Project description:We constructed a comparative proteome profile of female mouse fetal gonads at specific time points (11.5, 12.5, and 13.5 days post coitum), spanning a critical window for initiation of meiosis in female germ cells. We identified 3666 proteins, of which 473 were differentially expressed.
Project description:BackgroundMacrobrachium nipponense is an economically important species of freshwater shrimp in China. Unlike other marine shrimps, the ovaries in adult female M. nipponense can mature rapidly and periodically during the reproductive period, but the resulting high stocking densities and environmental deterioration can negatively impact the harvest yield and economic benefits. To better understand ovary development in female M. nipponense, we performed systematic transcriptome sequencing of five different stages of ovarian maturation.ResultsWe obtained 255,966 Gb of high quality transcriptome data from 15 samples. Of the 105,082 unigenes that were selected, 30,878 were successfully annotated. From these unigenes, we identified 17 differentially expressed genes and identified three distinct gene expression patterns related to different biological processes. We found that cathepins, legumains, and cystatin were enriched in the lysosome pathway, and they are related to vitellogenin hydrolysis. Additionally, we found that myosin heavy chain 67 participated in oocyte excretion.ConclusionsWe provide the first detailed transcriptome data relating to the ovarian maturation cycle in M. nipponense. Our results provide important reference information about the genomics, molecular biology, physiology, and population genetics of M. nipponense and other crustaceans. It is conducive to further solve the problem of M. nipponense rapid ovarian maturation from the aspects of energy supply and cell division.
Project description:This article describes a suite of global climate model output files that provide continental climatic conditions (monthly temperatures, precipitation, evaporation, precipitation minus evaporation balance, runoff) together with the calculated Köppen-Geiger climate classes and topography, for 28 evenly spaced time slices through the Phanerozoic (Cambrian to Quaternary, 540 Ma to 0 Ma). Climatic variables were simulated with the Fast Ocean Atmosphere Model (FOAM), using a recent set of open-access continental reconstructions with paleotopography and recent atmospheric CO2 and solar luminosity estimates. FOAM is a general circulation model frequently used in paleoclimate studies, especially in the Palaeozoic. Köppen-Geiger climate classes were calculated based on simulated temperature and precipitation fields using Wong Hearing et al.'s [1] implementation of Peel et al.'s [2] updated classification. This dataset provides a unique window onto changing continental climate throughout the Phanerozoic that accounts for the simultaneous evolution of paleogeography (continental configuration and topography), atmospheric composition and greenhouse gas forcing, and solar luminosity.
Project description:Here, we analyzed small RNA libraries derived from ovarian tissues heterozygous or mutant for the Tudor gene, Vreteno. In the absence of vret, Piwi-bound piRNAs are lost without changes in piRNA precursor transcript production, supporting a role for Vret in primary piRNA biogenesis. In the germline, piRNAs can engage in an Aub/Argonaute 3 (AGO3)-dependent amplification in the absence of Vret, suggesting that Vret function can distinguish between primary piRNAs loaded into Piwi/Aub complexes and piRNAs engaged in the amplification cycle. We propose that Vret acts at an early step in primary piRNA processing where it plays an essential role in transposon regulation. Keyword : Epigenetics 2 libraries were analyzed, with 1 being a control (heterozygote).
Project description:Organ shape and size, and, ultimately, organ function, relate in part to the cell and tissue spatial arrangement that takes place during embryonic development. Despite great advances in the genetic regulatory networks responsible for tissue and organ development, it is not yet clearly understood how specific gene functions are linked to the specific morphogenetic processes underlying the internal organ asymmetries found in vertebrate animals. During female chick embryogenesis, and in contrast to males where both testes develop symmetrically, asymmetrical gonad morphogenesis results in only one functional ovary. The disposition of paired organs along the left-right body axis has been shown to be regulated by the activity of the homeobox containing gene pitx2. We have found that pitx2 regulates cell adhesion, affinity, and cell recognition events in the developing gonad primordium epithelia. This in turn not only allows for proper somatic development of the gonad cortex but also permits the proliferation and differentiation of primordial germ cells. We illustrate how Pitx2 activity directs asymmetrical gonad morphogenesis by controlling mitotic spindle orientation of the developing gonad cortex and how, by modulating cyclinD1 expression during asymmetric ovarian development, Pitx2 appears to control gonad organ size. All together our observations indicate that the effects elicited by Pitx2 during the development of the female chick ovary are critical for cell topology, growth, fate, and ultimately organ morphogenesis and function.
