Is there a role for estrogen in follicular maturation in the primate

The present study focusses attention on the effects of blocking estrogen synthesis, during follicular phase, on follicular maturation in the adult female bonnet monkey (Macaca radiata). Administration of cycling females (n = 4) with an aromatase inhibitor CGS 16949A (AI) by Alzet mini-pump (2.5 mg/day) from day 3 of cycle resulted in significant reduction in basal (by 53%) and surge levels of estrogen (by 70%) but this had no effect on follicular maturation, ovulation and luteal function as assessed by serum hormone profiles as well as laparotomy. This lack of need for estrogen for completion of follicular maturation process was confirmed by administering cycling monkeys hFSH (25 IU/day) from day 3 till day 8 of the cycle along with (5 mg AI/day) or without Al (n = 3/group). Administration of Al resulted in suppression of FSH induced increase in serum estrogen (by 100%) and elevation in circulating androstenedione. Aromatase inhibitor treatment had no effect on either the number of follicles developed or their size relative to control. Testing the ability of both granulosa and thecal cells, removed on day 9 of treatment cycle, to respond to gonadotropins in vitro showed no change indicating that cellular development and maturation of follicular cells had occurred normally. It is concluded that follicular maturation in the primate can occur even when increase in estrogen synthesis is blocked.

Anatomical and developmental patterns of interleukin-2 gene expression in the mouse: analysis of IL-2 expressing cells and partial characterization of interactions involved in mediating IL-2 gene expression in vivo

Interleukin-2 (IL-2) is an important mediator in the vertebrate immune system. IL-2 is a potent growth factor that mature T lymphocytes use as a proliferation signal and the production of IL-2 is crucial for the clonal expansion of antigen-specific T cells in the primary immune response. IL-2 driven proliferation is dependent on the interaction of the lymphokine with its cognate multichain receptor. IL-2 expression is induced only upon stimulation and transcriptional activation of the IL-2 gene relies extensively on the coordinate interaction of numerous inducible and constitutive trans-acting factors. Over the past several years, thousands of papers have been published regarding molecular and cellular aspects of IL-2 gene expression and IL-2 function. The vast majority of these reports describe work that has been carried out in vitro. However, considerably less is known about control of IL-2 gene expression and IL-2 function in vivo.

To gain new insight into the regulation of IL-2 gene expression in vivo, anatomical and developmental patterns of IL-2 gene expression in the mouse were established by employing in situ hybridization and immunohistochemical staining methodologies to tissue sections generated from normal mice and mutant animals in which T -cell development was perturbed. Results from these studies revealed several interesting aspects of IL-2 gene expression, such as (1) induction of IL-2 gene expression and protein synthesis in the thymus, the primary site of T-cell development in the body, (2) cell-type specificity of IL-2 gene expression in vivo, (3) participation of IL-2 in the extrathymic expansion of mature T cells in particular tissues, independent of an acute immune response to foreign antigen, (4) involvement of IL-2 in maintaining immunologic balance in the mucosal immune system, and (5) potential function of IL-2 in early events associated with hematopoiesis.

Extensive analysis of IL-2 mRNA accumulation and protein production in the murine thymus at various stages of development established the existence of two classes of intrathymic IL-2 producing cells. One class of intrathymic IL-2 producers was found exclusively in the fetal thymus. Cells belonging to this subset were restricted to the outermost region of the thymus. IL-2 expression in the fetal thymus was highly transient; a dramatic peak ofiL-2 mRNA accumulation was identified at day 14.5 of gestation and maximal IL-2 protein production was observed 12 hours later, after which both IL-2 mRNA and protein levels rapidly decreased. Significantly, the presence of IL-2 expressing cells in the day 14-15 fetal thymus was not contingent on the generation of T-cell receptor (TcR) positive cells. The second class of IL-2 producing cells was also detectable in the fetal thymus (cells found in this class represented a minority subset of IL-2 producers in the fetal thymus) but persist in the thymus during later stages of development and after birth. Intrathymic IL-2 producers in postnatal animals were located in the subcapsular region and cortex, indicating that these cells reside in the same areas where immature T cells are consigned. The frequency of IL-2 expressing cells in the postnatal thymus was extremely low, indicating that induction of IL-2 expression and protein synthesis are indicative of a rare activation event. Unlike the fetal class of intrathymic IL-2 producers, the presence of IL-2 producing cells in the postnatal thymus was dependent on to the generation of TcR+ cells. Subsequent examination of intrathymic IL-2 production in mutant postnatal mice unable to produce either αβ or γδ T cells showed that postnatal IL-2 producers in the thymus belong to both αβ and γδ lineages. Additionally, further studies indicated that IL-2 synthesis by immature αβ -T cells depends on the expression of bonafide TcR αβ-heterodimers. Taken altogether, IL-2 production in the postnatal thymus relies on the generation of αβ or γδ-TcR^+ cells and induction of IL-2 protein synthesis can be linked to an activation event mediated via the TcR.

