Oocyte development and serum profiles of vitellogenin and steroid hormone levels in captive female Pacific herring Clupea pallasii during their first maturational cycle

ABSTRACT:   The present study investigates the relationship between oocyte development and serum steroid hormone levels in captive Pacific herring, Clupea pallasii , during the first reproductive cycle. The process of oocyte development in Pacific herring belongs to the group-synchronous type. Maturity of the ovary was divided into six periods based on histological observation (i.e. immature (April to September), onset of vitellogenesis (August to October), progress of vitellogenesis (October to December), completion of vitellogenesis (December to March), maturation and spawning (March to April) and spent (late April)). The pattern of seasonal change in the gonadosomatic index (GSI) well reflected the ovarian maturity. Serum vitellogenin levels showed good correlation with change in GSI, which increased from September to a peak (4.2 ± 0.3 mg/mL) in March. Serum estradiol-17β (E2) levels elevated from September and reached a peak (15.8 ± 4.2 ng/mL) in December, and remained comparatively high until March, suggesting that the active vitellogenin synthesis during vitellogenesis is controlled by the high E2 level. 17,20β-Dihydroxy-4-pregnen-3-one showed a single sharp peak (2.4 ± 0.28 ng/mL) in early April of the second year, suggesting it was a maturation-inducing steroid in this species.     

Delayed ovarian development and reduced serum steroid levels in female largemouth bass, Micropterus salmoides Lacépède, raised on pelleted feed

Female largemouth bass, Micropterus salmoides Lacépède, were raised on pelleted feed or a forage diet consisting of goldfish, Carassius auratus (L.). At the start of the present study, forage-fed bass were significantly smaller than pellet-fed bass. Forage-fed bass exhibited a significant increase in body weight, but not length, during the first month of the study; no further increases were seen in subsequent months. Pellet-fed bass did not grow significantly during the course of the study. In forage-fed bass, the gonadosomatic index (GSI) reached maximum levels in March; pellet-fed bass exhibited delayed ovarian recrudescence, reaching peak GSI approximately 4 weeks later than the forage-fed bass. There was no significant difference in the maximum GSI obtained in the two groups of fish. Serum levels of testosterone and oestradiol-17β reflected the pattern of GSI in both groups. Delayed ovarian recrudescence and suppressed serum steroid levels in pellet-fed bass might result from negative feedback effects of elevated levels of steroids found in the pelleted feed. These effects appear to be transient since pellet-fed bass resume ovarian development despite continuing to receive high steroid levels in their feed.     

养殖银鲳性腺发育规律和性类固醇激素变化

为掌握养殖银鲳的性腺发育规律及其与性类固醇激素水平的关系,本研究采用组织切片技术和酶联免疫吸附法(Elisa),观察养殖条件下银鲳的精巢与卵巢发育特征和血清中性类固醇激素的周年变化,并分析性腺成熟指数(GSI)、肝重指数(HSI)和肥满度(CF)与性腺发育的关系。结果显示,养殖银鲳雌鱼1龄即可发育成熟,成熟卵巢呈一对""形的囊状结构。11月份有近一半个体发育至III期,33.3%已发育至IV期,12月—翌年1月越冬期间约2/3退化至II期,3月又迅速发育至IV期,4月份达到V期并产卵,5月份产卵结束卵巢进入恢复期IV期。银鲳精巢属小叶型,III期精巢出现小叶腔,IV期精巢出现精子,精巢发育速率快于卵巢,即"雄性早熟",7月龄约20%个体可达到性成熟,越冬时退化至III期,3月重新发育,精子分批成熟排精。雌鱼GSI值为0.19%~12.89%,HSI为0.97%~2.30%,CF为2.30~3.08 g/cm~3,雄鱼的GSI为0.08%~2.62%,HSI值为0.73%~1.83%,CF为2.11~2.80 g/cm~3,GSI值在V期时达到最大,HSI和CF值则于IV期达到峰值。雌鱼血清中雌二醇(E2)周年表达水平为23.27~59.13 pg/m L,雄鱼为15.90~36.20 pg/m L,雌鱼的睾酮(T)为14.57~68.67 nmol/L,雄鱼的T为18.62~66.49 nmol/L,E2水平与HSI值呈现显著正相关关系,T表达水平与GSI值呈现显著正相关关系。研究表明,银鲳血清中性类固醇激素含量与精巢和卵巢的发育密切相关,可作为了解养殖银鲳性腺发育的重要指标。     

