Cellular development of the germinal epithelium during the gametogenic cycle of the golden mussel Limnoperna fortunei (Bivalvia: Mytilidae)

AbstractThe golden mussel Limnoperna fortunei is an invasive species that has quickly dispersed and colonized several potential different habitats distributed all over the world, causing environmental and economic impacts. Thus, in order to contribute to a better understanding of the reproductive aspects of L. fortunei, we described the cellular dynamic of the male and female germinal epithelium during the annual reproductive life history of this species, with the use of high resolution histology. An approximate of 1 200 specimens of L. fortunei were sampled periodically on the upper Paraná River floodplain (Brazil), from March 2010 to December 2012. Based on the activity of the germinal epithelium and consequent germ cell development, this study has resulted in the recognition of the following reproductive phases: Developing, Spawning Capable, Regressing and Regenerating. In the characterization of these phases, the following germ cells were described for males: spermatogonia, primary and secondary spermatocytes, spermatids and spermatozoa. Cell nests, oogonia, early prophase oocytes, previtellogenic oocytes and vitellogenic oocytes (early vitellogenic oocytes, middle vitellogenic oocytes and full-grown oocytes) were described for females. The morphological data and reproductive parameters obtained, showed the value of the cellular dynamics of the germinal epithelium, for the understanding of the cyclic gonadal events during the adult reproductive life of the mollusk in general. These results on the gametogenesis of this invasive species may be a fundamental tool for the development of control strategies and programs implementation, to reduce their proliferation and impacts in natural local environments. Rev. Biol. Trop. 64 (2): 521-536. Epub 2016 June 01.

ResumenEl mejillón dorado Limnoperna fortunei (Dunker, 1857) es una especie invasora que se ha dispersado rápidamente y ha colonizado diferentes hábitats potenciales distribuidos por todo el mundo, causando impactos ambientales y económicos. El control de esta especie depende del conocimiento de los aspectos reproductivos. Entre marzo 2010 y diciembre 2012 fueron muestreados periodicamente alrededor de 1 200 especímenes de L. fortunei en la planicie de inundación del Alto Rio Paraná, Brasil. Con el fin de contribuir con una mejor comprensión de los aspectos reproductivos de L. fortunei, se describió la dinámica celular del epitelio germinativo masculino y femenino durante la vida reproductiva anual de esta especie, mediante histología de alta resolución basada en la actividad del epitelio germinal y el consecuente desarrollo de las células germinales. En este estudio se reconocieron las siguientes fases reproductivas: desarrollo, capacidad de desove, regresión y regeneración. En los machos se describieron las siguientes células germinales: espermatogonias, espermatocitos primarios y secundarios, espermátidas y espermatozoides. Mientras que en las hembras se estudiaron: nidos celulares, ovogonias, ovocitos profásicos tempranos, ovocitos pre-vitelogénicos y ovocitos vitelogénicos (ovocitos vitelogénicos tempranos, ovocitos vitelogénicos y ovocitos completamente desarrollados).Los datos morfológicos y parámetros reproductivos que se muestran aquí confirman cómo la dinámica celular del epitelio germinal permite la comprensión de los acontecimientos cíclicos gonadales durante la vida reproductiva de adultos del molusco, en general. El conocimiento de la gametogénesis de esta especie invasora puede convertirse en una herramienta fundamental para el desarrollo de estrategias de control y aplicación de programas para disminuir su proliferación en ambientes naturales.

