Annual Cyclical Changes in the Histological Features and Surface Ultrastructure in Ovaries of the Freshwater Featherback Notopterus notopterus (Pallas, 1769)

The feather back, Notopterus notopterus is an important food fish. Its ovary is an extremely dynamic organ and the oocytes present an asynchronous development. Variations in ovary weight, GSI, diameter of oocytes were studied in different months of the year in this fish. Different developmental stages of female germ cells were identified on the basis of histological and ultrastructural characteristics in the ovary of N. notopterus (Pallas). In the present investigation the oocyte development of N. notopterus was divided into five stages (oogonia, perinucleolar oocyte, cortical alveolus, yolk granules stage and mature oocyte). The cytophysiological features like vitellogenesis, chorion formation and atresia of some follicles were also studied in the present investigation. The seasonal changes in the ovary have been described according to the variations in gonadosomatic index and the cytological changes of the female germ line cells.     

Ovarian development and histological observations of threatened dwarf snakehead fish,Channa gachua (Hamilton, 1822)

Channa gachua were monthly sampled throughout a year and the histological analysis of their ovaries was done to determine the changes occurring in ovarian development. Based on histological examination of the ovaries, the oogenic process of C. gachua undergoes distinct cyclic and seasonal morphological changes. Five different developmental stages were identified under three major categories: pre-spawning (immature, maturing, mature), spawning (ripe-running) and post-spawning (spent). The peak spawning period of C. gachua was noticed during December - February. The gonadosomatic index (GSI) and ova diameter ranged from 0.79 to 3.61% and 543–1123 μm respectively. The highest mean GSI (3.61 ± 0.16) and oocyte diameter (1123 ± 55 μm) were observed in December indicating that during this month the gonadal development reached maturity.     

Role of the tidal cycle in the gonadal development and spawning of the tropical wrasse Halichoeres trimaculatus

Many wrasses on coral reefs exhibit daily spawning that peaks around daytime high tides. In this study, we examined tidal-related ovarian development in the threespot wrasse, Halichoeres trimaculatus, a species common on coral reefs in the Indo-Pacific Ocean. When the fish were collected in the morning at different tidal phases, the gonadosomatic index (GSI) and ovarian histology changed; concomitant with increases in GSI towards high tide, a clutch of the most advanced oocytes developed from vitellogenic to maturation stages. Ovulated eggs and post-ovulated follicles (POF) existed in most ovaries around high tide, but only POF remained around ebb tide, suggesting that spawning occurred during or after high tide. We noticed that tidal-related spawning was considerable in the morning and that most ovaries collected on the afternoon high tide exhibited post-spawning features. This suggests that certain labrid species possess plasticity with regard to their spawning time and utilize potent environmental cues to ensure their reproductive success. When pieces of ovary were incubated with precursor steroids, high conversion of testosterone to 17beta-estradiol occurred during high and ebb tides, while that of 17alpha-hydroxyprogesterone to 17alpha,20beta-dihydroxy-4-pregnen-3-one and 17alpha,20beta,21-trihydroxy-4-pregnen-3-one was observed during low and flood tides. Incubation of pieces of ovary with human chorionic gonadotropin resulted in similar fluctuations in the steroid hormones with tidal phase. Production of these steroid hormones correlated with oocyte development in the ovaries and was probably regulated by gonadotropin. These results demonstrate that the daily cycle is fundamental for oocyte development, and that the tidal cycle is superimposed on this process.     

Seasonal changes in the pituitary- gonad axis of feral goldfish, Carassius auratus L., from Ontario, Canada

Apparently healthy, feral goldfish, Carassius auratus L., were collected from a pond in Cambridge, Ontario from March until November 1981, and during April 1982. Based on histological examination, oocyte development was divided into five stages and the reproductive cycle into five periods. The origins of oogonia and the atresia process were examined. The gonadosomatic index (GSI) was found to be an accurate indicator of ovarian activity in the feral population. Based on the GSI, proportion of various oocyte stages and egg size, females older than 2 years appeared to spawn during two periods, May to June and late August; 2-year-old fish spawned in the first period. There was a reduction in the density, abundance and nuclear diameter of presumptive pituitary gonadotrops in early August, after the completion of ovarian development and before the completion of spawning.     

