Chorion formation in panoistic ovaries requires windei and trimethylation of histone 3 lysine 9

Epigenetic modifications play key roles in transcriptional regulation. Trimethylation of histone 3 lysine 9 (H3K9me3) is one of the most widely studied histone post-translational modifications, and has been linked to transcriptional repression. In Drosophila melanogaster, Windei is needed for H3K9me3 in female germ line cells. Here, we report the occurrence of a D. melanogaster Windei (Wde) ortholog in the ovary of the hemimetabolous insect Blattella germanica, which we named BgWde. Depletion of BgWde by RNAi reduced H3K9me3 in follicular cells, which triggered changes in transcriptional regulation that led to the prevention of chorion gene expression. In turn, this impaired oviposition (and the formation of the ootheca) and, therefore, prevented reproduction. Windei and H3K9me3 have already been reported in follicular cells of D. melanogaster, but this is the first time that the function of these modifications has been demonstrated in the said cells. This is also the first time that an epigenetic marker is reported as having a key role in choriogenesis.Support for this research was provided by the Spanish Ministry of Science and Innovation (Grant nos. BFU2011-22404 to MDP and CGL2008-03517/BOS to XB), by the CSIC (Grant no. 2010TW0019, Formosa program) and from FEDER funds to XB. Support from Generalitat de Catalunya (Grant no. 2005 SGR 00053) and LINC-Global is also gratefully acknowledged. AH received a pre-doctoral research grant (JAE-LINCG program) from the CSIC, and The European Social Fund (ESF)

Chorion formation in panoistic ovaries requires windei and trimethylation of histone 3 lysine 9

Epigenetic modifications play key roles in transcriptional regulation. Trimethylation of histone 3 lysine 9 (H3K9me3) is one of the most widely studied histone post-translational modifications, and has been linked to transcriptional repression. In Drosophila melanogaster, Windei is needed for H3K9me3 in female germ line cells. Here, we report the occurrence of a D. melanogaster Windei (Wde) ortholog in the ovary of the hemimetabolous insect Blattella germanica, which we named BgWde. Depletion of BgWde by RNAi reduced H3K9me3 in follicular cells, which triggered changes in transcriptional regulation that led to the prevention of chorion gene expression. In turn, this impaired oviposition (and the formation of the ootheca) and, therefore, prevented reproduction. Windei and H3K9me3 have already been reported in follicular cells of D. melanogaster, but this is the first time that the function of these modifications has been demonstrated in the said cells. This is also the first time that an epigenetic marker is reported as having a key role in choriogenesis.Support for this research was provided by the Spanish Ministry of Science and Innovation (Grant nos. BFU2011-22404 to MDP and CGL2008-03517/BOS to XB), by the CSIC (Grant no. 2010TW0019, Formosa program) and from FEDER funds to XB. Support from Generalitat de Catalunya (Grant no. 2005 SGR 00053) and LINC-Global is also gratefully acknowledged. AH received a pre-doctoral research grant (JAE-LINCG program) from the CSIC, and The European Social Fund (ESF)

The notch pathway regulates both the proliferation and differentiation of follicular cells in the panoistic ovary of Blattella germanica

The Notch pathway is an essential regulator of cell proliferation and differentiation during development. Its involvement in insect oogenesis has been examined in insect species with meroistic ovaries, and it is known to play a fundamental role in cell fate decisions and the induction of the mitosis-to-endocycle switch in follicular cells (FCs). This work reports the functions of the main components of the Notch pathway (Notch and its ligands Delta and Serrate) during oogenesis in Blattella germanica, a phylogenetically basal species with panoistic ovary. As is revealed by RNAi-based analyses, Notch and Delta were found to contribute towards maintaining the FCs in an immature, non-apoptotic state. This ancestral function of Notch appears in opposition to the induction of transition from mitosis to endocycle that Notch exerts in Drosophila melanogaster, a change in the Notch function that might be in agreement with the evolution of the insect ovary types. Notch was also shown to play an active role in inducing ovarian follicle elongation via the regulation of the cytoskeleton. In addition, Delta and Notch interactions were seen to determine the differentiation of the posterior population of FCs. Serrate levels were found to be Notch-dependent and are involved in the control of the FC programme, although they would appear to play no crucial role in panoistic ovary oogenesis.This work was supported by the Ministry of Science and Innovation, Spain (grant no. BFU2011-22404), by the Catalan Regional Government (grant no. 2014 SGR 619), and by CONICYT + PAI Concurso Nacional Apoyo al Retorno de Investigadores desde el Extranjero, Convocatoria 2013 + 821320046. P.I. is the recipient of a postdoctoral research grant (BECAS CHILE from CONICYT. N.E. is the recipient of a JAE pre-doctoral research grant from the Consejo Superior de Investigaciones Científicas (CSIC

Comparative analysis of miRNA expression during the development of insects of different metamorphosis modes and germband types

BACKGROUND: Do miRNAs contribute to specify the germ-band type and the body structure in the insect embryo? Our goal was to address that issue by studying the changes in miRNA expression along the ontogeny of the German cockroach Blattella germanica, which is a short germ-band and hemimetabolan species. RESULTS: We sequenced small RNA libraries representing 11 developmental stages of B. germanica ontogeny (with especial emphasis on embryogenesis) and the changes in miRNA expression were examined. Data were compared with equivalent data for two long germ-band holometabolan species Drosophila melanogaster and Drosophila virilis, and the short germ-band holometabolan species Tribolium castaneum. The identification of B. germanica embryo small RNA sequences unveiled miRNAs not detected in previous studies, such as those of the MIR-309 family and 54 novel miRNAs. Four main waves of miRNA expression were recognized (with most miRNA changes occurring during the embryonic stages): the first from day 0 to day 1 of embryogenesis, the second during mid-embryogenesis (days 0-6), the third (with an acute expression peak) on day 2 of embryonic development, and the fourth during post-embryonic development. The second wave defined the boundaries of maternal-to-zygotic transition, with maternal mRNAs being cleared, presumably by Mir-309 and associated scavenger miRNAs. CONCLUSION: miRNAs follow well-defined patterns of expression over hemimetabolan ontogeny, patterns that are more diverse during embryonic development than during the nymphal stages. The results suggest that miRNAs play important roles in the developmental transitions between the embryonic stages of development (starting with maternal loading), during which they might influence the germ-band type and metamorphosis mode.This work was supported by the Spanish Ministry of Economy and Competitiveness (grants CGL2012–36251 and CGL2015–64727-P to X.B.), the Spanish Ministry of Science and Innovation (grants BFU2011–22404 and CGL2016–76011-R to M.D.P.) and the Catalan Government (2014 SGR 619). It also received financial assistance from the European Fund for Economic and Regional Development (FEDER funds to X.B. and M.D.P.