Variation in plasma oestrogens and androgens during the seasonal and semilunar spawning cycles of female gulf killifish, Fundulus grandis (Baird and Girard)

Variations in 17β-oestradiol, oestrone, testosterone, and 11-ketotestosterone were measured by radioimmunoassay in the plasma of female Gulf killifish, Fundulus grandis , during their seasonal and semilunar spawning cycles. Both 17β-oestradiol and testosterone exhibited distinct seasonal variation, peaking very early in the breeding season during March and April, decreasing gradually thereafter the cessation of spawning in late August and the seasonal regression of ovaries in September, and eventually falling below the detectable limits of our assays during the very early stages of seasonal ovarian recrudescence in November. Both steroids also exhibited distinct semilunar variation within the breeding season, with highest plasma concentrations immediately prior to, and during, each spring tide spawning. Such results suggest that these steroids have physiological roles in the generation and regulation of both seasonal and semilunar reproductive cycles: 17β-oestradiol by controlling development of vitellogenic oocytes; testosterone perhaps by acting as a precursor in the production of oestrogens and other steroids. In contrast, oestrone and 11-ketotestosterone were only rarely detected, implying that these particular oestrogens and androgens are probably not physiologically active in female killifish.     

Reproductive cycles in a reared strain of the mummichog, a daily spawner

Annual, lunar, and diel samplings were taken from a strain of mummichog (Arasaki strain) reared in outdoor tanks under natural conditions, to examine gonadal maturity. Gonads of yearling fish were quite immature in September. During late autumn and winter, a gradual increase in the GSI of both sexes was observed, and the growth of cortical alveolus phase oocytes in females and basal spermatogenesis in males progressed. In late February, a rapid increase in the GSI of both sexes, vitellogenesis in females, and active spermatogenesis in males, occurred. The spawning period of the yearling fish was from late March to August judging from the presence of milt-producing males and ovulated females. The spawning period of the underyearling fish started in the same month as the yearlings, but terminated 1 month earlier. Plasma levels of oestradiol-17 β (E₂) in females and testosterone in males were high during the spawning period in the yearlings. In the underyearlings, however, E₂ levels peaked in early spring, and declined in the latter part of the spawning period. Neither a lunar nor semilunar cycle was evident in the reproductive activity of this fish, which proved to be a typical daily spawner. Females showed an apparent daily reproductive cycle; oocyte maturation commenced at about 1200 hours, germinal vesicle breakdown (GVBD) occurred at about 2400 hours, and ovulation was completed by 2400 hours, 24 h after GVBD. Such clear annual and daily reproductive cycles make this strain of mummichog a suitable model for the study of environmental and endocrine regulation of reproductive cycles in marine and estuarine teleosts.     

Reproductive strategy of the European perch,Perca fluviatilis Linnaeus, 1758 (Osteichthyes: Percidae) in the Anzali wetland,southwest Caspian Sea

Temporal variation in reproductive traits of geographically distributed fish is supposed to take place in response to the spatial and environmental variations. With regard to the wide distribution of the European perch in the northern hemisphere, important reproductive traits such as the initiation and duration of the spawning activity are likely to vary in different latitudinal gradients. In this study, reproductive biology of the European perch, Perca fluviatilis, is described, based on 324 specimens caught in the Anzali wetland (southwest Caspian Sea) between June 2008 and May 2009. The gonadosomatic index, oocyte frequency distribution and histological examination suggested a long vitellogenic process (October to February) and a short spawning season (January and February). The size‐frequency distribution of the oocytes showed that this perch is a species with group‐synchronous ovarian development. Ovarian development occurred only in one clutch of oocytes (700–900 μm oocyte diameter) with no indication of maturation of any subsequent clutch in the spawning season. The average of (realized) fecundity (±SD) was estimated to be 16177 ± 5846 eggs in late vitellogenic stage, which was lower than the potential fecundity (17188 ± 6917 eggs). Histological examination of the gonads revealed the existence of atretic oocytes in early vitellogenic stages (October and November). This investigation highlights the temporal variation in the initiation and duration of the reproductive activity of the European perch in this region compared to other geographical regions. The results emphasize the necessity of specific temporal management in fishing of European perch based on spatial differences in reproductive biology.     

