RBM46 is essential for gametogenesis and functions in post-transcriptional roles affecting meiotic cohesin subunits

RBM46 is a germ cell-specific RNA-binding protein required for gametogenesis, but the targets and molecular functions of RBM46 remain unknown. Here, we demonstrate that RBM46 binds at specific motifs in the 3'UTRs of mRNAs encoding multiple meiotic cohesin subunits and show that RBM46 is required for normal synaptonemal complex formation during meiosis initiation. Using a recently reported, high-resolution technique known as LACE-seq and working with low-input cells, we profiled the targets of RBM46 at single-nucleotide resolution in leptotene and zygotene stage gametes. We found that RBM46 preferentially binds target mRNAs containing GCCUAU/GUUCGA motifs in their 3'UTRs regions. In Rbm46 knockout mice, the RBM46-target cohesin subunits displayed unaltered mRNA levels but had reduced translation, resulting in the failed assembly of axial elements, synapsis disruption, and meiotic arrest. Our study thus provides mechanistic insights into the molecular functions of RBM46 in gametogenesis and illustrates the power of LACE-seq for investigations of RNA-binding protein functions when working with low-abundance input materials.

Aspectos da biologia reprodutiva de Boa constrictor constrictor: um estudo histológico dos testículos nos períodos reprodutivos de quiescência e máxima atividade / Boa constrictor constrictor reproductive biology aspects: the testes histological study in the reproductive periods of quiescence and maximal activity

As serpentes pertencem ao segundo maior grupo dentro dos répteis, podendo apresentar sazonalidade quanto à espermatogênese, com produção descontínua ou contínua. O presente trabalho tem como objetivo caracterizar aspectos da biologia reprodutiva de Boa constrictor constrictor com base nos achados histológicos dos testículos nos períodos de máxima atividade (período de gametogênese) e quiescência. Os testículos de dois espécimes de Boa c. constrictor (7767 e 11752) foram cortados a uma espessura de 3µm em micrótomo, corados com azul de toluidina 1%, fotodocumentados e descritos. A presença de espermatozoides na luz do túbulo seminífero no indivíduo 7767 indica um período de máxima gametogênese, enquanto o lúmen dos túbulos seminíferos pouco evidentes, sem a presença de espermatozoides e de células gaméticas em divisão, caracteriza o indivíduo 11752 em período quiescente. Mediante os achados histológicos descritos no presente estudo, concluiu-se que Boa c. constrictor apresenta sazonalidade em relação à gametogênese, sendo esse padrão de sazonalidade associado ao período de cópulas relatado em literatura característico de serpentes com padrão pré-nupcial.(AU)

These snakes belong to the second largest group within the reptiles, being able to present seasonality regarding spermatogenesis, with discontinuous or continuous production. The present study aims to characterize Boa constrictor constrictor reproductive biology aspects from histological findings in testicles during periods of maximum activity (period of gametogenesis) and quiescence. The testicles of two specimens of Boa c. constrictor (7767 and 11752) were cut to a thickness of 3µm in microtome, stained with 1% toluidine blue, photodocumented and described. The spermatozoa presence in the seminiferous tubule lumen in individual 7767 indicates a period of maximum gametogenesis, whereas the seminiferous tubules lumen is not very evident without spermatozoa and the absence of dividing gametic cells characterizes individual 11752 in the quiescent period. Through the histological findings we concluded that Boa c. constrictor presents seasonality in relation to gametogenesis, and the pattern of reproductive seasonality observed along with the period of copulas reported in the literature resembles the pre-nuptial pattern.(AU)

Gonadal development of Peprilus medius (Peters, 1869) (Perciformes: Stromateidae) from Southeast of the Gulf of California, Mexico / Desarrollo gonádico de Peprilus medius (Peters, 1869) (Perciformes: Stromateidae) del Sureste del Golfo de California, México

The gonadal development assessment is a fundamental work in researching the reproduction of Peprilus medius, on the way to understand its population dynamics and to achieve a sustainable management of its fisheries. The diagnosis based on morphochromatic features is the quicker, more practical and cheaper method, and properly validated, it may be an accurate procedure. This is particularly useful in researches when it is required to provide prompt and accurate information to support decision-making for fisheries management, and during long-term fisheries monitoring to detect changes in reproductive patterns due to environmental anomalies. These are issues of concern in P. medius and other fishery resources. In this context, a scale that describes the gonadal development through morphochromatic and histological features is presented, and plus anatomical observations of the reproductive organs. Ovarian development was characterized through six phases: immature, development (sub-phases: early and late), mature, spawning (sub-phases: partial and partial post-spawning), post-spawning and resting. Testicular development was described through five phases: immature, development, ejaculation, ejaculated and resting. The position and shape of the reproductive organs are apparently defined by the shape of the fish and the visceral cavity. P. medius's gonads show a gonadal lobe at lower position and an upper lobe in the visceral cavity, and both lobes presented a horizontal placement from gonopore toward the back of the fish. In addition, gonadal deformities occurrence due to deformities in the visceral cavity was evidenced. We recommend to use presented descriptions as a guide in gonadal assessment.