Fecundidad de la medusa Stomolophus meleagris (Rhizostomeae: Stomolophidae) en el Golfo de California / Stomolophus meleagris fecundity (Rhizostomeae: Stomolophidae) in the Gulf of California

The cannonball jellyfish Stomolophus meleagris is regarded as a fishery resource with high commercial value, but with scarce biological information. With the aim to generate preliminary information on reproductive aspects, the present study analyzes its fecundity, based on the estimated number of vitellogenic oocytes in the gonad; in addition, we evaluated its relationship with the jellyfish body length, diameter and wet weight; and we established the relationship of the gonadosomatic index (IGS) with the jellyfish diameter and length. For this, a total of 30 specimens were collected, measured and weighed in a monthly basis, in Las Guásimas lagoon from January to May 2006. In order to estimate the gonad volume, 60 mature females were analyzed and their gonads were extracted and stained with hematoxylin-eosin. Gonad images from histological preparations were analyzed and the number of vitellogenic and previtellogenic oocytes obtained. We used catch data from The Guásimas for population recruitment analysis, that was estimated with FISAT II. Among results, we found that fecundity increased with jellyfish length, diameter and total wet weight. The lower fecundity rates occured on February and the highest during May (11 873 071 and 37 528 197 millions of vitellogenic oocytes per gonad, respectively). The IGS increased with length and diameter of the specimens, at its maximum value of 3.7% was observed in May. A continuous production of previtellogenic oocytes were observed during the full collection period. The maximum recruitment was observed in July and November (37.50% and 28.01% respectively). The high fecundity observed in S. meleagris, suggests a high population reproductive potential that may support the possibility of an important fishery consolidation on this jellyfish.

La medusa Stomolophus meleagris es considerada un recurso pesquero de alto valor comercial. Debido a la relevancia de conocer aspectos de importancia biológica actualmente desconocidos, el presente trabajo analiza la fecundidad con base en la estimación del número de ovocitos vitelogénicos en la gónada; su relación con la longitud, diámetro y peso húmedo; y la relación del índice gonadosomático (IGS) con su diámetro y longitud. De enero a mayo 2006 se recolectaron 30 ejemplares mensuales que fueron medidos y pesados, de los cuales se analizaron 60 hembras maduras para calcular el volumen de sus gónadas. Se extrajeron las gónadas, se tiñeron con hematoxilina-eosina y se seleccionaron las hembras con madurez gonadal. Mediante la digitalización de imágenes se estimó el número de ovocitos vitelogénicos y previtelogénicos. Se emplearon datos de captura en Las Guásimas para el análisis de reclutamiento poblacional, estimado con FISAT II. La fecundidad se incrementó con la longitud, el diámetro y el peso húmedo total. La menor fecundidad se observó en febrero y la mayor en mayo (11 873 071 y 37 528 197 millones de ovocitos vitelogénico por gónada, respectivamente). El IGS se incrementó con la longitud y diámetro de los ejemplares, con su máximo en mayo de 3.7%. Se observó una producción continua de ovocitos previtelogénicos en todo el periodo de recolecta. El máximo reclutamiento se observó en julio y noviembre (37.50% y 28.01% respectivamente). La alta fecundidad encontrada en S. meleagris sugiere un alto potencial reproductivo para la población y sustenta la posibilidad de la consolidación de una importante pesquería de esta medusa.

Reproduction of the non-native fish Lepomis gibbosus (Perciformes: Centrarchidae) in Brazil

Minas Gerais is the fourth largest Brazilian state, and has an estimate of 354 native fish species. However, these fish species may be threatened, as this state has the highest rank of fish introductions reported for Brazil and South America. As one from the total of 85 non-native species detected, Lepomis gibbosus was introduced in the 60s to serve both as foragefish and to improve sport fishing. In this study, we evaluated the establishment of L. gibbosus in a shallow lake in the city of Ouro Preto, Doce River basin, state of Minas Gerais, Southeastern Brazil. We collected fish with fishing rods every two months from March 2002-February 2003. Fragments of gonads from a total of 226 females and 226 males were obtained and processed following standard histological techniques; then 5-7μm thickness sections were taken and stained in hematoxylin-eosin. Besides, for each specimen, the biometric measurements included the standard length (SL) and body weight (BW); and the sex ratio was obtained. The reproductive cycle stages were confirmed by the distribution of oocytes and spermatogenic cells. The type of spawning was determined by the frequency distribution of the reproductive cycle stages and ovarian histology. Based on the microscopic characteristics of the gonads, the following stages of the reproductive cycle were determined: one=Rest, two=Mature, three=Spawned for females or Spent for males; males and females in reproduction were found throughout the study period. Post-spawned ovaries containing oocytes in stages one (initial perinucleolar), two (advanced perinucleolar), three (pre-vitellogenic), four (vitellogenic) and post-ovulatory follicles indicated fractionated-type spawning in this species. The smallest breeding male and female measured were 4.6 and 4.9cm standard length, respectively, suggesting stunting. The sex ratio did not vary between males and females along the year and bimonthly, being 1:1. Moreover, L. gibbosus appears to be at stage three of biological invasion: establishment through reproduction. We suggest to deliver information about “non-native species” through lectures in schools, colleges/ universities, NGOs, government and environmental agencies in the cities and villages, in order to try to prevent environmental degradation by the introduction of non-native fish such as L. gibbosus in the region. We also recommend high fines for redhanded, and the import ban of non-native fish species to the region.

