We aimed to analyze the oogenesis of adult females of the cichlid fish Laetacara araguaiae. The specimens’ gonads were removed and processed for light and transmission electron microscopy. Oogenesis in L. araguaiae showed the following characteristics: a germinal epithelium with three types of oogonia (A-undifferentiated, A-differentiated and B-oogonia), oocytes at meiotic prophase stage and ovarian follicle formation. Oocytes showing primary growth with pre-vitellogenic and cortical alveolus were observed. Similar to data for other cichlids, oocytes in secondary growth or vitellogenesis were characterized by the initial deposition of yolk microgranules. The event that characterizes the maturation stage is nucleolus migration, also called the germinal vesicle, to the oocyte periphery in the direction of the micropyle. The follicular complex undergoes several changes throughout the oocyte stages. To the best of our knowledge this study is the first to describe L. araguaiae oogenesis. Moreover, this study is the first step to better understand the reproductive biology of this species, which shows great potential for use as an ornamental fish.

Although oogenesis is the best-known stage of development in sponges of the subclass Calcinea (Porifera, Calcarea), several aspects of this process await further investigation. In the present study, we examined by light and electron microscopy the oogenesis of three species of the order Clathrinida (Clathrina aurea, Borojevia aspina and B. brasiliensis) from the coast of Rio de Janeiro state (Brazil). Oogenesis in these species was a rare and not synchronized event, neither in the individual nor in the population level. Oocytes grew between the choanoderm and the pinacoderm. They did not form any special structure for nourishing the egg and phagocytosis of somatic cells was not observed. The vitellogenesis of B. aspina and B. brasiliensis was performed in a mixed pathway, while in C. aurea it was not possible to observe the vitellogenesis. In the Borojevias, the surrounding choanocytes provided small clear vesicles at the same time that oocytes modified phagosomes to produce fibrous yolk inclusions. The phylogenetic character mapping analysis indicated that egg development in the mesohyl is the ancestral state for Calcinea, while the ancestral condition of the vitellogenesis is still uncertain.

The objective of this study was to elucidate the signalling pathways initiated by cAMP once inside the Xenopus laevis oocyte, where it triggers and maintains vitellogenin endocytic uptake. Our results showed the presence of Xepac transcripts at all stages of oogenesis and we demonstrated that a cAMP analogue that exclusively activates Xepac, 8-CPT, was able to rescue the endocytic activity in oocytes with uncoupled gap junctions. Inhibition experiments for the IP3/Ca2+ signalling pathway showed either a complete inhibition or a significant reduction of the vitellogenic process. These results were confirmed with the rescue capability of the A-23187 ionophore in those oocyte batches in which the IP3/Ca2+ pathway was inhibited. Taking our findings into account, we propose that the cAMP molecule binds Xepac protein enabling it to activate the IP3/Ca2+ pathway, which is necessary to start and maintain X. laevis vitellogenin uptake.

The aim of the present study was to show the participation and physiological role of calmodulin (CaM) and cAMP during vitellogenin endocytic uptake in the amphibian Xenopus laevis. The results showed a differential distribution of CaM in the ovary follicles during oogenesis. The CaM intracellular localization was not affected by gap junction's downregulation and CaM inhibition did not completely abolished the endocytic activity of oocytes. We showed that cAMP was able to completely rescue the endocytic competence in follicles in which gap junctional communication had been disrupted by octanol. Moreover cAMP was capable of restoring oocyte endocytic capability in the presence of octanol and stelazine, a CaM inhibitor. We propose that, in Vtg uptake regulation, cAMP is upstream of CaM during the endocytic signalling pathway.

