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.

A bird´s-eye view of chromosomes during meiotic prophase I / Una vista panorámica de los cromosomas en la profase I de la meiosis

ABSTRACT The present review aims to summarize the research carried out in relation to meiosis in birds, especially by observing the protein axes of the chromosomes in prophase I of meiosis. This line of research, initially developed in Argentina, has provided key data in the study of the evolution of sex chromosomes and the mechanisms involved in the frequency and distribution of crossing over in birds, among other topics. Some of these contributions, in addition to those made by other authors, are described also providing the general theoretical framework or the hypotheses that support them.

RESUMEN La presente revisión tiene por objetivo resumir las investigaciones realizadas en relación a la meiosis de las aves, especialmente mediante la observación de los ejes proteicos de los cromosomas en la profase I de la meiosis. Esta línea de investigación, desarrollada inicialmente en Argentina, ha aportado datos clave dentro del estudio de la evolución de los cromosomas sexuales y los mecanismos involucrados en la frecuencia y distribución del crossing over en las aves, entre otros temas. Algunas de estas contribuciones, además de las realizadas por otros autores, se describen proporcionando también el marco teórico general o las hipótesis que las sustentan.

Chromosomes of two Olyra l. species from Misiones, Argentina (Poaceae, Bambusoideae, Olyreae) / Cromosomas de dos especies de Olyra l. de Misiones, Argentina (Poaceae, Bambusoideae, Olyreae)

ABSTRACT The subfamily Bambusoideae comprises three monophyletic tribes, Arundinarieae, Bambuseae and Olyreae. Here we report the gametic number and the chromosomal meiotic behavior of two species belonging to the herbaceous tribe Olyreae, Olyra latifolia and Olyrahumilis. Accessions were collected in Misiones, at Northeastern Argentina. We report a new gametic number for O. humilis, n=18, and we confirmed n=11 for O. latifolia. Chromosomal features, like the basic and gametic chromosome number, are important in understanding the evolution of the Poaceae family, especially in delimiting clades and elucidate inter andintra-clades relationships, and therefore it is necessary to continue producing this type of data.

RESUMEN La subfamilia Bambusoideae comprende tres tribus monofiléticas, Arundinarieae, Bambuseae y Olyreae. Aquí reportamos el número gamético y el comportamiento meiótico de los cromosomas de dos especies pertenecientes a la tribu Olyreae de bambúes herbáceos, Olyra latifolia y Olyra humilis. Las introducciones se recolectaron en la provincia de Misiones, en el noreste argentino. Reportamos un nuevo número gamético para O. humilis, n=18, y confirmamos n=11 para O. latifolia. Los números cromosómicos básicos y gaméticos son importantes para comprender la evolución de la familia Poaceae, especialmente para delimitar sus clados y las relaciones existentes entre ellos, por lo que es necesario continuar produciendo este tipo de datos.

Molecular Mechanism of Aurora Kinase A Regulating the Meiosis of Oocyte / 中国医学科学院学报

Aurora kinase A (AURKA),a family member of aurora kinases,is involved in mitotic entry,maturation and separation of centrosome,assembly and stabilization of bipolar spindle,and condensation and separation of chromosome.Studies have demonstrated that AURKA plays a similar role in meiosis,while the specific mechanism and the similarities and differences in its role between meiosis and mitosis remain unclear.Therefore,we reviewed the studies about the localization and activation of AURKA in oocyte meiosis,and compared the role of AURKA in regulating spindle formation,activating spindle assembly checkpoint,and correcting the kinetochore-microtubule attachment between the meiosis of oocytes and the mitosis of somatic cells.This review will lay a theoretical foundation for revealing the mechanism of AURKA in the regulation of cell division and for the clinical research related to cancer and reproduction.

A novel homozygous frameshift mutation in / 亚洲男科学杂志(英文版)

Oligoasthenoteratozoospermia (OAT) refers to the combination of various sperm abnormalities, including a decreased sperm count, reduced motility, and abnormal sperm morphology. Only a few genetic causes have been shown to be associated with OAT. Herein, we identified a novel homozygous frameshift mutation in meiosis-specific nuclear structural 1 (MNS1; NM_018365: c.603_604insG: p.Lys202Glufs*6) by whole-exome sequencing in an OAT proband from a consanguineous Chinese family. Subsequent variant screening identified four additional heterozygous MNS1 variants in 6/219 infertile individuals with oligoasthenospermia, but no MNS1 variants were observed among 223 fertile controls. Immunostaining analysis showed MNS1 to be normally located in the whole-sperm flagella, but was absent in the proband's sperm. Expression analysis by Western blot also confirmed that MNS1 was absent in the proband's sperm. Abnormal flagellum morphology and ultrastructural disturbances in outer doublet microtubules were observed in the proband's sperm. A total of three intracytoplasmic sperm injection cycles were carried out for the proband's wife, but they all failed to lead to a successful pregnancy. Overall, this is the first study to report a loss-of-function mutation in MNS1 causing OAT in a Han Chinese patient.

