Alterations in transcript abundance of bovine oocytes recovered at growth and dominance phases of the first follicular wave

<p>Abstract</p> <p>Background</p> <p>Oocyte developmental competence is highly affected by the phase of ovarian follicular wave. Previous studies have shown that oocytes from subordinate follicles recovered at growth phase (day 3 after estrus) are developmentally more competent than those recovered at dominance phase (day 7 after estrus). However, the molecular mechanisms associated with these differences are not well elucidated. Therefore, the objective of this study was to investigate transcript abundance of bovine oocytes retrieved from small follicles at growth and dominance phases of the first follicular wave and to identify candidate genes related to oocyte developmental competence using cDNA microarray.</p> <p>Results</p> <p>Comparative gene expression analysis of oocytes from growth and dominance phases and subsequent data analysis using Significant Analysis of Microarray (SAM) revealed a total of 51 differentially regulated genes, including 36 with known function, 6 with unknown function and 9 novel transcripts. Real-time PCR has validated 10 transcripts revealed by microarray analysis and quantified 5 genes in cumulus cells derived from oocytes of both phases. The expression profile of 8 (80%) transcripts (ANAXA2, FL396, S100A10, RPL24, PP, PTTG1, MSX1 and BMP15) was in agreement with microarray data. Transcript abundance of five candidate genes in relation to oocyte developmental competence was validated using Brilliant Cresyl Blue (BCB) staining as an independent model. Furthermore, localization of mRNA and protein product of the candidate gene MSX1 in sections of ovarian follicles at days 0, 1, 3 and 7 of estrous cycle showed a clear fluorescent signal in both oocytes and cumulus cells with higher intensity in the former. Moreover, the protein product was detected in bovine oocytes and early cleavage embryos after fertilization with higher intensity around the nucleus.</p> <p>Conclusion</p> <p>This study has identified distinct sets of differentially regulated transcripts between bovine oocytes recovered from small follicles at growth and dominance phases of the first follicular wave. The validation with independent model supports our notion that many of the transcripts identified here may represent candidate genes associated with oocyte developmental competence. Further specific functional analysis will provide insights into the exact role of these transcripts in oocyte competence and early embryonic development.</p

Deciphering transcriptome profiles of peripheral blood mononuclear cells in response to PRRSV vaccination in pigs

List of DEGs in PBMCs of pigs at 6 hpv of PRRSV vaccination in pigs compared to control. (XLSX 57 kb

A global agenda for advancing freshwater biodiversity research

Global freshwater biodiversity is declining dramatically, and meeting the challenges of this crisis requires bold goals and the mobilisation of substantial resources. While the reasons are varied, investments in both research and conservation of freshwater biodiversity lag far behind those in the terrestrial and marine realms. Inspired by a global consultation, we identify 15 pressing priority needs, grouped into five research areas, in an effort to support informed stewardship of freshwater biodiversity. The proposed agenda aims to advance freshwater biodiversity research globally as a critical step in improving coordinated actions towards its sustainable management and conservation

Establishing a Large-Scale Field Experiment to Assess the Effects of Artificial Light at Night on Species and Food Webs

Artificial light at night (ALAN) is one of the most obvious hallmarks of human presence in an ecosystem. The rapidly increasing use of artificial light has fundamentally transformed nightscapes throughout most of the globe, although little is known about how ALAN impacts the biodiversity and food webs of illuminated ecosystems. We developed a large-scale experimental infrastructure to study the effects of ALAN on a light-naïve, natural riparian (i.e., terrestrial-aquatic) ecosystem. Twelve street lights (20 m apart) arranged in three rows parallel to an agricultural drainage ditch were installed on each of two sites located in a grassland ecosystem in northern Germany. A range of biotic, abiotic, and photometric data are collected regularly to study the short- and long-term effects of ALAN on behavior, species interactions, physiology, and species composition of communities. Here we describe the infrastructure setup and data collection methods, and characterize the study area including photometric measurements. None of the measured parameters differed significantly between sites in the period before illumination. Results of one short-term experiment, carried out with one site illuminated and the other acting as a control, demonstrate the attraction of ALAN by the immense and immediate increase of insect catches at the lit street lights. The experimental setup provides a unique platform for carrying out interdisciplinary research on sustainable lighting

Otimização do sistema de produção de clones por transferência nuclear de célula somática (NTSC)