Ovarian development in the Amazonian red discus, Symphysodon discus Heckel (Osteichthyes: Cichlidae).

The Neotropical red discus fish, Symphysodon discus, originates from the Amazonian basin and has a discoid body. Although this species is popularly used in aquaria and is exported as an ornamental fish, it has not been intensively studied. The purpose of this work was to study the morphological and histological aspects of the ovarian development in the red discus. Forty females of S. discus of varying body sizes and different stages of gonadal development were used in this study. The ovaries were weighed and examined macroscopically in order to observe the maturation stages. Histological staining of hematoxilin-eosin was used for microscopic observations of the ovaries. Macroscopic observations showed immature, maturing, mature and partially spent stages of ovarian development. Microscopic examination of the ovaries revealed five stages and five phases of ovarian development. The ovaries of the mature females showed all the different phases of oocyte development, indicating the multiple spawning habit of this species.     

Annual Variations of Gonadotropin Content and Ovarian Development of Feral Female Catfish, Silurus asotus, in Central China

The Japanese catfish Silurus asotus is widely distributed throughout the freshwaters reservoirs, lakes and rivers of China, Korea and Japan, and is a commercially valuable aquaculture fish in some regions of China and Japan. We studied seasonal variations of gonadotropin (GtH) content of the plasma and pituitary as well as ovarian development. Plasma GtH titres increase in April in the Pearl River and in May in the Liao River in central China. Annual cycles of plasma GtH levels in both rivers are the same, with a gradual decrease after ovulation with the lowest values observed in September (in the Liao River) or October (in the Pearl River). Plasma GtH levels increase gradually in the early stages of ovarian development, with the highest values observed during the late stages of development, and with the lowest GtH titres observed during ovarian regression in both rivers. GtH content of the pituitary remained very high through the spawning period, with the highest titres observed in March in the Liao River, and in July in the Pearl River. As with plasma GtH, the lowest concentrations of pituitary GtH were observed during stages when the ovary was regressed. We analyzed the relationships between plasma and pituitary GtH and reproductive conditions, such as gonadosomatic index (GSI), hepatosomatic index (HSI), oocyte diameter (OD) and oocyte weight (OW). The results indicate that annual changes of GtH content of the plasma and pituitary paralleled GSI, OD and OW, but were negatively correlated with HSI in feral female catfish.     

Ovarian development and plasma sex steroid levels in cultured female Senegalese sole Solea senegalensis

Ovarian development was studied in cultured female Senegalese sole Solea senegalensis. Females with regressed ovaries, mainly occupied by perinucleolar oocytes, predominated throughout summer exhibiting low condition factor (K), gonadosomatic index (I(G)), and plasma 17beta-estradiol and testosterone levels. Throughout autumn and winter (ovaries at early and intermediate maturation), oocytes progressed to cortical alveoli and vitellogenic stages accompanied by increasing K, I(G), and plasma 17beta-estradiol and testosterone levels. At late winter/early spring, ovarian development reached its maximum with the predominance of females at intermediate and final maturation (the latter occupied by late vitellogenic oocytes and few early maturation oocytes) and peak values of K, I(G), and 17beta-estradiol and testosterone concentrations. Steroid levels were lower (especially testosterone) than those for naturally-spawning females, which might cause extensive atresia without final oocyte maturation (no spawning was observed). This degenerative process reduced de size of the ovary (initial and intermediate phases of regression) in association with declining K, I(G), and plasma 17beta-estradiol and testosterone levels and increasing proportions of perinucleolar oocytes. The circulating 17,20beta-dihydroxy-4-pregnen-3-one levels, the proposed maturation-inducing steroid, remained relatively constant throughout the experimental period, suggesting that oocytes were unable to respond adequately to its stimulation. We propose the inadequate seasonal thermal regime as the main cause of such dysfunction.     

