Morphology and Biochemistry of Ovulation Morfologia e bioquímica da ovulação

Abstract The process of ovulation involves multiple and iterrelated genetic, biochemical, and morphological events: cessation of the proliferation of granulosa cells, resumption of oocyte meiosis, expansion of cumulus cell-oocyte complexes, digestion of the follicle wall, and extrusion of the metaphase-II oocyte. The present narrative review examines these interrelated steps in detail. The combined or isolated roles of the folliclestimulating hormone (FSH) and luteinizing hormone (LH) are highlighted. Genes indiced by the FSH genes are relevant in the cumulus expansion, and LH-induced genes are critical for the resumption ofmeiosis and digestion of the follicle wall. A nonhuman model for follicle-wall digestion and oocyte release was provided.

Resumo O processo de ovulação envolve modificações genéticas, bioquímicas e morfológicas múltiplas e interrelacionadas: suspensão da proliferação das células da granulosa, reinício da meiose do oócito, expansão das células do complexo cumulus-oócito, digestão da parede folicular, e extrusão do oócito. Esta revisão narrativa examina em detalhes cada um desses eventos e os principais genes e proteínas envolvidos. Mais importante, a ação combinada ou isolada do hormônio folículo-estimulante (HFE) e do hormônio luteinizante (HL) é destacada. Detalha-se o papel do HFE na expansão do cumulus e do HL na digestão da parede folicular, permitindo a extrusão do oócito na superfície ovariana. Proveu-se um modelo não humano para explicar a digestão da parede folicular.

Origens do(s) sexo(s) e a rainha dos problemas da biologia evolutiva / Origins of sex(es) and the queen of problems of evolutionary biology

A reprodução sexuada já foi considerada universal, e posteriormente, a forma mais perfeita de reprodução. Todavia, a partir de meados do século XIX, pesquisas no nível celular colocaram em xeque a ideia de que tipos de reprodução assexuadas fossem primitivos ou inferiores. Ao longo do século XX, e adentrando no XXI, hipóteses foram levantadas para explicar as vantagens da reprodução sexuada sobre a assexuada assim como o que permitiria a reprodução sexuada se manter quando seria mais vantajoso se reproduzir de forma assexuada. A mais importante e conhecida é a hipótese da Rainha Vermelha. Paralelamente, vários trabalhos procuraram entrever as pressões ecológicas que permitiram e favoreceram o aparecimento da reprodução sexuada em um cenário situado há cerca de dois bilhões de anos. O objetivo desse trabalho é revisar respostas históricas que marcaram o estudo da origem, da evolução e da manutenção da reprodução sexuada, identificando algumas das principais questões que a comunidade científica elaborou nos últimos duzentos anos.

Sexual reproduction has already been considered universal, and subsequently, the most perfect form of reproduction. However, since the mid-nineteenth century, research at the cellular level has questioned the idea that asexual reproduction types are primitive or inferior. During the twentieth century, and entering the XXI, hypotheses were raised to explain the advantages of sexual reproduction over the asexual as well as what would allow sexual reproduction to be maintained when it would be more advantageous to reproduce asexually. The most important and known is the Red Queen hypothesis. At the same time, several studies have sought to understand the ecological pressures that allowed and favored the appearance of sexual reproduction in a scenario that was around two billion years ago. The aim of this work is to review historical responses that marked the study of the origin, evolution and maintenance of sexual reproduction, identifying some of the main questions that the scientific community has elaborated over the last two hundred years.

Influence of epidermal growth factor (EGF) supplementation at different times of in vitro maturation of canine oocytes / Influência da suplementação do fator de crescimento epidermal (EGF) em diferentes momentos da maturação in vitro de oócitos caninos

The aim of this study was to evaluate the influence of epidermal growth factor (EGF) on in vitro maturation of canine oocytes at different times of the process. Ovaries were collected from 55 bitches considered healthy and aseptically isolated, immersed in physiological solution (0.9% NaCl) and transported under refrigeration. Grade 1 cumulus-oocyte complexes (COCs) were selected and divided into two groups: control group (CG) and treatment group (TG). In CG 698 grade I COCs were placed in 4-well plates containing TCM-199 medium supplemented with 25 mM HEPES, 100 IU/mL penicillin, 100 mg/mL streptomycin, 26 mM sodium bicarbonate, 1.5 mM sodium pyruvate, 2.9 mM sodium lactate pentahydrate, 0.6 mM cysteine, 0.03 IU/mL hCG, 0.5 µg/mL FSH, 20 µg/mL estrogen at 38.5ºC in a humidified atmosphere of 5% CO2 in times of 24 h, 48 h, and 72 h. In TG 547 COCs received the same maturation medium plus 10 ηg/mL EGF. Logistic regression models (SAS, 2011) were constructed in order to estimate the chances of oocytes being observed at nuclear maturation stages in different culture times (24 h, 48 h, and 72 h). Based on the results found EGF-supplemented medium showed 2.56 times more chances of having an oocyte at metaphase I (M-I) than medium without EGF (p < 0.0001). The results of this study demonstrated that the time of 72 h showed 5.88 times more chances of having an oocyte at metaphase II (M-II) compared to time of 24 h (p = 0.0001) and 7.69 times more chance than time of 48 h (p = 0.0001). The chances of finding an oocyte at M-II were also 9.09 times higher in medium supplemented with EGF than in medium without EGF (p = 0.0001). Thus, these results demonstrated the essential importance of EGF at different moments of oocyte maturation, being a key component for the acquisition of meiotic competence in bitches, increasing the M-I and M-II rates.(AU)

O objetivo deste estudo foi avaliar a influência do fator de crescimento epidermal (EGF) em diferentes momentos da maturação in vitro de oócitos caninos. Os ovários foram coletados de 55 cadelas consideradas sadias e isolados assepticamente, imersos em solução fisiológica e transportados refrigerados. Os complexos cumulus-oócito (COCs) grau 1 foram selecionados e divididos em dois grupos, denominados grupo controle (GC) e grupo tratamento (GT). No GC, 698 COCs grau I foram cultivados em placas de quatro poços contendo meio TCM-199 suplementado com 25 mM de HEPES, 100 UI/mL de penicilina, 100 mg/mL de estreptomicina, 26 mM de bicarbonato de sódio, 1,5 mM de piruvato de sódio, 2,9 mM de lactato de sódio penta hidratado, 0,6 mM de cisteína, 0,03 UI/mL de hCG, 0,5 µg/mL de FSH, 20 µg/mL de estrógeno em estufa úmida a 38ºC, 5% de CO2 nos períodos de 24h, 48 h e 72 h . Já no GT, 547 COCs receberam o mesmo meio de maturação acrescido de 10 ηg/mL do EGF. Modelos de regressão logística foram elaborados para estimar as chances do oócito ser observado nos estágios de maturação nuclear em diferentes tempos de cultivo. Com base nos resultados encontrados, o meio suplementado com EGF demonstrou 2,56 vezes mais chances de ter um oócito no estágio de metáfase I (M-I) do que o meio sem EGF (p < 0,0001). Os resultados desse estudo demonstraram também que o tempo de 72 h mostrou 5,88 vezes mais chances de ter um oócito no estágio de metáfase II (M-II) do que o tempo de 2 h (p = 0,0001) e 7,69 vezes mais chance do que o tempo de 48h (p = 0,0001). As chances de se encontrar um oócito em M-II também foram 9,09 vezes maiores no meio suplementado com EGF do que no meio sem EGF (p = 0,0001). Dessa forma, estes resultados demonstraram a importância essencial do EGF em diferentes momentos da maturação oocitária, sendo componente chave para a aquisição da competência meiótica nas cadelas, aumentando os índices de M-I e M-II.(AU)

