝和肾脏组织IGF-ImRNA的丰度显著高于幼鱼,对鲤成鱼和幼鱼腹腔注射促性腺激素释放激素类似物（LHRH-A,D-Ala^6-Pro^9-NEt-LHRH)使血清生长激素（GH）水平和肝组织IGF-ImRNA水平都显著升高,而成鱼生殖腺IGF-ImRNA的丰度比对照组显著增加,研究结果提示鲤在不同发育阶段肝组织IGF-ImRNA的丰度比对照组显著增加,研究结果提示鲤在不同发育阶段肝组织IGF-ImRNA的表达存在差别,其中2龄成鱼大于7月龄幼鱼;LHRH-A可能通过刺激垂体GH的释放间接促进肝组织IGF-ImRNA的表达,亦可能通过某种未知途径刺激生殖腺IGF-ImRNA的表达。     

Estimation of population fecundity of the bitterling,Rhodeus ocellatus,and ecological significance of its spawning habit into bivalves

Fluctuations of the population abundance of the rose bitterling,Rhodeus ocellatus (Kner) in a small pond, Shimizu-ike (700 m²), Osaka Prefecture, Japan, were estimated by the Petersen method from 1973 to 1977. The number of fish fluctuated between 12,600 and 46,700 during that period. In 1974, a large reproductive peak in May contributed mainly by 2- and 3-year-old spawners and a small peak in late July contributed by 1–2-year-old fish were observed. Average number of eggs laid in a bivalve,Anodonta woodiana Lea, in each month was estimated with field experiments from March to November, 1974. In total, 93,400 eggs were laid during the first reproductive peak, and 13,100 eggs during the second reproductive peak. The mortality of eggs and larvae incubated in the bivalve was less than 30%, and approximately 70 % of the larvae that had swum out from the host died in the following six months. Thus, it is estimated that approximately 20% of the eggs laid in the bivalves can survive and grow up to reach the first maturity. The high survival rate ofR. ocellatus among cyprinid fishes might be due to the fact that the eggs and larvae are protected from predation by being embedded in a bivalve, and to the fact that the larvae at the earliest free swimming stage have a good opportunity of surviving because they are much larger in size and more developed morphologically than those of other cyprinid fishes.     

Divergence in reproductive ecology of the three-spined stickleback,Gasterosteus aculeatus

An anadromous population (trachurus form) and three freshwater populations (leiurus form) of the three-spined stickleback,Gasterosteus aculeatus, in central Japan were compared with one another in the reproductive traits: body length at maturity, egg size, clutch size, the swell of abdomen and the number of eggs per nest. The anadromous fish which become larger in body size at maturity spawn eggs smaller in size and greater in number than the freshwater fish. The abdomen swell of gravid females expressed by the proportion of abdomen width to body length was greater in the freshwater fish. The anadromous male fish collected a mean of 2,638 eggs with a range of 1,119 to 4,052 eggs from about 6–7 females. In the three freshwater (the Yamayoke, the Tsuya and the Jizo) populations, males must have mated with about 9–22, 7–18 and 4–7 females respectively. It seems that theleiurus form increases its reproductive success by its much more mating opportunities and the parental efforts of nesting males as well as by spawning large eggs. Furthermore, among the freshwater populations, the Jizo one inhabiting the upper stream was clearly larger in body size, in egg size and in clutch size than the Yamayoke and the Tsuya ones which inhabit stable waterbodies with springs. It is possible that the Jizo population adopted the strategy of spawning a few large eggs as an adaptation to its habitat. The causal and functional explanations in reproductive characteristics among the four populations are discussed in regard to differences in the environmental conditions.     

诱导大鳍鱯和长吻鮠排卵过程中血清GTH水平的变化

繁殖期从嘉陵江收集性成熟的大鳍■ 和长吻 野生亲鱼,用Ｌｉｎｐｅ方法（即ＬＨＲＨ－Ａ加多巴胺Ｄ２受体拮抗剂地欧酮）或传统的ＬＨＲＨ—Ａ加脑垂体的方法进行催产,定时取血样,用放射免疫方法测定催产过程中血清ＧＴＨ水平的变化,进一步证实鲇形目鱼类ＧＴＨ的分泌受到下丘脑分泌的促性腺激素释放激素ＧｎＲＨ和多巴胺的双重调节;排卵和产卵也是以血清ＧＴＨ的急剧升高为先导的,而最终能否排卵还有赖于血清ＧＴＨ峰是否超过“排卵阈值”。尽管催产后的大鳍 和长吻 雄鱼血清ＧＴＨ水平也有一个高峰出现,但血清ＧＴＨ水平升高幅度都大大低于雌鱼,这种现象在硬骨鱼类可能具有普遍性。     

Reproductive cycle in female Waigieu seaperch (Psammoperca waigiensis) reared under different salinity levels and the effects of dopamine antagonist on steroid hormone levels