Project description:Most female ixodid ticks, once mated, feed to repletion within 6-10 days. Previous studies indicate that an engorgement factor (EF), passed to the female during copulation, may be the stimulus for engorgement. Here, we show that extracts of the testis/vas deferens of fed (but not unfed) male Amblyomma hebraeum contain EF bioactivity when injected into the hemocoel of feeding virgins. We have produced recombinant proteins (recproteins) from 28 feeding-induced genes in the male gonad and have identified a recombinant A. hebraeum engorgement factor (recAhEF) among these recproteins. recAhEF is a combination of two peptides, recAhEFalpha (16.1 kDa) and recAhEFbeta (11.6 kDa), neither of which has bioactivity on its own. recAhEF also stimulates salivary gland degeneration and partial development of the ovary, suggesting that it may be the same material as another male gonadal protein from this tick, male factor. We propose the name "voraxin" for the natural EF of ticks. When normal mated females were put on a rabbit immunized against recAhEF, 74% failed to feed beyond one-tenth the normal engorged weight within 14 days whereas all mated ticks put on a control rabbit engorged normally (mean duration of 8.8 +/- 0.8 days). This result constitutes preliminary evidence that an anti-tick vaccine might be developed from voraxin.
Project description:We present new global maps of the Köppen-Geiger climate classification at an unprecedented 1-km resolution for the present-day (1980-2016) and for projected future conditions (2071-2100) under climate change. The present-day map is derived from an ensemble of four high-resolution, topographically-corrected climatic maps. The future map is derived from an ensemble of 32 climate model projections (scenario RCP8.5), by superimposing the projected climate change anomaly on the baseline high-resolution climatic maps. For both time periods we calculate confidence levels from the ensemble spread, providing valuable indications of the reliability of the classifications. The new maps exhibit a higher classification accuracy and substantially more detail than previous maps, particularly in regions with sharp spatial or elevation gradients. We anticipate the new maps will be useful for numerous applications, including species and vegetation distribution modeling. The new maps including the associated confidence maps are freely available via www.gloh2o.org/koppen.
Project description:We used the MPSS technology to uncover gene expression profiling in a greater depth in the early embryonic gonads and primordial germ cells (PGCs) in the chicken. Total numbers of sequenced signatures were 1,012,533 and 995,676 for the PGCs and gonad. Using a false discovery rate cut-off of 0.05, we found 3.05 % of all signatures in the PCGs and 1.89 % of all signatures in the gonad signatures were significantly up-regulated compared to each sample. The MPSS result was very consistent with our previous result using EST data(http://chickgce.snu.ac.kr/). Keywords: cell type comparison Experimental animals provided for this experiment were maintained at the University Animal Farm, Seoul National University, and all experimental procedures were performed at the affiliated laboratories of the university. Gonadal cells were retrieved from the gonads of 6.5-day-old (stage 29) White Leghorn (WL) embryos by our standard procedure [26]. Embryos were freed from the yolk by rinsing with calcium- and magnesium-free PBS and the gonads were retrieved by dissection of embryo abdomen with sharp tweezers under a stereomicroscope. Embryonic gonads were collected from total 1,947 embryos by 10 highly-skilled persons through 8 separated experimental batches. Gonadal tissues were dissociated by gentle pipetting in 0.05% (v:v) trypsin solution supplemented with 0.53 mM EDTA. After centrifuged at 200xg for 5 min, total gonadal cells were loaded into MACS (Miltenyi Biotech, Germany), and the separated primordial germ cells (PGCs) were immediately stored in liquid nitrogen (-190ºC) until processed further. The numbers of PGCs in cell population before and after loading were counted. MACS treatment for chicken PGCs and counting PGC number Chicken gonadal cells were incubated with PGC-specific primary antibody, anti-stage specific embryo antigen (anti-SSEA)-1 antibody for chicken PGCs (mouse IgM isotype), for 20 min at the room temperature of 20-25?C. Anti-SSEA-1 antibody developed by [27] was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa, Development of Biological Science. After washing with 1mL buffer (PBS supplement with 0.5% BSA and 2mM EDTA), the supernatant was completely removed. The pellet was mixed with 100 ? buffer containing 20? of rat anti-mouse IgM microbeads for 15 min at 4?C. Treated cells were carefully washed by the addition of 500 ? buffer and subsequently loaded with MACS.[28] For counting cell number, chicken PGCs before or after MACS treatment were fixed with 1% (v:v) glutaraldehyde for 5 min and rinsed with 1x PBS twice. The anti-SSEA-1 ascites fluid diluted 1:1,000 in PBS was added and subsequent steps were carried out using DAKO universal LSAB® kit, Peroxidase (DAKO, USA) according to the manufacturer’s instruction. After 8 batches of cell preparation, total cell number of PGC-enriched fraction and gonadal stromal cells was 5.26X106 and 1.76X108, respectively. These cell populations were further used for total RNA isolation and massively parallel signature sequencing (MPSS) analysis.