LSM14B controls oocyte mRNA storage and stability to ensure female fertility.

Controlled mRNA storage and stability is essential for oocyte meiosis and early embryonic development. However, how to regulate mRNA storage and stability in mammalian oogenesis remains elusive. Here we showed that LSM14B, a component of membraneless compartments including P-body-like granules and mitochondria-associated ribonucleoprotein domain (MARDO) in germ cell, is indispensable for female fertility. To reveal loss of LSM14B disrupted primordial follicle assembly and caused mRNA reduction in non-growing oocytes, which was concomitant with the impaired assembly of P-body-like granules. 10× Genomics single-cell RNA-sequencing and immunostaining were performed. Meanwhile, we conducted RNA-seq analysis of GV-stage oocytes and found that Lsm14b deficiency not only impaired the maternal mRNA accumulation but also disrupted the translation in fully grown oocytes, which was closely associated with dissolution of MARDO components. Moreover, Lsm14b-deficient oocytes reassembled a pronucleus containing decondensed chromatin after extrusion of the first polar body, through compromising the activation of maturation promoting factor, while the defects were restored via WEE1/2 inhibitor. Together, our findings reveal that Lsm14b plays a pivotal role in mammalian oogenesis by specifically controlling of oocyte mRNA storage and stability.

Single-cell RNA sequencing of the Mongolia sheep testis reveals a conserved and divergent transcriptome landscape of mammalian spermatogenesis.

Spermatogenesis is a highly coordinated and complex process, and is pivotal for transmitting genetic information between mammalian generations. In this study, we investigated the conservation, differences, and biological functions of homologous genes during spermatogenesis in Mongolia sheep, humans, cynomolgus monkey, and mice using single-cell RNA sequencing technology. We compared X chromosome meiotic inactivation events in Mongolia sheep, humans, cynomolgus monkey, and mice to uncover the concerted activity of X chromosome genes. Subsequently, we focused on the dynamics of gene expression, key biological functions, and signaling pathways at various stages of spermatogenesis in Mongolia sheep and humans. Additionally, the ligand-receptor networks of Mongolia sheep and humans in testicular somatic and germ cells at different developmental stages were mapped to reveal conserved germ cell-soma communication using single-cell resolution. These datasets provided novel information and insights to unravel the molecular regulatory mechanisms of Mongolia sheep spermatogenesis and highlight conservation in gene expression during spermatogenesis between Mongolia sheep and humans, providing a foundation for the establishment of a large mammalian disease model of male infertility.

[Expression of T cell-specific transcription factors T-bet and GATA-3 in recurrent aphthous ulcerations].

OBJECTIVE: To investigate the expression of the T cell-specific transcription factors T-bet/GATA-3 in recurrent aphthous ulcerations (RAU). METHODS: Peripheral blood mononuclear cells (PBMC) were isolated by density gradient centrifugation from RAU patients and healthy controls. Expressions of T-bet and GATA-3 mRNA and protein in the PBMCs were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot respectively. The concentrations of gamma-interferon (gamma-IFN) and interleukin-4 (IL-4) in the serum were determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: The expressions of T-bet mRNA were 0.73 +/- 0.14 in the control group and 0.12 +/- 0.04 in the recurrent aphthous ulcerations group, the levels of T-bet protein expressions were 0.73 +/- 0.18 in the control group and 0.24 +/- 0.09 in the recurrent aphthous ulcerations group, there was a significant difference between the two groups (P < 0.01). The levels of GATA-3 mRNA expressions were 0.89 +/- 0.11 in the control group and 1.30 +/- 0.13 in the recurrent aphthous ulcerations group; GATA-3 protein expressions were 1.09 +/- 0.16 in the control group and 1.80 +/- 0.16 in the recurrent aphthous ulcerations group, there was a significant difference between the two groups (P < 0.01). In the control group, the concentration of gamma-interferon in the serum was (22.15 +/- 1.52) ng/L, which was significantly elevated compared with that in the recurrent aphthous ulcerations group (16.32 +/- 0.22) ng/L (P < 0.01), in the control group, the concentration of IL-4 in the serum was (25.58 +/- 2.59) ng/L, which was significantly lower than that of the recurrent aphthous ulcerations group (43.60 +/- 3.15) ng/L (P < 0.01). The ratio of gamma-IFN and IL-4 was positively correlated with the ratio of T-bet and GATA-3 mRNA and protein expression (r = 0.96, 0.94, P < 0.01). CONCLUSIONS: Imbalance of transcription factors T-bet and GATA-3 may be one of the key factors in immune dysregulation of recurrent aphthous ulcerations.