An exploratory analysis of the relationship between paternal age at pregnancy and difficulties symptomatic of specific learning disorders among Japanese undergraduate students / Journal of Rural Medicine

Objective: Causes and risk factors of neurodevelopmental disorders originate in the prenatal and perinatal periods. Several studies have demonstrated a relationship between prenatal and perinatal medical records, including maternal and paternal age at pregnancy, and the neurodevelopmental disorders, especially attention deficit/hyperactivity disorder and autism spectrum disorder. However, previous studies showed an association between specific learning disorders and environmental toxins such as lead and tobacco smoke, but not parental age.Patients and Methods: This study included 993 university freshmen, and their prenatal and perinatal medical data was collected from maternal and child handbooks. A mental health assessment questionnaire consisting of 24 items covering symptoms associated with neurodevelopmental disorders was administered, corresponding to aspects of attention deficit/hyperactivity disorder, autism spectrum disorder, and learning disorders. The relationship between prenatal and perinatal medical data and questionnaire results was statistically analyzed.Results: The number of available records was 881 (88.7%). Using Spearman’s rank correlation coefficient analysis and trend analysis, a weak but statistically significant relationship was confirmed between paternal age at pregnancy and the score for learning disorder difficulties.Conclusion: Error accumulation in meiosis during spermatogenesis may be one of the risk factors of learning disorders.

Generation of developmentally competent oocytes and fertile mice from parthenogenetic embryonic stem cells

Parthenogenetic embryos, created by activation and diploidization of oocytes, arrest at mid-gestation for defective paternal imprints, which impair placental development. Also, viable offspring has not been obtained without genetic manipulation from parthenogenetic embryonic stem cells (pESCs) derived from parthenogenetic embryos, presumably attributable to their aberrant imprinting. We show that an unlimited number of oocytes can be derived from pESCs and produce healthy offspring. Moreover, normal expression of imprinted genes is found in the germ cells and the mice. pESCs exhibited imprinting consistent with exclusively maternal lineage, and higher X-chromosome activation compared to female ESCs derived from the same mouse genetic background. pESCs differentiated into primordial germ cell-like cells (PGCLCs) and formed oocytes following in vivo transplantation into kidney capsule that produced fertile pups and reconstituted ovarian endocrine function. The transcriptome and methylation of imprinted and X-linked genes in pESC-PGCLCs closely resembled those of in vivo produced PGCs, consistent with efficient reprogramming of methylation and genomic imprinting. These results demonstrate that amplification of germ cells through parthenogenesis faithfully maintains maternal imprinting, offering a promising route for deriving functional oocytes and having potential in rebuilding ovarian endocrine function.

The formation and repair of DNA double-strand breaks in mammalian meiosis / 亚洲男科学杂志(英文版)

Programmed DNA double-strand breaks (DSBs) are necessary for meiosis in mammals. A sufficient number of DSBs ensure the normal pairing/synapsis of homologous chromosomes. Abnormal DSB repair undermines meiosis, leading to sterility in mammals. The DSBs that initiate recombination are repaired as crossovers and noncrossovers, and crossovers are required for correct chromosome separation. Thus, the placement, timing, and frequency of crossover formation must be tightly controlled. Importantly, mutations in many genes related to the formation and repair of DSB result in infertility in humans. These mutations cause nonobstructive azoospermia in men, premature ovarian insufficiency and ovarian dysgenesis in women. Here, we have illustrated the formation and repair of DSB in mammals, summarized major factors influencing the formation of DSB and the theories of crossover regulation.

The organization, regulation, and biological functions of the synaptonemal complex / 亚洲男科学杂志(英文版)

The synaptonemal complex (SC) is a meiosis-specific proteinaceous macromolecular structure that assembles between paired homologous chromosomes during meiosis in various eukaryotes. The SC has a highly conserved ultrastructure and plays critical roles in controlling multiple steps in meiotic recombination and crossover formation, ensuring accurate meiotic chromosome segregation. Recent studies in different organisms, facilitated by advances in super-resolution microscopy, have provided insights into the macromolecular structure of the SC, including the internal organization of the meiotic chromosome axis and SC central region, the regulatory pathways that control SC assembly and dynamics, and the biological functions exerted by the SC and its substructures. This review summarizes recent discoveries about how the SC is organized and regulated that help to explain the biological functions associated with this meiosis-specific structure.