Involvement of steroid hormones in the preovulatory gonadotropin surge in female goldfish

Goldfish,Carassius auratus, spawn several times within a spawning season. A gonadotropin (GtH) surge occurs at the time of ovulation in this cyprinid species. This ovulatory GtH surge mediates the processes of final oocyte maturation and ovulation, and occurs at the end of each spawning cycle. Within a cycle, there is a shift in the predominant plasma steroid from estradiol to testosterone, and finally 17α, 20β-dihydroxy-4-pregnen-3-one at the time of the ovulatory GtH surge. High levels of testosterone were always observed before ovulation. When ovariectomized or sexually regressed female goldfish were implanted with testosterone, they exhibited a GtH surge which was similar to those normally observed at ovulation. These results strongly suggest that elevated plasma testosterone is an important physiological requirement for the occurrence of the GtH surge.     

Testicular development and serum profiles of steroid hormone levels in captive male Pacific herring Clupea pallasii during their first maturational cycle

The present study investigates the relationship between testicular development and serum steroid hormone levels in captive Pacific herring Clupea pallasii during the first reproductive cycle. The maturity of the testis was divided into five periods based on histological observation. These are early spermatogenic stage (April to July), mid-spermatogenic stage (August to November), late spermatogenic stage (December to March), functional maturation stage (early April) and spent stage (late April). The pattern of seasonal change in gonadosomatic index (GSI) clearly reflected testicular maturity. 11-Ketotestosterone (11-KT) levels increased from October to a peak level (6.58 ± 1.87 ng/mL) in January, and were maintained at this level until March. In contrast, testosterone levels were consistently low, less than 1 ng/mL, at all times. These results suggest that 11-KT is the predominant androgen that controls spermatogenesis in this species. 17,20β-Dihydroxy-4-pregnen-3-one (DHP) showed a single sharp peak (3.38 ± 0.35 ng/mL) in early April of the second year, suggesting that milt production is induced by DHP as in some other teleost species.     

Serum steroid profiles in artificially maturing female Japanese eel,Anguilla japonica

To investigate whether steroid profiles in salmon pituitary homogenate (SPH)-induced artificially maturing Japanese eel, Anguilla japonica, resemble those in other, naturally maturing fishes, the daily changes in 11 steroids were analyzed for a 70-day period (average time needed to reach the maturational phase). Concentrations of most steroids were low and changed on a weekly basis, with maximum values 2–5 days after an SPH injection. Thus, pregnenolone, 17α-hydroxypregnenolone, dehydroepiandrosterone, progesterone, 17α-hydroxyprogesterone, 17α,20 β-dihydroxy-4-pregnen-3-one, androstenedione and estrone levels were barely or not detectable in serum throughout the experimental period, which is largely in keeping with what is known about oogenesis-related steroids in other fishes. In contrast, serum testosterone (T) levels were high, but fluctuated considerably with each SPH injection (about 0.3–8.3 ng/ml). The serum estradiol-17β (E₂) levels increased after SPH injections and gradually rose throughout the experiment, peaking at the end of the experimental period (about 0.2–7.8 ng/ml). Serum levels of 11-ketotestosterone (11-KT) before SPH treatment were higher (approximately 2 ng/ml) than those of the other steroid hormones (less than 0.5 ng/ml). 11-KT levels increased gradually over the experimental period, and, like E₂, levels peaked towards the end of the experimental period (about 15 ng/ml). The observed patterns for T, E₂ and 11-KT are unlike those in other fishes. Furthermore, the consistent elevations in levels of 11-KT, both before and after SPH treatment, are suggestive of an important role for this steroid in controlling oocyte growth.     

Plasma gastrin levels in horses with colic]

The plasma gastrin levels in fasted horses (21.1 +/- 15.6 pg/ml), in horses with spasmodic colic (7.3 +/- 5.4 pg/ml) and in horses with impaction of the left ventral large colon and/or pelvic flexure (11.4 +/- 3.1 pg/ml) were not significantly different. The plasma gastrin concentrations of horses with strangulation obstruction of the small intestine, large colon displacement or adynamic ileus, and which had no gastric reflux, were 12.9 +/- 8.7 pg/ml and did not differ from fasted gastrin levels. Horses which had 5-10 litres of stomach content reflux had a higher mean gastrin level (32.2 +/- 22.6 pg/ml) (range 8.7-83.0) than the fasted horses. The mean plasma gastrin level (69.0 +/- 32.2 pg/ml) (range 27.0-122.0 pg/ml) in horses which had gastric reflux and 11-20 litres of stomach content outflow through the nasogastric tube were significantly higher (P less than 0.0004) than in fasted horses or in horses with spasmodic colic, impaction of the left ventral large colon or in horses from which no gastric reflux could be obtained.     