Se analiza el establecimiento del pez introducido Lepomis gibbosus en una laguna natural de la ciudad de Ouro Preto, Cuenca del Rio Doce, provincia de Minas Gerais, región sureste de Brasil. Cada dos meses se realizaron muestreos con anzuelo y línea, entre marzo 2002-febrero 2003 y se capturaron 226 hembras y 226 machos. Se encontraron hembras y machos en actividad reproductiva durante todo el muestreo. Ovarios con signos de desove con oocitos de diferentes tallas y folículos post-ovulatorios indicaron la puesta parcial para L. gibbosus. La hembra y macho en reproducción más pequeños tenían entre 4.6cm y 4.9cm de longitud, que caracteriza el enanismo. La proporción sexual fue 1:1 y no presentó diferencias bimensuales ni anuales. De las cinco fases del proceso de bioinvasión, se confirmó que L. gibbosus se encuentra en la fase tres, llamada establecimiento a través de la reproducción. Se sugiere aclaración sobre la temática “especie foránea” para evitar la degradación del ambiente con la introducción de peces foráneos en la región.

Assessment of gonadal follicle size in the invading bivalve Limnoperna fortunei (Mytilidae)

Limnoperna fortunei is an invasive gonochoristic and byssate freshwater bivalve originary from Southeast Asia. It shows great adaptive-reproductive ability, so knowledge of the gonadal cycle is an important factor for the prevention and control of this bioinvasion. This species is highly damaging to natural and human environments. We analyzed the distribution and maturity state of reproductive follicles in the mantle of both male and females. Male results are not shown but, in general, they followed the same pattern as that of females. Routine histological techniques included serial longitudinal sections and transversal sections in three body regions (anterior, middle and psoterior). Oocytes with conspicuous nucleoli were measured on both types of sections to estimate the maturity stage in the different regions. ANOVA indicates that there were no significant differences in maturity ratio between the studied regions, so that a small number of sections would render precise results to assess maturity

Assessment of gonadal follicle size in the invading bivalve Limnoperna fortunei (Mytilidae)

Limnoperna fortunei is an invasive gonochoristic and byssate freshwater bivalve originary from Southeast Asia. It shows great adaptive-reproductive ability, so knowledge of the gonadal cycle is an important factor for the prevention and control of this bioinvasion. This species is highly damaging to natural and human environments. We analyzed the distribution and maturity state of reproductive follicles in the mantle of both male and females. Male results are not shown but, in general, they followed the same pattern as that of females. Routine histological techniques included serial longitudinal sections and transversal sections in three body regions (anterior, middle and psoterior). Oocytes with conspicuous nucleoli were measured on both types of sections to estimate the maturity stage in the different regions. ANOVA indicates that there were no significant differences in maturity ratio between the studied regions, so that a small number of sections would render precise results to assess maturity

Morphological and morphometric aspects of Crassostrea rhizophorae (Guilding, 1828) oocytes in three stages of the gonadal cycle / Aspectos Morfológicos y Morfométricos de Oocitos de Crassostrea rhizophorae (Guilding, 1828) en Tres Fases del Ciclo Gonadal