La vitamina A: reguladora de la espermatogénesis / Vitamin A: regulator of spermatogenesis

La vitamina A es esencial para el desarrollo de las células germinales masculinas, la fertili-dad y la espermatogénesis normal. El ácido retinoico, metabolito activo de la vitamina A, es necesario para la maduración de las espermatogonias y la entrada correcta de las células germinales en la profase meiótica en los testículos. La expresión de Stra8, fundamental en la meiosis y la espermatogénesis normal, está directamente relacionada con la disponibilidad del ácido retinoico. Este artículo se centra en el proceso de espermatogénesis y la interacción de la vitamina A con el ciclo seminífero, a fin de explicar la relación existente entre la disminución de los aportes de vitamina A, y las alteraciones presentadas en la espermatogénesis de humanos y algunos animales. También, se consideran los efectos de otros genes implicados en la síntesis de la vitamina A, su transporte y su degradación, y se discuten los posibles mecanismos celu-lares que pueden verse afectados por la falta de señalización de la vitamina A, en particular, la regulación del ciclo celular y la interacción célula-célula, puntos críticos para la esperma-togénesis normal. En esta revisión de los 42 artículos científicos más relevantes del tema, se hizo una búsqueda de información en bases de datos como Science Direct, Scielo, Medline, Redalyc y Pubmed, durante los meses de enero a abril de 2014. Se encontró que en diversos estudios se ha demostrado que se puede controlar in vitro la proliferación de células germinales y su dife-renciación, y además, que un deficiente consumo de vitamina A estaría relacionado con la disminución de la fertilidad masculina.

Vitamin A is essential for the development of male germ cells, male fertility and normal sper-matogenesis. Retinoic acid, an active metabolite of vitamin A, is necessary for the maturation of spermatogonia and the correct entry of germ cells in meiotic prophase in the testicles. Stra8expression, which is essential for normal spermatogenesis and meiosis, is directly related to the availability of retinoic acid. This review of 50 scientific articles relevant to the topic will focus on the process of sperma-togenesis and interaction of vitamin A with the seminiferous cycle, in order to explain the re-lationship between the decrease in vitamin A and alterations in spermatogenesis of humans and some animals. Also, it considers the effects of other genes involved in the synthesis of vitamin A, transport, and degradation. Similarly, potential cellular mechanisms that may be affected by the lack of signs of vitamin A will be discussed, in particular the regulation of cell cycle and cell-cell interactions, which are critical for normal spermatogenesis. In conclusion, studies have shown that you can control in vitro germ cell proliferation, and differentiation and poor intake of vitamin A can be related to a decrease in male fertility.

Dieta y cáncer gástrico en México y en el mundo / Diet and gastric cancer in Mexico and in the world

El cáncer gástrico (CG) es la cuarta causa de muerte por cáncer a nivel global. La dieta y el consumo de alcohol y tabaco, además de la infección por Helicobacter pylori determinan un gran número de casos de esta neoplasia. Algunos alimentos contienen sustancias que podrían influir en el proceso de carcinogénesis gástrica, aunque los mecanismos subyacentes no están completamente dilucidados. En México y el mundo, la disminución en el consumo de frutas, vegetales no feculentos y allium, leguminosas y alimentos fuente de selenio, así como el aumento en el consumo de sal, alimentos salados, salmuera y ahumados, chile, carnes procesadas y asadas o a la parrilla se han asociado respectivamente con un aumento de riesgo de CG. Con la evidencia disponible, se podrían desarrollar y evaluar programas para la prevención y control del CG.

Gastric cancer (GC) is the fourt leading cause of cancer death at global level. Diet, alcohol and tobacco, in addition to Helicobacter pylori infection, account for a large number of cases. Some substances contained in foods may influence GC carcinogenesis process; however, the underlying mechanisms have not been fully elucidated. In Mexico and worldwide, a low intake of fruits, non-starchy and allium vegetables, pulses, and foods containing selenium, as well as high intake of salt, salty, salted and smoked foods, chili pepper, processed and grilled/barbecued meats, have been respectively associated with an increased risk of GC. Based on the available evidence, programs for GC prevention and control could be developed and evaluated.

Esporogénesis, esporodermo y ornamentaciónde esporas maduras en Lycopodiaceae / Sporogenesis, sporoderm and mature spore ornamentation in Lycopodiaceae

Studies on reproductive aspects, spore morphology and ultrastructure of Lycopodiaceae are not very common in the scientific literature, and constitute essential information to support taxonomic and systematic relationships among the group. In order to complete existing information, adding new and broader contributions on these topics, a comparative analysis of the sporogenesis ultrastructure, with emphasis on cytological aspects of the sporocyte coat development, tapetum, monoplastidic and polyplastidic meiosis, sporoderm ontogeny and ornamentation of the mature spores, was carried out in 43 taxa of eight genera of the Lycopodiaceae: Austrolycopodium, Diphasium, Diphasiastrum, Huperzia (including Phlegmariurus), Lycopodium, Lycopodiella, Palhinhaea and Pseudolycopodiella growing in the Andes of Colombia and the Neotropics. For this study, the transmission electron microscopy (TEM) samples were collected in Cauca and Valle del Cauca Departments, while most of the spores for scanning electron microscopy (SEM) analysis were obtained from herbarium samples. We followed standard preparation procedures for spore observation by TEM and SEM. Results showed that the sporocyte coat is largely composed by primary wall components; the sporocyte develop much of their metabolic activity in the production of their coat, which is retained until the spores release; protective functions for the diploid cells undergoing meiosis is postulated here for this layer. The abundance of dictyosomes in the sporocyte cytoplasm was related to the formation and development of the sporocyte coat. Besides microtubule activity, the membrane of sporocyte folds, associated with electrodense material, and would early determine the final patterns of spore ornamentation. Monoplastidic condition is common in Lycopodium s.l., whereas polyplastidic condition was observed in species of Huperzia and Lycopodiella s. l.. In monoplastidic species, the tapetum presents abundant multivesicular bodies, while in polyplastidic species, the secretory activity of the tapetum is less intense. Sporoderm development is centripetal, exospore is the first formed layer, then the endospore and, if present, perispore is the final deposited layer. Adult spores of the Lycopodiaceae showed two patterns of ornamentation: negative or caviform (foveolate spores) and positive or muriform ornamentation, the latter with two subtypes (rugate and reticulate spores). The spores of Huperzia are characteristically foveolate, the rugate spores were found in a few species of Huperzia and in all of the Lycopodiella s. l. taxa studied, while Lycopodium s.l. spores bear reticulate ornamentation. Numerous ornamentation traits are diagnostic at the specific level. The types of ornamentation found do not support the recent extreme fragmentation of the family in several genera, but could match, a priori, with the idea of three subfamilies. The findings of sporogenesis, extremely similar in all taxa studied, point more to consider fewer genera, more comprehensive, than the recent, markedsplitting of the family. Rev. Biol. Trop. 62 (3): 1161-1195. Epub 2014 September 01.