The effect of growth rate,size, and season on oocyte development and maturity of Atlantic cod (Gadus morhua L.)

The effect of growth rate, body weight, age, and season on ovarian development and maturation was investigated for Atlantic cod (Gadus morhua L.) reared in the laboratory over 10 months, for each of two consecutive years, 1978–1980. Cod were also collected from the Gulf of St. Lawrence, the Scotian Shelf, Georges Bank, the Flemish Cap, and the N.E. Newfoundland Shelf.The state of maturity was recognized by oocyte size. Stage I oocytes did not vary in size with growth rate, season, age, or maturity, while the size of stage II oocytes was positively correlated with maturity and negatively correlated with maximum stage of development achieved. Ovarian wall thickness was positively correlated with age and maturity.The frequency distribution of stage I and II oocytes distinguished the state of maturation, with cod that would mature by the next spawning season having a minimum of 20% ($\text{x}\text{?}\text{= 42\%}$) of their oocytes at stage II or greater level of development.Maturing 3-yr-old cod had greater life specific growth rates than immature 3-yr-olds, but growth rates during the third year itself were not significantly different. A hypothesis of a three-part density-dependent mechanism controlling fecundity is postulated. Future reductions in partial recruitment and total fecundity are predicted for the Gulf of St. Lawrence cod stock based on calculated growth rates for Gulf cod in 1979.     

The dynamics of oocyte growth during vitellogenesis in the rainbow trout (Oncorhynchus mykiss)

This report describes the dynamics of oocyte growth during vitellogenesis in a population of virgin female rainbow trout. Indices of ovarian development increased dramatically during the period of study: the gonadosomatic index (GSI) increased over 50-fold, reaching a peak of 20 just before ovulation; the mean oocyte diameter increased from less than 1 mm to 5.4 mm; and plasma levels of vitellogenin increased from less than 1.5 mg/ml to 25 mg/ml. There were no changes in the numbers of developing oocytes (measuring 0.5 mm or greater in diameter) from the time when the majority of oocytes undergoing secondary development had entered vitellogenesis in August to ovulation in February (averaging 4000 oocytes per fish). The increase in ovary weight during vitellogenesis was, therefore, due to an increase in the size of oocytes rather than to recruitment of more maturing oocytes. The numbers of vitellogenic oocytes in the ovary during the entire study also suggested that atresia of vitellogenic oocytes does not play a prominent role in determining fecundity. During early vitellogenesis, the volume of maturing oocytes within an ovary varied by as much as 250-fold. From September onwards, when all oocytes to be ovulated that season had entered vitellogenesis, a gradual uniformity in size began to develop, such that at ovulation, in February, all the eggs were very similar in size (there was less than a 2-fold variation in volume). The pattern of growth of oocytes in an ovary during vitellogenesis suggests that growth between oocytes is closely coordinated.     

Stages of oocyte development in the zebrafish,Brachydanio rerio

Oocyte development has been divided into five stages in the zebrafish Brachydanio rerio, based on morphological criteria and on physiological and biochemical events. In stage I (primary growth stage), oocytes reside in nests with other oocytes (Stage IA) and then within a definitive follicle (Stage IB), where they greatly increase in size. In stage II (cortical alveolus stage), oocytes are distinguished by the appearance of variably sized cortical alveoli and the vitelline envelope becomes prominent. In stage III (vitellogenesis), yolk proteins appear in oocytes and yolk bodies with crystalline yolk accrue during this major growth stage. Ooctes develop the capacity to respond in vitro to the steroid 17α, 20β-dihydroxy-4-pregnen-3-one (DHP) by undergoing oocyte maturation. In stage IV (oocyte maturation), oocytes increase slightly in size, become translucent, and their yolk becomes non-crystalline as they undergo final meiotic maturation in vivo (and in response to DHP in vitro). In stage V (mature egg), eggs (approx. 0.75 mm) are ovulated into the ovarian lumen and are capable of fertilization. This staging series lays the foundation for future studies on the cellular processes occurring during oocyte development in zebrafish and should be useful for experimentation that requires an understanding of stage-specific events. © 1993 Wiley-Liss, Inc.     