Reproductive biology of the prawn Melicertus kerathurus (Decapoda: Penaeidae) in Thermaikos Gulf (N. Aegean Sea)

The reproductive biology including insemination frequency, ovarian maturation, gonadosomatic index (GSI), size at first maturity, and fecundity of Melicertus kerathurus were investigated using monthly samples from Thermaikos Gulf. Insemination was recorded by the presence of a spermatophore in the thelycum, and ovarian development was based on macroscopic ovarian staging (ST1–ST5). Inseminated females were found throughout the year with high percentages recorded from April to July. Although all ovarian stages were represented in mated females, insemination increased with size and ovarian maturation. High percentages of vitellogenic or mature ovarian stages were observed from May to July, while immature and developing ovaries were predominant mainly in winter. Spawned ovaries occurred from May to October. Carapace length at first maturity based on the presence of a spermatophore (CL_50sp) was estimated at 39.20 mm, while that based on the presence of vitellogenic and mature ovaries (CL_50ov) at 40.70 mm. The seasonal peak in the proportion of mature females (ST4) varied with size. Inseminated females at ST4 and GSI peaked in June–July. GSI varied in relation to insemination status and ovarian stage. In large females (>50 mm CL), the decline in mature ovaries and GSI increment with size indicates a relative reduction in the reproductive output. The number of oocytes ranged from 62,742 to 602,947 (mean ± SD: 268,000 ± 113,000). As the prawns are targeted during the spawning season, mainly by the artisanal fishery, and female size at first maturity is selected by artisanal net size, managerial measures toward artisanal fishery should be implemented.     

Reproductive biology ofAwaous guamensis, an amphidromous Hawaiian goby

Synopsis Spawning season, size at first reproduction, oocyte maturation, and fecundity ofAwaous guamensis, an amphidromous Hawaiian goby, were studied from June 1989 through May 1991 in the Wainiha River, Kau'ai, Hawai'i. Female fish larger than 73 mm standard length (SL) had mature gonads from August through December in 1989 and 1990. Gonadosomatic index (GSI) values for mature females ranged from 0.2 to 14.5 during the spawning season. Male fish larger than 64 mm SL had elevated GSI values from June 1989 through December 1989 and from August 1990 through December 1990. Mature sperm were found in two male fish collected in January and February. GSI values for mature males ranged from less than 0.01 to 4.0 in the spawning season. Size-frequency distributions of measurements of vitellogenic oocyte diameters and microscopic observations of oocytes indicated this species has group-synchronous oocyte development. Ovarian maturation stages examined over a 29-month period suggest that members of the stock spawned at different times within the spawning season, although mass spawning events have been documented for this species. Estimates of clutch sizes from nests measured in situ were comparable to estimates of potential fecundity from in vitro examination of ovaries, and indicated that female fish deposited an entire clutch during a spawning event. No evidence for multiple spawning by an individual fish in a single season was found. However, microscopic observations of brown bodies in some ovaries suggested that individual fish probably spawn more than once in a lifetime.     

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.     

Oogenesis: chromatin and microtubule dynamics during meiotic prophase.

Changes in the organization of germinal vesicle chromatin in mouse oocytes have been analyzed by fluorescence microscopy with respect to progressive stages of follicular development and the disposition of oocyte cytoplasmic microtubules. Four discrete patterns of chromatin organization exist in germinal vesicle (GV)-stage oocytes isolated from the ovaries of 21-25-day-old gonadotropin-primed mice. Analysis of ovarian cryosections stained with the DNA-binding fluorochrome Hoechst 33258 indicates that sequential changes in GV chromatin occur during folliculogenesis that result in the formation of a continuous perinucleolar chromatin sheath at the time of antrum formation. Specific alterations in the cytoplasmic microtubule complex of GV-stage oocytes were observed that correlate with chromatin patterns. The extensive cytoplasmic microtubule complex seen in oocytes of preantral follicles initially localizes to perinuclear areas of the ooplasm. This is followed by a progressive reduction in cytoplasmic microtubules and the appearance of prominent microtubule-organizing centers at the nuclear periphery. Coordinated nuclear and microtubular alterations also occur under in vitro conditions prior to progression of meiosis to prometaphase-1. The results are discussed with respect to the ongoing differentiation of the oocyte nucleus and the microtubule cytoskeleton during folliculogenesis in preparation for the resumption of meiosis.     

