鱼类卵子发育与卵泡闭锁的研究进展

鱼类种群繁衍离不开优质卵子的产出.卵子质量问题限制了许多海洋和淡水鱼类养殖业的发展.卵子质量主要受卵黄形成与卵母细胞发育的影响.亲本营养、管理以及其他内源或外源因素可能会影响卵黄组成与卵泡发育.与哺乳动物相比,鱼类卵母细胞发育与调控机制的研究尚不清晰.本文分析了鱼类卵子发育特点,探讨了亲本来源、育龄、环境因子以及营养等...     

福建省东山岛西埔湾港养斑鰶的繁殖生物学

本文研究了福建省东山岛西埔湾港养斑鰶性腺周期发育、成熟系数周年变化、个体生殖力以及雌雄同体。港养斑鰶能在半封闭式湾内发育成熟而且自然产卵。其卵细胞发育分为6个时相,精巢发育分为6期。卵膜由透明带、外放射带和内放射带组成,内有孔道结构。第4时相中期卵母细胞的内放射带有7～8层纤维状片层。产卵后鞘膜细胞和颗粒细胞分裂增殖变成游走的吞噬细胞。西埔湾港养斑鰶Ⅰ龄鱼初次性成熟,生殖季节从12月下旬至翌年3月下旬,生殖盛期为2月,雌鱼成熟系数高达17.88％。个体绝对生殖力为7575～72621粒,平均 35310粒,系属于分批产卵类型,第1批生殖力大于第2批。     

Oocyte maturation and ovulation in the Atlantic halibut, Hippoglossus hippoglossus (L.), examined using ultrasonography

High resolution ultrasound scanning was tested as a non-invasive technique for monitoring oocyte maturation and ovulalion in the Atlantic halibut Hippoglossus hippoglossus (L.). Female broodstock halibut were examined using a 7.5-Mhz linear ultrasound transducer, prior to and during spawning. Representative images of halibut ovaries are presented and discussed. Individual oocytes of spawning females were discernible during oocyte final maturation, due to the large increase in volume caused by water uptake. The yolky (vitellogenic) cocytes of pre-spawning fish were more reflective to ultrasound than hydrating oocytes. The resulting differences in depth of ultrasound penetration permitted easy distinction of pre-spawning from spawning females. In addition, short-term changes in the depth of ultrasound penetration were observed during repeated scanning of a spawning female, owing to progressive hydration of the oocyte batch destined for ovulation. Although of similar diameter, hydrating oocytes in the ovarian tissue could be discerned from eggs in the ovarian lumen because of the different acoustic properties of the surrounding media. The findings of the present study are considered promising for the future routine use of ultrasonography in halibut broodstock management.     

Multiple vitellogenins and product yolk proteins in striped bass,Morone saxatilis: molecular characterization and processing during oocyte growth and maturation

The multiple vitellogenin (Vtg) system of striped bass, a perciform species spawning nearly neutrally buoyant eggs in freshwater, was investigated. Vitellogenin cDNA cloning, Western blotting of yolk proteins (YPs) using Vtg and YP type-specific antisera, and tandem mass spectrometry (MS/MS) of the YPs revealed the complex mechanisms of yolk formation and maturation in this species. It was discovered that striped bass possesses a tripartite Vtg system (VtgAa, VtgAb, and VtgC) in which all three forms of Vtg make a substantial contribution to the yolk. The production of Vtg-derived YPs is generally similar to that described for other perciforms. However, novel amino-terminal labeling of oocyte YPs prior to MS/MS identified multiple alternative sites for cleavage of these proteins from their parent Vtg, revealing a YP mixture far more complex than reported previously. This approach also revealed that the major YP product of each form of striped bass Vtg, lipovitellin heavy chain (LvH), undergoes limited degradation to smaller polypeptides during oocyte maturation, unlike the case in marine fishes spawning buoyant eggs in which LvHAa undergoes extensive proteolysis to osmotically active free amino acids. These differences likely reflect the lesser need for hydration of pelagic eggs spawned in freshwater. The detailed characterization of Vtgs and their proteolytic fate(s) during oocyte growth and maturation establishes striped bass as a freshwater model for investigating teleost multiple Vtg systems.     