Comparative study of reproductive biology in single- and multiple-spawner cyprinid fish. I. Morphological and histological features

To clarify the dynamics and regulation of oogenesis in single- and multiple-spawning cyprinid fish with group-synchronous oocyte development, a multidisciplinary approach to their reproduction was undertaken using three species from the River Meuse (Belgium): the roach Rutilus rutilus as a single spawner, and the bleak Alburnus alburnus and the white bream Blicca bjoerkna as multiple spawners. The gonadosomatic index (GSI) and histomorphometric changes (distribution of oocyte size, relative proportion of the various oocyte stages) in the ovary are compared. Different patterns of GSI and oocyte growth were observed both between the single- and multiple-spawner fish and between the two multiple spawners. Maximum GSIs were higher in roach (21%) than in bleak and white bream (17.7 and 14.5%, respectively), and compared to the rapid decline of GSI in the roach population, the GSI of multiple spawners decreased progressively during the spawning season. In roach, a short gonadal quiescent period and an early onset of vitellogenesis was recorded from late summer onwards whereas, in bleak and white bream, exogenous vitellogenesis was not systematically observed before winter. A protracted spawning season and/or a low water temperature in autumn are hypothesized to explain this long period of gonadal quiescence. In bleak, during the spawning season, the oocytes recruited arose from the stock of endogenous vitellogenesis and attained the final maturation stage very rapidly. This recruitment occurred during the whole spawning season. In white bream, the differentiation of vitellogenic oocytes from smaller oocytes was completed before the onset of the spawning season. During the spawning period, the proportion of vitellogenic oocytes decreased progressively whereas the percentage of oocytes in the final maturation stage remained approximately constant.     

Lipid Content and Composition during the Oocyte Development of Two Gorgonian Coral Species in Relation to Low Temperature Preservation

Our previous studies have suggested that chilling sensitivity of coral oocytes may relate to their relatively high lipid intracellular content and lipid composition. The distribution of lipids during the oocyte development was determined here for the first time in two gorgonian species (Junceella juncea and Junceella fragilis). The main lipid classes in the two gorgonian oocytes were total lipid, wax ester, triacylglycerol, total fatty acid, phosphatidylethanolamine and phosphatidylcholine. The results indicated that early stage oocytes of J. juncea and J. fragilis were found to have increased lipid content than late stage oocytes. The content of wax ester was significantly higher in the early stage oocytes of two gorgonian corals (51.0±2.5 and 41.7±2.9 μg/mm(3)/oocyte) than those of late stage oocytes (24.0±1.4 and 30.4±1.2 μg/mm(3)/oocyte, respectively). A substantial amount of phosphatidylethanolamine and total fatty acid was detected at each stage of oocyte development in two gorgonian ranges from 107 to 42 μg/mm(3)/oocyte and 106 to 48 μg/mm(3)/oocyte, whilst low levels of phosphatidylcholine were found in two gorgonian oocytes. The levels of total lipid in the late stage oocytes of J. juncea were significantly higher than those of J. fragilis. The observed differences may partially be related to different habitat preferences as higher lipid levels in J. juncea, a deeper-water coral species exposed to lower temperature seawater, might relate to adjustments of cell membranes in order to increase membrane fluidity.     

Breeding ecology of an amphidromous goby of the genusRhinogobius

Field observations of markedRhinogobius sp. DA (Dark type) individuals and monthly sampling in the Kashiwa River, Shikoku Island, Japan, indicated the breeding season of the species to be from mid-April to early July, peaking in May. Mark-and-recapture data showed that at least half of the females spawned more than once in one breeding season. Although eggs in male-guarded nests were all at the same developmental stage, their mean number exceeded that of pre-spawning yolk stage oocytes per female, suggesting that at least some guarding males received eggs from more than one female. The physical condition of both sexes deteriorated considerably during the breeding season, the hepatosomatic index of guarding males decreasing concurrently with the development of eggs. The decline in physical condition of guarding males was attributed mainly to their restricted feeding opportunities.     