We conducted a histological investigation of the ovaries and testes during the gametogenesis and spawning in the giant jellyfish Nemopilema nomurai, which has bloomed in East Asian marginal seas almost annually since 2002. Oocytes arising from the ovarian epithelium make intimate contact with special epithelial cells, called trophocytes, which have microvilli on the subgenital sinus side, Golgi complexes and vesicles in the cytoplasm. In the early vitellogenic stage, yolk bodies occur in the ooplasm adjacent to the trophocytes, suggesting that the trophocytes transfer nutrients from the subgenital sinus to the oocyte. Later, yolk bodies are formed by the Golgi complexes in the entire ooplasm and accumulate until the oocyte matures. In the late vitellogenic stage, the oocyte separates from the trophocytes and forms microvilli on its surface, indicating nutrient uptake from the surrounding mesoglea. Nutrient support from the trophocytes in the early vitellogenic stage may make the oocytes mature rapidly after medusae are physically damaged. Microvilli-rich epithelial cells also associate with sperm follicles where spermatocytes arise from the follicle cells and accumulate, but their function in nutrient uptake is possibly less than that of trophocytes according to their morphology. During ovulation, which takes 1.5 hours after light exposure, trophocytes separate from each other and make an ovulation pit where the oocyte passes out to the subgenital sinus with the surrounding basal lamina. Spermiation occurs 5–20 minutes after light exposure, and spermatozoa are liberated through the spermiation pit that was formed by which the microvilli-rich cells dissociate. The trophocytes in ovaries and microvilli-rich cells in testes have important roles not only in the gametogenesis but also in the spawning of N. nomurai.

Fine structure observations were performed by means of electron microscopy on oogenesis and vitellogenesis and the special functions of follicular cells in the Chinese soft-shelled turtle (Pelodiseus sinensis). Histological examination of the ovary showed a well developed lacunae system containing fine granules, fibres or gelatiniform materials with one or two germinal beds dispersed on the dorsal surface of the ovarian cortex. The process of oogenesis comprised 10 consecutive phases according to the morphology of the yolk platelets, follicular cells and zona pellucida (ZP). Electron microscopy of vitellogenesis revealed some of the mitochondria gradually being transformed into yolk granules. In the advanced stage of vitellogenesis, large amounts of rough endoplasmic reticula, Golgiosomes and other cell organelles that are involved in synthesis and secretion were observed in follicular cells. The ZP was formed by microvilli, thus increasing the absorptive surface of the oocyte and facilitating transport of nutrients from the follicular epithelium to the ooplasm. This study demonstrated that the ovaries of members of the Testudinidae share more features with Archosaurs than with Squamates, indicating that these features were phylogenetically conserved in the Archosauria. The present observations suggest that the accumulation of yolk materials was controlled by the intrinsic and extrinsic pathways as well as by the activity of follicular cells. These results might also support a sibling relationship of the Testudinidae with the Archosauria and not with all extant reptiles.

Aspects of vitellogenesis, known to be controlled by juvenile hormone, are adversely affected by Hymenolepis diminuta infection of Tenebrio molitor, in spite of circulating titres of the hormone remaining unchanged. It has therefore been proposed that juvenile hormone binding is disrupted at the tissue site level. Juvenile hormone III binding sites were located in the nuclear, microsomal and post-microsomal supernatant fractions of the follicle cells of Tenebrio molitor. When JH-III binding was quantified for both control and Hymenolepis diminuta-infected beetles, binding in the nucleus and cytosol were found to be largely unaffected. However, microsomal binding was severely disrupted; on days 3 and 6 post-infection, binding was greatly diminished, on day 9 post-infection, binding was slightly reduced and, by day 15, binding was ‘restored’ to that of control insects. Using follicle cell microsomes at day 3 post-infection, previous Scatchard analysis revealed the presence of at least two JH-III binding sites. The first is of higher affinity, Kd = 5·3 x 10·5 x 10−11 mol/mg protein and the second of lower affinity Kd = 7·7 x 10−7 M, Bmax = 9·75 x 10−11 mol/mg protein. A comparison with microsomal binding parameters of follicle cells from non-infected Tenebrio indicated that although the Bmax values were unchanged, the Kd value of the higher affinity site was increased by approximately 5-fold. These data are indicative of a parasite-induced competitive binding inhibitor.