Reproductive parameters of the chub mackerel <Emphasis Type="Italic">Scomber japonicus</Emphasis> estimated from human chorionic gonadotropin-induced final oocyte maturation and ovulation in captivity

ABSTRACT:   Final oocyte maturation and ovulation of captive chub mackerel Scomber japonicus with fully yolk-accumulated oocytes were induced by a single injection of human chorionic gonadotropin. Reproductive parameters, including spawning frequency and batch fecundity, which are required to estimate spawning biomass in pelagic fish by the daily egg production method, were analyzed. Germinal vesicle migration (GVM) occurred at 18–24 h post-injection, and the hydration and ovulation of oocytes were completed at 30 and 36 h post-injection, respectively. The results of the maturation process suggest that fish with GVM-stage ovaries captured in the daytime from the field are capable of spawning on the night following their capture. The oocytes used in the oocyte size-frequency distribution method for batch fecundity estimates should be at late GVM and more advanced stages. The results of sequential artificial insemination showed that the quality of ovulated eggs held in the ovarian lumen rapidly deteriorated as time progressed after ovulation. This indicates that the fertilization window for the ovulated eggs of chub mackerel lasts only a few hours, and spawning behavior should be performed within a few hours after ovulation in the wild population.     

细鳞鱼的卵黄发生

从发育生物学—卵黄发生角度,通过光镜和透射电镜对细鳞鱼(Brachymystax lenok)卵黄发生进行观察。结果表明,细鳞鱼的卵黄发生主要集中在初级卵母细胞阶段完成,可分为4个特征时期:初级生长期,皮质泡期,卵黄积累期,卵黄积累完成期。当卵母细胞进入初级生长早期,卵子发生特征物——类核周体出现;进入皮质泡期,其主要特征是皮质泡、卵黄外膜和巴尔比亚尼体结构;处于卵黄积累期,位于其外周细胞质附近会出现卵黄颗粒;进入卵黄积累完成期,卵黄颗粒不断融合增大,占据细胞绝大部分。以上结果为细鳞鱼的卵巢发育分期提供了理论基础,并深入探讨其发育的规律性、特殊性和复杂性,从而指导细鳞鱼的人工繁殖、选育种工作及野生群体的资源保护和合理利用。     

Oogenesis and relevant changes in egg quality of abalone Haliotis discus hannai during a single spawning season

Oogenesis of abalone Haliotis discus hannai was examined histologically during a single spawning season using broodstock of various maturation conditions, which were controlled by effective accumulative temperature (EAT). The quality of eggs spawned was determined in relation to oogenesis. For histological examinations, three to five females were sacrificed at 300, 600, 850, 1050, and 1150 °C days EAT, without induction of artificial spawning. Other females were successfully induced to spawn at 700 °C days EAT and were reared following spawning. Three of these females were then sacrificed every 200 °C days EAT until 1300 °C days EAT. Gonad histology showed that two oocyte cohorts matured in H. discus hannai ovaries during a single spawning season. One mature oocyte cohort could be spawned in multiple times. The second oocyte cohort started developing after the first oocyte cohort had been spawned or reabsorbed, and became fully mature 400 °C days EAT after the first cohort was depleted. For egg quality measurements, three to five females were successfully induced to spawn at 850, 1050, 1150, 1900, and 2350 °C days EAT (Experiment 1). Three females were induced to spawn twice, at 700 and 1500 °C days EAT, resulting in two batches of eggs from the same individuals (Experiment 2). Total lipid and protein content of eggs were measured and were greater in eggs from the second cohort than in eggs from the first cohort. No carbohydrates were detected in eggs and there was no difference in cytoplasm volume between the two cohorts. In hatcheries producing H. discus hannai, it is important to increase post-larval starvation tolerance by increasing the quality of eggs, to yield higher and more consistent survival. The results of this study suggest that H. discus hannai hatcheries should use eggs from the second oocyte cohort, which are of higher quality, rather than eggs from the first oocyte cohort.     