Estudios sobre aspectos reproductivos, morfología y ultraestructura de las esporas de Lycopodiaceae no son abundantes en la literatura científica y constituyen información esencial para apoyar las relaciones taxonómicas y sistemáticas en el grupo. Con el fin de completar la información existente, añadiendo contribuciones nuevas y más amplias sobre estos temas, se realizó un análisis comparado de la ultraestructura de la esporogénesis, con énfasis en aspectos citológicos que tienen que ver con la formación de la cubierta de los esporocitos, el tapete, las meiosis monoplastidial y poliplastidial, la ontogenia del esporodermo y la ornamentación de las esporas maduras en 43 táxones de ocho géneros de Lycopodiaceae: Austrolycopodium, Diphasium, Diphasiastrum, Huperzia (incluyendo Phlegmariurus), Lycopodium, Lycopodiella, Palhinhaea y Pseudolycopodiella que crecen en los Andes de Colombia y el Neotrópico. Para estudios con microscopía electrónica de trasmisión (MET) las muestras se recolectaron en los departamentos de Cauca y Valle del Cauca, mientras que la mayoría de las muestras para microscopía electrónica de barrido (MEB) provienen de material herborizado de colecciones. Para la observación de las muestras con MET y MEB se utilizaron protocolos estándar para el procesamiento de esporas. La cubierta de los esporocitos está formada por pared primaria; los esporocitos invierten gran parte de su actividad metabólica en la producción de esa cubierta, que es mantenida hasta la liberación de las esporas y tiene funciones de protección de las células que harán meiosis. La abundancia de dictiosomas en los esporocitos se relacionó con la formación y desarrollo de la cubierta. Además de la actividad de los microtúbulos, la presencia de sinuosidades y plegamientos asociados con material electro denso en la membrana de los esporocitos determinarían tempranamente los patrones de ornamentación de las esporas. La condición monoplastidial es común en Lycopodium s.l.y la poliplastidial se observó en Huperzia y Lycopodiella s. l. En especies monoplastidiales el tapete presenta abundantes cuerpos plurivesiculares, en las poliplastidiales la actividad secretora del tapete es menos intensa. El desarrollo del esporodermo es centrípeto, el exosporio se forma primero, seguido del endosporio y el perisporio, si está presente, se deposita de último. En las esporas adultas de Lycopodiaceae se encontraron dos patrones de ornamentación: negativo o caviforme (esporas foveoladas) y positivo o muriforme (esporas rugadas y reticuladas). Las esporas foveoladas son características de Huperzia; las rugadas de unas pocas especies de Huperzia y las especies de Lycopodiella s. l., mientras que las reticulada son típicas de Lycopodium s. l.. Numerosos caracteres de la ornamentación resultan diagnósticos en el nivel específico. Los tipos principales no apoyan la extrema fragmentación reciente de la familia en varios géneros, aunque podría coincidir, a priori, con la idea de tres subfamilias. Los hallazgos de la esporogénesis, extremadamente similar en todos los táxones estudiados, apuntan más a la unificación de los géneros en la familia que a su segregación.

Los cromosomas de las tortugas tropicales: Kinosternon leucostomum, Trachemys scripta y Staurotypus triporcatus (Testudines: Kinosternidae/Emydidae) / Tropical turtles chromosomes: Kinosternon leucostomum, Trachemys scripta and Staurotypus triporcatus (Testudines: Kinosternidae/Emydidae)

Mexico is a biodiverse country in several taxa as reptiles, that include several species of freshwater and marine turtles. Eventhough most of this group species are under protection, Tabasco State has nine native freshwater turtles, like Kinosternon leucostomum, Trachemys scripta and Staurotypus triporcatus that are very important in traditional dishes. This has resulted in a critical level of their populations, together with little biological knowledge for their conservation. Therefore, this study was dedicated to turtle cytogenetics. The study was conducted using the conventional methods for cytogenetics. The results showed the modal diploid and haploid number for K. leucostomum of 2n=56 (2n=56+3 microchromosomes “B”) and 1n=28 chromosomes in mitosis and meiosis, respectively. In T. scripta 2n=50 chromosomes (2n=50+2 microchromosomes “B”) and 1n=25 chromosomes were also characterized. Whereas in S. triporcatus we only report the 2=54 chromosomes (2n=54+2 microchromosomes “B”). The karyological formula for K. leucostomum was integrated by 12 metacentric-submetacentric chromosomes “msm”/”A”+22 subtelocentrictelocentric chromosomes “stt”/”B”+22 telocentric chromosomes “T”/”C” with fundamental number (FN) of 90 chromosome arms. While T. scripta karyotype was integrated by 32 “msm/”A”+10 “stt”/”B”+8”T/”C” chromosomes, with FN of 92 arms. S. triporcatus karyotype formula was built up by 20 chromosomes “msm”/”A”+34 chromosomes “T”/”C” with FN of 74. The variation in chromosome classification, the fundamental number and the presence of supernumerary microchromosomes “B” in the studied species, were evidence of a particular chromosome cytotypes in Tabasco. We considered that the presence of microchromosomes “B” probably has different origins, and they may be very important as a pattern for the formation or separation of new species. This study also showed the absence of heterologous chromosomes between the females and males karyotypes from the studied species. Rev. Biol. Trop. 62 (2): 671-688. Epub 2014 June 01.