Waigieu seaperch (Psammoperca waigiensis) is a tropical marine finfish species, which may inhabit wide range of salinity during the entire life cycle. Regardless of the wide salinity tolerance, little is known about how salinity may influence the reproductive endocrinology of this important tropical species. In the present study, we investigated the seasonal variations in steroid hormone levels, oocyte maturation (OM) and ovulation in fish reared under different salinity levels. In addition, we investigated the effects a dopamine antagonist (domperidone: DOM) during the peak spawning period. Mature brood fish at 3 years old were cultured in four different salinities of 5, 10, 20 and 30‰ (part per thousand, ppt) from March to December 2007. Blood samples were collected monthly and key steroid hormones (testosterone (T), 11-ketotestosterone (11-KT), 17β-estradiol (E2) and progesterone (P)) levels in female fish plasma were analyzed using enzyme immunoassay method. OM, ovulation and several reproductive performance indexes were evaluated twice per month during the breeding season. Plasma hormone analysis showed significant differences in fish groups cultured in different salinities during the seasonal cycle. The gonadosomatic index (GSI) gradually increased from March, peaking in July at 10 and 30‰, and then decreased thereafter until December. Plasma steroid levels (E2, T, 11-KT and P) were observed in variable concentrations during the spawning period and showed monthly fluctuations that were apparently salinity dependent during on and off-seasons. Interestingly, 11-KT levels were found in small amounts in female fish and peaked in September at salinities of 10 and 20‰. DOM exposure produced significant differences in steroid hormone levels that were apparently dependent on dose, exposure time and the individual hormone. The present study indicated that holding Waigieu seaperch brood fish in different salinities during the breeding season produced significant effects on gonadal development and spawning incidences. The first spawning was observed on 8th of April for all groups after 100% water exchange at night. No significant differences of absolute and relative fecundity were observed between groups. Fish held at 10 ppt salinity during the breeding season spawned naturally, but maturity and spawning rates were reduced and no hatching was observed at the same salinity. Fish holding at 5‰ resulted to 100% mortality in brood fish. This indicated that salinity limits for oocyte maturation or ovulation and subsequent reduction in spawning, but embryonic development requires a higher salinity. The effect of salinity on embryo development and hatching rate suggests that in breeding season the brood fish spawned in areas where the salinity and other parameters are favorable.     

Induced ovulation and egg deposition in the direct developing anuran <Emphasis Type="Italic">Eleutherodactylus coqui</Emphasis>

This study investigates ovulation and egg deposition behaviors in the anuran Eleutherodactylus coqui from Puerto Rico in response to stimulation with gonadotropin and gonadotropin releasing hormones. Five hormones were tested by injection over a range of doses, including mammalian LHRH, avian LHRH, fish LHRH, D-Ala6, des-Gly10 ethylamide LHRH and hCG. We report a low level of ovulation and egg deposition in response to all hormones, with the most complete and consistent results from the non-natural D-Ala6, des-Gly10 ethylamide LHRH derivative. To confirm the viability of eggs produced in this manner we performed in vitro fertilization experiments that resulted in the development of normal frogs. Reproductive behaviors in E. coqui are apparently not controlled by a mammalian form of LHRH as reported in other common laboratory anuran species. D-Ala6, des-Gly10 ethylamide LHRH induces ovulation and deposition of mature and fertilizable eggs in E. coqui.     

Carry-over effect of captive breeding reduces reproductive fitness of wild-born descendants in the wild

Supplementation of wild populations with captive-bred organisms is a common practice for conservation of threatened wild populations. Yet it is largely unknown whether such programmes actually help population size recovery. While a negative genetic effect of captive breeding that decreases fitness of captive-bred organisms has been detected, there is no direct evidence for a carry-over effect of captive breeding in their wild-born descendants, which would drag down the fitness of the wild population in subsequent generations. In this study, we use genetic parentage assignments to reconstruct a pedigree and estimate reproductive fitness of the wild-born descendants of captive-bred parents in a supplemented population of steelhead trout (Oncorhynchus mykiss). The estimated fitness varied among years, but overall relative reproductive fitness was only 37 per cent in wild-born fish from two captive-bred parents and 87 per cent in those from one captive-bred and one wild parent (relative to those from two wild parents). Our results suggest a significant carry-over effect of captive breeding, which has negative influence on the size of the wild population in the generation after supplementation. In this population, the population fitness could have been 8 per cent higher if there was no carry-over effect during the study period.     

Effects of luteinizing hormone releasing hormone on gonadotrophins in the hamster

The changes in serum gonadotrophins in male hamsters following one injection of 15 μg luteinizing hormone releasing hormone (LHRH) (Group A) were compared with those following the last injection of LHRH in animals receiving an injection approximately every 12 hr for 4 days (Group B) or 12 days (Group C). Peak follicle stimulating hormone (FSH) levels (ng/ml) were 1776±218 (Group A), 2904±346 (Group B), and 4336±449 (Group C). Peak luteinizing hormone (LH) values (ng/ml) were 1352±80 (Group A), 410±12 (Group B), and 498±53 (Group C). Serum FSH:LH ratios, calculated from the concentrations measured 16 hr after the last LHRH injections, were higher in Groups B and C than in Group A. Similar injections of LHRH (100 ng or 15 μg/injection) for 6 days elevated the serum FSH:LH ratio in intact males. Five such LHRH injections (100 ng/injection) blunted the rise in serum LH in orchidectomized hamsters. Direct effects of LHRH on gonadotrophin secretory dynamics or altered brain-pituitary-testicular interactions may alter the ratio of FSH to LH in the hamster.     

Growth,reproduction, and feeding oF Fundulus heteroclitus (L.) on a North Carolina salt marsh

Fundulus heteroclitus (L.) from a North Carolina Spartina marsh teed largely on small crustaceans (amphipods, tanaids and copepods) and polychaetes. Fish longer than 30 mm standard length also ingested considerable amounts of living plant material. Smaller individuals were distinctly carnivorous. Recognizable particles of Spartina detritus occurred in less than 15% of the guts examined. The relationship of weight to length changed significantly during the year. Although females were larger than males of the same age. males were heavier than females of the same length, except for a brief period at the peak of the spawning season in the early spring. An average second season fish may lay up to 512 eggs from March through August, but first season fish did not reach reproductive size by the end of the spawning season. Growth of first season fish in mid-summer averaged 5% of their total weight per day. These significant seasonal changes in ecological properties of killifish populations are important in any estimates of growth, reproduction, and production.