A técnica de transferência nuclear é uma ferramenta que possibilita a produção de embriões clones que podem ser utilizados tanto na clonagem reprodutiva como no modelo para o estudo de diversos mecanismos fisiológicos durante o desenvolvimento embrionário. Neste sentido, embriões clones bovinos foram produzidos por transferência nuclear, com o objetivo de estabelecer a técnica de clonagem, bem como otimizála para as condições do Laboratório de Embriologia e Biotécnicas de Reprodução da UFRGS. Durante este estudo, 1.123 estruturas foram reconstruídas em diferentes condições, sendo que 95 blastocistos foram produzidos em 56 replicações. O primeiro blastocisto foi produzido na 5ª rotina. Após a transferência de parte destes embriões, 13 prenhezes foram estabelecidas, entretanto, a maioria delas foi interrompida no terço inicial da gestação. Uma prenhez gemelar alcançou 260 dias, momento em que os fetos foram abortados. Outra gestação foi a termo, com o nascimento de um clone vivo, porém, o animal veio a óbito 42 horas após o nascimento

Uso de oócitos bovinos como citoplasma receptor na produção de embriões por transferência nuclear de célula somática interespécie (NTSCi) Use of bovine oocytes as recipient cytoplasm in the production of embryos through nuclear transfer of interspecies so

ABSTRACT Interspecies embryo clones have been produced by research groups with relative success in some species. Bovine oocytes matured in vitro and enucleated by micromanipulation were used in three experiments as recipeient cytoplasm in nuclear transfer of ovine, caprine and porcine fibroblasts. The fibroblasts were cultivated until the third passage before being frozen and used. The electrofusion was induced by an application of a 20V pulse during 45 ms. The activation was done with 5 mM ionomycin and subsequently 2 mM 6DMAP. NTSC bovine embryos, NTSCi caprine and ovine embryos were cultivated in SOF medium and NTSCi porcine embryos were cultivated in NCSU23 medium. The fusion rates of the reconstructed complexes with bovine cells did not differ from those observed with ovine cells (88.2%), caprine cells (74.1%) and porcine cells (79.4%). The cleavage rates in ovine (60.3%), caprine (68.4%) and porcine (57.1%) NTSCi groups did not differ from the control group NTSC bovine. The blastocyst rate observed in the group of NTSCi ovine embryos (10.3%) was similar to the group of NTSC bovine embryos (12.7%). In NTSCi caprine embryos, 5.3% of the embryos developed up to the blastocyst stage, while in the NTSCi porcine group there was no development up to the blastocyst stage. In conclusion, the bovine cytoplasm was able to support the embryo development in NTSCi up to the blastocyst stage using ovine and caprine fibroblasts as donor cells. Keywords: Embryo, Clones, Interspecies, Nuclear Transfer, In vitro. RESUMO Embriões clones interespécie vêm sendo produzidos por diferentes grupos de pesquisa, com relativo sucesso em algumas espécies. Oócitos bovinos maturados in vitro e enucleados por micromanipulação foram utilizados em três experimentos como citoplasma receptor na transferência nuclear de fibroblastos ovinos, caprinos e suínos. Os fibroblastos foram cultivados até a terceira passagem antes de serem congelados e utilizados. A eletrofusão foi induzida pela aplicação de um pulso de 20 V durante 45 ms. A ativação foi realizada com 5 mM de ionomicina e 2 mM de 6DMAP. Embriões NTSC bovinos, NTSCi caprinos e ovinos foram cultivados em meio SOF, e embriões NTSCi suínos foram cultivados em NCSU23. As taxas de fusão dos complexos reconstruídos com células bovinas não diferiram daquelas observadas com células ovinas (88,2%), caprinas (74,1%) e suínas (79,4%). As taxas de clivagem nos grupos NTSCi ovino (60,3%), caprino (68,4%) e suína (57,1%) não diferiram dos grupos controles NTSC bovino. A taxa de blastocisto observada nos embriões NTSCi ovinos (10,3%) foi semelhante à taxa observada no grupo NTSC bovino (12,7%). No grupo NTSCi caprino, 5,3% dos embriões chegaram ao estádio de blastocisto, enquanto que no grupo NTSCi suíno não houve desenvolvimento até o estádio de blastocisto. O citoplasma bovino foi capaz de suportar o desenvolvimento de embriões NTSCi até o estádio de blastocisto utilizando-se núcleo de fibroblatos ovinos e caprinos

Worldwide variations in artificial skyglow

Despite constituting a widespread and significant environmental change, understanding of artificial nighttime skyglow is extremely limited. Until now, published monitoring studies have been local or regional in scope, and typically of short duration. In this first major international compilation of monitoring data we answer several key questions about skyglow properties. Skyglow is observed to vary over four orders of magnitude, a range hundreds of times larger than was the case before artificial light. Nearly all of the study sites were polluted by artificial light. A non-linear relationship is observed between the sky brightness on clear and overcast nights, with a change in behavior near the rural to urban landuse transition. Overcast skies ranged from a third darker to almost 18 times brighter than clear. Clear sky radiances estimated by the World Atlas of Artificial Night Sky Brightness were found to be overestimated by ~25%; our dataset will play an important role in the calibration and ground truthing of future skyglow models. Most of the brightly lit sites darkened as the night progressed, typically by ~5% per hour. The great variation in skyglow radiance observed from site-to-site and with changing meteorological conditions underlines the need for a long-term international monitoring program