México es un país biodiverso en varios grupos taxonómicos incluyendo a los reptiles, por ello en el país existen varias especies de tortugas dulceacuícolas y marinas. Las especies que integran dicho grupo se encuentran dentro del listado de especies sujetas a protección. El estado de Tabasco cuenta con nueve especies de tortugas de agua dulce, de las cuales Kinosternon leucostomum, Trachemys scripta y Staurotypus triporcatus son de las más importantes dentro de la tradición culinaria, hecho que las ha llevado a niveles críticos en sus poblaciones; aunado al poco conocimiento biológico que sobre dichas especies existe para conservarlas. Por lo anterior, el presente estudio de citogenética es el primero en tortugas de agua dulce en la región. El estudio se realizó, empleando el método convencional de citogenética. Los resultados muestran, el número modal diploide y haploide de K. leucostomum de 2n=56 (2n=56+3 microcromosomas “B”) y 1n=28 cromosomas 686 en mitosis y meiosis, respectivamente. En T. scripta de 2n=50 cromosomas (2n=50+2 microcromosomas “B”) y 1n=25 cromosomas. Mientras que en S. triporcatus solo se reporta el 2n=54 cromosomas (2n=54+2 microcromosomas “B”). La fórmula cromosómica en K. leucostomum, fue de 12 cromosomas metacéntricos submetacéntricos “msm”/“A”+22 cromosomas subtelocéntricos-telocéntricos “stt”/“B”+22 cromosomas telocéntricos “T”/“C”, con número fundamental (NF) de 90 brazos cromosómicos. En T. scripta fue de 32 cromosomas “msm”/“A”+10 cromosomas “stt”/“B”+8 cromosomas “T”/“C”, con NF de 92 y en S. triporcatus 20 cromosomas “msm”/“A”+34 cromosomas “T”/“C” con NF de 74. La variación en la clasificación cromosómica, el número fundamental y la presencia de microcromosomas “B” supernumerarios en las tres especies, son evidencia de citotipos cromosómicos particulares de las tortugas de Tabasco. Se argumenta que la presencia de los microcromosomas “B” tiene diferentes orígenes y de su importancia como pauta para la formación o separación de nuevas especies. En el estudio se descarta la presencia de cromosomas heterólogos entre las hembras y los machos de las especies estudiadas.

Ontogenia de los esporangios, formación y citoquímica de esporas en licopodios (Lycopodiaceae) colombianos / Ontogeny of the sporangium, spore formation and cytochemistry in Colombian Lycopodials (Lycopodiaceae)

Studies on reproductive aspects of Lycopodiaceae are not very abundant in the scientific literature, and constitute essential information to support taxonomic and systematic relationships among the group. Here we present a detailed study of the ontogeny of sporangia and sporogenesis, and the chemical determination of several compounds generated during spore formation. The analyses were performed in 14 taxa of six genera of the family, Diphasiastrum, Diphasium, Huperzia (a genus which is treated here including Phlegmariurus), Lycopodiella, Lycopodium and Palhinhaea. Specimens were collected in three departments from the Colombian Andes between 1 454-3 677m altitude. Ontogeny was studied in small, 1cm long pieces of strobili and axis, which were fixed in glutaraldehyde or FAA, dehydrated in alcohol, embedded in LR White, sectioned in 0.2-0.5μm and stained with toluidine blue (TBO), a metachromatic dye that allows to detect both sporopollenin and lignin or its precursors, during these processes. For other studies, paraplast plus-embedded sections (3-5μm) were stained with safranin-fast green and alcian blue-hematoxylin. Chemical tests were also conducted in sections of fresh sporangia at different stages of maturity using alcian blue (mucopolysaccharides), Lugol solution (starch), Sudan III (lipids), phloroglucinol (lignin) and orcein (chromosomes). Sections were observed with photonic microscope equipped with differential interference contrast (DIC) and fluorescence microscopy (for spore and sporangium walls unstained). Strobili and sporangia were dehydrated with 2.2 dimethoxypropane, critical point dried and coated with gold for scanning electron microscopy (SEM). Our results indicated that the ontogeny of sporangia and sporogenesis were very similar to the previously observed in Huperzia brevifolia. Cutinisation occurs in early stages of development of sporangium cell walls, but in their final stages walls become lignified. As for the sporoderm development, the exospore is the first layer formed, composed by sporopollenin. The endospore deposits as a thin inner layer composed of cellulose, pectin and carboxylated polysaccharides. The perispore, if present, deposits at last. Mucopolysaccharides were found on the sporocyte coat and its abundance in sporangial cavity persists up to the immature tetrads stage, and then disappears. The lipids were abundant in the sporocytes, tetrads and spores, representing the main source of energy of the latter. In contrast, starch is not detected in the spores, but is abundant in premeiotic sporocytes and immature tetrads, developmental stages of high cellular metabolic activity. Intrinsic fluorescence corroborates the presence of lignin and cutin in the sporangium wall, while the sporopollenin is restricted to the exospore. The transfusion cells and the perispore are not always present. However, the processes of ontogeny and sporogenesis are extremely similar throughout the taxa studied, suggesting that they represent conservative family traits, nonspecific or generic.

Los estudios sobre aspectos reproductivos no son muy abundantes en la literatura científica sobre los taxones de Lycopodiaceae y constituyen información esencial para apoyar la taxonomía y relaciones sistemáticas en el grupo. Por lo tanto, se presenta aquí un análisis detallado de la ontogenia de los esporangios y esporogénesis, así como determinaciones químicas de varios compuestos generados durante la formación de las esporas. Los análisis se llevaron a cabo en 14 taxones de seis géneros de la familia: Diphasiastrum, Diphasium, Huperzia (un género que se trata aquí, incluyendo Phlegmariurus), Lycopodiella, Lycopodium y Palhinhaea. Las muestras fueron recolectadas en tres departamentos de los Andes de Colombia entre 1 454-3 677m de altitud. La ontogenia se estudió en trozos de estróbilos y ejes, de 1cm de largo, que se fijaron en glutaraldehido o FAA, se deshidrataron en alcohol, se incluyeron en LR White, se seccionaron en cortes de 0.2-0.5μm y se colorearon con azul de toluidina (TBO), un colorante metacromático que permite detectar tanto esporopolenina como lignina o sus precursores. Para estudios adicionales, secciones de 3-5μm de material incluido en paraplast plus se colorearon con safranina-verde rápido y azul alciánhematoxilina. Las pruebas químicas se llevaron a cabo en secciones de esporangios sin fijar en diferentes etapas de madurez utilizando azul alcián (mucopolisacáridos), solución de Lugol (almidón), Sudán III (lípidos), fluoroglucinol (lignina) y orceína (cromosomas). Las observaciones se efectuaron con microscopio fotónico equipado con contraste diferencial de interferencia (DIC) y microscopía de fluorescencia (para esporas y pared de los esporangios sin colorear). Para observaciones con microscopía electrónica de barrido (MEB), los estróbilos y esporangios se deshidrataron con 2,2 dimetoxipropano, se desecaron a punto crítico y se metalizaron con oro. Los resultados indican que la ontogenia de los esporangios y esporogénesis es muy similar a la observada previamente en Huperzia brevifolia. En las primeras etapas de desarrollo, las paredes celulares de la epidermis del esporangio se cutinizan y en las finales se lignifican. En el desarrollo del esporodermo, la primera capa que se forma es el exosporio, compuesto por esporopolenina. El endosporio es una capa interna delgada compuesta de celulosa, pectina y polisacáridos carboxilados. El perisporio, si está presente, es la última capa que se deposita. Los mucopolisacáridos se encontraron en la cubierta del esporocito, son abundantes en la cavidad esporangial hasta la etapa de tétradas inmaduras y luego desaparecen. Los lípidos son abundantes en esporocitos, tétradas y esporas, y representan la principal fuente de energía de estas. En contraste, el almidón no se detecta en las esporas pero es abundante en esporocitos premeióticos y tétradas inmaduras, ambos con gran actividad metabólica. La fluorescencia intrínseca corrobora la presencia de lignina y cutina en la pared del esporangio, mientras que la esporopolenina se limita al exosporio. Las células de transfusión y el perisporio no siempre están presentes. Sin embargo, los procesos de la ontogenia y esporogénesis son extremadamente similares en todos los taxones estudiados, lo que sugiere que representan rasgos típicos de familia, no específicos ni genéricos.