With regard to tissue specificity of IL-2 gene expression in vivo, analysis of whole body sections obtained from normal neonatal mouse pups by in situ hybridization demonstrated that IL-2 mRNA^+ cells were found in both lymphoid and nonlymphoid tissues with which T cells are associated, such as the thymus (as described above), dermis and gut. Tissues devoid of IL-2 mRNA^+ cells included brain, heart, lung, liver, stomach, spine, spinal cord, kidney, and bladder. Additional analysis of isolated tissues taken from older animals revealed that IL-2 expression was undetectable in bone marrow and in nonactivated spleen and lymph nodes. Thus, it appears that extrathymic IL-2 expressing cells in nonimmunologically challenged animals are relegated to particular epidermal and epithelial tissues in which characterized subsets of T cells reside and thatinduction of IL-2 gene expression associated with these tissues may be a result of T-cell activation therein.

Based on the neonatal in situ hybridization results, a detailed investigation into possible induction of IL-2 expression resulting in IL-2 protein synthesis in the skin and gut revealed that IL-2 expression is induced in the epidermis and intestine and IL-2 protein is available to drive cell proliferation of resident cells and/or participate in immune function in these tissues. Pertaining to IL-2 expression in the skin, maximal IL-2 mRNA accumulation and protein production were observed when resident Vγ_3^+ T-cell populations were expanding. At this age, both IL-2 mRNA^+ cells and IL-2 protein production were intimately associated with hair follicles. Likewise, at this age a significant number of CD3ε^+ cells were also found in association with follicles. The colocalization of IL-2 expression and CD3ε^+ cells suggests that IL-2 expression is induced when T cells are in contact with hair follicles. In contrast, neither IL-2 mRNA nor IL-2 protein were readily detected once T-cell density in the skin reached steady-state proportions. At this point, T cells were no longer found associated with hair follicles but were evenly distributed throughout the epidermis. In addition, IL-2 expression in the skin was contingent upon the presence of mature T cells therein and induction of IL-2 protein synthesis in the skin did not depend on the expression of a specific TcR on resident T cells. These newly disclosed properties of IL-2 expression in the skin indicate that IL-2 may play an additional role in controlling mature T-cell proliferation by participating in the extrathymic expansion of T cells, particularly those associated with the epidermis.

Finally, regarding IL-2 expression and protein synthesis in the gut, IL-2 producing cells were found associated with the lamina propria of neonatal animals and gut-associated IL-2 production persisted throughout life. In older animals, the frequency of IL-2 producing cells in the small intestine was not identical to that in the large intestine and this difference may reflect regional specialization of the mucosal immune system in response to enteric antigen. Similar to other instances of IL-2 gene expression in vivo, a failure to generate mature T cells also led to an abrogation of IL-2 protein production in the gut. The presence of IL-2 producing cells in the neonatal gut suggested that these cells may be generated during fetal development. Examination of the fetal gut to determine the distribution of IL-2 producing cells therein indicated that there was a tenfold increase in the number of gut-associated IL-2 producers at day 20 of gestation compared to that observed four days earlier and there was little difference between the frequency of IL-2 producing cells in prenatal versus neonatal gut. The origin of these fetally-derived IL-2 producing cells is unclear. Prior to the immigration of IL-2 inducible cells to the fetal gut and/or induction of IL-2 expression therein, IL-2 protein was observed in the fetal liver and fetal omentum, as well as the fetal thymus. Considering that induction of IL-2 protein synthesis may be an indication of future functional capability, detection of IL-2 producing cells in the fetal liver and fetal omentum raises the possibility that IL-2 producing cells in the fetal gut may be extrathymic in origin and IL-2 producing cells in these fetal tissues may not belong solely to the T lineage. Overall, these results provide increased understanding of the nature of IL-2 producing cells in the gut and how the absence of IL-2 production therein and in fetal hematopoietic tissues can result in the acute pathology observed in IL-2 deficient animals.