Crassostrea rhizophorae (Guilding, 1828) es una de las especies de ostras tropicales cultivadas en la costa brasilera, que presenta alto valor comercial, no existiendo información sobre las características gonadales, durante las fases reproductivas de esta especie. El objetivo de este trabajo fue analizar la morfología de los ovocitos y de los folículos gonadales de Crassostrea rhizophorae, en tres fases del ciclo gonadal. Analizando los cortes histológicos de las gónadas y considerando los ovocitos que presentaban núcleo y nucléolo evidentes, fueron determinadas tres fases del ciclo gonadal: gametogénesis inicial, crecimiento y maduración. En la fase de gametogénesis inicial, el diámetro de los folículos y de los ovocitos era de 180,29 (+ - 41,91) e 18,66 (+ - 6,85) µm, respectivamente; también identificamos gran cantidad de tejido conjuntivo interfolicular y un número mayor de ovocitos previtelogénicos. En la fase de crecimiento, el diámetro de los folículos y de los ovocitos era de 218,02 (+ - 43,19) y 25,92 (+ - 9,94) µm, respectivamente. Este fase fue caracterizada por una pequeña cantidad de tejido conjuntivo interfolicular y predominio de ovocitos vitelogénicos. En la fase de maduración, el diámetro de los folículos y de los ovocitos era de 298,16 ( + - 99,24) y 35,27 (+ - 6,2) µm, respectivamente, existiendo gran número de ovocitos maduros. De esta manera, concluimos que durante la implantación del cultivo, Crassostrea rhizophorae tolera la influencia de los factores intrínsecos y extrínsecos y no presenta alteraciones significativas en su actividad reproductiva.

Crassostrea rhizophorae (Guilding, 1828) is one of the tropical species of oysters cultivated on the Brazilian shore. Despite its high commercial value, there is no information on the gonadal characteristics during the reproductive stages of this species. The objective of this work was to analyze the morphology and morphometry of Crassostrea rhizophorae oocytes and follicles in three stages of the gonadal cycle. Were analized histological sections of gonads considering that the oocytes presented visible nuclei and nucleoli, it was observed three gonadal cycle stages: early gametogenesis, growth and maturation. In the early gametogenesis stage, follicles and oocytes presented diameters of 180.29 (± 41.91) and 18.66 (± 6.85) µm, respectively, and presence of large amount of connective tissue and previtellogenic oocyte. In the growth stage, follicles and oocytes presented diameters of 218.02 (± 43.19) and 25.92 (± 9.94) µm, respectively, this stage was characterized by a small amount of interfollicular connective tissue with vitellogenic oocyte predominate. In the maturation stage, follicles and oocytes presented diameters of 298.16 (± 99.24) and 35.27 (± 6.2) µm, respectively, and presence of large number of mature oocytes. We concluded that during culture Crassostrea rhizophorae tolerates the influence of intrinsic and extrinsic factors and does not undergo significant changes in reproductive activity.

Gametogenesis and reproduction of the matrinxã Brycon orthotaenia (Günther, 1864) (Pisces: Characidae) in the São Francisco river, Minas Gerais, Brazil

Estudaram-se a gametogênese e a reprodução de Brycon orthotaenia do rio São Francisco, na região de Pirapora, Minas Gerais, no período de dezembro/1998 a novembro/1999. O matrinxã B. orthotaenia é uma espécie endêmica da bacia do rio São Francisco, Brasil e importante na pesca comercial. O tamanho de 1ª maturação gonadal, indicado pelo comprimento total do menor macho espermiado e da menor fêmea desovada, foi de 32,0 e 40,5 cm, respectivamente. A reprodução ocorreu de outubro a janeiro, quando observam-se altas freqüências de fêmeas e machos no estádio maturação avançada/maduro e fêmeas desovadas. O curto período de desova e as características histológicas de ovários desovados, contendo somente ovócitos nos estádios iniciais de desenvolvimento ao lado de folículos pós-ovulatórios e atrésicos, indicam que a desova de B. orthotaenia é do tipo total.