The role of retinoic acid in the commitment to meiosis / 亚洲男科学杂志(英文版)

Male meiosis is a complex process whereby spermatocytes undergo cell division to form haploid cells. This review focuses on the role of retinoic acid (RA) in meiosis, as well as several processes regulated by RA before cell entry into meiosis that are critical for proper meiotic entry and completion. Here, we discuss RA metabolism in the testis as well as the roles of stimulated by retinoic acid gene 8 (STRA8) and MEIOSIN, which are responsive to RA and are critical for meiosis. We assert that transcriptional regulation in the spermatogonia is critical for successful meiosis.

The molecular control of meiotic double-strand break (DSB) formation and its significance in human infertility / 亚洲男科学杂志(英文版)

Meiosis is an essential step in gametogenesis which is the key process in sexually reproducing organisms as meiotic aberrations may result in infertility. In meiosis, programmed DNA double-strand break (DSB) formation is one of the fundamental processes that are essential for maintaining homolog interactions and correcting segregation of chromosomes. Although the number and distribution of meiotic DSBs are tightly regulated, still abnormalities in DSB formation are known to cause meiotic arrest and infertility. This review is a detailed account of molecular bases of meiotic DSB formation, its evolutionary conservation, and variations in different species. We further reviewed the mutations of DSB formation genes in association with human infertility and also proposed the future directions and strategies about the study of meiotic DSB formation.

Crossover patterns under meiotic chromosome program / 亚洲男科学杂志(英文版)

Repairing DNA double-strand breaks (DSBs) with homologous chromosomes as templates is the hallmark of meiosis. The critical outcome of meiotic homologous recombination is crossovers, which ensure faithful chromosome segregation and promote genetic diversity of progenies. Crossover patterns are tightly controlled and exhibit three characteristics: obligatory crossover, crossover interference, and crossover homeostasis. Aberrant crossover patterns are the leading cause of infertility, miscarriage, and congenital disease. Crossover recombination occurs in the context of meiotic chromosomes, and it is tightly integrated with and regulated by meiotic chromosome structure both locally and globally. Meiotic chromosomes are organized in a loop-axis architecture. Diverse evidence shows that chromosome axis length determines crossover frequency. Interestingly, short chromosomes show different crossover patterns compared to long chromosomes. A high frequency of human embryos are aneuploid, primarily derived from female meiosis errors. Dramatically increased aneuploidy in older women is the well-known "maternal age effect." However, a high frequency of aneuploidy also occurs in young women, derived from crossover maturation inefficiency in human females. In addition, frequency of human aneuploidy also shows other age-dependent alterations. Here, current advances in the understanding of these issues are reviewed, regulation of crossover patterns by meiotic chromosomes are discussed, and issues that remain to be investigated are suggested.

Cytotaxonomy of Dipetalogaster maxima Uhler, 1894 (Hemiptera, Reduviidae, Triatominae) / Citotaxonomia de Dipetalogaster maxima Uhler, 1894 (Hemiptera, Reduviidae, Triatominae)

Abstract The Triatomini tribe consists of ten genera and is regarded as one of the most important tribes from epidemiological point of view. The genus Dipetalogaster Usinger, 1939 is composed only by the species Dipetalogaster maxima Uhler, 1894. This triatomine is exclusive of the Mexico and is a potential vector for Chagas disease. Besides the epidemiological importance, the insects of the Triatominae subfamily are important biological models for cytogenetic studies. Therefore, in order to contribute to the knowledge on the reproductive biology and assist in citotaxonomy of D. maxima, this study aimed to describe spermatogenesis, as well as confirm the karyotype and heterochromatic patterns of this Mexican triatomine species. The seminiferous tubules were torn, fixed to a cover slip and underwent the cytogenetic technique of Lacto-acetic orcein and C-banding. Through the cytogenetics analysis of testicular material D. maxima it was possible to confirm the karyotype (2n = 22), describe the stages of spermatogenesis and characterize the heterochromatic pattern (restricted to sex chromosome Y) of the species. D. maxima showed the same arrangement of heterochromatin described for Triatoma lecticularia (Stål, 1859) (a species that occur in United States of American and Mexico and is phylogenetically related with D. maxima), highlighting the importance of this analysis as an optimization tool to explore phylogenetic correlations.