Oogenesis in the common Japanese conger Conger myriaster

ABSTRACT:    We investigated the process and characteristics of oogenesis in the common Japanese conger Conger myriaster . Young fish caught in November 1996 were reared for use in this experiment. Fish were sampled monthly from December 1997 to August 1998. Some were injected with human chorionic gonadotropin to stimulate ovarian maturation from May to August 1998. Oocytes from the chromatin nucleolus stage to the secondary yolk globule stage were obtained from non-hormone-treated fish; those of more advanced stages were obtained from hormone-treated ones. We divided oocyte development into eight stages from the chromatin nucleolus stage to the maturation stage. The yolk vesicle stage was not separated because yolk vesicles began to appear just after appearance of yolk globules. Oocyte, oil droplet, yolk globule and nucleus diameters all increased concomitant with oocyte development. Oil droplet and yolk globule diameters increased remarkably at the maturation stage. However, zona radiata thickness peaked at the secondary yolk globule stage, decreasing gradually thereafter. Increased gonadosomatic index was related to oocyte development as found in European and Japanese eels receiving hormone treatment to mature. The present study is the first report describing oogenesis characteristics in congrid eels. It indicates that oogenesis is almost identical to that of other anguillid eels.     

漂流性鱼卵的孵化特性及水动力需求研究进展

鱼卵运动规律研究是鱼类产卵场位置推算、水库生态调度和鱼类资源保护的基础。对于产漂流性卵鱼类,当流水环境不足以维持鱼卵漂流时,鱼卵便下沉死亡而无法孵化成幼鱼。通过对照中国内陆鱼类名录,查阅国内重要流域及地方鱼类志中有关产漂流性卵鱼类的繁殖条件,梳理了典型鱼类的产卵水温、产卵时间和孵化时长等相关信息。针对现有鱼类志中对于鱼卵基本物理性质及鱼卵孵化所需水动力条件信息缺失的情况,补充查阅了国内外相关文献,归纳总结了漂流性鱼卵安全漂流的水动力条件。一种观点认为鱼卵在水体中是否能安全漂流取决于水体的流速,另一种观点认为控制漂流性鱼卵悬浮或下沉的关键参数是湍流状态或剪切速度,而不是水流的线速度,现有漂流性鱼卵孵化的关键水动力条件和阈值尚有争议,仍需加强鱼卵运动规律的相关试验研究。目前国内外对鱼卵的漂流输移研究方法和手段,主要采用野外监测、室内实验和模型模拟。已有研究大多只是针对鱼卵运动规律本身进行单一试验或模型验证,如何通过水库调度最大程度保障漂流性鱼卵的漂流孵化条件是未来鱼类保护的重要方向。     

Rapid, nongenomic steroid actions initiated at the cell surface: lessons from studies with fish

In addition to the classic mechanism of steroid action mediated by binding to nuclear receptors, there is a growing body of evidence that steroids also exert rapid, nongenomic actions that are initiated at the cell surface by binding to specific steroid membrane receptors. However, the lack of information on the molecular structures of any steroid membrane receptors has impeded development of this alternative model of steroid hormone action. One of the best characterized models of nongenomic steroid action is the progestin induction of oocyte maturation (OM) in fish and amphibians via activation of plasma membrane progestin receptors (mPRs) on oocytes. Investigations of the marked changes in mPR abundance during OM in fishes have provided the first clear evidence of hormonal regulation of steroid membrane receptors and its physiological importance. Recently, a novel gene was discovered in the ovaries of spotted seatrout whose protein has the characteristics of an mPR and the intermediary in the progestin induction of oocyte maturation in this species. The putative mPR is also detected on seatrout sperm by Western blot analysis and is likely the mPR previously characterized in this species that mediates progestin initiation of sperm hyperactivity. Both structural and functional studies suggest the seatrout mPR is a G-protein coupled receptor (GPCR). Subsequently, thirteen structurally-related cDNAs were identified in other vertebrate species, and several of them were also shown to have characteristics of mPRs. The discovery of the molecular structure and likely orientation in the plasma membrane of this new class of steroid membrane receptors provides a plausible mechanistic explanation of how steroids acting at the cell surface can cause rapid intracellular responses. Membrane receptors for estrogens (mER) and androgens (mAR) have also been characterized in teleost gonads. In addition, the first clear evidence for endocrine disruption of a nongenomic steroid action by binding to a membrane steroid receptor was obtained with the seatrout mPR. Thus fish are valuable models for investigating nonclassical steroid actions and their interference by environmental contaminants.     