La evaluación del desarrollo gonádico es una labor fundamental para investigar la reproducción de Peprilus medius, en vías a comprender la dinámica de sus poblaciones y lograr un manejo sustentable de sus pesquerías. El diagnóstico a partir de características morfocromáticas es el método más rápido, práctico y barato, y adecuadamente validado puede ser un procedimiento preciso. Este es particularmente útil en investigaciones cuando se requiere aportar información rápida y precisa que apoye la toma de decisiones de manejo pesquero, y durante los monitoreos pesqueros de largo plazo, que permiten detectar cambios en los patrones reproductivos debido a anomalías ambientales. Estos son temas preocupantes en P. medius y otros recursos pesqueros. En ese contexto se presenta una escala que describe el desarrollo gonádico a través de características morfocromáticas e histológicas, y además se presentan observaciones anatómicas de los órganos reproductores. El desarrollo ovárico fue caracterizado a través de seis fases: inmadura, desarrollo (subfases: inicial y avanzado), maduro, desove (subfases: parcial y postdesove parcial), postdesove y reposo. El desarrollo testicular fue descrito a través de cinco fases: inmadura, desarrollo, eyaculación y reposo. La posición y forma de los órganos reproductores están aparentemente definidos por la forma del pez y su cavidad visceral. Las gónadas de P. medius muestran un lóbulo gonádico en posición inferior y uno superior en la cavidad visceral, y ambos lóbulos presentan una colocación horizontal desde el gonoporo hacia la parte posterior del pez. Además se evidencia la ocurrencia de deformidades en las gónadas debido a deformidades en la cavidad visceral. Recomendamos utilizar las descripciones presentadas como una guía en la evaluación de las gónadas.

Reproductive periods of Lucina pectinata (BIVALVE; LUCINIDAE) in the Paranaguá Estuarine Complex, Paraná - Brazil / Período reprodutivo de Lucina pectinata (BIVLAVE; LUCINIDAE) no Complexo Estuarino de Paranaguá, PR - Brasil

Abstract The objectives of the present study is to describe the size for exploitation and gonadal maturation of L. pectinata in Peças Island in Paraná coast. Twenty two individuals were sampled every month, from an intertidal flat of the Peças River, Municipality of Guaraqueçaba, during two periods – summer/autumn and winter of the 2009. Environmental parameter data was obtained at the same time of the biological sampling: temperature, salinity and seawater transparency at the Peças River mouth. The animals’ height, length, width, gross weight, wet weight of meat and dry weight of meat were measured at the lab. The gonad development stages (GDS), condition index (CI) and yield index (Y) were calculated. Sex identification was conducted by direct observation of the gonads and gametes under an optic microscope. Individuals were classified as males (M), females (F) or undetermined (U). The medium height observed was of 49.62 ± 6.84 mm. Period of more expressive gonad repletion were observed in summer when water temperature was higher. Results of GDS in both periods showed a predominance of partially filled gonad stages. This may be related with the fact that L. pectinata has intense gametogenesis activity, that is, the organisms do not have a resting interval, but go back to gonad restructuring rapidly after releasing the gametes. The continuity of the reproductive cycle of the specimens found in both studied periods could demonstrate a possible continuous reproduction of the species and a size for exploitation above 40.00 mm of height.

Resumo O presente estudo tem como objetivo descrever o tamanho de exploração e a maturação gonadal de L. pectinata na Ilha das Peças no litoral do Paraná. Vinte e dois indivíduos foram amostrados mensalmente na região entre-marés no Rio das Peças, Município de Guaraqueçaba, durante 2 períodos – verão/outono e inverno de 2009. Paralelamente às amostragens, foram obtidos dados dos parâmetros ambientais: temperatura, salinidade e transparência da água do mar na foz do Rio das Peças. A altura, comprimento, largura, peso bruto, peso úmido da carne e peso seco da carne dos animais foram medidos no laboratório. Foram calculados os estágios de desenvolvimento gonadal (EDG), índices de condição (IC) e de rendimento (R). A Identificação do sexo foi realizada por observação direta das gônadas e gametas sob microscópio óptico. Os indivíduos foram classificados como: machos (M), fêmeas (F) ou indeterminados (I). A altura média observada foi de 49,62 ± 6,84 mm. O período de maior repleção gonadal foi observado no verão, quando a temperatura da água foi mais elevada. Resultados dos EDG em ambos os períodos mostrou um predomínio de gônadas parcialmente cheias. Isso pode estar relacionado com o fato de que L. pectinata tem atividade gametogênese intensa, ou seja, os organismos não têm um intervalo de repouso, mas voltam a reestruturação das gônadas rapidamente após liberação dos gametas. A continuidade do ciclo reprodutivo dos espécimes encontrados em ambos os períodos estudados, pode demonstrar possível reprodução contínua da espécie e tamanho de exploração superior a 40,00 mm de altura.