Ontogenia de los estróbilos, desarrollo de los esporangios y esporogénesis de Equisetum giganteum (Equisetaceae) en los Andes de Colombia

Ontogeny of strobili, sporangia development and sporogenesis in Equisetum giganteum (Equisetaceae) from the Colombian Andes. Studies on the ontogeny of the strobilus, sporangium and reproductive biology of this group of ferns are scarce. Here we describe the ontogeny of the strobilus and sporangia, and the process of sporogenesis using specimens of E. giganteum from Colombia collected along the Rio Frio, Distrito de Sevilla, Piedecuesta, Santander, at 2 200m altitude. The strobili in different stages of development were fixed, dehydrated, embedded in paraffin, sectioned using a rotatory microtome and stained with the safranin O and fast green technique. Observations were made using differential interference contrast microscopy (DIC) or Nomarski microscopy, an optical microscopy illumination technique that enhances the contrast in unstained, transparent. Strobili arise and begin to develop in the apical meristems of the main axis and lateral branches, with no significant differences in the ontogeny of strobili of one or other axis. Successive processes of cell division and differentiation lead to the growth of the strobilus and the formation of sporangiophores. These are formed by the scutellum, the manubrium or pedicel-like, basal part of the sporangiophore, and initial cells of sporangium, which differentiate to form the sporangium wall, the sporocytes and the tapetum. There is not formation of a characteristic arquesporium, as sporocytes quickly undergo meiosis originating tetrads of spores. The tapetum retains its histological integrity, but subsequently the cell walls break down and form a plasmodium that invades the sporangial cavity, partially surrounding the tetrads, and then the spores. Towards the end of the sporogenesis the tapetum disintegrates leaving spores with elaters free within the sporangial cavity. Two layers finally form the sporangium wall: the sporangium wall itself, with thickened, lignified cell walls and an underlying pyknotic layer. The mature spores are chlorofilous, morphologically similar and have exospore, a thin perispore and two elaters. This study of the ontogeny of the spore-producing structures and spores is the first contribution of this type for a tropical species of the genus. Fluorescence microscopy indicates that elaters and the wall of the sporangium are autofluorescent, while other structures induced fluorescence emitted by the fluorescent dye safranin O. The results were also discussed in relation to what is known so far for other species of Equisetum, suggesting that ontogenetic processes and structure of characters sporoderm are relatively constant in Equisetum, which implies important diagnostic value in the taxonomy of the group. Rev. Biol. Trop. 59 (4): 1845-1858. Epub 2011 December 01.

Estudios sobre la ontogenia del estróbilo, los esporangios y la biología reproductiva de Equisetum son escasos, por lo tanto, para la especie E. giganteum, se estudiaron estos aspectos en especímenes recolectados a orillas del Río Frío, Santander, Colombia (2 200m). Los estróbilos en diferentes etapas de maduración fueron fijados, deshidratados, embebidos en parafina, seccionados en micrótomo rotatorio y teñidos con safranina O-fast green. Las observaciones se efectuaron mediante un microscopio óptico de alta resolución con contraste diferencial de interferencia (DIC) y microscopio de fluorescencia. Los estróbilos se inician a partir del meristemo apical, tanto en el eje principal como en los laterales, sin diferencias en el proceso de ontogenia y esporogénesis entre estróbilos de diferentes ejes. Sucesivas mitosis y diferenciación celular conducen al crecimiento del estróbilo, y a la formación de los esporangióforos peltados, formados por el manubrio, o porción basal con aspecto de pedicelo, el escutelo, o porción apical aplanada y las iniciales del esporangio, los cuales se diferenciarán para formar la pared del esporangio, los esporocitos y el tapete. No se forma arquesporio y los esporocitos experimentan meiosis para formar tétradas de esporas. El tapete mantiene la integridad histológica hasta la formación de las tétradas y en esa etapa forma un plasmodio que invade la cavidad esporangial la cual rodea parcialmente las tétradas y luego las esporas, y aparecen las cámaras plasmodiales, un término propuesto aquí para las formaciones designadas en inglés "tapetal gaps". La pared del esporangio queda reducida a dos capas celulares: una externa con engrosamientos lignificados en todas las paredes celulares y una interna picnótica. Al finalizar la esporogénesis, el tapete degenera, y las esporas, con exosporio, perisporio delgado, casi membranáceo y eláteres quedan libres en la cavidad esporangial. El esporodermo, los núcleos y nucléolos presentan fluorescencia roja, inducida por coloración con safranina O, mientras que los eláteres y las células de la pared del esporangio presentan autofluorescencia amarillo-naranja.

Comportamento meiótico durante a microsporogênese em espécies da família meliaceae / Comportamiento meiótica durante la microsporogénesis en especies de la familia meliaceae / Meiotic behavior during microsporogenesis of some species of meliaceae family

A família Meliaceae inclui espécies de grande interesse agronômico, ecológico, econômico e ornamental de alto potencial madeireiro no mundo. No Brasil ocorrem cerca de seis gêneros e 100 espécies. Considerando a falta de estudos citológicos para esse grupo, este trabalho descreve o comportamento meiótico durante a microsporogênese de seis espécies (Trichilia pallida, Trichilia elegans, Trichilia catigua, Cedrela fissilis, Cabralea canjerana, Guarea guidonea) da família Meliaceae, representando quatro dos seis gêneros que estão presentes no Brasil. Inflorescências foram coletadas e fixadas em etanol/ácido acético (3:1 v/v) por 24 horas, transferidos para álcool a 70%, e acondicionadas sob refrigeração. As lâminas foram preparadas pela técnica de esmagamento, coradas com carmim acético a 1%. A análise citogenética revelou poucas irregularidades meióticas, sendo estas relacionadas a segregação dos cromossomos, fusos irregulares e conexão citoplasmática. Como consequência da segregação irregular dos cromossomos na meiose I e II e da organização irregular dos fusos na meiose II, o padrão de citocinese também foi irregular originando tétrades com micrócitos e tríades, resultando em micrósporos desbalanceados e de núcleo restituído (2n).