Effect of dietary protein and lipid levels on gonadal development of female redclaw crayfish Cherax quadricarinatus

The effect of two crude dietary protein (26% and 32% P) and two lipid levels (4.7% and 12% L) on gonadal development of female redclaw crayfish Cherax quadricarinatus was determined. The gonadosomatic index (GSI), hepatosomatic index (HSI), biochemical composition of gonad and hepatopancreas, oocyte diameter and frequency of the developmental stage of oocytes were analysed. The GSI was significantly less using 26% P to 4.7% L. A significant interaction between P and L content was found for GSI. A significantly higher HSI value was obtained when L was 12%. P content in the hepatopancreas was significantly higher using 32% P and 4.7% L. The maximum L content in the hepatopancreas was obtained from the 32% P and 12% L diet. A significant interaction between P and L content was found for L content in the hepatopancreas. A higher frequency of post‐vitellogenic oocytes was obtained with diets containing 32% P. There was an interaction between dietary P and L level causing lipid accumulation in the hepatopancreas and changes in GSI. For an optimum 32% P, excess dietary lipid is bioaccumulated in the hepatopancreas, indicating that there is excess available energy. When the P and L levels are restricted, there is a limited gonad development, thus affecting overall reproduction in female C. quadricarinatus.     

Evaluation of practical diets containing different protein levels on gonad development of female redclaw crayfish Cherax quadricarinatus

The effect of five experimental diets with different crude protein content (220, 270, 330, 390 and 450 g kg⁻¹) on gonad development of female Cherax quadricarinatus was tested under laboratory conditions. After 70 days, a significant linear relationship indicated that higher concentrations of protein and carbohydrates in the hepatopancreas were produced as the dietary crude protein increased ( P  < 0.05). There were significant responses of the gonadosomatic index, hepatosomatic index, biochemical composition of the gonad (protein, lipids, carbohydrates and energy) and frequency of secondary vitellogenic oocytes to dietary protein level, as indicated by significant fits of the quadratic equation to the observed experimental data. The optimal response of the criteria parameters corresponded to levels of crude protein in the range 284–355 g kg⁻¹. Overall, 330 g kg⁻¹ crude protein with a protein : energy ratio of 15.6 mg kJ^–1 was considered the most adequate concentration of dietary protein for gonad development and biochemical composition in female redclaw crayfish.     

Sex differences in circulating steroid hormone levels in the red drum, Sciaenops ocellatus L.

Steroid profiles of cultured and captive red drum (Sciaenops ocellatus L.) were investigated to evaluate the potential use of circulating sex steroid levels as a tool for gender identification in this species. Cultured 18‐month‐old fish were maintained on a 120‐day shortened photothermal cycle to induce precocious maturation. Additionally, wild‐caught fish were maintained in captivity under simulated natural photothermal conditions from late spring to early fall. Circulating 11‐ketotestosterone (11‐KT) levels were significantly higher in males compared with females during the early stages of gonadal growth in both cultured and captive fish. Plasma testosterone (T) levels showed a similar trend; however, the differences were significant only when males were already producing sperm. 17β‐estradiol (E₂) concentrations were low in males and females before gonadal recrudescence but increased significantly with the progression of vitellogenesis in females. These results show that a test using a minimum concentration of circulating 11‐KT could be developed to differentiate between sexes in the early stages of gonadal maturation in red drum. Moreover, plasma E₂ concentrations could be used to identify vitellogenic females. The two steroids considered together could help avoid possible error in gender identification due to unusually high levels of certain steroids encountered in some individuals.     