Citizen science’s transformative impact on science, citizen empowerment and socio-political processes

Citizen science (CS) can foster transformative impact for science, citizen empowerment and socio-political processes. To unleash this impact, a clearer understanding of its current status and challenges for its development is needed. Using quantitative indicators developed in a collaborative stakeholder process, our study provides a comprehensive overview of the current status of CS in Germany, Austria and Switzerland. Our online survey with 340 responses focused on CS impact through (1) scientific practices, (2) participant learning and empowerment, and (3) socio-political processes. With regard to scientific impact, we found that data quality control is an established component of CS practice, while publication of CS data and results has not yet been achieved by all project coordinators (55%). Key benefits for citizen scientists were the experience of collective impact (“making a difference together with others”) as well as gaining new knowledge. For the citizen scientists’ learning outcomes, different forms of social learning, such as systematic feedback or personal mentoring, were essential. While the majority of respondents attributed an important value to CS for decision-making, only few were confident that CS data were indeed utilized as evidence by decision-makers. Based on these results, we recommend (1) that project coordinators and researchers strengthen scientific impact by fostering data management and publications, (2) that project coordinators and citizen scientists enhance participant impact by promoting social learning opportunities and (3) that project initiators and CS networks foster socio-political impact through early engagement with decision-makers and alignment with ongoing policy processes. In this way, CS can evolve its transformative impact

Neurocalcin Delta Suppression Protects against Spinal Muscular Atrophy in Humans and across Species by Restoring Impaired Endocytosis

This document is the Accepted Manuscript version of the following article: Riessland et al., 'Neurocalcin Delta Suppression Protects against Spinal Muscular Atrophy in Humans and across Species by Restoring Impaired Endocytosis', The American Journal of Human Genetics, Vol. 100 (2): 297-315, first published online 26 January 2017. The final, published version is available online at doi: http://dx.doi.org/10.1016/j.ajhg.2017.01.005 © 2017 American Society of Human Genetics.Homozygous SMN1 loss causes spinal muscular atrophy (SMA), the most common lethal genetic childhood motor neuron disease. SMN1 encodes SMN, a ubiquitous housekeeping protein, which makes the primarily motor neuron-specific phenotype rather unexpected. SMA-affected individuals harbor low SMN expression from one to six SMN2 copies, which is insufficient to functionally compensate for SMN1 loss. However, rarely individuals with homozygous absence of SMN1 and only three to four SMN2 copies are fully asymptomatic, suggesting protection through genetic modifier(s). Previously, we identified plastin 3 (PLS3) overexpression as an SMA protective modifier in humans and showed that SMN deficit impairs endocytosis, which is rescued by elevated PLS3 levels. Here, we identify reduction of the neuronal calcium sensor Neurocalcin delta (NCALD) as a protective SMA modifier in five asymptomatic SMN1-deleted individuals carrying only four SMN2 copies. We demonstrate that NCALD is a Ca(2+)-dependent negative regulator of endocytosis, as NCALD knockdown improves endocytosis in SMA models and ameliorates pharmacologically induced endocytosis defects in zebrafish. Importantly, NCALD knockdown effectively ameliorates SMA-associated pathological defects across species, including worm, zebrafish, and mouse. In conclusion, our study identifies a previously unknown protective SMA modifier in humans, demonstrates modifier impact in three different SMA animal models, and suggests a potential combinatorial therapeutic strategy to efficiently treat SMA. Since both protective modifiers restore endocytosis, our results confirm that endocytosis is a major cellular mechanism perturbed in SMA and emphasize the power of protective modifiers for understanding disease mechanism and developing therapies.Peer reviewedFinal Accepted Versio

A global agenda for advancing freshwater biodiversity research

Global freshwater biodiversity is declining dramatically, and meeting the challenges of this crisis requires bold goals and the mobilisation of substantial resources. While the reasons are varied, investments in both research and conservation of freshwater biodiversity lag far behind those in the terrestrial and marine realms. Inspired by a global consultation, we identify 15 pressing priority needs, grouped into five research areas, in an effort to support informed stewardship of freshwater biodiversity. The proposed agenda aims to advance freshwater biodiversity research globally as a critical step in improving coordinated actions towards its sustainable management and conservation.Peer reviewe