大弹涂鱼性腺发育的组织学观察

于光镜下对大弹涂鱼性腺切片作了组织学观察,对大弹涂鱼卵细胞和精子发育规律进行研究。结果表明:大弹涂鱼在一个生殖季节中只能产卵1次,大弹涂鱼属于一次性产卵类型。大弹涂鱼3月卵母细胞进入大生长期发育阶段,4—6月为繁殖盛期,7—8月为繁殖末期。10月卵巢基本修整完毕,进入Ⅱ′恢复期。卵细胞发育可分为6个时相:卵原细胞、卵母细胞单层滤泡、卵母细胞出现脂滴和卵黄、卵母细胞卵黄充满、卵母细胞核极化、卵母细胞退化时相。卵母细胞膜单层,由具有辐射纹的放射带构成,滤泡膜细胞分泌而成的次级卵膜成为成熟卵子的附着丝。大弹涂鱼2月精巢开始发育,5月GSI值达最高值,平均成熟系数达0.70%,排精量最旺盛,出现高峰。7—9月GSI值明显下降。11月至翌年2月GSI值波动于0.08%—0.20%之间,变辐小,此期间精巢处于静止发育状态。大弹涂鱼的精巢属于小叶型结构。精子发育分为6个时期:精原细胞期、精原细胞增殖期、精母细胞生长成熟期、精子细胞变态期、精子成熟期和退化吸收期。繁殖季节精小叶内充满精子,精小囊消失。       

Ultrastructure of oogenesis in the bluefin tuna, Thunnus thynnus

Ovarian ultrastructure of the Atlantic bluefin tuna (Thunnus thynnus) was investigated during the reproductive season with the aim of improving our understanding of the reproductive biology in this species. The bluefin, like the other tunas, has an asynchronous mode of ovarian development; therefore, all developmental stages of the oocyte can be found in mature ovaries. The process of oocyte development can be divided into five distinct stages (formation of oocytes from oogonia, primary growth, lipid stage, vitellogenesis, and maturation). Although histological and ultrastructural features of most these stages are similar among all studied teleosts, the transitional period between primary growth and vitellogenesis exhibits interspecific morphological differences that depend on the egg physiology. Although the most remarkable feature of this stage in many teleosts is the occurrence of cortical alveoli, in the bluefin tuna, as is common in marine fishes, the predominant cytoplasmic inclusions are lipid droplets. Nests of early meiotic oocytes derive from the germinal epithelium that borders the ovarian lumen. Each oocyte in the nest becomes surrounded by extensions of prefollicle cells derived from somatic epithelial cells and these form the follicle that is located in the stromal tissue. The primary growth stage is characterized by intense RNA synthesis and the differentiation of the vitelline envelope. Secondary growth commences with the accumulation of lipid droplets in the oocyte cytoplasm (lipid stage), which is then followed by massive uptake and processing of proteins into yolk platelets (vitellogenic stage). During the maturation stage the lipid inclusions coalesce into a single oil droplet, and hydrolysis of the yolk platelets leads to the formation of a homogeneous mass of fluid yolk in mature eggs.     

Timecourse of oocyte development in saithe Pollachius virens

Wild caught North Sea saithe Pollachius virens were monitored for growth, sex steroid profiles and oocyte development pre‐spawning and measured for egg size and group fecundity during the spawning season in the laboratory. Vitellogenesis commenced in late October–early November, at a leading cohort size (C_L) of c. 250 µm, after which oocytes grew rapidly in size until spawning started in February. Notably, a distinct cortical alveoli stage was virtually absent with yolk granules observed in developing oocytes at the very beginning of vitellogenesis. Little atresia was observed pre‐spawning, but atretic re‐absorption of remnant oocytes containing yolk granules was found in all females immediately post‐spawning. As expected, concentrations of sex steroids, oestradiol‐17β (females), testosterone (both sexes) and 11‐ketotestosterone (both sexes), increased pre‐spawning before dropping post‐spawning. The present experiment provides the first validation of sex steroid levels in P. virens. Post‐ovulatory follicles were visible in histological sections from female gonads 9–11 months post‐spawning, but then disappeared. Spawning commenced around a C_L of c. 750 µm (700–800 µm). Hydrated oocytes (eggs) measured between 1·04 and 1·31 mm (mean = 1·18 mm) with decreasing sizes towards the end of spawning. The average estimated realized fecundity was c. 0·84 million eggs (median female total length, L_T = 60 cm). Spawning lasted from 13 February to 29 March.     