Nutrition et fausses couches spontanées

Les fausses couches spontanées (FCS) constituent environ 15 % des grossesses et les fausses couches à répétition (FCSR) touchent environ 1 % des femmes.De nombreuses études soulignent l'impact de la nutrition sur la gamétogenèse, le développement embryonnaire et l'évolution des grossesses. Ainsi, le bilan étiologique des FCS, et en particulier des FCSR, doit tenir compte de la nutrition qui semble représenter une voie prometteuse dans l'amélioration de la prise en charge des pathologies liées à la reproduction entre autres les fausses couches et qui reste encore mal explorée.Evaluer le rôle de la nutrition comme facteur favorisant la survenue de FCS est d'autant plus important qu'elle représente un facteur modulable sur lequel on peut agir pour améliorer le pronostic de la grossesse

Histological validation of morphochromatically-defined gonadal maturation stages of green abalone (Haliotis fulgens) Philippi,1845 and pink abalone (Haliotis corrugata) Wood, 1828 / Validación histológica de las fases morfocromáticas de maduración gonádica del abulón azul (Haliotis fulgens) Philippi, 1845 y del abulón amarillo (Haliotis corrugata) Wood, 1828

Identifying the maturity stage of specimens is essential for the determination of the spawning season, which in turn facilitates the formulation of adequate fisheries management measures for the sustainable use of sea resources, such as abalone. This study aims to validate, based on histological evidence, a gonad maturation scale that encompasses four morphochromatically-defined stages (developing, maturity, spawning and resting) for females and males, plus an additional stage for reproductively inactive (undifferentiated) specimens, to be used as an identification key. Given the similarity between Haliotis fulgens and Haliotis corrugata as to their morphochromatic features, the gonadal maturation scale can be used for both species.

La determinación de la fase de madurez es importante para identificar la temporada de desove y permitir establecer medidas de manejo pesquero para el aprovechamiento responsable de los recursos como el abulón. El presente trabajo valida histológicamente una escala de maduración gonádica a manera de guía de identificación, que consiste de cuatro fases morfocromáticas para hembras y machos (desarrollo, madurez, desove y reposo), y un estadio para los organismos reproductivamente inactivos (indiferenciado). Dada la similitud entre las características morfocromáticas de Haliotis fulgens y Haliotis corrugata, la escala de maduración gonádica puede ser utilizada indistintamente.

Gametogenesis and weight change of body organs of the sea cucumber Holothuria (Halodeima) grisea (Aspidochirotida: Holothuriidae) in Southern Brazil / Aspectos reproductivos y la relación entre los órganos y la gametogénesis en Holothuria (Halodeima) grisea (Echinodermata: Holothuroidea)

Abstract The sea cucumber Holothuria (Halodeima) grisea is a widely distributed and economically important species that has been harvested in Brazil for several years, where there are no studies on its reproduction. We studied a population near Guaratuba Bay in Southern Brazil. Four sampling campaigns were made, in February, April, June and October 2008. We examined the macroscopic and microscopic features of the gonads to describe the gametogenesis. In addition, we analyzed the weight change of body organs. A total of 94 individuals were examined and no evidence of external sexual dimorphism was found. In April there were more males than females. Biometrically the body wall and gonads are the most important organs. The body wall increases in the June and the gonads during October and February; there might be an exchange of energy between these organs. Mature gonads are elongated and thick (red in females, cream in males). There were mature individuals year round, with an increase of sexual activity in February when oocyte mature diameter ranges from 70 to 160 µm. Spawning was in winter (June). Maturation period and body component variations were similar between sexes. Rev. Biol. Trop. 63 (Suppl. 2): 285-296. Epub 2015 June 01.