Ancylidae (Mollusca, Heterobranchia, Pulmonata, Basommatophora) do Estado do Rio de Janeiro, Brasil: morfologia, sistemática e distribuição geográfica

Os moluscos pateliformes de água doce da região Neotropical são comumente atribuídos a família Ancylidae sensu latum, abrangendo sete gêneros com pelo menos 13 espécies válidas e sete com identificação duvidosa. Os ancilídeos possuem pequenas dimensões, alcançam no máximo 15 mm de comprimento. Sua concha é frágil, composta por duas regiões, a protoconcha e a teleoconcha, as quais apresentam caracteres relevantes para a sistemática. Na parte mole as impressões musculares, a pigmentação do manto, o sistema reprodutor e a rádula são importantes para o estudo da família. Apesar de existirem vários registros de ocorrência para ancílideos no Estado do Rio de Janeiro (ERJ), existem poucos dados morfológicos. O principal objetivo deste estudo foi fornecer e ampliar as informações sobre a morfologia e distribuição geográfica das espécies de Ancylidae encontradas no ERJ. Os materiais utilizados foram procedentes de coletas próprias, material depositado em Coleções Científicas e dados de revisão bibliográfica. O estudo da morfologia comparada das conchas foi realizado com o auxílio de imagens de microscópio óptico e de varredura. Para a comparação das partes moles, os espécimes foram corados e dissecados sob lupa. Através da conquiliometria analisamos as diferenças inter e intrapopulacionais. Com este trabalho, a riqueza conhecida para Ancylidae no ERJ, aumentou de cinco para sete espécies: Burnupia sp., Ferrissia sp., Gundlachia radiata (Guilding, 1828),G. ticaga (Marcus & Marcus, 1962), Gundlachia sp., Hebetancylus moricandi (d`Orbigny, 1837) e Uncancylus concentricus (d`Orbigny, 1837). Gundlachia radiata e U. concentricus constituem novos registros para o ERJ, e G. radiata para a Região Sudeste. As três espécies com o maior número de registros de ocorrência no ERJ foram: G. ticaga (66%), Ferrissia sp. (37%) e Gundlachia sp. (18%). A ampla distribuição de G. ticaga pode ser devido à capacidade de suportar ambientes impactados. Em relação à morfologia, Burnupia sp. difere de Burnupia ingae Lanzer, 1991, única espécie deste gênero descrita para o Brasil, por apresentar diferenças na microescultura da concha e também na forma das impressões musculares. Ferrissia sp. difere de F. gentilis Lanzer, 1991, devido a diferenças na microescultura apical e número de cúspides no dente central da rádula. Gundlachia sp. é diferente de G. ticaga e G. radiata, por apresentar a abertura da concha mais arredondada, ápice mais recurvado, ultrapassando a borda da concha, pontuações irregulares em toda a protoconcha e forma dos músculos adutores anterior direito e posterior mais elíptica. A morfologia interna também mostra diferenças entre Gundlachia sp. e G. ticaga, como o apêndice terminal do útero e o número de folículos do ovoteste. Através das análises conquiliométricas, constatamos para os gêneros Burnupia, Ferrrissia e Gundlachia, que os índices morfométricos se mostraram melhores que as medidas lineares para a discriminação das espécies, provavelmente porque esses índices diminuem o efeito da amplitude de tamanho das conchas, que são fortemente influenciadas pelas variações ecofenotípicas. Contudo, os caracteres diagnósticos das conchas e das partes moles (impressões musculares) são indispensáveis para a identificação dos gêneros e espécies de Ancylidae. Palavras-chave: Mollusca. Moluscos de água doce. Morfologia. Distribuição geográfica. Estado do Rio de Janeiro.uscos.