Semilunar spawning cycles ofFundulus similis (Cyprinodontidae)

Synopsis The longnose killifish,Fundulus similis (Cyprinodontidae), spawns with a semilunar periodicity during an extended breeding season (March through August) along the Alabama (U.S.A.) Gulf coast. Spawning characteristically occurs during the 3–6 days of the ascending and spring tides of each semilunar tidal cycle. Such spawning cycles appear to be a relatively common adaptation within the reproductive strategies of intertidal cyprinodontids.     

Factors affecting follicular population, oocyte yield and quality in camels (Camelus dromedarius) ovary with special reference to maturation time in vitro

Three experiments were conducted to study a series of factors affecting in vitro reproductive parameters in camels. In Experiment 1, the effect of season and presence of a corpus luteum (CL) on ovarian follicular populations, oocyte yield and quality was studied using a total of 252 and 208 ovaries collected during the breeding and non-breeding season, respectively. Small, medium, large and the total number of ovarian follicles, oocyte yield and quality were measured. In Experiment 2, the effect of methods of oocyte retrieval and needle gauge on oocyte yield and quality was evaluated with oocytes recovered using slicing and aspiration with 18-, 19- or 20-gauge needle. Oocytes were evaluated microscopically and classified into three categories. The objective of Experiment 3 was to identify the optimum time for oocyte maturation in the dromedary camel. Oocytes were cultured in CR1aa medium at 38.5 degrees C under 5% CO(2) for 24, 32, 36, 48 and 72h. Maturation was calculated as the percentage of cumulus expansion and oocytes reaching metaphase II (MII). The number of small, medium, large and the total number of ovarian follicles were higher (P<0.01) during the breeding than non-breeding season. The recovery of total number of oocytes and Category I oocytes were also greater (P<0.01) during the breeding season. Ovaries without a CL possessed significantly (P<0.01) more ovarian follicles and more (P<0.05) small and large follicles. The total number of oocytes and Category I oocytes were also greater (P<0.01) in ovaries without CL. Slicing of camel ovaries increased (P<0.01) the yield of oocytes as compared to aspiration. The aspiration of follicles using a 20-gauge needle had greater yields of the total number of oocytes and Category I oocytes than when using 19- (P<0.05) and 18-gauge needle (P<0.01). The culture of camel oocytes for 36h produced higher (P<0.01) percentages of cumulus expansion and oocytes at MII. Increasing culture times up to 48 or 72h increased (P<0.01) the percentage of degenerated oocytes.In conclusion, the growth and development of ovarian follicles in the camel as well as yields of Category I oocyte were greater during the breeding season. Slicing or aspirations using a 20-gauge needle yielded greater numbers of total and Category I oocytes. Finally, maturation of oocytes in CR1aa medium for 36h produced higher percentages of cumulus expansion and oocytes at MII stage.     

Effect of age and breeding season on the developmental capacity of oocytes from unstimulated and follicle-stimulating hormone-stimulated rhesus monkeys

Effects of age and season on the developmental capacity of oocytes from unstimulated and FSH-stimulated rhesus monkeys were examined. Immature cumulus-oocyte complexes were matured in vitro in modified CMRL-1066 medium containing 20% bovine calf serum and subjected to in vitro fertilization followed by embryo culture. After fertilization, ova from unstimulated prepubertal monkeys displayed lower development to morula (4%) than those from unstimulated adult females (18% in breeding season and 22% in nonbreeding season). No developmental difference was found between ova from unstimulated adult monkeys in breeding and nonbreeding seasons. However, ova from FSH-primed prepubertal monkeys displayed greater development to blastocyst stage (54%) than those from adult monkeys in the breeding season (16%) and nonbreeding season (0%); and ova from FSH-primed adult females in the breeding season had significantly (P < 0.05) greater developmental competence than those obtained in the nonbreeding season (> or = morula stage, 54% vs. 3%; blastocyst stage, 16% vs. 0%). These data indicate that 1) rhesus monkey oocytes acquire developmental competence in a donor age-dependent manner, and 2) animal age and breeding season modulate the effect of FSH on oocyte developmental competence in the rhesus monkey.     