The Meliaceae family includes the most important species with great agronomic, ecological, economic and ornamental value and high timber potential in the world. In Brazil, approximately six genera and 100 species are found. Considering the lack of cytological studies for its genera, the present study describes the meiotic behavior during microsporogenesis of six species of the Meliaceae family, representing four of the six genera that are present in Brazil; these species are: Trichilia pallida, Trichilia elegans, Trichilia catigua, Cedrela fissilis, Cabralea canjerana, Guarea guidonea. Inflorescences were collected and fixed in ethanol/acetic acid (3:1 v/v) for 24 hours, transferred to alcohol at 70%, and stored under refrigeration. The slides were prepared by squashing, and stained with acetic carmine 1% and observed under optical microscopy. Cytogenetic analysis revealed few meiotic irregularities, which are related to segregation of chromosome spindles and irregular cytoplasmic connection. As a result of irregular segregation of chromosomes during meiosis I and II and the irregular organization of the spindles in meiosis II, the pattern of cytokinesis was also irregular resulting in microcytic tetrads and triads, unbalanced microspores and restored nuclei (2n).

La familia Meliaceae incluye especies de gran interés agronómico, ecológico, económico y ornamental de alto potencial maderero en el mundo. En Brasil ocurren cerca de seis géneros y 100 especies. Considerando la falta de estudios citológicos para ese grupo, este trabajo describe el comportamiento meiótica durante la microsporogénesis de seis especies (Trichilia pallida, Trichilia elegans, Trichilia catigua, Cedrela fissilis, Cabralea canjerana, Guarea guidonea) de la familia Meliaceae, representando cuatro de los seis géneros que están presentes en Brasil. Inflorescencias fueran colectadas y fijadas en etanol/ácido acético (3:1 v/v) por 24 horas, transferidos para alcohol a 70%, y acondicionadas bajo refrigeración. Las láminas fueron preparadas por la técnica de aplastamiento, coloradas con carmín acético a 1%. El análisis citogenética reveló pocas irregularidades meióticas, siendo éstas relacionadas a la segregación de los cromosomas, fusos irregulares y conexión citoplasmática. Como consecuencia de segregación irregular de cromosomas en la meiosis I y II y de la organización irregular de los fusos en la meiosis II, el estándar de citocinesis también fue irregular originando tétradas con micrófitos y tríades, resultando en micros poros desbalanceados y de núcleo restituido (2n).

Caracterización cariotípica en mitosis y meiosis del robalo blanco Centropomus undecimalis (Pisces: Centropomidae)

Karyotypic characterization in mitosis and meiosis of the common snook Centropomus undecimalis (Pisces: Centropomidae). The common snook Centropomus undecimalis inhabits marine, brackish and freshwater habitats in the Western Central Atlantic Ocean, including the Gulf of Mexico. Common snook is an economically important fish in many localities, nevertheless the number of studies on its biology and genetics are still few. The present study attempts to establish the cytogenetic profiles of the specimens collected in Paraiso Municipality Tabasco, Mexico. Tissue of five females and eight male organisms were processed by conventional cytological techniques to obtain chromosome slides of high quality in order to assemble the karyotype. The results from the kidney tissue analysis showed that 85.1% of 288 mitosis had a 2n=48 chromosomes, and 52.8% of 104 meiosis exhibited the haploid number 1n=24. The diploid karyotype showed 48 monoarmed chromosomes of the telocentric (T) type. There was no chromosome heteromorphism between females and males. The diploid karyotype was very similar to that observed in the majority of marine fishes. Rev. Biol. Trop. 59 (2): 683-692. Epub 2011 June 01.

El robalo blanco Centropomus undecimalis, vive en hábitats marinos, salobres y dulceacuícolas en el océano Atlántico occidental, incluyendo el golfo de México. La especie, es económicamente importante en varias localidades, no obstante los estudios sobre su biología y genética son hasta el momento pocos. El presente estudio tiene como propósito, la caracterización citogenética de especímenes recolectados en el municipio de Paraíso, Tabasco, México. Cinco hembras y ocho machos fueron procesados por técnicas citológicas convencionales para la obtención de preparaciones cromosómicas de buena calidad para elaborar el cariotipo. Los resultados del análisis del tejido del riñón, mostraron que 85.1% de 288 mitosis tienen 2n=48 cromosomas y 52.8% de 104 meiosis exhiben el número haploide de 1n=24. El cariotipo diploide mostro 48 cromosomas monorrámeos de tipo telocéntrico (T). No se observó heteromorfismo cromosómico entre hembras y machos. El cariotipo diploide fue similar a los observados en la mayoría de peces marinos.

Bovine parthenogenotes produced by inhibition of first or second polar bodies emission

Parthenogenetic embryos are an ethically acceptable alternative for the derivation of human embryonic stem cells. In this work, we propose a new strategy to produce bovine parthenogenetic embryos inhibiting the emission of the first polar body during in vitro maturation, and allowing the extrusion of the second polar body during oocyte activation. Cytochalasin B, an inhibitor of actin microfilaments, was employed during in vitro maturation to inhibit first polar body emission or during parthenogenetic activation to block second polar body emission. Only one polar body was inhibited in each strategy in order to keep the diploid chromosome set. In experiment 1, the effect of cytochalasin B on in vitro maturation of bovine oocytes was evaluated. Most oocytes (77%) were arrested at a meiotic stage characterized by the presence of a large internal metaphase plate and absence of polar body. In experiment 2, development of embryos exposed to cytochalasin B during in vitro maturation (CytoB-IVM) or during activation (CytoB-ACT) was compared. Developmental rates did not differ between diploidization strategies, even when three agents were employed to induce activation. Both groups, CytoB-IVM and CytoB-ACT, tended to maintain diploidy. CytoB-IVM parthenogenesis could help to obtain embryos with a higher degree of homology to the oocyte donor.

Karyotype and male pre-reductional meiosis of the sharpshooter Tapajosa rubromarginata (Hemiptera: Cicadellidae)

Cicadellidae in one of the best represented families in the Neotropical Region, and the tribe Proconiini comprises most of the xylem-feeding insects, including the majority of the known vectors of xylem-born phytopathogenic organisms. The cytogenetics of the Proconiini remains largely unexplored. We studied males of Tapajosa rubromarginata (Signoret) collected at El Manantial (Tucumán, Argentina) on native spontaneous vegetation where Sorghum halepense predominates. Conventional cytogenetic techniques were used in order to describe the karyotype and male meiosis of this sharpshooter. T. rubromarginata has a male karyological formula of 2n=21 and a sex chromosome system XO:XX (♂:♀). The chromosomes do not have a primary constriction, being holokinetic and the meiosis is pre-reductional, showing similar behavior both for autosomes and sex chromosomes during anaphase I. For this stage, chromosomes are parallel to the acromatic spindle with kinetic activities in the telomeres. They segregate reductionally in the anaphase I, and towards the equator during the second division of the meiosis. This is the first contribution to cytogenetic aspects on proconines sharpshooters, particularly on this economic relevant Auchenorrhyncha species. Rev. Biol. Trop. 59 (1): 309-314. Epub 2011 March 01.

