RESUMO
In a free-living flatworm, Macrostomum lignano, an S-phase kinase-associated protein 1 (SKP1) homologous gene was identified as enriched in proliferating cells, suggesting that it can function in the regulation of stem cells or germline cells since these are the only two types of proliferating cells in flatworms. SKP1 is a conserved protein that plays a role in ubiquitination processes as a part of the Skp1-Cullin 1-F-box (SCF) ubiquitin ligase complex. However, the exact role of Mlig-SKP1 in M. lignano was not established. Here, we demonstrate that Mlig-SKP1 is neither involved in stem cell regulation during homeostasis, nor in regeneration, but is required for spermatogenesis. Mlig-SKP1(RNAi) animals have increased testes size and decreased fertility as a result of the aberrant maturation of sperm cells. Our findings reinforce the role of ubiquitination pathways in germ cell regulation and demonstrate the conserved role of SKP1 in spermatogenesis.
Assuntos
Platelmintos , Animais , Masculino , Platelmintos/genética , Sêmen/metabolismo , Células Germinativas/metabolismo , Células-Tronco/metabolismo , Ubiquitinação , Proteínas Ligases SKP Culina F-Box/metabolismo , Proteínas Quinases Associadas a Fase S/genética , Proteínas Quinases Associadas a Fase S/metabolismoRESUMO
Regeneration-capable flatworms are informative research models to study the mechanisms of stem cell regulation, regeneration, and tissue patterning. The free-living flatworm Macrostomum lignano is currently the only flatworm where stable transgenesis is available, and as such it offers a powerful experimental platform to address questions that were previously difficult to answer. The published transgenesis approach relies on random integration of DNA constructs into the genome. Despite its efficiency, there is room and need for further improvement and diversification of transgenesis methods in M. lignano. Transposon-mediated transgenesis is an alternative approach, enabling easy mapping of the integration sites and the possibility of insertional mutagenesis studies. Here, we report for the first time that transposon-mediated transgenesis using piggyBac can be performed in M. lignano to create stable transgenic lines with single-copy transgene insertions.
Assuntos
Elementos de DNA Transponíveis/genética , Técnicas de Transferência de Genes , Platelmintos/genética , Animais , TransgenesRESUMO
The genus Haplotropis Sauss. is a relatively small genus in the family Pamphagidae (Orthoptera: Caelifera). Historically, there has been a discussion on the placement of this genus, whether it belongs in the subfamily Pamphaginae or in the subfamily Thrinchinae. Here we present a phylogenetic analysis of nucleotide sequences of two mitochondrial genes (COI, COII) and the ITS2 rRNA nuclear region of multiple species of the family Pamphagidae and related taxa. Our results clearly support the placement of the genus Haplotropis, and other species of the tribe Haplotropidini, in the subfamily Thrinchinae.