来宾地区雌性克氏原螯虾卵巢发育分析

为探究克氏原螯虾在来宾地区生长繁殖周期,于2019年5—8月,采用石蜡包埋法,对当年4月下旬放入的20 g左右雌虾的卵巢进行切片研究。结果表明,在5月中下旬雌虾卵巢已经开始发育,6月卵巢发育缓慢,7月其发育呈现不同步性,7—8月卵巢发育迅速,8月下旬已经出现大量成熟卵母细胞,9月中旬开始大规模抱卵,来宾地区4月放养的种虾,在9月便可达到性成熟并进行繁殖。     

条斑星鲽卵巢发育规律和性类固醇激素周年变化研究

采用组织学方法、形态测量法、放射免疫法(RIA)等技术方法,研究了人工养殖条件下条斑星鲽雌性亲鱼卵巢发育成熟和性类固醇激素的年周期变化规律.结果表明,条斑星鲽属于非同步分批产卵类型.卵母细胞按其组织学特点可分为6个时相,卵巢发育依据其形态特征可划分为相应的6个时期.卵巢年周期发育过程中,性腺指数(GSI)、肝脏指数(HSI)和肥满度(CF)发生变化,表现出与性腺发育协同一致的变化规律.统计分析表明,卵巢年周期发育过程中,GSI和HSI值与CF值都存在显著的正相关关系,GSI值的变化与水温调控呈显著的负相关关系.亲鱼血浆中雌二醇(E2)在6月份开始升高,9月份达到年周期中的第1个小高峰,翌年2月份,E2表达水平达到最高峰值,4月产卵结束后,E2表达水平降低并维持较低水平至下一个生殖季节.血浆中睾酮(T)含量在7月达到第1个峰值,并在翌年1月份达到第2个峰值,在4月份产卵后降至最低水平并维持至下一个生殖季节.统计分析表明,E2与T存在正相关关系,E2与光周期调控存在正相关关系,同时与GSI和CF呈弱的正相关关系.     

Sperm production and quality in brill Scophthalmus rhombus L.: relation to circulating sex steroid levels

The aims of the present study were to characterize sperm quality and to quantify seasonal changes in sexual hormone (testosterone [T], 11-ketotestosterone [11-KT] and 17,20β-dihydroxypregn-4-en-3-one [17,20β-P]) levels in male brill (Scophthalmus rhombus) plasma, as well as to test a more intensive sampling strategy to establish relationships between sex steroid levels and sperm production parameters. Sperm concentration ranged from 0.5 to 3.1 × 10⁹ spermatozoa mL^?1, and changes in sperm quality parameters depending on sampling date were observed. Plasma sexual steroid levels remained high and changed in parallel during the spawning season and afterwards decreased to very low levels in summer. The analysis of annual changes of 11-KT and T ratios suggests that 11-KT can be the main circulating androgen for stimulating spermatogenesis in S. rhombus and that T could be involved in the beginning of spermatogenesis through the positive feedback on brain-pituitary-gonad axis. Finally, daily 11-KT and T levels showed similar patterns of variation in males sampled, whereas 17,20β-P amounts showed somewhat opposite trends. These differences could be related with the different role of androgens and progestin during the spermatogenesis.     

Effect of dietary protein levels on reproductive development and distribution of amino acids in the body tissues of female Pangasianodon hypophthalmus (Sauvage, 1878) broodstock in captivity

The present study investigated the optimum dietary protein level for the maturation of adult Pangasianodon hypophthalmus broodstock. Four isocaloric diets containing 250, 300, 350 and 400 g kg^?1 of protein levels were prepared and presented to triplicate groups of fish. The fish (mean weight 770 ± 17.23 g and 712 ± 23.42 g for females and males respectively) were stocked in outdoor canvas tanks (4 m × 1 m × 1 m) at a stocking density of 20 fish/tank with a male: female ratio of 1:4. The fish were fed the test diets to satiation twice daily for 6 months. Gonadosomatic index (GSI) and fecundity were similar among fish fed dietary protein levels, higher than those fed on the 250 g kg^?1. Final weight, weight gain, oocyte weight were significantly highest (P < 0.05) for the fish fed 350 and 400 g kg^?1 dietary protein treatments. Only the 350 g kg^?1 dietary protein treatment resulted in significantly best ovipositor diameter and % ripe egg. Amino acid levels were highest in the muscle followed by the oocyte and liver of fish fed 350 and 400 g kg^?1 dietary protein levels. The present results suggested that a dietary protein level of 350 g kg^?1 can be recommended for the development of P. hypophthalmus broodstock.     