Resumen El pepino de mar Holothuria (Halodeima) grisea es una especie ampliamente distribuida y económicamente importante que se ha recolectado en Brasil desde hace varios años. No hay estudios previos realizados sobre la reproducción de esta especie en Brasil. Además, son aún poco conocidos los aspectos bio-ecológicos de esta especie. En este estudio, se investigaron algunos aspectos de la reproducción del pepino de mar Holothuria (Halodeima) grisea en una población cerca de la bahía de Guaratuba en el sur de Brasil. Se hicieron cuatro salidas de muestreo, en febrero, abril, junio y octubre de 2008, abarcando las cuatro estaciones. Examinamos las características macroscópicas y microscópicas de las gónadas para describir la gametogénesis, adicionalmente, se analizó el cambio de peso de cada componente de los órganos del cuerpo (pared corporal, gónadas, sistema haemal, tracto digestivo y árbol respiratorio). Se examinó un total de 94 individuos y no se encontraron evidencias de dimorfismo sexual externo. Los resultados del análisis estadístico de la prueba x² rechazan la hipótesis nula (H0) con una relación de sexos igual a 1, sin embargo, el análisis de cada mes, esta diferencia de género se observó sólo en el mes de abril, cuando el número de los males es mucho mayor que la de las hembras. Las medidas biométricas mostraron que la pared del cuerpo y las gónadas son los órganos más importantes, con aumentos de la pared corporal en el junio y de las gónadas durante octubre y febrero, que pueden indicar que hay un intercambio de energía entre estos órganos. Macroscópicamente, las gónadas maduras muestran túbulos largos, gruesos y ramificados siendo que las hembras presentan coloración rojiza e los machos tonos de crema. Sin embargo, durante el crecimiento y agotado, no se observaron diferencias morfológicas macroscópicas entre los sexos. Microscópicamente, se observó la presencia constante de individuos maduros entre meses, con un aumento de la actividad sexual en febrero, cuando los ovocitos maduros tuvieron un diámetro oscila entre 70 a 160 μ. El período de desove ocurrió en los meses de invierno (junio). El periodo de maduración y la variación del cuerpo de los componentes fueron similares entre los sexos. La similitud en las demandas metabólicas de la población podría estar confirmando una estrategia para maximizar la sincronía reproductiva.

Impact of glycosylation on the unimpaired functions of the sperm / 대한생식의학회지

One of the key factors of early development is the specification of competence between the oocyte and the sperm, which occurs during gametogenesis. However, the starting point, growth, and maturation for acquiring competence during spermatogenesis and oogenesis in mammals are very different. Spermatogenesis includes spermiogenesis, but such a metamorphosis is not observed during oogenesis. Glycosylation, a ubiquitous modification, is a preliminary requisite for distribution of the structural and functional components of spermatids for metamorphosis. In addition, glycosylation using epididymal or female genital secretory glycans is an important process for the sperm maturation, the acquisition of the potential for fertilization, and the acceleration of early embryo development. However, nonemzymatic unexpected covalent bonding of a carbohydrate and malglycosylation can result in falling fertility rates as shown in the diabetic male. So far, glycosylation during spermatogenesis and the dynamics of the plasma membrane in the process of capacitation and fertilization have been evaluated, and a powerful role of glycosylation in spermatogenesis and early development is also suggested by structural bioinformatics, functional genomics, and functional proteomics. Further understanding of glycosylation is needed to provide a better understanding of fertilization and embryo development and for the development of new diagnostic and therapeutic tools for infertility.

Artificial gametes from stem cells / 대한생식의학회지

The generation of artificial gametes is a real challenge for the scientific community today. In vitro development of human eggs and sperm will pave the way for the understanding of the complex process of human gametogenesis and will provide with human gametes for the study of infertility and the onset of some inherited disorders. However, the great promise of artificial gametes resides in their future application on reproductive treatments for all these people wishing to have genetically related children and for which gamete donation is now their unique option of parenthood. This is the case of infertile patients devoid of suitable gametes, same sex couples, singles and those fertile couples in a high risk of transmitting serious diseases to their progeny. In the search of the best method to obtain artificial gametes, many researchers have successfully obtained human germ cell-like cells from stem cells at different stages of differentiation. In the near future, this field will evolve to new methods providing not only viable but also functional and safe artificial germ cells. These artificial sperm and eggs should be able to recapitulate all the genetic and epigenetic processes needed for the correct gametogenesis, fertilization and embryogenesis leading to the birth of a healthy and fertile newborn.

Rhox in mammalian reproduction and development / 대한생식의학회지

Homeobox genes play essential roles in embryonic development and reproduction. Recently, a large cluster of homeobox genes, reproductive homeobox genes on the X chromosome (Rhox) genes, was discovered as three gene clusters, alpha, beta, and gamma in mice. It was found that Rhox genes were selectively expressed in reproduction-associated tissues, such as those of the testes, epididymis, ovaries, and placenta. Hence, it was proposed that Rhox genes are important for regulating various reproductive features, especially gametogenesis in male as well as in female mammals. It was first determined that 12 Rhox genes are clustered into alpha (Rhox1-4), beta (Rhox5-9), and gamma (Rhox10-12) subclusters, and recently Rhox13 has also been found. At present, 33 Rhox genes have been identified in the mouse genome, 11 in the rat, and three in the human. Rhox genes are also responsible for embryonic development, with considerable amounts of Rhox expression in trophoblasts, placenta tissue, embryonic stem cells, and primordial germ cells. In this article we summarized the current understanding of Rhox family genes involved in reproduction and embryonic development and elucidated a previously unreported cell-specific expression in ovarian cells.