A influência do fumo sobre a morfologia ovariana de camundongos Swiss

Considerando que nos tempos atuais o hábito de fumar atingiu uma parcela significativa das adolescentes brasileiras e apesar de seus conhecidos efeitos deletérios sobre diversos órgãos, pouco se sabe da sua ação sobre os ovários. Este estudo teve como objetivo avaliar os efeitos da exposição à fumaça de cigarro sobre o ciclo estral e a morfologia ovariana. Para tal, foram utilizados camundongos da linhagem Swiss, cujo estudo teve início com o estudo do perfil das características da maturação sexual. Após o desmame, aos 21 dias de idade, a partir da abertura vaginal, o 1 estro e o início da ciclicidade foram acompanhados através da citologia vaginal. As características do ciclo estral foram determinadas no decorrer de doze semanas. O efeito da exposição à fumaça de cigarro 3R4F utilizou fêmeas Swiss de 35 dias de idade que foram subdivididos em dois grupos expostos à fumaça de cigarro (grupo 15E) e animais controles livres de fumaça (grupo 15C). A exposição ocorreu por 15 dias e ao final deste período, metade dos animais de cada grupo foi sacrificada e ovários direitos foram coletados. A outra metade permaneceu em observação durante 30 dias, sem exposição à fumaça, originando os grupos 45Ex e 45C. A citologia vaginal foi avaliada durante todo o período experimental. Ao final dos 30 dias, sangue e ovários direitos foram coletados. Estes foram pesados e processados por técnica de rotina histológica para análise morfológica. A caracterização dos eventos da puberdade estabeleceu o tempo de abertura vaginal com média de 33,60,24 de idade, o primeiro estro 39,42,58 dias de idade e o início da ciclicidade, com média de 39,51,19 dias de idade, concomitante com o primeiro estro. Além disso, os ciclos estrais apresentaram períodos de cinco dias com freqüência baixa da fase diestro. Com relação à exposição da fumaça de cigarro ocorreu aumento significativo na extensão dos ciclos estrais e uma forte tendência ao aumento de estros nos animais 45Ex, apesar de não ser significativa. O número e o diâmetro de folículos grandes foram maiores no 15E, enquanto o de corpos lúteos foi menor. Em relação ao grupo 45C, o 45Ex não se alterou, porém, mostrou uma discreta redução da massa ovariana, do número de folículos pequenos, do número e do diâmetro dos folículos médios, dos corpos lúteos e aumento de folículos atrésicos. A comparação entre controles, 15C e 45C e expostos, 15E e 45Ex, mostrou uma redução no diâmetro de folículos médios e grandes. O estudo do perfil das características reprodutivas de fêmeas Swiss é indispensável para modelos experimentais em pesquisa que fazem uso desta linhagem. Permitiu verificar que a exposição à fumaça de cigarro promove alteração do ciclo estral, da massa ovariana e antecipa alterações morfológicas tempo dependente que sinaliza a finalização da vida reprodutiva

The effect of temperature on the duration of spawning markers—migratory-nucleus and hydrated oocytes and postovulatory follicles—in the multiple-batch spawner Japanese flounder (Paralichthys olivaceus)

The duration of spawning markers (e.g. signs of previous or imminent spawnings) is essential information for estimating spawning frequency of fish. In this study, the effect of temperature on the duration of spawning markers (i.e., oocytes at early migratory nucleus, late migratory nucleus, and hydrated stages, as well as new postovulatory follicles) of an indeterminate multiple-batch spawner, Japanese f lounder (Paralichthys olivaceus), was evaluated. Cannulation was performed to remove samples of oocytes, eggs, and postovulatory follicles in individual females at 2–4 hour intervals over 27–48 hours. The duration of spawning markers was successfully evaluated in 14 trials ranging between 9.2° and 22.6°C for six females (total length 484–730 mm). The durations of spawning markers decreased exponentially with temperature and were seen to decrease by a factor of 0.16, 0.36, 0.30, and 0.31 as temperature increased by 10°C for oocytes at early migratory nucleus, late migratory nucleus, and hydrated stages, and new postovulatory follicles, respectively. Thus, temperature should be considered when estimating spawning frequency from these spawning markers, especially for those fish that do not spawn synchronously in the population.

