Function and Distribution of the Wamide Neuropeptide Superfamily in Metazoans

This is the final version. Available on open access from Frontiers Media via the DOI in this recordThe Wamide neuropeptide superfamily is of interest due to its distinctive functions in regulating life cycle transitions, metamorphic hormone signaling, and several aspects of digestive system function, from gut muscle contraction to satiety and fat storage. Due to variation among researchers in naming conventions, a global view of Wamide signaling in animals in terms of conservation or diversification of function is currently lacking. Here, I summarize the phylogenetic distribution of Wamide neuropeptides based on current data and describe recent findings in the areas of Wamide receptors and biological functions. Common trends that emerge across Cnidarians and protostomes are the presence of multiple Wamide receptors within a single organism, and the fact that Wamide signaling likely functions across an extensive variety of biological systems, including visual, circadian, and reproductive systems. Important areas of focus for future research are the further identification of Wamide-receptor pairs, confirmation of the phylogenetic distribution of Wamides through largescale sequencing and mass spectrometry, and assignment of different functions to specific subsets of Wamide-expressing neurons. More extensive study of Wamide signaling throughout larval development in a greater number of phyla is also important in order to understand the role of Wamides in hormonal regulation. Defining the evolution and function of neuropeptide signaling in animal nervous systems will benefit from an increased understanding of Wamide function and signaling mechanisms in a wider variety of organisms, beyond the traditional model systems.Biotechnology & Biological Sciences Research Council (BBSRC

Distinctive growth requirements and gene expression patterns distinguish progenitor B cells from pre-B cells.

Long-term bone marrow cultures have been useful in determining gene expression patterns in pre-B cells and in the identification of cytokines such as interleukin 7 (IL-7). We have developed a culture system to selectively grow populations of B lineage restricted progenitors (pro-B cells) from murine bone marrow. Pro-B cells do not grow in response to IL-7, Steel locus factor (SLF), or a combination of the two. c-kit, the SLF receptor, and the IL-7 receptor are both expressed by pro-B cells, indicating that the lack of response is not simply due to the absence of receptors. Furthermore, SLF is not necessary for the growth of pro-B cells since they could be expanded on a stromal line derived from Steel mice that produces no SLF. IL-7 responsiveness in pre-B cells is associated with an increase in n-myc expression and is correlated with immunoglobulin (Ig) gene rearrangements. Although members of the ets family of transcription factors and the Pim-1 kinase are expressed by pro-B cells, n-myc is not expressed. Pro-B cells maintain Ig genes in the germline configuration, which is correlated with a low level of recombination activating genes 1 and 2 (Rag-1 and 2) mRNA expression, but high expression of sterile mu and terminal deoxynucleotidyl transferase. Pro-B cells are unable to grow separated from the stromal layer by a porous membrane, indicating that stromal contact is required for growth. These results suggest that pro-B cells are dependent on alternative growth signals derived from bone marrow stroma and can be distinguished from pre-B cells by specific patterns of gene expression

Editorial: The Evolution of Neuropeptides - a Stroll Through the Animal Kingdom: Updates From the Ottawa 2019 ICCPB Symposium and Beyond

This is the final version. Available on open access from Frontiers Media via the DOI in this recordBiotechnology & Biological Sciences Research Council (BBSRC

Myoinhibitory peptide regulates feeding in the marine annelid Platynereis

This is the final version of the article. Available from the publisher via the DOI in this record.BACKGROUND: During larval settlement and metamorphosis, marine invertebrates undergo changes in habitat, morphology, behavior and physiology. This change between life-cycle stages is often associated with a change in diet or a transition between a non-feeding and a feeding form. How larvae regulate changes in feeding during this life-cycle transition is not well understood. Neuropeptides are known to regulate several aspects of feeding, such as food search, ingestion and digestion. The marine annelid Platynereis dumerilii has a complex life cycle with a pelagic non-feeding larval stage and a benthic feeding postlarval stage, linked by the process of settlement. The conserved neuropeptide myoinhibitory peptide (MIP) is a key regulator of larval settlement behavior in Platynereis. Whether MIP also regulates the initiation of feeding, another aspect of the pelagic-to-benthic transition in Platynereis, is currently unknown. RESULTS: Here, we explore the contribution of MIP to the regulation of feeding behavior in settled Platynereis postlarvae. We find that in addition to expression in the brain, MIP is expressed in the gut of developing larvae in sensory neurons that densely innervate the hindgut, the foregut, and the midgut. Activating MIP signaling by synthetic neuropeptide addition causes increased gut peristalsis and more frequent pharynx extensions leading to increased food intake. Conversely, morpholino-mediated knockdown of MIP expression inhibits feeding. In the long-term, treatment of Platynereis postlarvae with synthetic MIP increases growth rate and results in earlier cephalic metamorphosis. CONCLUSIONS: Our results show that MIP activates ingestion and gut peristalsis in Platynereis postlarvae. MIP is expressed in enteroendocrine cells of the digestive system suggesting that following larval settlement, feeding may be initiated by a direct sensory-neurosecretory mechanism. This is similar to the mechanism by which MIP induces larval settlement. The pleiotropic roles of MIP may thus have evolved by redeploying the same signaling mechanism in different aspects of a life-cycle transition.The research leading to these results received funding from the European Research Council under the European Union’s Seventh Framework Programme (FP7/2007-2013)/European Research Counci