Biología Reproductiva de Mytilopsis leucophaeata (Conrad, 1831) (Bivalvia: Dreissenidae) de la Laguna de Tampamachoco, Túxpam-Veracruz / Reproductive Biology of Mytilopsis leucophaeata (Conrad, 1831) (Bivalvia: Dreissenidae) of Tampamachoco Lagoon, Tuxpan-Veracruz

Se caracterizó el ciclo reproductivo de Mytilopsis leucophaeata (Conrad, 1831) en organismos colectados mensualmente durante un ciclo anual en la laguna de Tampamachoco, Túxpam, Veracruz, que se ubica en la costa del Golfo de México en la Región Terrestre Prioritaria (RTP-103) de CONABIO considerada sitio RAMSAR 1602 en México. Los resultados histológicos indican que M. leucophaeata presenta cuatro etapas gametogénicas: gametogénesis, madurez, desove total para hembras y expulsión parcial para machos. Uno de los principales factores que influyen en el desove sincrónico de gametos tanto en machos como en hembras es la salinidad, en los meses de enero a marzo.

The reproductive cycle of Mytilopsis leucophaeata (Conrad, 1831) was characterized in organisms collected monthly during an annual cycle in Tampamachoco lagoon Tuxpam-Veracruz, located on the coast of the Gulf of Mexico in the Terrestre Prioritaria Region (RTP-103) of CONABIO considered a RAMSAR 1602 site in Mexico. Histological results indicated that M. leucophaeata has four gametogenic stages: gamete, maturity, female's total spawning and partial expulsion inmales. One of the main factors that influence the synchronous spawning gametes in both males and females is salinity, in the months of January to March.

Reproductive cycle and embryonic development of the gastropod Melampus coffeus (Linnaeus, 1758) (Ellobiidae) in the Brazilian Northeast / Ciclo reprodutivo e desenvolvimento embrionário do gastrópode Melampus coffeus (Linnaeus, 1758) (Ellobiidae) no Nordeste do Brasil

Melampus coffeus belongs to a primitive group of pulmonate mollusks found mainly in the upper levels of the marine intertidal zone. They are common in the neotropical mangroves. Little is known about the biology of this species, particularly about its reproduction. The aim of this study was to 1) characterize the morphology and histology of M. coffeus' gonad; 2) describe the main gametogenesis events and link them to a range of maturity stages; 3) chronologically evaluate the frequency of the different maturity stages and their relation to environmental factors such as water, air and sediment temperatures, relative humidity, salinity and rainfall; and 4) characterize M. coffeus' spawning, eggs and newly hatched veliger larvae. Samples were collected monthly between February, 2007 and January, 2009 from the mangroves of Praia de Arpoeiras, Acaraú County, State of Ceará, northeastern Brazil. The characterization of the gonad development stages was carried out using routine histological techniques. The results of this study show that Melampus coffeus is a simultaneous hermaphrodite. The follicles have masculine and feminine elements, interleaved within the gonad. M. coffeus presents a well-defined synchronous reproductive cycle, showing successive maturation, release and resting periods. The average diameter of the oocytes was negatively correlated with salinity and positively correlated with rainfall. The results show that no reproductive activity occurs during periods of drought. After the dry season, the increasing rainfall levels and reduced salinity lead to the appearance of very dense populations, predominantly composed of small individuals.

Melampus coffeus encontra-se em um grupo primitivo de moluscos pulmonados, primordialmente marinhos, habitantes dos níveis superiores da zona entremarés, comum em manguezais do Oceano Atlântico. Pouco se sabe sobre a biologia da espécie, em especial, sobre a sua reprodução. Os objetivos deste trabalho foram: 1) caracterizar morfo-histologicamente a gônada de M. coffeus; 2) descrever os principais eventos da gametogênese e associá-los a uma escala de estágios de maturidade; 3) avaliar temporalmente a frequência dos estágios de maturidade e sua possível relação com os fatores ambientais, como a temperatura da água, do ar e do sedimento, a umidade relativa, a salinidade e a pluviosidade, e 4) caracterizar a postura, os ovos e a véliger recém-eclodida de M. coffeus. Foram realizadas coletas mensais de fevereiro de 2007 a janeiro de 2009, no manguezal da Praia de Arpoeiras, município de Acaraú-Ceará, na Região Nordeste do Brasil. A caracterização das fases de desenvolvimento da gônada foi realizada utilizando-se técnicas histológicas de rotina. Os resultados deste estudo mostram que Melampus coffeus é um hermafrodita simultâneo. Os folículos apresentam elementos masculinos e femininos, distribuindo-se na gônada intercalados radialmente. M. coffeus apresenta um ciclo reprodutivo sincrônico bem definido, evidenciando-se períodos sucessivos de amadurecimento, de liberação e de repouso. O diâmetro médio dos oócitos apresentou correlação negativa com a salinidade e positiva com a pluviosidade. Os resultados indicam que, nos períodos de estiagem, não há atividade reprodutiva. Após essa estação, quando os valores de pluviosidade aumentam e a salinidade diminui, as populações são bastante densas e predominam indivíduos pequenos.