Estimates of body sizes at maturation and at sex change, and the spawning seasonality and sex ratio of the endemic Hawaiian grouper (Hyporthodus quernus, F. Epinephelidae)

A case study of the reproductive biology of the endemic Hawaiian grouper or hapu’upu’u (Hyporthodus quernus) is presented as a model for comprehensive future studies of economically important epinephelid groupers. Specimens were collected throughout multiple years (1978–81, 1992–93, and 2005–08) from most reefs and banks of the Northwestern Hawaiian Islands. The absence of small males, presence of atretic oocytes and brown bodies in testes of mature males, and both developed ovarian and testicular tissues in the gonads of five transitional fish provided evidence of protogynous hermaphroditism. No small mature males were collected, indicating that Hawaiian grouper are monandrous (all males are sex-changed females). Complementary microscopic criteria also were used to assign reproductive stage and estimate median body sizes (L50) at female sexual maturity and at adult sex change from female to male. The L50 at maturation and at sex change was 580 ±8 (95% confidence interval [CI]) mm total length (TL) and 895 ±20 mm TL, respectively. The adult sex ratio was strongly female biased (6:1). Spawning seasonality was described by using gonadosomatic indices. Females began ripening in the fall and remained ripe through April. A February–June main spawning period that followed peak ripening was deduced from the proportion of females whose ovaries contained hydrated oocytes, postovulatory follicles, or both. Testes weights were not affected by season; average testes weight was only about 0.2% of body weight—an order of magnitude smaller than that for ovaries that peaked at 1–3% of body weight. The species’ reproductive life history is discussed in relation to its management.

Reproductive biology of blue marlin (Makaira nigricans) in the western Pacific Ocean

The reproductive biology of blue marlin (Makaira nigricans) was assessed from 1001 fish (ranging from 121 to 275 cm in eye-to-fork length; EFL) caught by Taiwanese offshore longliners in the western Pacific Ocean from September 2000 to December 2001 and from 843 gonad samples from these fish, The overall sex ratio of the catch was approximately 1:1 dur ing the sampling period, but blue marlin are sexually dimorphic; females are larger than males. Reproductive activity (assessed by histology), a gonadosomatic index, and the distribution of oocyte diameters, indicated that spawning occurred predominantly from May to September. The estimated sizes-at-maturity (EFL50) were 179.76 ±1.01 cm (mean ±standard error) for females and 130 ±1 cm EFL for males. Blue marlin are multiple spawners and oocytes develop asynchronously. The proportion of mature females with ovaries containing postovulatory follicles (0.41) and hydrated oocytes (0.34) indicated that the blue marlin spawned once every 2–3 days on average. Batch fecundity (BF) for 26 females with the most advanced oocytes (≥1000 μm), but without postovulatory follicles, ranged from 2.11 to 13.50 million eggs (6.94 ± 0.54 million eggs). The relationships between batch fecundity (BF, in millions of eggs) and EFL and round weight (RW, kg) were BF = 3.29 × 10 –12 EFL5.31 (r2 = 0.70) and BF = 1.59 × 10–3 RW 1.73 (r2= 0.67), respectively. The parameters estimated in this study are key information for stock assessments of blue marlin in the western Pacific Ocean and will contribute to the conservation and sustainable yield of

Advances in methods for determining fecundity: application of the new methods to some marine fishes

Estimation of individual egg production (realized fecundity) is a key step either to understand the stock and recruit relationship or to carry out fisheries-independent assessment of spawning stock biomass using egg production methods. Many fish are highly fecund and their ovaries may weigh over a kilogram; therefore the work time can be consuming and require large quantities of toxic fixative. Recently it has been shown for Atlantic cod (Gadus morhua) that image analysis can automate fecundity determination using a power equation that links follicles per gram ovary to the mean vitellogenic follicular diameter (the autodiametric method). In this article we demonstrate the precision of the autodiametric method applied to a range of species with different spawning strategies during maturation and spawning. A new method using a solid displacement pipette to remove quantitative fecundity samples (25, 50, 100, and 200 milligram [mg]) is evaluated, as are the underlying assumptions to effectively fix and subsample the ovary. Finally, we demonstrate the interpretation of dispersed formaldehyde-fixed ovarian samples (whole mounts) to assess the presence of atretic and postovulatory follicles to replace labor intensive histology. These results can be used to estimate down regulation (production of atretic follicles) of fecundity during maturation.