Ovarian development of albacore (Thunnus alalunga Bonnaterre, 1788) in the Eastern Mediterranean Sea: a histological characterisation

In order to investigate the stages of oogenesis in the albacore Thunnus alalunga, 129 females (64.0?83.5 cm fork length‐FL) were captured by gill nets from April to August 2002–2008, in the eastern Mediterranean Sea. Based on histological examination of the ovaries, a total of seven ovarian developmental stages were identified: perinucleolar, lipid, early vitellogenesis, late vitellogenesis, migratory nucleus, pre‐hydrated and hydrated. The monthly gonadosomatic index values and the results of the histological analysis indicated that the females mature in May. The spawning stage becomes predominant in June and reaches a maximum in July. The evidence reported here indicates that the size of females at first sexual maturity was lower for the Mediterranean albacore stock than for the Pacific and Atlantic stocks.     

Spawning season, ovarian development and fecundity of female Trichomycterus corduvense (Osteichthyes, Siluriformes)

We studied female reproductive biology of Trichomycterus corduvense. The study was carried out in the Anizacate River (Córdoba, Argentina). Samples were collected throughout one year on a monthly basis. Seven oocyte phases were identified. Five ovarian stages were described based on the macroscopic analysis of the oocyte phases. The spawning season for the species ranges from October to February; it was determined based on variations in GSI and the existence of mature females. The dynamics of oocyte development of T. corduvense, indicated the asynchronism of maturation. The correlation coefficient between fecundity and the individual weight was very low. Therefore, we suggest that this coefficient cannot be used to predict T. corduvense fecundity.     

A histological study of seasonal ovarian development in freshwater drum in the Red Lakes, Minnesota

Female freshwater drum Aplodinotus grunniens Rafinesque, from the Red Lakes of north central Minnesota, U.S.A. were sampled periodically over a 13-month period. Red Lakes drum mature at approximately age 9 and can live to be over 60 years old. The ovarian development of drum from its northern limit of distribution is described histologically. The stages of oocyte growth and the seasonal change in the proportion of oocyte stages within the ovaries are presented. Drum ovaries contained a continuum of oocyte sizes and at spawning, only the largest oocytes are ovulated. A large percentage of the oocytes became atretic mid-way through the spawning season, while gonadosomatic indexes were still high. This may have reduced the spawning potential of Red Lakes drum.     

雅鲁藏布江黑斑原鮡繁殖生物学研究

对2004-2006年采集于雅鲁藏布江拉萨河的190尾黑斑原鮡进行了繁殖生物学研究。雄性最小性成熟(精巢Ⅳ期)个体体长141.7mm,体重45.2g,性体指数1.09%,雌性最小性成熟(卵巢Ⅳ期)个体体长146.8mm,体重66.7g性体指数11.52%,相应年龄均为5龄。初次性成熟年龄(L50):♂,170.1mm相应年龄为7龄;♀,150.2mm,相应年龄5龄。通过组织切片法和GSI的周年变化分析,繁殖时间集中在5-6月,每年繁殖一次,繁殖之后的6-8月卵巢从Ⅵ期回复到Ⅲ期,9月卵巢发育到Ⅳ期越冬。卵径频率分布显示,卵巢发育类型为分批同步型,卵巢中至少存在2批卵径,每年成熟一批卵并同时产出,产卵类型为完全同步产卵。卵黏性,成熟卵卵径在2.04-3.37mm之间,平均(2.83±0.16)mm。对19尾产卵前夕(体长为151.0-210.0mm)的标本进行统计,其绝对繁殖力范围在525-2058粒之间,平均为(1244±346)粒,相对繁殖力为(14.7±5.8)粒/g。绝对繁殖力与体长呈直线正相关,表达式为F=13.624L-1187。       

Apoptosis, cell proliferation and vitellogenesis during the folliculogenesis and follicular growth in teleost fish