Gestación uterina y sus limitaciones / Uterine gestation and their limitations

Se describe sumariamente los pasos que secuencialmente tienen lugar para lograr una gestación uterina. Cuando la pareja por razones diversas es incapaz de alcanzar el desarrollo embrionario requerido para un embarazo uterino, se utilizan protocolos que permiten determinar las causas de la infertilidad. Se describen las principales modalidades de la fecundación asistida: la inseminación artificial, la fecundación in vitro y los diversos pasos que permiten lograrla. Se presentan las modalidades de microinyección espermática incluidas las de aplicación más reciente (inyección de espermatozoides morfológicamente seleccionadas y examen de la morfología de las organelas que participan en la motilidad espermática). Se describe la transferencia embrionaria, especialmente los logros en el diagnóstico genético preimplantacional y en cuales enfermedades es factible. La fertilización en casos de cáncer requiere un enfoque multidisciplinario. Finalmente se enumeran las otras causas de infertilidad y su tratamiento. A modo de conclusión, se presenta la significación de los avances logrados desde el Reimer "bebe probeta" en 1984 y algunos datos de interes indicativos de la actividad que se desarrolla en el Servicio de Medicina de la Reproducción en el Instituto Universitario Dexeus de Barcelona, España

Sequential steps to seek uterine gestation are brielly and sequentially described. When a couple, due to diverse causes, is incapable to concieve and develop an embryo and hence a normal intrauterine gestation, specifit protocols are used aimed to determine the causes of infertility. The main modalities of assisted fertilization are presented: artificial insemination, in vitro fertilization including the steps needed to achieve embryogenesis. Spermatic modalities of microinjection, including the most up to date modalities (intracytoplasmatic morphologicaly selected sperm inyection and motile sperm organelle morphology examination) are presented. Embryonic transference, and in special, data related to preimplantation genetic diagnosis are analyzed, including a listing of those disease in which useful results are now available. Finally, other causes of infertility, including its treatment, are briefty mentioned. Fertilization in women with cancer needs a multidisciplinary approach. In closing remarks, the significance of advances achieved since the first test-tube baby in 1984 is emphasized, and some of the significant advances in this field at Institute Dexeus (Barcelona-Spain) are summarized

Roles of gangliosides in mouse embryogenesis and embryonic stem cell differentiation

Gangliosides have been suggested to play important roles in various functions such as adhesion, cell differentiation, growth control, and signaling. Mouse follicular development, ovulation, and luteinization during the estrous cycle are regulated by several hormones and cell-cell interactions. In addition, spermatogenesis in seminiferous tubules of adult testes is also regulated by several hormones, including follicle-stimulating hormone (FSH) and luteinizing hormone (LH) and cell-cell interactions. The regulation of these processes by hormones and cell-cell interactions provides evidence for the importance of surface membrane components, including gangliosides. During preimplantation embryo development, a mammalian embryo undergoes a series of cleavage divisions whereby a zygote is converted into a blastocyst that is sufficiently competent to be implanted in the maternal uterus and continue its development. Mouse embryonic stem (mES) cells are pluripotent cells derived from mouse embryo, specifically, from the inner cell mass of blastocysts. Differentiated neuronal cells are derived from mES cells through the formation of embryonic bodies (EBs). EBs recapitulate many aspects of lineage-specific differentiation and temporal and spatial gene expression patterns during early embryogenesis. Previous studies on ganglioside expression during mouse embryonic development (including during in vitro fertilization, ovulation, spermatogenesis, and embryogenesis) reported that gangliosides were expressed in both undifferentiated and differentiated (or differentiating) mES cells. In this review, we summarize some of the advances in our understanding of the functional roles of gangliosides during the stages of mouse embryonic development, including ovulation, spermatogenesis, and embryogenesis, focusing on undifferentiated and differentiated mES cells (neuronal cells).