Segmenting subregions of the human hippocampus on structural magnetic resonance image scans: An illustrated tutorial

BACKGROUND: The hippocampus plays a central role in cognition, and understanding the specific contributions of its subregions will likely be key to explaining its wide-ranging functions. However, delineating substructures within the human hippocampus in vivo from magnetic resonance image scans is fraught with difficulties. To our knowledge, the extant literature contains only brief descriptions of segmentation procedures used to delineate hippocampal subregions in magnetic resonance imaging/functional magnetic resonance imaging studies. // METHODS: Consequently, here we provide a clear, step-by-step and fully illustrated guide to segmenting hippocampal subregions along the entire length of the human hippocampus on 3T magnetic resonance images. // RESULTS: We give a detailed description of how to segment the hippocampus into the following six subregions: dentate gyrus/Cornu Ammonis 4, CA3/2, CA1, subiculum, pre/parasubiculum and the uncus. Importantly, this in-depth protocol incorporates the most recent cyto- and chemo-architectural evidence and includes a series of comprehensive figures which compare slices of histologically stained tissue with equivalent 3T images. // CONCLUSION: As hippocampal subregion segmentation is an evolving field of research, we do not suggest this protocol is definitive or final. Rather, we present a fully explained and expedient method of manual segmentation which remains faithful to our current understanding of human hippocampal neuroanatomy. We hope that this 'tutorial'-style guide, which can be followed by experts and non-experts alike, will be a practical resource for clinical and research scientists with an interest in the human hippocampus

Inspiring to inspire: Developing teaching in higher education

Following a three-year staff development initiative within one faculty in a UK university, the authors reflected on inspiring teaching and the role that staff development can play in enhancing individual practice. Teaching is a core component of Higher Education and is complex and multi-faceted both theoretically and in practice. Through individual reflections to a set of pre-determined questions, a group of Higher Education teachers (n = 5) with a responsibility for the development of learning, teaching and assessment, share their thoughts, feelings and beliefs on inspiring teaching. The interpretive analysis of the data shows from a staff perspective that the notion of inspiring teaching has three main components which are all interrelated, those being; the actual teaching and learning experience; the design of the curriculum and the teacher/student relationship. Staff development initiatives were found to help people explore and develop their own teaching philosophy, to develop new practices and to share and learn from others. However, individual’s mindset, beliefs and attitudes were found to be a challenge. Teachers can frame their development around the different aspects of inspiring teaching and with support from senior leadership as well as a positive culture, teaching communities can work together towards inspiring teaching

Object-based representation and analysis of light and electron microscopic volume data using Blender

This is the final version of the article. Available from the publisher via the DOI in this record.BACKGROUND: Rapid improvements in light and electron microscopy imaging techniques and the development of 3D anatomical atlases necessitate new approaches for the visualization and analysis of image data. Pixel-based representations of raw light microscopy data suffer from limitations in the number of channels that can be visualized simultaneously. Complex electron microscopic reconstructions from large tissue volumes are also challenging to visualize and analyze. RESULTS: Here we exploit the advanced visualization capabilities and flexibility of the open-source platform Blender to visualize and analyze anatomical atlases. We use light-microscopy-based gene expression atlases and electron microscopy connectome volume data from larval stages of the marine annelid Platynereis dumerilii. We build object-based larval gene expression atlases in Blender and develop tools for annotation and coexpression analysis. We also represent and analyze connectome data including neuronal reconstructions and underlying synaptic connectivity. CONCLUSIONS: We demonstrate the power and flexibility of Blender for visualizing and exploring complex anatomical atlases. The resources we have developed for Platynereis will facilitate data sharing and the standardization of anatomical atlases for this species. The flexibility of Blender, particularly its embedded Python application programming interface, means that our methods can be easily extended to other organisms.The research leading to these results received funding from the European Research Council under the European Union’s Seventh Framework Programme (FP7/2007-2013)/European Research Council Grant Agreement 260821