Reproductive biology of female Rikuzen sole (Dexistes rikuzenius)*

The annual ovarian cycle, mode of maturation, age at maturity, and potential fecundity of female Rikuzen sole (Dexistes rikuzenius) from the North Pacific Ocean off the coast of Japan were studied by 1) histological examination of the gonads, 2) measurement and observation of the oocytes, and 3) by otolith aging. The results indicated that ovulation occurs from September to December and peaks between September and October. Vitellogenesis began again soon after the end of the current season. Maturity was divided into eight phases on the basis of oocyte developmental stages. Mature ovaries contained developing oocytes and postovulatory follicles but no recruiting oocytes, indicating that this species has group-synchronous ovaries and is a multiple spawner. Almost all females matured first at an age of 1+ year and spawned every year until at least age 8+ years. Potential fecundity increased exponentially with body length and the most fecund fish had 15 times as many oocytes as the least fecund fish. Potential fecundity and relative fecundity were both positively correlated with age from 1 to 6+ years, but were negatively correlated, probably because of senescence, in fish over 7 years. These results emphasize that the total productivity of a D. rikuzenius population depends not only on the biomass of females older than 1+ but also on the age structure of the population.

Movements and spawning of white marlin (Tetrapturus albidus) and blue marlin (Makaira nigricans) off Punta Cana, Dominican Republic

With a focus on white marlin (Tetrapturus albidus), a concurrent electronic tagging and larval sampling effort was conducted in the vicinity of Mona Passage (off southeast Hispaniola), Dominican Republic, during April and May 2003. Objectives were 1) to characterize the horizontal and vertical movement of adults captured from the area by using pop-up satellite archival tags (PSATs); and 2) by means of larval sampling, to investigate whether fish were reproducing. Trolling from a sportfishing vessel yielded eight adult white marlin and one blue marlin (Makaira nigricans); PSAT tags were deployed on all but one of these individuals. The exception was a female white marlin that was unsuitable for tagging because of injury; the reproductive state of its ovaries was examined histologically. Seven of the PSATs reported data summaries for water depth, temperature, and light levels measured every minute for periods ranging from 28 to 40 days. Displacement of marlin from the location of release to the point of tag pop-up ranged from 3l.6 to 267.7 nautical miles (nmi) and a mean displacement was 3.4 nmi per day for white marlin. White and blue marlin mean daily displacements appeared constrained compared to the results of other marlin PSAT tagging studies. White marlin ovarian sections contained postovulatory follicles and final maturation-stage oocytes, which indicated recent and imminent spawning. Neuston tows (n=23) yielded 18 istiophorid larvae: eight were white marlin, four were blue marlin, and six could not be identified to species. We speculate that the constrained movement patterns of adults may be linked to reproductive activity for both marlin species, and, if true, these movement patterns may have several implications for management. Protection of the potentially important white marlin spawning ground near Mona Passage seems warranted, at least until further studies can be conducted on the temporal and spatial extent of reproduction and associated adult movement.

The reproductive cycle of the thorny skate (Amblyraja radiata) in the western Gulf of Maine

The thorny skate (Amblyraja radiata) is a large species of skate that is endemic to the waters of the western north Atlantic in the Gulf of Maine. Because the biomass of thorny skates has recently declined below threshold levels mandated by the Sustainable Fisheries Act, commercial harvests from this region are prohibited. We have undertaken a comprehensive study to gain insight into the life history of this skate. The present study describes and characterizes the reproductive cycle of female and male thorny skates, based on monthly samples taken off the coast of New Hampshire, from May 2001 to May 2003. Gonadosomatic index (GSI), shell gland weight, follicle size, and egg case formation, were assessed for 48 female skates. In general, these reproductive parameters remained relatively constant throughout most of the year. However, transient but significant increases in shell gland weight and GSI were obser ved during certain months. Within the cohort of specimens sampled monthly throughout the year, a subset of females always had large preovulatory follicles present in their ovaries. With the exception of June and September specimens, egg cases undergoing various stages of development were observed in the uteri of specimens captured during all other months of the year. For males (n=48), histological stages III through VI (SIII−SVI) of spermatogenesis, GSI, and hepatosomatic index (HSI) were examined. Although there appeared to be monthly fluctuations in spermatogenesis, GSI, and HSI, no significant differences were found. The production and maintenance of mature spermatocysts (SVI) within the testes was observed throughout the year. These findings collectively indicate that the thorny skate is reproductively active year round.