Changes in oocyte and blood plasma osmotic components of ayu, Plecoglossus altivelis Temminck & Schlegel during oocyte maturation

Oocyte maturation in teleosts is characterized by a complex series of cytoplasmic and nuclear changes. The purpose of this study is to present sequential changes in the oocyte constituents and regulatory mechanisms involved in the hydration and osmotic regulation of oocytes during final maturation of ayu, Plecoglossus altivelis Temminck & Schlegel. Oocyte volumes and water contents in ayu (P. altivelis) markedly increased from 0.372 mm³ and 60.5% at the mature stage to 0.712 mm³ and 79.4% at the time of oviposition. In the same period, net increases in total free amino acids and electrolytes were noted, and involvements of these constituents in the water uptake by oocytes are discussed. Moderate oocyte hydration in ayu is accompanied by steady increases in Na⁺, but not K⁺, and these observations are in contrast to results for all other marine teleosts reported elsewhere. In the present study, a steady increase in oocyte Na⁺ during maturation until ovulation pre‐dominantly results from diffusional leakage due to concentration gradients, while at the time of oviposition, both active transport and diffusional pathways play important roles in osmotic regulation in ayu oocytes.     

Spawning grounds and reproductive traits of anadromous and resident pond smelt, Hypomesus nipponensis, in Lake Ogawara, Japan

SUMMARY: The objective of this study was to reveal the spawning grounds and reproductive characteristics of anadromous and resident pond smelt, Hypomesus nipponensis , coexisting in Lake Ogawara. Life history styles of females shedding in spawning grounds in the lake and its inflowing rivers were differentiated by otolith increment analysis. Size, dry weight, and water content of mature oocytes and fecundity of fish were compared between resident and anadromous fish. Both anadromous and resident fish spawned in the lake. In contrast, no resident fish were found in any of the inflowing rivers, where only anadromous fish spawned. Regression of fecundity against standard length was discontinuous with an inflection point at 63.8 mm, which is the body size that differentiates large anadromous spawning groups from small resident spawning groups. Mean oocyte diameters were not significantly different between resident and anadromous fish. The eggs of resident fish had significantly more water content and a significantly lower dry weight than those of anadromous fish. These differences might influence the growth and developmental processes of progeny.     

Oocyte Growth in the Striped Mullet Mugil cephalus L. Maturing at Different Salinities

Profiles of oocyte growth were obtained from female striped mullet Mugil cephalus L. held in salinities ranging from fresh water to seawater during two consecutive spawning seasons (1988–1990) in Hawaii. Females underwent vitellogenesis at all salinities (i.e., 32–35, 13–20, and 0%‰) tested. Females maturing in fresh water exhibited a slower rate of oocyte growth, and a significantly lower number completed vitellogenesis. All females were induced to spawn in full seawater. The number of fertilized eggs per spawning was highest from females maturing in brackish water. More females were able to be induced to spawn twice in brackish water during the 1989–1990 season than in fresh or seawater. The results from the current investigation suggest that salinities ranging from 13–35 ppt are adequate for ovarian maturation in captive striped mullet females.     