Reproductive biology of Echinometra lucunter (Echinodermata: Echinoidea) in a northeast Brazilian sandstone reef

The edible sea urchin Echinometra lucunter (Linnaeus, 1758) is a very common species on the sublittoral-midlittoral in Brazilian rocky shores. The aim of this work was to describe the gametogenesis and reproductive strategy of the E. lucunter population at Muro Alto beach in the Northeast coast of Brazil from August 2004 to August 2005. A total of 240 specimens were collected on the sandstone reef flat from a tidepool during spring low tides. The overall sex ratio was1.12:1,withoutsignificanttemporalvariationexceptinOctober2004. Firsts sexual maturity ocurred in individuals from a diameter of 20.8 mm. There was not a significant difference in gonad index between females and males during the sampling period. The female's gonad index variation was associated with a well-defined spawning, corroborated by the histological analysis of the gonads, which demonstrates sex differences of the gamete production. By contrast, the males showed no clear pattern. It is suggested that continuous reproduction with seasonal peaks in the E. lucunter population occurs at Muro Alto beach.

O ouriço-do-mar comestível Echinometra lucunter (Linnaeus, 1758) é uma espécie muito comum no infralitoral e mediolitoral do Brasil. O objetivo deste trabalho foi descrever a gametogênese e a estratégia reprodutiva da população de E. lucunterna praia de Muro Alto entre agosto de 2004 e agosto de 2005. Um total de 240 espécimes foi capturado de uma poça de maré situada no topo recifal, durante as marés baixas de sizígia. A razão sexual total foi de 1,12:1 sem variação temporal significativa, exceto em outubro de 2004. A primeira maturidade sexual ocorreu em indivíduos a partir de 20,8 mm de diâmetro. Não houve nenhuma diferença significativa no índice gonadal entre fêmeas e machos durante o período de amostragem. A variação do índice gonadal das fêmeas foi associada a um período de desova bem definido, corroborado pela análise histológica das gônadas, que demonstra diferenças sexuais na produção de gametas. Ao contrário, os machos não apresentaram nenhum padrão. Sugere-se que a reprodução da população de E. lucunter na praia de Muro Alto é contínua, compicos sazonais.

Producing recombinant mTEX101; a murine testis specific protein

Production of antibodies against specific proteins of testis germ cells is of great significance for the investigation of processes involved in spermatogenesis, study of infertility problems and determination of the probable role of these proteins as cancer-testis antigens. Murine Testis Specific Recombinant Protein 101 [mTEX101] is a 38kDa, GPI-anchored protein which is expressed in testis germ cells of adult mice but it seems to be absent in other tissues. The structure and function of mTEX101 is not completely understood yet, but it is speculated that it may transduce biochemical signals into the cytoplasm since mTEX101 does not have an intracellular domain but the precise mechanisms are still ambiguous. RNA was extracted from three adult mice testis. The RNA was used in RT-PCR, employing a pair of specific primers for mTEX101 ORF region. TA-cloning technique was performed by the insertion of mTEX101 into a pGEM-T Easy Vector, followed by its subcloning into a His-tagged expression vector, pET-28a [+]. The recombinant mTEX101 was then produced by transfection of the expression vector into BL 21 [DE3] E. coli strain. A recombinant protein, weighing 27kDa, was produced upon IPTG-induction of the bacterial host. The presence of mTEX101 protein was detected through Western blot analysis by anti-mTEX101 peptide antibodies. We produced mTEX101 recombinant protein that could be used for the production of mono and polyclonal antibodies

Gametogênese: estágio fundamental do desenvolvimento para reprodução humana / Gametogenesis: fundamental stage the development for human reproduction

A reprodução é o mecanismo essencial para perpetuação e diversidade das espécies, assim como para a continuidade da vida. Os gametas são os veículos de transferência dos genes para as próximas gerações. A gametogênese pode ser caracterizada por três etapas distintas denominadas multiplicação (mitose), crescimento e maturação (meiose), que se diferenciam em vários aspectos na espermatogênese e na oogênese. A oogênese diferencia-se da espermatogênese na ocorrência do processo ao longo da vida do indivíduo, no número de gametas formados, nas etapas de multiplicação, crescimento e maturação, assim como nas diferenciações e maturações citoplasmáticas e nucleares dos gametas masculinos e femininos. O processo da oogênese em relação à maturação oocitária ainda merece mais estudos, pois os aspectos bioquímicos e genéticos deste processo precisam ser mais bem esclarecidos. Por meio da gametogênese masculina e feminina a natureza proporcionou a fertilidade e a capacidade de gerar filhos a casais de diferentes espécies que se reproduzem sexuadamente incluindo a espécie humana. Quando existem falhas neste mecanismo por diferentes motivos como oligospermia, teratozoospermia, astenoospermia, azoospermia, ou distúrbios da ovulação, endometriose, fatores tubários, esterilidade sem causa aparente, casais podem recorrer àstecnologias de Reprodução Assistida como inseminação intra-uterina, fertilização in vitro, injeção intra-citoplasmática de espermatozóides, e de maturação oocitária in vitro para chegarem à plenitude da reprodução.