Reproductive biology of carpenter seabream (Argyrozona argyrozona) (Pisces: Sparidae) in a marine protected area

The carpenter seabream (Argyrozona argyrozona) is an endemic South African sparid that comprises an important part of the handline fishery. A three-year study (1998−2000) into its reproductive biology within the Tsitsikamma National Park revealed that these fishes are serial spawning late gonochorists. The size at 50% maturity (L50) was estimated at 292 and 297 mm FL for both females and males, respectively. A likelihood ratio test revealed that there was no significant difference between male and female L50 (P>0.5). Both monthly gonadosomatic indices and macroscopically determined ovarian stages strongly indicate that A. argyrozona within the Tsitsikamma National Park spawn in the astral summer between November and April. The presence of postovulatory follicles (POFs) confirmed a six-month spawning season, and monthly proportions of early (0−6 hour old) POFs showed that spawning frequency was highest (once every 1−2 days) from December to March. Although spawning season was more highly correlated to photoperiod (r = 0.859) than temperature (r = −0.161), the daily proportion of spawning fish was strongly correlated (r= 0.93) to ambient temperature over the range 9−22oC. These results indicate that short-term upwelling events, a strong feature in the Tsitsikamma National Park during summer, may negatively affect carpenter fecundity. Both spawning frequency and duration (i.e., length of spawning season) increased with fish length. As a result of the allometric relationship between annual fecundity and fish mass a 3-kg fish was calculated to produce fivefold more eggs per kilogram of body weight than a fish of 1 kg. In addition to producing more eggs per unit of weight each year, larger fish also produce significantly larger eggs.

Efeito da restrição alimentar materna durante a lactação sobre a função ovariana da prole fêmea na puberdade e vida adulta

α.O objetivo deste estudo foi avaliar se a restrição alimentar materna durante a lactação altera a função ovariana da prole na puberdade e na vida adulta. No dia do nascimento da ninhada, as ratas foram separadas aleatoriamente nos seguintes grupos: (C) controle = dieta com 23% de proteína; (PER) restrição protéica calórica = dieta com 8% de proteína. Após o desmame, as proles tiveram livre acesso a dieta com 23% proteína e apenas os animais na fase diestro foram sacrificados com uma dose letal de pentobarbital aos 40 e 90 dias de idade. O sangue foi retirado por punção cardíaca e o soro armazendo para posterior dosagem dos níveis hormonais por radioimunoensaio. O ovário direito foi excisado, pesado e armazenado a 80C para subseqüente avaliação do receptor de androgênio (AR), das isoformas dos receptores de estrogênio (ERα, ERβ1 e ERβ2), do receptor do hormônio folículo estimulante (FSHR), do receptor do hormonio luteinizante (LHR), das isoformas dos receptores de leptina (Ob-R, Ob-Ra and Ob-Rb), e da enzima aromatase pela técnica de RT-PCR. O ovário esquerdo foi incluido em parafina, seccionado em cortes de 5 μm de espessura e processado por análise histológica de rotina, para classificação dos foliculos ovarianos. Na puberdade, o grupo PER apresentou um aumento significativo (p<0,05) no número dos folículos pré-antrais e antrais, enquanto o número de folículos primordiais, folículos de Graaf e corpo lúteo foram reduzidos significativamente (p<0,05). As concentrações sericas de estradiol (pg/ml) foram significativamente aumentadas (p<0,05). Houve aumento significativo na expressão do RNAm dos FSHR (p<0.05) e LHR (p<0.05), e diminuição significativa na expressão do RNAm dos AR (p<0.05), ERα (p<0.05), ERβ1 (p<0.05) e ERβ2 (p<0.05). Na vida adulta, a restrição alimentar causou uma diminuição no número total de folículos ovarianos, entretanto esta redução só foi significativa no número dos folículos primordiais, primários e de Graaf (p<0.05). Houve redução significativa da expressão da enzima aromatase (p<0.01), do FSHR (p<0.05), LHR (p<0.05), Ob-R (p<0.05) e Ob-Rb (p<0.05). Podemos concluir que a restrição alimentar materna durante a lactação causa uma programação metabólica no ovário da prole, alterando a foliculogênese, expressão das diferentes isoformas dos receptores de estrogênio, leptina, androgênio e gonadotropinas e da enzima aromatase. Essa programação, possivelmente decorrente dos baixos níveis de leptina nos primeiros dias de vida, pode contribuir para uma senescência precoce e redução da fertilidade já descrita na literatura.