大宁河龙溪段鱼类早期资源研究

为了解十年禁渔背景下大宁河鱼类早期资源变化状况,2020和2021年5-7月在大宁河龙溪断面开展了鱼类早期资源现状的调查。结果表明,龙溪江段产浮性卵鱼类1种、漂流性卵鱼类4种、粘沉性卵鱼类19种;中华纹胸鮡（Glyptothorax sinensis）和大眼鳜（Siniperca kneri）的产卵规模相对较大,其卵占比分别达48%和30%以上;2020年的鱼类卵苗资源量和种类都少于2021年。RDA分析结果显示,中华纹胸鮡（Glyptothorax sinensis）、张氏?(Hemiculter tchangi)鱼卵漂流密度与水温变化呈现一定相关性,适宜水温促使中华纹胸鮡（Glyptothorax sinensis）、张氏?(Hemiculter tchangi)产卵;随着流速和流量增加,多鳞白甲鱼(Scaphesthes macrolepis)和云南盘鮈(Discogobio yunnanensis)卵漂流密度均相应增加;大眼鳜（Siniperca kneri）和峨眉后平鳅(Metahomaloptera omeiensis)产卵期间江水酸碱度相对较低。十年禁渔的实施对大宁河鱼类资源的恢复可能产生积极影响。     

Effects of Phase-Shifted Photoperiod Regimes on Oocyte Growth and Hormonal Profiles in Female Striped Bass Morone saxatilis

Abstract.— The use of 12–mo long, but phase-shifted advanced and delayed photoperiod cycles in the regulation of the reproductive cycle was investigated in captive-reared female striped bass Morone saxatilis during the 3-yr study in an attempt to control the timing of sexual maturation under simulated photoperiod conditions. Phase-shifted photoperiod cycles did not induce a full shift in oogenesis during the first year cycles, but did in the following years. Spawning time, indicated by maximum oocyte diameters, was advanced up to 4 mo in females maintained under the phase-shifted advanced photoperiod, and delayed up to 4 mo when they exposed to the phase-shifted delayed photoperiod, compared to the natural spawning time in Spring (March-May). Phase-shifted photoperiod regimes shifted the profiles of plasma testosterone (T) and estradiol (E₂), corresponding to the shift of oogenesis in the respective groups. Significant increases in T and E₂ levels occurred during the vitellogenic phase, and these levels peaked before the occurrence of maximum oocyte diameters. The studies demonstrate that phase-shifted photoperiod regimes can be used to control oogenesis, and have implications for ensuring the year-round supply of mature female striped bass, particularly in domesticated striped bass.     

Changes in fatty acid and sterol composition during oogenesis in the pearl oyster Pinctada margaritifera

The fatty acid and sterol composition of the oyster Pinctada margaritifera during oogenesis and in eggs was analysed. No major differences were observed during oogenesis, but the egg composition was significantly different from that of gonads. The amount of saturated fatty acids was the highest in eggs and the C16:0 predominant (P<5%); by contrast, the amount of 22:6(n‐3) was significantly lower (P<5%) than in gonads. No major differences were observed for the polar lipid (PL) composition during oogenesis. The main free sterols in gonads and eggs were cholesterol and brassicasterol. Among free sterols, the proportion of cholesterol diminished continuously from the beginning to the end of gonad maturation, and this decrease persisted in eggs after spawning. Cholesterol represented 40% to 55% of the sterol ester encountered in gonad and eggs. This study allowed us to investigate the fatty acid and sterol composition during oogenesis of the pearl oyster P. margaritifera, leading to a clearer understanding of the nutritional requirements of pearl oyster during the reproduction process.     

三峡库区支流磨刀溪产粘沉性卵鱼类早期资源现状

研究磨刀溪回水区的鱼类早期资源,可为三峡库区重要生境、水生生物多样性保护提供科学依据。2019年的4-6月,在三峡库区支流磨刀溪回水区江段（新津乡-龙角镇）开展鱼类资源监测,以了解该区域鱼类早期资源种类组成、仔稚鱼密度的时间动态以及与环境因子的关系。结果表明,采集到7种产粘沉性卵鱼类的鱼苗中,以贝氏?(Hemiculter bleekeri)、鲤(Cyprinus carpio)、鲫(Carassius auratus)为主;密度高峰期集中在4月下旬至5月中旬。磨刀溪新津口至龙角镇江段共分布有鲤、鲫集中产卵场4处。冗余分析(RDA)表明,饵料丰度、溶解氧和pH是影响鱼苗分布的主要因素。磨刀溪变动回水区江段是粘沉性鱼类产卵场的分布区域,三峡水库4-6月水位的快速消落,很可能导致鲤、鲫等产粘沉性卵鱼类资源的损失。建议通过实施生态调度减小水位日变幅或在回水区放置人工鱼巢、恢复水生植被来减缓其影响。     