The reproduction is the essential mechanism for perpetuation and diversity of the species, as well as for the continuity of the life. The gametes are the vehicles of transfer of the genes for the next generations. The gametogenesis can be characterized by three stages denominated different multiplication (mitosis), growth and maturation (meiosis), which differ inseveral aspects in the spermatogenesis and in the oogenesis. The oogenesis differs of the spermatogenesis in several aspects as in the occurrence of the process along the individual’s lifesuch as in the number of formed gametes, in the multiplication stages, growth and maturation, aswell as in the differentiations and cytoplasmtic and nuclear maturations of the masculine and feminine gametes. The process of the oogenesis in relation to the oocyte maturation still deserve many studies, because the biochemical and genetic aspects of this process need to be better explained. Through the masculine and feminine gametogenesis the nature provided the fertility and the capacity of generating children to you marry of different species that sexually reproduce including the human species. When flaws exist in this mechanism for different reasons as oligospermy, teratozoospermy, astenoospermy, azoospermy or ovulation disturbs, endometriosis, tubal factors, or unexplained infertility couples can fall back upon the technologies of Assisted Reproduction as in vitro fertilization, intracytoplasmic sperm injections, artificial insemination, invitro maturation of oocytes to reach the fullness of the reproduction.

From embryonic stem cells to functioning germ cells: science, clinical and ethical perspectives.

Embryonic stem cells have been well recognized as cells having a versatile potential to differentiate into all types of cells in the body including germ cells. There are many research studies focusing on the differentiation processes and protocols to derive various types of somatic cells from embryonic stem cells. However, germ cells have unique differentiation process and developmental pathway compared with somatic cells. Consequently, they will require different differentiation protocols and special culture techniques. More understanding and established in vitro systems for gametogenesis will greatly contribute to further progression of knowledge and technology in germ cell biology, reproductive biology and reproductive medicine. Moreover if oocytes can be efficiently produced in vitro, this will play an important role on progression in nuclear transfer and nuclear reprogramming technology. The present article will provide concise review on past important discoveries, current ongoing studies and future views of this challenging research area. An ethical perspective has also been proposed to give comprehensive summary and viewpoint for future clinical application.

Microsporogenesis in sexual Brachiaria hybrids (Poaceae)

Three sexual interspecific hybrids of Brachiaria (HBGC076, HBGC009, and HBGC014) resulting from crosses between B. ruziziensis (female genitor) and B. decumbens and B. brizantha (male genitors) produced by Embrapa Beef Cattle in the 1980s were cytologically analyzed by conventional methods for meiotic studies. The cytogenetic analysis showed the occurrence of common meiotic abnormalities among them. The most frequent abnormalities were those related to irregular chromosome segregation due to polyploidy. Other abnormalities, such as chromosome stickiness, absence of cytokinesis, irregular cytokinesis, abnormal spindle orientation, and abnormal nucleolus disintegration, were found in the three hybrids, while, chromosome disintegration was detected only in HBGC014. All the abnormalities, except for abnormal nucleolus disintegration, can cause unbalanced gamete formation, leading to pollen sterility. Multivalent chromosome association at diakinesis revealed genome affinity between the two parental species in the hybrids, suggesting some possibility for gene introgression. Presently, the Brachiaria breeding program has the objective of releasing, primarily, apomictic hybrids as new cultivars since they do not segregate but preserve the genetic makeup indefinitely. Besides, they result in homogeneous pastures which are easier to manage. The sexual hybrids, however, are paramount in the breeding program: they work as ‘bridges’ to introgress traits of interest into the apomictic genotypes. The cytogenetic analyses of these three hybrids substantiate their maintenance in the breeding program due to low frequency of meiotic abnormalities, complemented by interesting agronomic traits. They may be used in crosses to generate new cultivars in the future.

Microsporogenesis in inbred line of popcorn (Zea mays L)

Endogamy places genes for several characteristics in homozygosis, which include those related to meiosis causing abnormalities that may impair gamete viability. An original population (S0) of popcorn (CMS-43) produced by Embrapa Maize and Sorghum was self-pollinated for seven years, generating inbred lines (S1 to S7). Conventional studies of microsporogenesis revealed that meiotic abnormalities did not increase with endogamy. Univalent chromosomes, irregular chromosome segregation, abnormal cell shape, partial asynapsis, cell fusion, absence of cytokinesis, abnormal spindle orientation, and chromosome stickiness were recorded in low frequency in meiocytes. Since the frequency of abnormalities was low, mainly in S7, inbred lines from CMS-43 have a high potential for hybridization.