Aerococcus agrisoli sp. nov., isolated from paddy soil.

A non-spore-forming, Gram-stain-positive, short rod-shaped strain, designated SJQ22T, was isolated from a paddy soil sample collected in Shanghai, PR China. A comparative analysis of 16S rRNA gene sequences showed that strain SJQ22T fell within the genus Aerococcus, forming a clear cluster with the type strains of Aerococcus viridans (98.6 % sequence similarity) and Aerococcus urinaeequi (98.5 % sequence similarity). Strain SJQ22T grew at 30-45 °C (optimum, 30 °C), pH 6.0-8.0 (optimum, pH 7.0) and with a NaCl concentration of 0-4 % (optimum, 1 %). Cells were negative for oxidase and catalase activity. Chemotaxonomic analysis showed that strain SJQ22T possessed C16:0 and C18:1 ω9c as the predominant fatty acids. The DNA G + C content was 39.0 mol%. Strain SJQ22T exhibited DNA-DNA relatedness levels of 13±2 % with A. viridans ATCC 11563T and 9±2 % with A. urinaeequi IFO 12173T. Based on the data obtained, strain SJQ22T represents a novel species of the genus Aerococcus, for which the name Aerococcus agrisoli sp. nov. is proposed. The type strain is SJQ22T (=JCM 33111T=CCTCC AB 2018283T).

Retinoic acid-induced differentiation of porcine prospermatogonia in vitro.

Spermatogenesis is an intricate developmental process occurring in testes by which spermatogonial stem cells (SSCs) self-renew and differentiate into mature sperm. The molecular mechanisms for SSC self-renewal and differentiation, while have been well studied in mice, may differ between mice and domestic animals including pigs. To gain knowledge about the molecular mechanisms for porcine SSC self-renewal and differentiation that have so far been poorly understood, here we isolated and enriched prospermatogonia from neonatal porcine testes, and exposed the cells to retinoic acid, a direct inducer for spermatogonial differentiation. We then identified that retinoic acid could induce porcine prospermatogonial differentiation, which was accompanied by a clear transcriptomic alteration, as revealed by the RNA-sequencing analysis. We also compared retinoic acid-induced in vitro porcine spermatogonial differentiation with the in vivo process, and compared retinoic acid-induced in vitro spermatogonial differentiation between pigs and mice. Furthermore, we analyzed retinoic acid-induced differentially expressed long non-coding RNAs (lncRNAs), and demonstrated that a pig-specific lncRNA, lncRNA-106504875, positively regulated porcine spermatogonial proliferation by targeting the core transcription factor ZBTB16. Taken together, these results would help to elucidate the roles of retinoic acid in porcine spermatogonial differentiation, thereby contributing to further knowledge about the molecular mechanisms underlying porcine SSC development and, in the long run, to optimization of both long-term culture and induced differentiation systems for porcine SSCs.

Sertoli cell and spermatogonial development in pigs.

BACKGROUND: Spermatogenesis is an intricate developmental process during which undifferentiated spermatogonia, containing spermatogonial stem cells (SSCs), undergo self-renewal and differentiation to generate eventually mature spermatozoa. Spermatogenesis occurs in seminiferous tubules within the testis, and the seminiferous tubules harbor Sertoli and germ cells. Sertoli cells are an essential somatic cell type within the microenvironment that support and steer male germ cell development, whereas spermatogonia are the primitive male germ cells at the onset of spermatogenesis. While the developmental progression of Sertoli cells and spermatogonia has been well established in mice, much less is known in other mammalian species including pigs. RESULTS: To acquire knowledge of Sertoli cell and spermatogonial development in pigs, here we collected as many as nine ages of Duroc porcine testes from the neonate to sexual maturity, i.e., testes from 7-, 30-, 50-, 70-, 90-, 110-, 130-, 150- and 210-day-old boars, and performed histological and immunohistochemical analyses on testis sections. We first examined the development of spermatogenic cells and seminiferous tubules in porcine testes. Then, by immunofluorescence staining for marker proteins (AMH, SOX9, DBA, UCHL1, VASA, KIT, Ki67 and/or PCNA), we delved into the proliferative activity and development of Sertoli cells and of spermatogonial subtypes (pro-, undifferentiated and differentiating spermatogonia). Besides, by immunostaining for ß-catenin and ZO-1, we studied the establishment of the blood-testis barrier in porcine testes. CONCLUSIONS: In this longitudinal study, we have systematically investigated the elaborate Sertoli cell and spermatogonial developmental patterns in pigs from the neonate to sexual maturity that have so far remained largely unknown. The findings not only extend the knowledge about spermatogenesis and testicular development in pigs, but also lay the theoretical groundwork for porcine breeding and rearing.