The annual reproductive cycle of the snail Megalobulimus abbreviatus (Bequaert, 1948) (Gastropoda, Pulmonata)

Variações morfológicas nos órgãos do sistema reprodutor dos gastrópodes pulmonados são observadas ao longo do ano e podem ser correlacionadas a fases de seu ciclo reprodutivo. A partir dessa observação, a massa dos órgãos do sistema reprodutor do caracol Megalobulimus abbreviatus foi tomada em cada estação do ano e foram obtidas secções histológicas das gônadas. Os valores de massa foram utilizados para a obtenção do índice organo-somático e as secções, para calcular o diâmetro médio dos ovócitos e o índice de maturação ovocitária. Concluímos que M. abbreviatus é um caracol "iteroparous", apresentando ciclo reprodutivo anual caracterizado por acasalamento e oviposição durante a primavera e o início do verão e pela preparação do sistema reprodutor para um nova fase reprodutiva, que se inicia em janeiro e se estende até o final do inverno.

Estudo citogenético das gônadas em pacientes com amenorréia primária / Gonadal cytogenetic analysis in patients with primary amenorrhea

INTRODUÇAO: pacientes em amenorréia primária com disgenesia gonadal têm níveis elevados de gonadotrofinas e necessitam de avaliação cromossômica. O estudo citogenético (cariótipo) pode ser realizado na gônada ou no sangue periférico. Nos casos de amenorréia primária sem sinal de virilização, a necessidade de investigação adicional do cariótipo da gônada não está estabelecida. OBJETIVO: revisar os cariótipos de gônadas (ovários) de mulheres com amenorréia primária e compará-los com os resultados do cariótipo no sangue periférico, relacionando-os às características fenotípicas das pacientes. MÉTODOS: foram analisados retrospectivamente os dados clínicos e os cariótipos de doze pacientes atendidas no período de janeiro de 1997 a dezembro de 2003 no Hospital de Clínicas de Porto Alegre. RESULTADOS: nas pacientes incluídas, quando o motivo da investigação foi amenorréia primária sem sinal de virilização, o cariótipo da gônada foi concordante com o cariótipo do sangue periférico nos oito casos avaliados (sete pacientes com cariótipo 46XX e uma paciente com cariótipo 46XY). A paciente oito foi a única com sinal de virilização (hipertrofia de clitóris) e o único caso de cariótipos discordantes. CONCLUSAO: este estudo sugere que o cariótipo de gônada não traz informação adicional ao cariótipo do sangue periférico nas pacientes com amenorréia primária sem sinais de virilização. Até o momento todos os trabalhos publicados tiveram número pequeno de pacientes. A análise da relação custo-benefício pode permitir redução de estresse psicológico para paciente e familiares, bem como redução de custos para as instituições.

Differentiation and morphogenesis of Triatoma infestans (Klug 1834) female gonads. I- Post embryonic development

The post-embryonic development of the female gonads in Triatoma infestans (Hemiptera, Heteroptera), insects of importance in health affairs as harbors and vectors of different tripanosomatidea flagellates, is presented in a complete follow-up since insect hatches from the egg up to the last molt in the fifth instar stage. The detailed description of the morphological changes which occur in each instar as well as careful measurements evaluating its size increase have been analyzed by stereomicroscopy, phase contrast, dark field, and oblique ilumination, in order to optimize the observations as well the photographic register of gonad morphology and structure. The analysis was performed on gonad specimens obtained from broods no less than twenty (20) nymph bugs, reared at constant temperature and fed-up regularly. According to the results of our study we can assert that gonad differentiation takes place in early phases of the insect development. Such is tge case that first instars nymph´s present absolutely and easily recognizably male and female gonads. From the third instar on beside the filament region, the three zones in each ovariole body is distinguished, a differentiation that is more noticeable during the fourth instar where a definite organization is present at the vitelarium. Such a clear cut zone development continues intensively during the fifth instar. Finally at the end of such fifth nymph stage and when the last molt toward adults is prepared, clear signs of ovariole maturation take place since oocytes in early vitelogenesis are found.