We have previously described the major yolk protein, vitellin, in the British blackfly Simulium ornatum Meigen. Here we demonstrate that vitellogenin, synthesized in the fat body and secreted into the haemolymph, is composed of subunits with the same approximate molecular weight as vitellin, namely 200 and 68 kDa. Simulium ornatum is the natural vector for the cattle filarial nematode Onchocerca lienalis Stiles, which induces host fecundity depletion. A significant reduction in ovarian vitellin content was associated with infection by intrathoracic injection of 20 O. lienalis microfilariae immediately after blood-feeding. Fat body synthesis of vitellogenin was significantly reduced as early as 8 h post-infection in comparison with sham-injected flies. When total haemolymph protein from infected and sham injected flies was compared, titres were significantly depressed 6 h post-infection. However, later in the infection, titres were elevated by 30%, the major component being vitellogenin. The injection of dead microfilariae had no effect. An infection burden of a single parasite caused a significant reduction in ovarian protein content in comparison with shams, but no further significant decrease was observed as the parasite burden was increased from 5 to 20. Possible mechanisms underlying the disturbance of Simulium reproductive physiology are proposed.

Some organisms rely on stored energy to fuel reproductive expenditure (capital breeders) whereas others use energy gained during the reproductive bout itself (income breeders). Most species occupy intermediate positions on this continuum, but few experimental data are available on the timescale over which food intake can affect fecundity. Mark–recapture studies of free-ranging female aspic vipers Vipera aspis have suggested that reproductive output relies not only on the energy in fat bodies accumulated in previous years, but also on food intake immediately before ovulation. A simple experiment was conducted to test this hypothesis, maintaining female snakes in captivity throughout the vitellogenic period and controlling their food intake. The energy input of a female strongly influenced the amount of mass that she gained and the number of ova that she ovulated. Multiple regression showed that litter size in these snakes was affected both by maternal body condition in early spring (an indicator of foraging success over previous years) and by food intake in the spring before ovulation. Our experimental data thus reinforce the results of descriptive studies on free-ranging snakes, and emphasize the flexibility of energy allocation patterns among vipers. Reproducing female vipers may combine energy from ‘capital’ and ‘income’ to maximize their litter sizes in the face of fluctuating levels of prey abundance.

The present study analyses, by transmission electron microscopy, vitellogenesis in two anuran amphibian families: Leptodactilidae (Ceratophrys cranwelli) and Bufonidae (Bufo arenarum). These differ in the type of stimulus that sets off their reproductive period, pluvial changes being the trigger in C. cranwelli and temperature increase in B. arenarum. We found that vitellogenesis follows an endocytic pathway that involves membranous structures (coated pits, coated vesicles, endosomes and multivesicular bodies). This process results in a fully grown yolk platelet of similar structure in both species. Despite the above similarity, a distinctive feature in B. arenarum was that the multivesicular bodies exhibited condensed proteins together with lipid droplets, the latter remaining as such even in the primordial yolk platelet. In C. cranwelli, however, lipids droplets were only found attached to the primordial yolk platelet. The coexistence of lipid droplets together with proteins in the nascent precursor yolk platelets observed in B. arenarum is similar to that found in B. marinus. This fact might constitute a characteristic feature of the Bufonidae family.