Resumo A tribo Triatomini consiste em dez gêneros e é considerada uma das tribos mais importantes do ponto de vista epidemiológico. O gênero Dipetalogaster Usinger, 1939 é composto apenas pela espécie Dipetalogaster maxima Uhler, 1894. Este triatomíneo é exclusivo do México e é um vetor potencial da doença de Chagas. Além da importância epidemiológica, os insetos da subfamília Triatominae são importantes modelos biológicos para estudos citogenéticos. Portanto, a fim de contribuir para o conhecimento da biologia reprodutiva e complementar o conceito específico de D. maxima, este trabalho objetivou descrever a espermatogênese, bem como confirmar o padrão cariotípico e heterocromático desta espécie mexicana, com foco citotaxonômico. Os túbulos seminíferos foram dilacerados, fixados em uma lamínula e submetidos à técnica citogenética de Orceína lacto-acética e Bandamento-C. Por meio da análise citogenética do material testicular de D. maxima foi possível confirmar o cariótipo (2n = 22), descrever os estágios da espermatogênese e caracterizar o padrão heterocromático (restrito ao cromossomo Y sexual) da espécie. D. maxima apresentou o mesmo arranjo de heterocromatina descrito para Triatoma lecticularia (Stål, 1859) (espécie que ocorre no México e nos Estados Unidos da América, filogeneticamente relacionada com D. maxima), destacando a importância desta técnica como ferramenta para explorar correlações filogenéticas.

Male and female meiosis evince differential patterns in chiasma formation: a case study of ornamental plant, Delphinium ajacis L.

Chromosomal behaviour during megasporogenesis and microsporogenesis has been studied in ornamental Delphinium ajacis L. Meiosis in female sex cell initiates later than male. The floral buds which carry egg mother cell (EMC) at diplotene stage has pollen mother cells (PMCs) at tetrad stage of meiosis suggesting protandry. Although the 16 chromosomes formed regular eight bivalents in both the sex cells, they differed in overall chiasma frequency which was 32.95% higher in EMCs and found to be 18.52 ± 2.12 per cell. In PMCs, the average chiasma frequency recorded was 13.93 ± 1.40 per cell. Interestingly, this variation in chiasma frequency was largely confined to the two large bivalents which shared 42.61% chiasma per EMC. The use of Q–Q plot, Box plot and Whisker plot showed departure in the chiasma frequency distributions in EMCs and PMCs from the normal distribution pattern. The difference in chiasma frequency in the two sex cells was significant at all levels as indicated by the low P values of 3.094 9 10-11 obtained from nonparametric test, i.e. Wilcoxon rank-sum test. It is suggested that the two different mechanisms of recombination are operational in the two sex cells, and the sex differences of chiasma frequency could have arisen due to differential epigenetic modifications of the chromatin which pattern the double-strand breaks, and the position and frequency of crossing over visible as chiasmata.

Cytomorphological and molecular characterization of inter-specific hybrid between cultivated sunflower and Helianthus argophyllus

Aim: To study useful variability generated in an inter-specific hybrid between cultivated sunflower inbred (ARM-243B) and silver leaf sunflower, H. argophyllus (ARG-2126) through cytomorphological traits and hybrid confirmation by molecular markers.Methodology: The present study on hybridization was undertaken using cultivated sunflower inbred (ARM-243B) and silver leaf sunflower H. argophyllus (ARG-2126). Thirty one morphological characters were recorded of cultivated sunflower (ARM-243B), H. argophyllus (ARG-2126) and inter-specific hybrid. A total of 50 PMCs were analysed for cytological observations. 62 SSR primers were used for molecular characterisation and hybridity confirmation in inter-specific hybrid. Results: Considerable variability was observed in inter-specific hybrid, with common traits including branched stems, anthocyanin coloration in petiole and disk florets, leaf petioles, and stems etc. Cytological abnormalities and reduction in pollen fertility indicated that generated inter-specific hybrid was true type. Out of sixty two SSR primers, nineteen showed parental polymorphism. ORS-1021 showed 10 bp allelic variation in inter-specific hybrid in a co-dominant manner and, hence, was used for hybrid confirmation. Interpretation: The results of this investigation showed that inter-specific hybridization between cultivated sunflower and H. argophyllus and transfer of desirable traits into cultivated sunflower is possible.

