Male mouse meiosis has been traditionally studied using descriptive methods like histological sections and spreading or squashing techniques, which allow the observation of fixed meiocytes in either wildtype or genetically modified mice. For these studies, the sacrifice of the males and the extraction of the testicles are required to obtain the material of study. Other functional in vivo studies include the administration of intravenous or intraperitoneal drugs, or the exposure to mutagenic agents or generators of DNA damage, in order to study their impact on meiosis progression. However, in these studies, the exposure times or drug concentration are important limitations to consider when acknowledging animal welfare. Recently, several approaches have been proposed to offer alternative methodologies that allow the in vitro study of spermatocytes with a considerable reduction in the use of animals. Here we revisit and validate an optimal technique of organotypic culture of fragments of seminiferous tubules for meiotic studies. This technique is a trustable methodology to develop functional studies that preserve the histological configuration of the seminiferous tubule, aim homogeneity of the procedures (the use of the same animal for different study conditions), and allow procedures that would compromise the animal welfare. Therefore, this methodology is highly recommendable for the study of meiosis and spermatogenesis, while it supports the principle of 3R\'s for animal research.

Homologous recombination plays pivotal roles in physical attachments and genetic diversity. In the past, it was studied among individuals from different populations. However, only few gametes from individual could generate offspring, which limits its exploration in nature selection. In the last few years, preimplantation blastocysts based on trio SNP-chip data were available in individuals for preimplantation genetic testing (PGT). In this protocol, we demonstrate how to detect meiotic recombination events and construct the genetic map based on trio SNP-chip data, obtained from biopsied blastocysts and their related individuals in PGT cycles, which may allow better understanding of recombination events in nature selection.

Acute myeloid leukemia (AML) is a notably lethal disease, characterized by malignant clonal proliferation of hematopoietic stem cells in the bone marrow. This study seeks to unveil potential therapeutic targets for AML, using a combined approach of microarray analysis and Mendelian randomization (MR). We collected data samples from the Gene Expression Omnibus (GEO) database and extracted pQTL data from genome-wide association studies (GWAS) to identify overlapping genes between the DEGs and GWAS data. Gene enrichment and pathway annotation analyses were performed on these genes. Furthermore, we validated gene expression levels and assessed their clinical relevance. By taking the intersection of these gene sets, we obtained a list of co-expressed genes, including four upregulated genes (REC8, TPM2, ZMIZ1, CD82) and two downregulated genes (IFNAR1, TMCO3). MR analysis demonstrated that genetically predicted protein levels of CD82, REC8, ZMIZ1, and TPM2 were significantly associated with increased odds of AML, while IFNAR1 and TMCO3 showed a protective effect. Gene ontology and KEGG pathway analyses revealed significant enrichment in functions related to female gamete generation, meiosis, p53 signaling pathway, and cardiac muscle contraction. Differences in immune cell profiles were observed between AML survivors and those with poor prognosis, including lower levels of neutrophils and higher levels of follicular helper T cells in the latter group. This study identifies a causal relationship between gene expression and AML and highlights the potential role of REC8 in leukemogenesis, possibly through its impact on gametocyte meiotic abnormalities. The findings provide new insights into the prevention and treatment of leukemia.