Augmentation of free amino acids in eggs of red snapper,Lutjanus campechanus as part of the induced spawning protocol

An attempt was made to increase free amino acids (FAA) concentrations in eggs of red snapper Lutjanus campechanus as part of an induced spawning protocol. Mature female red snapper were given intramuscular injections of human chorionic gonadotropin (HCG) at a 1100 IU kg^?1 of female body weight. Immediately after the HCG injection, one set of females received an intramuscular injection of FAA in a buffered saline solution at a dose of 20 mg FAA cocktail kg^?1 of body weight. The FAA cocktail was 25% isoleucine, 25% leucine, 25% lysine and 25% valine on a molecular weight basis. A second set of fish did not receive the FAA supplement. Both sets of fish were equally successful in spawning with ovulation occurring 21–32.5 h post injection. Injection of FAA into brooders as eggs began final maturation and hydration resulted in an increase in leucine (nmoles mg egg^?1) and a slight increase in isoleucine (nmoles egg^?1) in recently ovulated eggs. FAA supplementation via brood injection altered FAA utilization rates by embryos and during larval development, resulted in a greater yolk sac and oil globule reserve in larvae as they approached the time of first feeding.     

Hormone Induced Spawning of Summer Flounder Paralichthys dentatus

During their first year in captivity, summer flounder Paralicthys denratus were induced to spawn with gonadotropin releasing hormone analogue (GnRHa) implants, injected carp pituitary extract (CPE) or human chorionic gonadotropin (hCG) injections. The percentage of fertile eggs was greatest (69%) in CPE-treated females. CPE, but not GnRHa or hCG, was capable of stimulating oocyte growth (increased follicle diameter during vitellogenesis) followed by ovulation. Fish with maximum ovarian follicle diameters between 180 and 435 μm at the initiation of CPE injections produced the greatest percentage of fertile eggs. For most females, fertilization rate was greatest for the first batch of eggs ovulated. The mean fertilization rate for the first spawn of CPE-treated fish was 42% compared with 14% for the second spawn from the same fish. Fish with maximum initial follicle diameters of 585 40 μm that were implanted with GnRHa ovulated the greatest number of eggs, but fertility was low and variable. Approximately 35% of females injected with hCG ovulated a limited number of eggs, but only one hCG-treated female produced fertile eggs. Only a limited number of spermiating males were available for spawning trials. Hormone treatments used on females were ineffective for inducing or maintaining spermiation in male summer flounder.     

促性腺激素调控半滑舌鳎(Cynoglossus semilaevis)卵母细胞孕酮受体膜组分1的表达特征

采用qRT-PCR方法分析性成熟半滑舌鳎(Cynoglossus semilaevis)不同发育时期卵巢卵母细胞的孕酮受体膜组分1 (PGRMC1)mRNA的表达,研究发现,在发育Ⅳ期的卵巢,处于第Ⅳ时相的卵母细胞PGRMC1 mRNA表达量最高(P＜0.05);在发育Ⅴ期的卵巢,成熟期卵母细胞的PGRMC1mRNA表达量最高(P＜0.05).采用不同浓度促性腺激素(HCG)处理卵巢发育Ⅴ期半滑舌鳎不同时相的卵母细胞,并通过qRT-PCR和Western blotting技术对其表达量变化进行检测.结果显示,20IU/mlHCG对PGRMC1 mRNA和蛋白表达的调控作用比10 IU/ml HCG作用更明显,表明PGRMC1 mRNA和蛋白表达对HCG调控作用存在剂量依存关系.HCG对半滑舌鳎不同时相的卵母细胞的调控作用效果不同,对第Ⅴ时相卵母细胞作用最明显(P＜0.05),表明PGRMC1主要在卵母细胞成熟阶段发挥作用.PGRMC1对HCG调控作用的正向应答效应预示其参与了卵母细胞成熟调控,为进一步探讨PGRMC1在半滑舌鳎繁殖过程的功能提供重要基础资料.