Karyotype description of Pomacea patula catemacensis (Caenogastropoda, Ampullariidae), with an assessment of the taxonomic status of Pomacea patula

Mitotic chromosomes of the freshwater snail Pomacea patula catemacensis (Baker 1922) were analyzed on gill tissue of specimens from the type locality (Lake Catemaco, Mexico). The diploid number of chromosomes is 2n = 26, including nine metacentric and four submetacentric pairs, therefore, the fundamental number is FN = 52. No sex chromosomes could be identified. The same chromosome number and morphology were already reported for P. flagellata, i.e., the other species of the genus living in Mexico. The basic haploid number for family Ampullariidae was reported to be n = 14 in the literaure; so, its reduction to n = 13 is probably an apomorphy of the Mexican Pomacea snails. Lanistes bolteni, from Egypt, also shows n = 13, but its karyotype is much more asymmetrical, and seems to have evolved independently from P. flagellata and P. patula catemacencis. The nominotypical subspecies, P. patula patula (Reeve 1856), is a poorly known taxon, whose original locality is unknown. A taxonomical account is presented here, and a Mexican origin postulated as the most parsimonious hypothesis.

Post-Transcription Regulation of mRNA and Hormone Synthesis and Release at the Anterior Pituitary Gland: Information derived from the recovery of pituitary gonadotrope cell function after with a GnRH agonist

This review presents a summary of post-transcription regulation of mRNAs with a focus on the anterior pituitary gland. The control of gene transcription and production of mRNAs is the predominant form of regulation of hormone synthesis. However, post-transcription regulation of mRNAs provides another level of control of hormone synthesis. Examples of how hormone synthesis can be controlled at the level of mRNA include mRNA nuclear export and subcellular localization, mRNA stability and turnover, and regulation of mRNA translation. The gonadotrope cells of the anterior pituitary have multiple internal effector systems and provide an ideal model cell to study post-transcription regulation of mRNAs. Gonadotrope cells are stimulated to release LH and FSH by hypothalamic GnRH that binds to GnRH receptors. GnRH receptors are coupled to G-proteins and second messenger signaling pathways that involve cAMP and IP3. These signaling pathways are associated with the release of LH and FSH and transcription of mRNAs for LH and FSH. The stability of these mRNAs can be influenced by androgens, estrogens and progestagens. Therapy with a GnRH agonist leads to desensitization of gonadotrope cells to GnRH and a depletion of cellular stores of LH and FSH mRNAs, and content of LH and FSH. After discontinuation of therapy with GnRH agonist, levels of LH and FSH mRNAs return to normal some time before LH and FSH content and secretion are restored. This is indicative of post-transcription regulation of LH and FSH mRNAs. Future studies on post-transcription regulation of mRHAs will provide new molecular insights into how gonadotrope cells balance and integrate stimulation by GnRH with feedback modulation by the gonads.

Disgenesia gonadal 46,XX con ausencia de cromatina X en citología gonadal / Gonadal dysgenesis 46,XX with abscense of chromatin X in gonadal cytology

Se presenta el caso de 21 años de edad con fenotipo femenino, quien consultó por amenorrea secundaria y presentaba ligero desarrollo mamario y de vello púbico. El estudio hormonal reveló hipoestrenismo e hipegonadotropismo. En la laparoscopía se encontraron cintillas gonadales. Aunque el cariotipo practicado en linfocitos de sangre periférica resultó normal 46,XX, se encontró disparidad en los estudios citogenéticos de la cromatina X, la cual fue positiva en la mucosa oral negativa en el tejido gonadal. Se enfatiza el hecho de que algunas pacientes con disgenesia gonadal y cariotipo 46,XX pueden corresponder a mosaicismos cromasómicos, los cuales no se detectan generalmente porque no se realizan estudios de cariotipo en otros tejidos, sobre todo en el esbozo de tejido gonadal.