Chuanxiong Rhizoma is a well-known Sichuan-specific herbal medicine. Its original plant, Ligusticum chuanxiong, has been cultivated asexually for a long time. L. chuanxiong has sexual reproductive disorders, which restricts its germplasm innovation. However, there is little research on the reproductive system of L. chuanxiong. This study is based on a comparative anatomical research approach, using morphological dissection, paraffin sectioning, staining and compression, and combined with scanning electron microscopy technology, to observe and compare the flowers, fruits, and seeds at various stages of reproductive growth of L. chuanxiong and its wild relative L. sinense. The results showed that the meiosis of pollen mother cells is abnormal in L. chuanxiong anthers, and the size and number of microspores are uneven and inconsistent in the tetrad stage. tapetum cells are not completely degenerated during anther development. During the pollen ripening stage, there are fine cracks in the anther wall, while most anthers could not release pollen normally. The surface of mature pollen grains is concave and partially deformed, and the pollens are all inactive and cannot germinate in vitro. The starch, polysaccharides, and lipids in the pollen were insufficient. The filaments of L. chuanxiong are short at the flowering stage and recurved downward. Double-hanging fruits were observed in the fruiting stage, being wrinkled; with shriveled seeds. Compared with L. sinense at the same stage, the anthers of L. sinense developed normally, and the pollen grains are vigorous and can germinate in vitro. The double-hanging fruits of L. sinense are full and normal; at the flowering period, the filaments are long and erect, significantly higher than the stigma. Mature blastocysts are visible in the ovary of both L. chuanxiong and L. sinense, and there is no significant difference in stigmas. The conclusion is that during the development of L. chuanxiong stamens, the meiosis of pollen mother cells is abnormal, and tetrad, tapetum, filament and other pollen structures develop abnormally. L. chuanxiong has the characteristic of male infertility, which is an important reason for its sexual reproductive disorders.

Xenopus has proven to be a remarkably versatile model organism in the realm of biological research for numerous years, owing to its straightforward maintenance in laboratory settings and its abundant provision of ample-sized oocytes, eggs, and embryos. The cell cycle of these oocytes, eggs, and early embryos exhibits synchrony, and extracts derived from these cells serve various research purposes. Many fundamental concepts in biochemistry, cell biology, and development have been elucidated through the use of cell-free extracts derived from Xenopus cells. Over the past few decades, a wide array of cell-free extracts has been prepared from oocytes, eggs, and early embryos of different Xenopus species at varying cell cycle stages. Each of these extracts possesses distinct characteristics. This review provides a concise overview of the Xenopus species employed in laboratory research, the diverse types of cell-free extracts available, and their respective properties. Furthermore, this review delves into the extensive investigation of spindle assembly in Xenopus egg extracts, underscoring the versatility and potency of these cell-free systems in the realm of cell biology.

Germ cells as the means for the transmission of genetic information between generations have been a hot topic of research for decades. The analysis of the transcriptomes, that is of the RNA transcripts produced by the genotype at a given time, of germ cells and the surrounding somatic cells, is essential to unravel the cellular and molecular processes regulating gametogenesis. However, the asynchronized differentiation of germ cells and high cellular heterogeneity in the developing ovary or testis represent two unsurmountable challenges for delineating the transcription regulation mechanism of germ cells using traditional bulk RNA sequencing. By performing single-cell RNA sequencing (scRNA-seq), it is now possible to dissect the transcriptome of germ cell development at single-cell resolution, and apply powerful bioinformatics methods to translate raw sequencing data into meaningful information. Here, using the 10× Genomic platform and the most widely cited bioinformatics tools, we describe how to analyze early female germ cell development using scRNA-seq data generated from mouse E11.5 to E14.5 ovaries. This pipeline will provide a guide for exploring the processes of early germ cell development at single-cell resolution.

Among the 50,474 spider species, only 849 have chromosomal data available in the literature. Fifty spider families remain unknown from a cytogenetic perspective. The aim of this study was to analyze chromosomally selected araneomorph spiders from Brazil, to contribute to the cytotaxonomy and chromosome evolution of this group. The karyotypes of 12 species belonging to families Corinnidae, Linyphiidae, Oonopidae, Palpimanidae, Theridiidae, Theridiosomatidae, Trachelidae, and Zodariidae were analyzed, including the first chromosomal record for the first two families. Specimens (with the abdomen content partially exposed by perforation) were subjected to colchicine, hypotonization, and fixation. In most cases, the total content of the abdomen was used to prepare slides by spreading of cell suspension and subsequent Giemsa staining. The results were as follows: Cinetomorpha simplex Simon, 1892 (Oonopidae) 2n♂ = 9, X0; Neotrops sp. and Neoxyphinus termitophilus (Oonopidae), 2n♂ = 7, X0; Otiothops birabeni (Palpimanidae); Agyneta sp. (Linyphiidae), 2n♂ = 24, X1X20; Coleosoma floridanum, Thymoites sp.1 and Thymoites sp.2 (Theridiidae), 2n♂ = 22, X1X20; Naatlo sp. (Theridiosomatidae), 2n♂ = 30, X1X20; Orthobula sp. (Trachelidae) 2n♂ = 21, X0; Falconina sp. (Corinnidae), 2n♂ = 28, X1X20; Epicratinus sp. (Zodariidae) 2n♂ = 42, X1X20. The chromosomal morphology was determined for all the samples except for Oonopidae. Most species exhibited telocentric chromosomes, with the exception of Palpimanidae and Theridiosomatidae. The main findings: 1) support an hypothesis on ancestral karyotype of Zodariidae and Oonopidae; 2) reveal a relatively high chromosome number in Palpimanidae that supports an idea on relatively high ancestral chromosome number (2n♂ = 42) of entelegyne spiders (Palpimanoidea is sister group of entelegyne spiders); 3) show that the karyotype found in Theridiosomatidae is exceptional within the Araneoidea.