Los Cicadellidae son una de las familias mejor representadas en la región neotropical. La tribu Proconiini incluye a muchos de los insectos que se alimentan de xilema y la mayoría de los vectores de organismos fitopatógenos asociados con dicho tejido de conducción. La citogenética de los Proconiini es prácticamente inexplorada. Por lo tanto, se utilizaron técnicas citogenéticas convencionales para describir el cariotipo y la meiosis en los machos de Tapajosa rubromarginata Signoret. Este cicadélido presenta el complemento cromosómico diploide de 2n=20A+X0 en los machos. Los cromosomas no presentan constricción primaria, son holocinéticos, y la meiosis es pre-reduccional, muestra un comportamiento similar tanto en los cromosomas sexuales como en los autosómicos durante la anafase I. En ese estado, los cromosomas se orientan de manera paralela a las fibras del huso acromático con actividad cinética en los telómeros y segregan de manera reduccional en la fase I y ecuacional en la fase II de la meiosis.

Los cromosomas meióticos de la rana arborícola Smilisca baudinii (Anura: Hylidae)

Meiotic chromosomes of the tree frog Smilisca baudinii (Anura: Hylidae). The Mexican tree frog Smilisca baudinii, is a very common frog in Central America. In spite their importance to keep the ecological equilibrium of the rainforest, its biology and genetics are poorly known. In order to contribute with its biological knowledge, we described the typical meiotic karyotype based in standard cytogenetic protocols to specimens collected in Tabasco, Mexico. The study was centered in the analysis of 131 chromosome spreads at meiotic stage from two adults of the species (one female and one male). The metaphase analysis allowed the establishment of the modal haploid number of 1n=12 bivalent chromosomes. The chromosomic formulae from the haploid bivalent karyotype was integrated by 12 biarmed chromosomes characterized by twelve pairs of metacentric-submetacentric (msm) chromosomes. The meiotic counting gives the idea that diploid chromosome number is integrated by a complement of 2n=24 biarmed chromosomes. The presence of sex chromosomes from female and male meiotic spreads was not observed. Current results suggest that S. baudinii chromosome structure is well shared among Hylidae family and "B" chromosomes are particular structures that have very important evolutionary consequences in species diversification. Rev. Biol. Trop. 59 (1): 355-362. Epub 2011 March 01.

La rana arborícola mexicana Smilisca baudinii, es una especie de rana común en Centroamérica. Sin embargo, la biología y genética de la especie, es pobremente conocida a pesar de su importancia para mantener en equilibrio ecológico las selvas tropicales. Con el propósito de contribuir con el conocimiento biológico de esta especie, establecimos el cariotipo típico en meiosis en especímenes recolectados en Tabasco, México, mediante procedimientos citogenéticos estándares. El estudio, se fundamentó en el análisis de 131 dispersiones cromosómicas en estadio meiótico de dos adultos de la especie (una hembra y un macho). El análisis de las metafases, permitió establecer el número modal haploide de 1n=12 cromosomas bivalentes. La fórmula cromosómica del cariotipo haploide, se integró por 12 cromosomas birrámeos caracterizado por 12 pares de cromosomas bivalentes metacéntricos-submetacéntricos (msm). Los conteos en meiosis, hacen suponer como número diploide de cromosomas a un complemento integrado por 2n=24 cromosomas birrámeos. No fue posible observar presencia de cromosomas sexuales, entre las dispersiones meióticas del espécimen hembra y macho. Los resultados sugieren que la estructura cromosómica de S. baudinii, es compartida ampliamente entre las especies de la familia Hylidae y los cromosomas "B" son estructuras importantes en la diversificación de las especies.

Influência da diminuição da temperatura sobre o fuso meiótico de oócitos de camundongas e de mulheres maturados in vitro / Low temperature influence on meiotic oocyte spindle of mice and humans after maturation in vitro

Introdução: O fuso meiótico dos oócitos de mamíferos pode se despolimerizar quando exposto a pequenas variações de temperatura. Este fato já está bem estabelecido e estudado em oócitos maduros em metáfase II (MII). No entanto, pouco se sabe a respeito da influência da diminuição da temperatura sobre o fuso meiótico dos oócitos imaturos. Desse modo, este estudo tem como objetivos: 1) avaliar a influência da diminuição da temperatura sobre o fuso meiótico de oócitos de camundongas maturados in vitro e 2) avaliar o fuso meiótico em oócitos humanos maturados in vitro submetidos à criopreservação pela técnica de congelação lenta ou por vitrificação quando em estágio de vesícula germinativa. Métodos: Realizaram-se dois experimentos, denominados 1 e 2, sendo o primeiro em oócitos de camundongas e o segundo em oócitos humanos. No experimento 1 oócitos imaturos de camundongas nos estágios de metáfase I (MI), telófase I(TI) e MII foram cultivados nas seguintes temperaturas: 37º C (controle), temperatura ambiente (22oC) e 4º C por 0, 10, 30 e 60 minutos. Após este período de tempo o fuso meiótico oocitário foi avaliado por meio de microscopia de luz polarizada (MLP) (LC-Polscope-Oosight image software) e imunocitoquímica (IC). No experimento 2 oócitos em estágio de vesícula germinativa (GV) coletados de pacientes submetidas à indução da ovulação e fertilização in vitro, foram divididos de forma randômica em três grupos: oócitos a fresco (A), oócitos congelados pela técnica de congelação lenta (B) e oócitos congelados pela técnica de vitrificação (C). Os oócitos a fresco, os descongelados e os aquecidos foram maturados in vitro até estágio de (MII). A análise do fuso meiótico foi realizada por microscópio invertido equipado com uma câmera de vídeo analógica e um sistema de imagens que combina luz polarizada em cristal líquido (ICSI Guard Octax)...

Introduction: The meiotic spindle of most mammals is sensitive to cooling and depolymerizes even after a slight reduction in temperature. This is well described and studied on matured oocytes at metaphase II (MII). However, little is known about the influence of low temperatures under meiotic spindle of imature oocytes. In this way, we sougth to evaluate: 1) the influence of low temperatures on mice oocyte meiotic spindle matured in vitro e 2) the oocyte meiotic spindle from human oocytes matured in vitro and cryopreserved by slow-rate freezing or vitrification at GV stage. Methods: Two experiments were done: the first one on mice and the second one on women.At experiment 1, immature mice oocytes at metaphase I (MI), telophase I (TI) and MII were cultured at 37º C (control), room temperature (22oC) and 4º C for 0, 10, 30 and 60 minutes and then spindle analysis was made with polarized light microscopy (PLM) (LC-Polscope-Oosight image software) or immunocytochemistry (ICC). At experiment 2, GV oocytes retrieved from women submitted to ovulation induction and in vitro fertilization were randomly divided in three groups: fresh oocytes (A), cryopreserved by slow-freezing (B) and cryopreserved by vitrification (C). Fresh, thawed and warmed oocytes were matured in vitro to metaphase II oocytes (MII). A meiotic spindle analysis was done by polarized light microscopy (ICSI Guard Octax). Results: Experiment 1: At time 0 min and 37º C, all oocytes had polymerized spindles both at PLM or ICC. At 4º C, the number of MI oocytes with detectable spindles at PLM was smaller than those analysed by ICC, and it decreased with time, which had also occured with TI oocytes at a smaller proportion. However, at 4º C, TI meiotic spindle recognition with polarized light microscopy and ICC was comparable...