Aiming to better understand folliculogenesis, this study evaluated cell death and proliferation of ovarian cells, besides cathepsin-D expression in Prochilodus argenteus captured in two sites of the São Francisco River downstream from the Três Marias Dam, Brazil. In the site immediately following the Dam (S1), low levels of dissolved oxygen were registered in the rainy period. The water temperature was higher in the São Francisco River immediately after the confluence with the Abaeté River (S2), regardless of the period. In S1, the ovaries showed smaller oocytes, high caspase-3 enzymatic activity and apoptosis, lower cells in proliferation and GSI, as well as a lesser quantity of cathepsin-D when compared to females captured from S2. Regarding relative frequency of ovarian structures, in the dry period, only oogonia and perinucleolar oocytes were found in fish ovaries from both sites. On the other hand, in the rainy period, the relative frequency of oogonia and perinucleolar oocytes decreased and the vitellogenic oocytes increased in S2. Postovulatory follicles were observed only in S2, whereas atretic follicles occurred at a higher frequency in S1. Our results showed that apoptosis, cell proliferation and cathepsina-D evaluation can be used as biomarkers of environmental impact.     

Formation of the hamster zona pellucida in relation to ovarian differentiation and follicular growth

Using an immunofluorescence technique on ovarian sections, zona-immunoreactive components were detected in the cytoplasm of the oocyte from the beginning of its growth, when it is surrounded by only a thin squamous follicular cell layer, up to the end of its growth. In parallel with oocyte growth, the staining intensity decreased in the ooplasm. No staining was observed in the cytoplasm of the granulosa cells during normal follicular development in adult cyclic females. However, staining of the granulosa cells was observed at some stages of follicular development in immature females. This staining was especially evident in the ovaries of immature females (22 or 26 days old) stimulated with PMSG. In addition, the staining of the granulosa cells was consistently observed in ovaries showing an abnormal histology. Increased staining of the zona at its outer and inner regions could be distinguished in normal follicles, but when staining occurred on the granulosa cells no such pattern was observed over the zona matrix. These studies indicate that the oocyte itself but not the granulosa cells elaborates the native immunogenic material of the zona pellucida. The administration of PMSG at particular stages of ovarian differentiation interferes with follicular development leading to an abnormal extracellular assembly of the zona and its degradation (phagocytosis) by the surrounding granulosa cells.     

Age, growth, and reproductive biology of the blackbelly rosefish from the Carolinas, U.S.A.

Otoliths and gonads of blackbelly rosefish Helicolenus dactylopterus dactylopterus were collected from the commercial fishery off the Carolinas in 1994–1997. Opaque bands on transverse sections of otoliths were determined to be annuli by analysis of marginal increments. Opaque zone formation occurs between July and January. Ages ranged from 7 to 30 years. Blackbelly rosefish have intraovarian gestation. Fertilization is internal, as free spermatozoa were found primarily in resting ovaries from July through early December with peak occurrence in September through November. There was a delay of 1–3 months before fertilization, as oocyte development did not begin until December. Occurrence during January through April of early-celled embryos, the most advanced stage observed, and postovulatory follicles indicated that oocyte development was rapid. Egg development occurs in a clear gelatinous matrix secreted into the ovarian cavity. The reproductive mode is a zygoparous form of oviparity, intermediate between oviparity and viviparity. Population sex ratio departed markedly from 1: 1 for most length intervals. Males were more abundant at lengths >250 mm L _T and the overall male: female ratio was 1 : 0·60.     

拟目乌贼卵子发生与卵巢发育

为了丰富拟目乌贼(Sepia lycidas)生物学资料, 为人工育苗与养殖提供理论依据, 采用解剖学和组织学的方法, 对水泥池养殖条件下拟目乌贼卵子发生和卵巢发育进行了研究。结果表明: 经过6个月水泥池养殖, 平均体重为256.34 g, 最大体重达到457.08 g, 个别发育成熟, 绝大部分未达性成熟。卵子发生不同步, 根据细胞形态、细胞大小、滤泡细胞形态和卵黄形成情况可分为卵原细胞阶段(卵原细胞期)、原生质生长阶段(无滤泡期、单层滤泡期和双层滤泡期)、间质生长阶段(滤泡内折早期、滤泡内折中期和滤泡内折晚期)和营养质生长阶段(卵黄发生早期、卵黄发生晚期和成熟期), 共4个阶段10个时期。卵巢发育根据外观形态、性腺指数变化和切面上各期细胞所占的比例, 可分为形成前期、形成期、小生长期、大生长期、成熟前期和成熟期6个时期。拟目乌贼繁殖周期为一年。