Triticale (× Triticosecale Wittmack) is of great interest as an insurance crop that can ensure the stability of the gross harvest of feed and food grains at a lower cost. In Western Siberia, only winter triticale varieties are cultivated, however, spring triticales are important for cultivation in regions not suitable for winter crops. To create spring varieties with high yields and good grain quality, it is necessary to study and enrich the gene pool, identify donors of economically valuable traits. One of the possible ways to solve this problem can be through the production of secondary hexaploid triticales with the involvement of the tetraploid wild-growing species of emmer wheat Triticum dicoccum (Schrank) Schuebl. The aim of this work was to create and study hybrids of emmer T. dicoccum (Schrank) Schuebl. with hexaploid triticale using genomic in situ hybridization for staining of meiotic chromosomes and analysis of plant productivity elements in F4-F8. DT4, DT5, DT6 plants and the prebreeding F6 forms obtained from them - DT 4/168, DT 5/176 and DT 6/186 - were selected according to the characteristics of the productivity and the nature of the grain in the F4 hybrid population. The offspring of hybrids DT4 and DT5 and prebreeding forms DT 4/168 and DT 5/176 had an increased grain nature (over 750 g/l), but low productivity. The hybrid DT6 and the breeding form DT 6/186 obtained from it had high grain productivity (785 ± 41 and 822 ± 74 g/m2, respectively), but, like the paternal form of triticale UK 30/33, had a reduced nature of the grain. In F8 DT 6/186 plants, 7 homologous pairs of rye chromosomes and from 27 to 30 wheat chromosomes were found in meiosis, which indicates the presence of a complete rye genome and two wheat ААВВ genomes. Rye chromosomes showed stable formation of bivalents in contrast to wheat chromosomes, which caused the presence of aneuploids in plant populations. Thus, hexaploid forms DT 4/168 and DT 5/176 with well-made smooth grain and high grain size were obtained, which can be used as a source of this trait for selection of food-grade triticale. DT 6/186 is a promising form for further breeding in order to obtain high-yielding forms of triticale.
Тритикале (× Triticosecale Wittmack) представляет большой интерес как страховая культура, способная обеспечить стабильность валового сбора фуражного и продовольственного зерна с более низкими затратами. В Западной Сибири возделываются сорта только озимых тритикале, однако яровые тритикале являются значимыми для выращивания в регионах, не пригодных для озимых культур. Для создания яровых сортов с высокой урожайностью и хорошим качеством зерна необходимо изучение и обогащение генофонда, выделение доноров хозяйственно ценных признаков. Одним из возможных путей решения этой задачи может быть получение вторичных гексаплоидных тритикале с привлечением тетраплоидного дикорастущего вида пшеницы полба Triticum dicoccum (Schrank) Schuebl. Целью данной работы было создание и изучение гибридов полбы T. dicoccum (Schrank) Schuebl. с гексаплоидной тритикале с использованием геномной in situ гибридизации при окрашивании мейотических хромосом и анализ элементов продуктивности растений в F4–F8. По признакам продуктивности и натуры зерна в гибридной популяции F4 были отобраны растения ДТ4, ДТ5, ДТ6 и выявленные в их потомстве пребридинговые формы F6 – ДТ 4/168, ДТ 5/176 и ДТ 6/186. Потомства гибри- дов ДТ4 и ДТ5 и форм ДТ 4/168 и ДТ 5/176 имели повышенную натуру зерна (свыше 750 г/л), но невысокую продуктивность. Гибрид ДТ6 и полученная от него форма ДТ 6/186 отличались высокими показателями продуктивности зерна (785 ± 41 и 822 ± 74 г/м2 соответственно), но, как и отцовская форма тритикале УК 30/33, имели пониженную натуру зерна. У растений потомства F8 ДТ 6/186 в мейозе обнаружено 7 гомологичных пар хромосом ржи и от 27 до 30 хромосом пшеницы, что свидетельствует о наличии полного генома ржи и двух геномов пшеницы AABB. Хромосомы ржи демонстрировали стабильное формирование бивалентов в отличие от хромосом пшеницы, что вызвало анеуплоидию в популяциях растений. Таким образом, получены гексаплоидные формы ДТ 4/168 и ДТ 5/176 с хорошо выполненным гладким зерном и высокой натурой зерна, которые можно использовать в качестве источника этого признака для селекции тритикале пищевого направления. Форма ДТ 6/186 перспективна для дальнейшего селекционного процесса с целью получения высокоурожайных форм тритикале.