A facile method for the stain-free visualization of hierarchical structures with electron microscopy

Diblock copolymers form hierarchical morphologies with numerous applications in drug delivery and as nanoreactors. Yet, the visualization of these structures by electron microscopy can be extremely difficult, requiring complex staining techniques with associated health risks and the potential to alter structural morphology. Reported here is the synthesis of diblock copolymers by RAFT containing 3,5-bis(trifluoromethyl)phenyl functionality allowing for facile visualization of their resulting hierarchical structures by TEM with no further sample preparation.P.E.W. thanks the AWE and E.A. thanks Schlumberger for financial support, and J.d.B is grateful for a Marie Curie Intraeuropean Fellowship (project # 273807). This work was also supported by an ERC Starting Investigator Grant (ASPiRe) and a Next Generation Fellowship provided by the Walters-Kundert Foundation.This is the accepted manuscript. The final published version is available from Wiley at http://onlinelibrary.wiley.com/doi/10.1002/pola.27517/abstract

Bacteria of the Burkholderia cepacia complex are cyanogenic under biofilm and colonial growth conditions

<p>Abstract</p> <p>Background</p> <p>The <it>Burkholderia cepacia </it>complex (Bcc) is a collection of nine genotypically distinct but phenotypically similar species. They show wide ecological diversity and include species that are used for promoting plant growth and bio-control as well species that are opportunistic pathogens of vulnerable patients. Over recent years the Bcc have emerged as problematic pathogens of the CF lung. <it>Pseudomonas aeruginosa </it>is another important CF pathogen. It is able to synthesise hydrogen cyanide (HCN), a potent inhibitor of cellular respiration. We have recently shown that HCN production by <it>P. aeruginosa </it>may have a role in CF pathogenesis. This paper describes an investigation of the ability of bacteria of the Bcc to make HCN.</p> <p>Results</p> <p>The genome of <it>Burkholderia cenocepacia </it>has 3 putative HCN synthase encoding (<it>hcnABC</it>) gene clusters. <it>B. cenocepacia </it>and all 9 species of the Bcc complex tested were able to make cyanide at comparable levels to <it>P. aeruginosa</it>, but only when grown surface attached as colonies or during biofilm growth on glass beads. In contrast to <it>P. aeruginosa </it>and other cyanogenic bacteria, cyanide was not detected during planktonic growth of Bcc strains.</p> <p>Conclusion</p> <p>All species in the Bcc are cyanogenic when grown as surface attached colonies or as biofilms.</p

The neuropeptide complement of the marine annelid Platynereis dumerilii.

This is the final version of the article. Available from BioMed Central via the DOI in this record.BACKGROUND: The marine annelid Platynereis dumerilii is emerging as a powerful lophotrochozoan experimental model for evolutionary developmental biology (evo-devo) and neurobiology. Recent studies revealed the presence of conserved neuropeptidergic signaling in Platynereis, including vasotocin/neurophysin, myoinhibitory peptide and opioid peptidergic systems. Despite these advances, comprehensive peptidome resources have yet to be reported. RESULTS: The present work describes the neuropeptidome of Platynereis. We established a large transcriptome resource, consisting of stage-specific next-generation sequencing datasets and 77,419 expressed sequence tags. Using this information and a combination of bioinformatic searches and mass spectrometry analyses, we increased the known proneuropeptide (pNP) complement of Platynereis to 98. Based on sequence homology to metazoan pNPs, Platynereis pNPs were grouped into ancient eumetazoan, bilaterian, protostome, lophotrochozoan, and annelid families, and pNPs only found in Platynereis. Compared to the planarian Schmidtea mediterranea, the only other lophotrochozoan with a large-scale pNP resource, Platynereis has a remarkably full complement of conserved pNPs, with 53 pNPs belonging to ancient eumetazoan or bilaterian families. Our comprehensive search strategy, combined with analyses of sequence conservation, also allowed us to define several novel lophotrochozoan and annelid pNP families. The stage-specific transcriptome datasets also allowed us to map changes in pNP expression throughout the Platynereis life cycle. CONCLUSION: The large repertoire of conserved pNPs in Platynereis highlights the usefulness of annelids in comparative neuroendocrinology. This work establishes a reference dataset for comparative peptidomics in lophotrochozoans and provides the basis for future studies of Platynereis peptidergic signaling.This work was supported by Max Planck Society Sequencing Grant M.IF.A.ENTW8050 to GJ. The research leading to these results was supported by the European Research Council under European Union Seventh Framework Program FP7/2007–2013 and European Research Council Grant Agreement 260821