Two specific Enzyme Linked lmmunosorbent Assays (ELISA) for vitellogenin were developed using two different antisera anti-plasma vitellogenin purified by SDS polyacrilamide gel electrophoresis and electroelution. Purified vitellogenins migrated as single bands under SDS conditions with an apparent molecular mass of 154 and 152 kDa for Chrysichthys nigrodigitatus and Heterobranchus longifilis respectively.The ELISA was performed on 96 well microtiter plates using specific antibodies raised on rabbits and a second antibody (Goat anti-rabbit) labeled with peroxidase. Parallel displacement curves were obtained between purified vitellogenin and female plasma as well as plasma from estradiol treated fish. Both antibodies cross-reacted with the heterologous vitellogenin but the curves were not parallel, indicating different antigenic structures of the two vitellogenin molecules from the two catfish species. The reliability and the specificity of these assays allowed us to quantify vitellogenin in plasma samples during vitellogenesis of the two species. The annual vitellogenin plasma levels presented a good correlation with mean oocyte diameter and reproduction period in Chrysichthys nigrodigitatus. In Heterobranchus longifilis the mean oocyte diameter is almost constant all year round, but vitellogenin levels presented significant variations with higherlevels corresponding mainly to the low gonado-somatic index values observed during the dry seasons.

Infection with developing metacestodes of the rat tapeworm, Hymenolepis diminuta, is known to retard the accumulation of the yolk protein, vitellin, in the terminal ovarian follicles of the intermediate host, Tenebrio molitor. It is probable that this is the result of competitive inhibition of juvenile hormone binding at a microsomal binding site in the beetle follicular epithelium. Experiments were designed to test the hypothesis that inhibitor molecules were circulating in the haemolymph of infected beetles. Whole haemolymph, collected from male or female beetles at various stages post-infection, was injected into non-infected female recipients 2 days post-emergence. Ovaries were removed 3 days later and the vitellin content of the same sized follicles measured using an ELISA. The vitellin content of follicles from recipients of haemolymph from females infected with metacestodes at stage 1 and stage 3–4 was significantly reduced (24 and 27·9%) compared to sham-infected females. However, haemolymph from females infected with mature metacestodes did not affect the vitellin content. Results were thus comparable to those obtained by monitoring ovarian vitellin levels in female T. molitor with bona fide infections. Haemolymph from infected males did not affect ovarian vitellin content. These results indicate that molecules that can modulate vitellogenesis may be present in the haemolymph of females infected with developing metacestodes but that these factors disappear later in infection.

L'examen macroscopique, histologique et cytologique des ovaires de Barbus holotaenia adultes récoltés dans le bassin de la Luki entre juillet 1986 et septembre 1988 a permis de décrire six stades de développement des ovaires et sept stades au niveau des ovocytes et des follicules. Quelques conclusions biologiques et écologiques sont tirées de ces différentes observations. La fécondité individuelle partielle est comprise entre 0,005 et 0,018 ovocytes par kg de la masse du poisson.

Chrysichthys nigrodigitatus and Heterobranchus longifilis are two commercially important African catfish species that have been studied for their reproduction and breeding possibilities in the lagoon waters of Ivory Coast. C. nigrodigitatus is characterized by an annual reproductive cycle with a vitellogenesis period from March to June. The reproductive season coincides with the short rainy season which is characterized by lower water temperature and salinity at the beginning (September) and by a progressive rise of these parameters at the end of the season (November). The previtellogenic oocytes enter vitellogenesis at an average diameter of 320 μm. This initial oocyte stock is reconstituted progressively as soon as the spawning is achieved and lasts for a period of approximately 3 months. Fully vitellogenic oocytes reach an average diameter of 2.4–2.8 mm by the end of June. The maturation is possible only if a male and a female are confined in artificial spawning receptacles. Maturation and spawning generally occur within the following 3 to 5 weeks for 30 to 80% of the females. When the non-spawning females are kept in the nests they maintain fully vitellogenic oocytes for 3 months with a continuous atresia of the larger oocytes, while the ovaries of the non- spawning females, placed out of the receptacles, exhibit a complete atresia of all the vitellogenic oocytes. After an HCG-induced ovulation, a new reproductive cycle of H. longifilis can be achieved in less than a month. The previtellogenic oocytes enter vitellogenesis at an oocyte diameter of about 380 μm and become fully vitellogenic after 15 days (1.2–1.6 mm in diameter). If maturation is not artificially stimulated the females remain at the same stage of vitellogenesis while larger oocytes undergo atresia and are continuously replaced by growing vitellogenic oocytes.