Changes in brain and pituitary GnRH levels during ovarian maturation in wild female Japanese flounder

To elucidate the role of gonadotropin-releasing hormone (GnRH) in gonadal maturation in wild female Japanese flounder Paralichthys olivaceus, we monitored changes in the levels of seabream GnRH (sbGnRH) in the olfactory bulb, telencephalon, hypothalamus, and pituitary during ovarian development together with changes in plasma levels of testosterone (T), estradiol-17β (E₂), and 17α, 20β-dihydroxy-4-pregnen-3-one (DHP). Fish were caught offshore of the northern mainland of Japan in the Pacific Ocean at 3- to 4-week intervals between April and September by gill net. The netted fish were categorized into six groups based on ovarian stages: previtellogenic (April–early May), early yolk (April–late May), late yolk (late May–June), early spawning (June–August), late spawning (September), and termination (September) stages. The gonadosomatic index significantly increased from the previtellogenic to early spawning stages and decreased thereafter. In the olfactory bulb, no significant differences were observed in sbGnRH levels among the developmental stages. In contrast, sbGnRH levels in the telencephalon and hypothalamus were very high in the previtellogenic stage, lower in the early spawning stage, and relatively high in latter stages. sbGnRH levels in the pituitary were high in the previtellogenic stage and low in the early spawning stage. In addition, the relatively high levels of pituitary sbGnRH were found together with high plasma T, E₂, and DHP levels in fish in the late yolk stage. These results indicate that sbGnRH in the telencephalon, hypothalamus, and pituitary is involved in ovarian maturation and that sbGnRH may play an important role in the initiation of ovarian recrudescence in wild Japanese flounder.     

Plasma levels of insulin,glucagon and glucagon-like peptide in salmonids of different weights

Plasma levels of insulin in rainbow trout,Oncorhynchus mykiss, Atlantic salmon,Salmo salar, and Pacific coho salmon,Oncorhynchus kisutch and plasma circulating levels of glucagon and glucagon-like peptide, in rainbow trout and Atlantic salmon, were measured by homologous radioimmunoassays. Hormonal levels were compared against the average body weight of the same group of fish. Plasma insulin levels were significantly correlated (r=0.56, 0.46 and 0.42 respectively) with body weight in all three salmonid species. Moreover, rainbow trout from fast-growing families had significantly higher (p<0.005) plasma insulin levels than did fish from slow-growing families. Plasma titres of glucagon and glucagon-like peptide were always lower than insulin titres and did not correlate with body weight.Reported in part at Satellite Symposium on Applications of Comparative Endocrinology to Fish Culture, Almunecar, Spain (Sundby, 1989).     

Diurnal rhythm of serum steroid hormone levels in the Japanese whiting,Sillago japonica,a daily-spawning teleost

Ovarian developmental stages and serum steroid hormone levels were examined at six different times of day (0100, 0600, 1000, 1300, 1600, 2000 h) in a marine teleost, the Japanese whiting Sillago japonica, which has an asynchronous-type ovary containing oocytes at various stages of development and spawns every day during a period ranging up to three months. The largest oocytes in the ovaries at the active vitellogenic or post-vitellogenic stages were found between 0100 and 1300 h. Oocyte maturation indicated by germinal vesicle breakdown (GVBD) occurred at 1600 h, and ovulated oocytes were observed in the ovaries collected at 2000 h. These processes were accompanied by a significant daily change in serum steroid hormone levels. The serum level of estradiol-17β showed a peak in fish with mature oocytes sampled at 1600 h. In these fish, the second-largest oocytes in the ovaries were at the initial stage of vigorous vitellogenesis, the secondary yolk stage. Therefore the highest level of serum estradiol-17β was considered to be due to the second-largest oocytes. Testosterone levels remained low and constant throughout the experimental period. The serum levels of 17α,20β-dihydroxy-4-pregnen-3-one (17α,20β-diOHprog) peaked at 1600 h at which time all fish had mature oocytes. These results indicate that the Japanese whiting possesses a diurnal rhythm of oocyte development including vitellogenesis, oocyte maturation and ovulation, and further suggest that daily cycles in oocyte growth and maturation which simultaneously take place in an ovary are regulated by diurnal secretions of estradiol-17β and the maturation-inducing steroid, 17α,20β-diOHprog.