Reproduction in the Apodid Sea Cucumber Patinapta ooplax: Semilunar Spawning Cycle and Sex Change

Many fertile individuals of the apodid holothurian Patinata ooplax, living in the intertidal area of the Ichiki Fishing Port in southern Kyushu, Japan, spawned during the two days after every full and new moon, probably the first and second days, in the period from the middle of July to the end of August during 1990, 1992 and 1993. Matured individuals were divided into three sexual types: males, hermaphroditic males with an early stage of oocytes, and females, using a dissecting microscope. The distribution frequency and gonadal histology of these sexual types indicate that some individuals changed from male to female or in the reverse direction at two-week intervals between spawnings, and suggest that some change first from male to female, and then back to male during the main breeding season. In addition, it was found that during the main breeding season, synchronous gametogenesis occurred in association with the sex changes, and that the period from the initiation of spermatogonia proliferation to sperm release during the same season was two weeks, and the period from the initiation of oocyte growth to egg shedding was probably slightly longer than two weeks.     

The maternal gene product D7 is not required for early Xenopus development.

The Xenopus D7 gene codes for a novel protein whose expression is restricted to early development. D7 protein is synthesized for the first time during oocyte maturation (1988, Genes Dev. 2, 1296-1306). Injection of D7 RNA into the full-grown oocyte and its subsequent translation into D7 protein neither induced oocyte maturation nor affected the kinetics of hormone-induced maturation. Overexpression of D7 protein by 20-fold in the early Xenopus embryo by injection of D7 RNA into fertilized eggs did not affect subsequent development. Oocytes specifically lacking D7 mRNA were generated by oligodeoxynucleotide-mediated RNA destruction within the oocyte. Unfertilized eggs generated from such oocytes lacked detectable D7 protein, but nevertheless could be activated and fertilized. Embryos generated from such eggs, estimated to contain less than 5% of wildtype levels of D7 protein, developed normally up to the tailbud stage. Thus the D7 protein, the product of a maternal mRNA that is under strict translational repression in oocytes, appears not to be required for oocyte maturation, activation, fertilization or early embryonic development in Xenopus.     

Disappearance of a novel protein component of the 26S proteasome during Xenopus oocyte maturation

We have prepared polyclonal antibodies against Xenopus 20S proteasomes. The antibodies cross-react with several proteins that are common to 20S and 26S proteasomes and with at least two proteins that are unique to 26S proteasomes. The antibodies were used to analyze changes in the components of proteasomes during oocyte maturation and early development of Xenopus laevis. A novel protein with a molecular weight of 48 kDa, p48, was clearly detected in immature oocytes, but was found at very low levels in mature oocytes and ovulated eggs. p48 was reduced to low levels during oocyte maturation, after maturation-promoting factor was activated. The amount of p48 in eggs remained low during early embryonic development, but increased again after the midblastula transition. These results show that at least one component of 26S proteasomes changes during oocyte maturation and early development and suggest that alterations in proteasome function may be important for the regulation of developmental events, such as the rapid cell cycles, of the early embryo.     

Annual reproductive cycle based on histological changes in the ovary of the female mosquitofish,Gambusia affinis, in central Japan

To clarify the annual reproductive cycle of wild female mosquitofish,Gambusia affinis, in Mie Prefecture, central Japan, changes in ovarian histology were investigated. Female mosquitofish kept in aquaria under constant temperature (25°C) and photoperiod (16L: 8D) conditions produced successive broods at intervals of 22.1±0.46 days (n=7). Between days 0–3 following parturition, females began active vitellogenesis. Between days 3–5, fully grown oocytes matured and were fertilized, and embryonic development began in the follicles. By day 10, as fertilized eggs continued embryonic development, some oocytes at the oil-droplet stage had begun to accumulate yolk globules for the next gestation. Thus, vitellogenesis of the succeeding batch of oocytes overlaps with gestation during reproduction in the mosquitofish. A rearing experiment showed the annual reproductive cycle of mosquitofish breeding in Nagashima to be as follows. Although oocytes had not at that point developed to the yolk globule stage, copulation occurred in February. Females began vitellogenesis in early May, the first pregnancy of the year commencing in mid-May. From mid-May to August, females repeated the gestation cycle (vitellogenesis, maturation, fertilization, pregnancy and parturition) at around one month intervals. In September, oocyte recruitment from the oil-droplet to the yolk globule stage ceased. After the final parturition, the ovaries contained only non-vitellogenic oocytes. Spermatozoa in the ovarian cavity were scare from November to January.