This work examines the effects of constant long and short days on the hormonal profile of plasma17β-oestradiol (E2) levels and the ovarian vitellogenic events, as well as on the fecundity, time of spawning and egg quality of sea bass. Constant short days first administered in April advanced spawning (by 45 days on average) whereas exposure to constant long days from the same date induced a delay (averaging 38 days) compared to controls reared on an ambient light regime (Lat. 40°N). These light regimes produced shifts in the profiles of plasma 17β-oestradiol and in the percentage of the vitellogenic oocytes according to the spawning time of the respective groups. Relative fecundity of the short day group was similar to controls (257,000 versus 230,000 eggs/kg spawned female, respectively). However, in the long day group, relative fecundity was reduced to one half that of control fish (124,000). Plasma oestradiol showed a bimodal pattern under long days and vitellogenetic and atretic oocytes were present for a longer period (6-7 months compared to 4-5 months in controls). In summary, in this paper we describe, for the first time in the sea bass, the effect of constant photoperiod regimes on the 17β-oestradiol plasma profiles, the timing of spawning and the quality of the eggs.

Changes in the morphology of the ovary are described during maturation and it is established that it is a homogeneous structure for sampling purposes at maturity stages four and five. Several aspects of vitellogenic oocyte growth and recruitement were examined as criteria to ascertain whether the annual fecundity can be determined in Solea solea just prior to spawning. The first criterion was whether a hiatus develops in the size frequency distribution between the previtellogenic and vitellogenic oocytes just prior to spawning. We were unable to demonstrate this feature in sole from division VIId (N = 5) or IVc (N = 5) but it was present in two out of five fish from division VIIe and all fish examined from division VIIa (N = 5) and IVb west (N = 5). The hiates occurred between 175 and 250 µm (cohort size 25 m) in the size frequency distribution and measured between 25 and 75 µm. Oocyte size frequency distribution from mid spawning and late spawning-spent sole from division IVc showed a hiatus in 9 out of 10 fish which became wider as the residual annual potential fecundity declined. Next the rate of recruitment and growth of vitellogenic oocytes was studied in sole sampled at monthly intervals (July 1993 to February 1994) from division VIId in relation to the start and duration of the annual spawning season. The growth rate of the leading oocyte cohort was 2.73 × 10−5 mm3 per day on 20 September and increased to 3.91 × 10−3 at a mean diameter of 839 µm by 10 February just prior to spawning. However, the observed maximum growth rate, even when adjusted for the positive influence of temperature, allowed little scope for previtellogenic oocytes to complete maturation during a spawning period of 60 days. The recruitment of viltellogenic oocytes was 3013 oocytes per day on 3 August and declined rapidly as spawning approached. In the three months prior to spawning no significant increase was found in the annual potential fecundity. An analysis of a sample of spawning fish from IVc demonstrated that 88% had less than 50% of their predicted potential fecundity remaining in the ovary. The mean batch fecundity was 8 400 (SE ± 13 363) and in conjunction with existing data on spawning frequency and duration gave a realised fecundity close to the potential fecundity. It is concluded that for all practical purposes the sole can be regarded as having a determinate fecundity in all the areas studied.

The regulation of potential fecundity by follicular atresia was assessed in pre-spawning and spawning sole. In ICES areas IV and VII the prevalence of atretic oocytes in pre-spawning fish varied between 0.04 and 0.69 and the relative intensity varied between 0.017 and 0.076. During spawning in division IVc the prevalence of atresia varied between 0.45 and 0.57 and the relative intensity between 0.022 and 0.058. If we assume a turnover rate of 9 days the loss from the potential annual fecundity, during spawning, in division IVc would be 12.4%.