MIME-Mitosis instead of meiosis and its application in crop apomixis / 生物工程学报

Apomixis has been widely concerned because of its great potential in heterosis fixation. Artificial apomixis is an important direction of current apomixis research. Mitosis instead of Meiosis (MIME) produces diploid gametes that is identical with the maternal genetic composition and is a key step in the artificial creation of apomixes. This paper reviews the occurrence of MIME and its application in crop apomixis and the problems encountered, in an aim to provide reference for expanding the application of MIME in crop apomixis.

Pollen viability and meiotic irregularities in a collection of Stevia rebaudiana (Bertoni) Bertoni cultivated in Tucumán, Argentina / Viabilidad de polen e irregularidades meióticas en una colección de Stevia rebaudiana (Bertoni) Bertoni cultivada en Tucumán, Argentina

Stevia rebaudiana (Bertoni) Bertoni (2n=2x=22) (Asteraceae family) is a species of economic value due to the presence of steviol glycosides in leaves -mainly stevioside and rebaudioside A- which are non-caloric sugars. In 2013, a collection of plants (genotypes) from four contrasting environments was established in Tucuman, Argentina, for evaluation under local conditions and, eventually, breeding purposes. As a first step, pollen viability and meiosis was studied in a sample of 56 plants. The percentage of pollen viability varied from medium (69.4%) to high (99.6%) in 52 of them, but from low (36.5%) to relatively low (51.5%) in the remaining four. The latter four plants also presented pollen grains of heterogeneous size, which were classified as n (normal, the most frequent size in the sample),

Stevia rebaudiana (Bertoni) Bertoni (2n=2x=22) (familia Asteraceae) es una especie de valor economico debido a la presencia de glucosidos de esteviol en sus hojas, principalmente esteviosido y rebaudiosido A, que son azucares no caloricos. En 2013, se establecio una coleccion de plantas (genotipos) provenientes de cuatro ambientes contrastantes en Tucuman, Argentina, para su evaluacion en condiciones locales y, eventualmente, con fines de mejoramiento genetico. Como primer paso, se estudio la viabilidad del polen y la meiosis en una muestra de 56 plantas. El porcentaje de viabilidad del polen fue de medio (69,4%) a alto (99,6%) en 52 de ellas, pero de bajo (36,5%) a relativamente bajo (51,2%) en las cuatro restantes. Las ultimas cuatro plantas tambien presentaron granos de polen de tamano heterogeneo, que se clasificaron como n (normal, el tamano mas frecuente en la muestra),

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.

LIMK1 takes part in the regulation of Aurora-A-localization on spindle during mouse oocyte meiosis / 基础医学与临床

Objective To investigate the sub-cellular distribution correlation between activated LIMK1 (pLIMK1Thr508) and Aurora-A in mouse oocyte meiosis,and changes in Aurora-A location and spindle structure in condition of LIMK1 inhibition.Methods Immunofluorescence staining was employed to detect the sub-cellular localization of pLIMK1Thr508and its spatial-temporal correlation with spindle organizing regulator Aurora-A in mouse oocyte meiosis; BMS-3, the specific inhibitor to LIMK1 activity, was applied to analyze the effects of LIMK1 inhibition on Aurora-A distribution and spindle formation. Results At meiotic prophase,pLIMK1Thr508was weakly detected and concentrated in the germinal vesicle(GV) in oocytes,with no signal of Aurora-A across the cytoplasm and nuclear area;as meiotic assumption approaching,pLIMK1Thr508left nuclear,aggregating as a single dense dote in the vicinity of nuclear, and being co-localized with the emerging Aurora-A; After germinal vesicle broke down (GVBD), pLIMK1Thr508and Aurora-A remained overlapped and concentrated as multi foci around the condensed chromosomes;at metaphase Ⅰ(MⅠ) and metaphase Ⅱ(MⅡ), pLIMK1Thr508was co-localized with Aurora-A on spindle poles;During anaphase Ⅰ(AⅠ) to telophase Ⅰ(Tel Ⅰ) progression, pLIMK1Thr508was detached from spindle poles and mainly concentrated on the cleavage furrow,while Aurora-A loosely congressed on spindle. In ad-dition, LIMK1 inhibition with BMS- 3 destroyed Aurora-A polar location and spindle formation. Conclusions pLIMK1Thr508is a microtubule organizing center (MTOC)-associated protein, may participate in spindle assembly and maintenance through regulating Aurora-A in mouse oocytes during meiotic progression.