Abnormal behavior of spindle during microsporogenesis of passiflora (passifloraceae) / Comportamento anormal de fuso durante a microsporogênese de passiflora (passifloraceae)ien

Passiflora genus, Passifloraceae family, has more than 500 species and 120 of them are native species of Brazil. All species produce fruits that are used as food, medicine and decoration. Floral buttons of five species were collected and fixed in a mixture of ethanol and acetic acid (3:1). The slides were prepared by squashing and staining with 1% propionic carmine. Results showed that during microsporogenesis there were few irregularities, mostly frequently related to chromosome irregular segregation as: precocious migration to poles in metaphase I and II, non-oriented bivalent chromosomes at metaphase I and II, and laggard chromosomes in anaphase I and II, forming micronuclei in telophases I and II and tetrad with microcyte. Another observed irregularity is related to the organization of spindle fibers in meiosis II as they organize themselves in T and V shapes and in sequential spindle. However, in the V-shaped spindle configuration, there was fusion between two nuclei that were close, forming triads instead of tetrads. Irregular chromosome segregation, abnormal spindles and irregularities in the cytokinesis process were responsible for the formation of monads, dyads, triads and polyads. However, the pollen grain viability was not harmed, presenting an 83.98% to 98.59% fertility variation.

O gênero Passiflora, família Passifloraceae, apresentam mais de 500 espécies, havendo no Brasil aproximadamente 120 espécies nativas. Todas as espécies produzem frutos que são utilizados como produtos alimentícios, medicinais e ornamentais. Botões florais de cinco espécies foram coletados e fixados em etanol/acido acético (3:1). As lâminas foram preparadas utilizando a técnica de esmagamento e coradas com carmim propiônico a 1%. Como resultado, observou-se que durante a microsporogênese poucas irregularidades foram encontradas, as mais frequentes estão relacionadas à segregação irregular dos cromossomos, tais como: migração precoce para os pólos em metáfase I e II, bivalente não orientado em metáfase I e II, e cromossomos retardatários em anáfase I e II, levando a formação de micronúcleos em telófases I e II, e micrócito em tétrades. Outra irregularidade observada esta relacionada a organização das fibras dos fusos em meiose II, que se organizam na forma em T, em V e fuso sequencial. Na configuração de fuso na forma de V ocorreu fusão entre dois núcleos que estavam próximos, formando tríade ao invés de tétrade. A segregação irregular dos cromossomos, a formação de fusos anormais e as irregularidades no processo de citocinese foram responsáveis pela formação de mônades, díades, tríades e políades como produtos final da meiose. Porém, a viabilidade dos grãos de pólen não foi comprometida, apresentado uma variação de 83,98% a 98,59% de fertilidade.

Mecanismos fisiológicos e bioquímicos envolvidos na ovogênese / Physiological and biochemical mechanism involved in ovogenesis

O artigo refere-se a uma atualização sobre os mecanismos fisiológicos e bioquímicos envolvidos na ovogênesis de mamífero.(AU)

The article concern with an up to date about physiologic and biochemical mechanism of ovogenesis in mammals.(AU)

Bloqueio da meiose sem aumento da incidência de anomalias meióticas / Blockage of meiosis without increase of meiotic abnormalities incidence

Objetivo: investigar o efeito do pré-maturação com o inibidor de maturação nuclear, butirolactona-I (BLI), sobre o fuso meiótico e distribuição cromossômica de oócitos bovinos, o que poderia melhorar a qualidade oocitária e embriogênese. Material e métodos: oócitos imaturos, obtidos de vacas abatidas em matadouro (n igual 610) foram submetidos nos grupos: Controle (n igual 208) submetidos à maturação in vitro (MI) em TCM 199 por 24 horas; BLI -18 horas (n igual 208) submetidos à pré-maturação com 100 miuM de BLI por 24 horas e posterior indução da MI em TCM 199, por 18 horas; BLI 24 horas (n igual 195), pré-maturados com 100 miuM de BLI, por 24 horas, seguida por 24 horas de MIV em TCM 199. Em seguida, os oócitos foram fixados, corados por imunofluorescência e avaliados. Resultados: o bloqueio meiótico ocorreu em 88,8% dos oócitos pré-maturados. As taxas de maturaçao foram similares (79,3; 73,5 e 82%, respectivamente, para Controle, BLI 18 horas e BLI 24 horas). Observou-se 82,5% oócitos normais em metáfase II no Controle e 80 e 82% nos grupos BLI 18 horas e BLI 24 horas, respectivamente. A incidência de anomalias meióticas não diferiu (17,5; 20 e 18%, respectivamente, para Controle, BLI 18 horas e BLI 24 horas) - p menor que 0,05, teste do x2. Conclusões: a BLI bloqueia a meiose sem promover danos ao fuso meiótico e distribuição cromossômica oocitária após a MIV

Cytogenetic studies of three Lycosidae species from Argentina (Arachnida, Araneae)

Cytogenetic studies of the family Lycosidae (Arachnida: Araneae) are scarce. Less than 4% of the described species have been analyzed and the male haploid chromosome numbers ranged from 8+X1 X 2 to 13+X1 X 2. Species formerly classified as Lycosa were the most studied ones. Our aim in this work was to perform a comparative analysis of the meiosis in "Lycosa" erythrognatha Lucas, "Lycosa" pampeana Holmberg and Schizocosa malitiosa (Tullgren). We also compared male and female karyotypes and characterized the heterochromatin of "L." erythrognatha. The males of the three species had 2n = 22, n = 10+X1X2, all the chromosomes were telocentric and there was generally a single chiasma per bivalent. In "Lycosa" pampeana, which is described cytogenetically for the first time herein, the bivalents and sex chromosomes showed a clustered arrangement at prometaphase I. The comparison of the male/female karyotypes (2n = 22/24) of "Lycosa" erythrognatha revealed that the sex chromosomes were the largest of the complement and that the autosomes decreased gradually in size. The analysis of the amount, composition and distribution of heterochromatin with C-banding and staining with DAPI- and CMA3 - showed that "Lycosa" erythrognatha had little GC-rich heterochromatin in the pericentromeric region of all chromosomes. In addition, the actual occurrence of the genus Lycosa in the Southern Hemisphere is discussed.