A functional link between bone morphogenetic proteins and insulin-like peptide 3 signaling in modulating ovarian androgen production

Bone morphogenetic proteins (BMP) are firmly implicated as intra-ovarian regulators of follicle development and steroidogenesis. Here we report a microarray analysis showing that treatment of cultured bovine theca cells (TC) with BMP6 significantly (>2-fold; P<0.01) up- or down-regulated expression of 445 genes. Insulin-like peptide 3 (INSL3) was the most heavily down-regulated gene (-43-fold) with CYP17A1 and other key transcripts involved in TC steroidogenesis including LHCGR, INHA, STAR, CYP11A1 and HSD3B1 also down-regulated. BMP6 also reduced expression of NR5A1 encoding steroidogenic factor-1 known to target the promoter regions of the aforementioned genes. Real-time PCR confirmed these findings and also revealed a marked reduction in expression of INSL3 receptor (RXFP2). Secretion of INSL3 protein and androstenedione were also suppressed suggesting a functional link between BMP and INSL3 pathways in controlling androgen synthesis. RNAi-mediated knockdown of INSL3 reduced INSL3 mRNA and secreted protein level (75 and 94%, respectively) and elicited a 77% reduction in CYP17A1 mRNA level and 83% reduction in androstenedione secretion. Knockdown of RXFP2 also reduced CYP17A1 mRNA level (81%) and androstenedione secretion (88%). Conversely, treatment with exogenous (human) INSL3 increased androstenedione secretion ~2-fold. The CYP17 inhibitor abiraterone abolished androgen secretion and reduced expression of both INSL3 and RXFP2. Collectively, these findings indicate a positive autoregulatory role for INSL3 signaling in maintaining thecal androgen production, and visa versa. Moreover, BMP6-induced suppression of thecal androgen synthesis may be mediated, at least in part, by reduced INSL3-RXFP2 signaling

Cell-free protein synthesis as a tool to study RXFP3- Relaxin-3 protein interactions

With the discovery of the relaxin family peptide receptors there is interest in obtaining a clearer understanding of the structure of these proteins and the molecular mechanism of receptor-ligand interaction. As G-protein coupled receptors, obtaining milligram quantities for structural investigations is hampered by the inherent instability of these integral membrane proteins. In the current context, understanding of GPCR structural biology has increased dramatically with crystal structures of several inactive and now active forms solved. In addition, the first nuclear magnetic resonance structure of a GPCR was obtained which is of crucial importance to studying these receptors in a more “biologically relevant” setting. However despite this expansion in the field, most structures have been solved on modified systems so as to increase stability and are not necessarily representative of the native receptors. In relation to the relaxin family peptide receptors, we chose to investigate relaxin-family peptide receptor-3 expressed by cellfree protein synthesis. In contrast to in-vivo expression, cell-free was capable of producing large amounts of native receptor which makes it amenable to demanding structural studies

Benefits of a College STEM Faculty Development Initiative: Instructors Report Increased and Sustained Implementation of Research-Based Instructional Strategies

The Summer Institutes on Scientific Teaching (SI) is a faculty development workshop in which science, technology, engineering, and mathematics (STEM) instructors, particularly from biology, are trained in the Scientific Teaching (ST) pedagogy. While participants have generally reported positive experiences, we aimed to assess how the SI affected participants’ teaching practices. Building on a previously developed taxonomy of ST practices, we surveyed SI participants from the 2004–2014 SI classes regarding specific ST practices. Participants’ self-reported use and implementation of ST practices increased immediately after SI attendance as well as over a longer time frame, suggesting that implementation persisted and even increased with time. However, instructors reported implementation gains for some practices more than others. The practices with the highest gains were engaging students in their own learning, using learning goals in course design, employing formative assessment, developing overarching course learning goals, representing science as a process, and facilitating group discussion activities. We propose that the ST practices showing the greatest gains may serve as beneficial focal points for professional development programs, while practices with smaller gains may require modified dissemination approaches or support structures

INSL3 in the Ruminant: A Powerful Indicator of Gender- and Genetic-Specific Feto-Maternal Dialogue

The hormone Insulin-like peptide 3 (INSL3) is a major secretory product of the Leydig cells from both fetal and adult testes. Consequently, it is a major gender-specific circulating hormone in the male fetus, where it is responsible for the first phase of testicular descent, and in the adult male. In most female mammals, circulating levels are very low, corresponding to only a small production of INSL3 by the mature ovaries. Female ruminants are exceptional in exhibiting high INSL3 gene expression by the thecal cells of antral follicles and by the corpora lutea. We have developed a specific and sensitive immunoassay to measure ruminant INSL3 and show that, corresponding to the high ovarian gene expression, non-pregnant adult female sheep and cows have up to four times the levels observed in other female mammals. Significantly, this level declines during mid-pregnancy in cows carrying a female fetus, in which INSL3 is undetectable. However, in cows carrying a male fetus, circulating maternal INSL3 becomes elevated further, presumably due to the transplacental transfer of fetal INSL3 into the maternal circulation. Within male fetal blood, INSL3 is high in mid-pregnancy (day 153) corresponding to the first transabdominal phase of testicular descent, and shows a marked dependence on paternal genetics, with pure bred or hybrid male fetuses of Bos taurus (Angus) paternal genome having 30% higher INSL3 levels than those of Bos indicus (Brahman) paternity. Thus INSL3 provides the first example of a gender-specific fetal hormone with the potential to influence both placental and maternal physiology

Simulating impacts on UK air quality from net-zero forest planting scenarios

The UK proposes additional bioenergy plantations and afforestation as part of measures to meet net-zero greenhouse gas emissions, but species and locations are not yet decided. Different tree species emit varying amounts of isoprene and monoterpene volatile organic compounds that are precursors to ozone and secondary organic aerosol (SOA) formation, the latter of which is a component of PM2.5. The forest canopy also acts as a depositional sink for air pollutants. All these processes are meteorologically influenced. We present here a first step at coupling information on tree species planting suitability and other planting constraints with data on UK-specific BVOC emission rates and tree canopy data to simulate via the WRF-EMEP4UK high spatial resolution atmospheric chemistry transport model the impact on UK air quality of four potential scenarios. Our ‘maximum planting’ scenarios are based on planting areas where yields are predicted to be ≥50 % of the maximum from the Ecological Site Classification Decision Support System (ESC-DSS) for Eucalyptus gunnii, hybrid aspen (Populus tremula), Italian alder (Alnus cordata) and Sitka spruce (Picea sitchensis). The additional areas of forest in our scenarios are 2.0 to 2.7 times current suggestions for new bioenergy and afforestation landcover in the UK. Our planting scenarios increase UK annual mean surface ozone concentrations by 1.0 ppb or 3 % relative to the baseline landcover for the highest BVOC emitting species (e.g., E. gunni). Increases in ozone reach 2 ppb in summer when BVOC emissions are greatest. In contrast, all the additional planting scenarios lead to reductions in UK annual mean PM2.5 – ranging from -0.2 µg m-3 (-3 %) for Sitka spruce to -0.5 µg m-3 (-7 %) for aspen – revealing that PM2.5 deposition to the additional forest canopy area more than offsets additional SOA formation. Relative decreases in annual mean PM2.5 are greater than the relative increases in annual mean ozone. Reductions in PM2.5 are least in summer, coinciding with the period of maximum monoterpene emissions. Although only a first step in evaluating the impact of increased forest plantation on UK air quality, our study demonstrates the need for locally relevant data on landcover suitability, emissions and meteorology in model simulations