Meiosis is usually described as 4 essential and sequential processes: (1) homolog pairing; (2) synapsis, mediated by the synaptonemal complex; (3) crossing over; and (4) segregation. In this canonical model, the maturation of crossovers into chiasmata plays a vital role in holding homologs together and ensuring their segregation at the first meiotic division. However, Lepidoptera (moths and butterflies) undergo 3 distinct meiotic processes, only one of which is canonical. Lepidoptera males utilize 2 meiotic processes: canonical meiosis that produces nucleated fertile sperm, and a noncanonical meiosis that produces anucleated nonfertile sperm which are nonetheless essential for reproduction. Lepidoptera females, which carry heteromorphic sex chromosomes, undergo a completely achiasmate (lacking crossovers) meiosis, thereby requiring an alternative mechanism to ensure proper homolog segregation. Here, we report that the development of a molecular cell biology toolkit designed to properly analyze features of meiosis, including the synaptonemal complex structure and function, in the silkworm Bombyx mori. In addition to standard homology searches to identify Bombyx orthologs of known synaptonemal complex encoding genes, we developed an ortholog discovery app (Shinyapp) to identify Bombyx orthologs of proteins involved in several meiotic processes. We used this information to clone genes expressed in the testes and then created antibodies against their protein products. We used the antibodies to confirm the localization of these proteins in normal male spermatocytes, as well as using in vitro assays to confirm orthologous interactions. The development of this toolkit will facilitate further study of the unique meiotic processes that characterize meiosis in Lepidoptera.

Interactions between the meiosis-expressed gene 1 (MEIG1) and Parkin co-regulated gene (PACRG) protein are critical in the formation of mature sperm cells. Targeting either MEIG1 or PACRG protein could be a contraceptive strategy. The W50A and Y68A mutations on MEIG1 are known to interrupt the MEIG1-PACRG interactions resulting in defective sperm cells. However, the details about how the mutants disrupt the protein-protein binding are not clear. In this study, we reveal insights on MEIG1 and PACRG protein dynamics by applying Gaussian-accelerated molecular dynamics (GaMD) simulations and post-GaMD analysis. Our results show that the mutations destabilize the protein-protein interfacial interaction. The effect of the Y68A mutation is more significant than W50A as Y68 forms stronger polar interactions with PACRG. Because both human and mouse models demonstrate similar dynamic properties, the findings from mouse proteins can be applied to the human system. Moreover, we report a potential ligand binding pocket on the MEIG1 and PACRG interaction surface that could be a target for future drug design to inhibit the MEIG1-PACRG interaction. PACRG shows more qualified pockets along the protein-protein interface, implying that it is a better target than MEIG1. Our work provides a fundamental understanding of MEIG1 and PACRG protein dynamics, paving the way for drug discovery in male-based contraception.