Reproductive dynamics of female spotted seatrout (Cynoscion nebulosus) in South Carolina

We describe reproductive dynamics of female spotted seatrout (Cynoscion nebulosus) in South Carolina (SC). Batch fecundity (BF), spawning frequency (SF), relative fecundity (RF), and annual fecundity (AF) for age classes 1−3 were estimated during the spawning seasons of 1998, 1999, and 2000. Based on histological evidence, spawning of spotted seatrout in SC was determined to take place from late April through early September. Size at first maturity was 248 mm total length (TL); 50% and 100% maturity occurred at 268 mm and 301 mm TL, respectively. Batch fecundity estimates from counts of oocytes in final maturation varied significantly among year classes. One-year-old spotted seatrout spawned an average of 145,452 oocytes per batch, whereas fish aged 2 and 3 had a mean BF of 291,123 and 529,976 oocytes, respectively. We determined monthly SF from the inverse of the proportion of ovaries with postovulatory follicles (POF) less than 24 hours old among mature and developing females. Overall, spotted seatrout spawned every 4.4 days, an average of 28 times during the season. A chronology of POF atresia for water temperature >25°C is presented. Length, weight (ovary-free), and age explained 67%, 65%, and 58% of the variability in BF, respectively. Neither RF (number of oocytes/g ovary-free weight) nor oocyte diameter varied significantly with age. However, RF was significantly greater and oocyte diameter was smaller at the end of the spawning season. Annual fecundity estimates were approximately 3.2, 9.5, and 17.6 million oocytes for each age class, respectively. Spotted seatrout ages 1−3 contributed an average of 29%, 39%, and 21% to the overall reproductive effort according to the relative abundance of each age class. Ages 4 and 5 contributed 7% and 4%, respectively, according to predicted AF values.

Factors influencing the timing and frequency of spawning and fecundity of the goldlined seabream (Rhabdosargus sarba) (Sparidae) in the lower reaches of an estuary

We have studied the reproductive biology of the goldlined seabream (Rhabdosargus sarba) in the lower Swan River Estuary in Western Australia, focusing particularly on elucidating the factors influencing the duration, timing, and frequency of spawning and on determining potential annual fecundity. Our results demonstrate that 1) Rhabdosargus sarba has indeterminate fecundity, 2) oocyte hydration commences soon after dusk (ca. 18:30 h) and is complete by ca. 01:30−04:30 h and 3) fish with ovaries containing migratory nucleus oocytes, hydrated oocytes, or postovulatory follicles were caught between July and November. However, in July and August, their prevalence was low, whereas that of fish with ovaries containing substantial numbers of atretic yolk granule oocytes was high. Thus, spawning activity did not start to peak until September (early spring), when salinities were rising markedly from their winter minima. The prevalence of spawning was positively correlated with tidal height and was greatest on days when the tide changed from flood to ebb at ca. 06:00 h, i.e., just after spawning had ceased. Because our estimate of the average daily prevalence of spawning by females during the spawning season (July to November) was 36.5%, individual females were estimated to spawn, on average, at intervals of about 2.7 days and thus about 45 times during that period. Therefore, because female R. sarba with total lengths of 180, 220, and 260 mm were estimated to have batch fecundities of about 4500, 7700, and 12,400 eggs, respectively, they had potential annual fecundities of about 204,300, 346,100 and 557,500 eggs, respectively. Because spawning occurs just prior to strong ebb tides, the eggs of R. sarba are likely to be transported out of the estuary into coastal waters where salinities remain at ca. 35‰. Such downstream transport would account for the fact that, although R. sarba exhibits substantial spawning activity in the lower Swan River Estuary, few of its early juveniles are recruited into the nearshore shallow waters of this estuary.