Discovery, cloning, and analysis of novel fluorescent proteins from various color morphs of Corynactis californica

Although the number of fluorescent protein (FP) genes cloned from the GFP family continues to increase, few studies of GFP-type pigments in non-bioluminescent, non-symbiotic organisms have been attempted. The first goal of this study was to locate, clone, characterize, and analyze fluorescent proteins from an organism exhibiting these traits in order to better understand their evolution and function. I successfully cloned two full-length GFP homologs by applying a FACS-based screening method to a cDNA library constructed from a temperate corallimorpharian, Corynactis californica. The full-length coding regions of each gene were subcloned into an expression vector and bacterial cultures were used to express the proteins. Spectral properties of purified proteins were characterized and chromophore maturation behavior was examined. Phylogenetic methods were also used to analyze the new gene sequences in relation to homologous GFP family members. After discovering two GFP-like proteins in a single red morph, I investigated six additional morphs of Corynactis californica, and found indications of a variety of fluorescent pigments based on fluorescence emission spectra from live specimens. The second goal of this study was to identify and describe the variation in fluorescent pigments among morphs of C. californica and to relate the in vivo emission patterns and colors to FP genes cloned from and expressed in each morph. Specifically, I found that all morphs express a similar suite of GFP-like proteins, generated by at least three to four genetic loci, which code for three colors: green, orange, and red. The genes exhibit tissue-specific expression patterns that differ by morph, and two major expression patterns emerged. Sequence and phylogenetic analyses comparing the new FP genes from C. californica to one another and to homologous members of the GFP family indicate that FP genes from this species are most closely related to one another, but that FP genes arose in an ancestor to the Anthozoa before speciation events separating anthozoan subclasses, including the Corallimorpharia. Possible ecological roles of variations in fluorescent pigmentation among morphs of C. californica are also discussed

Meeting report of Ctenopalooza : the first international meeting of ctenophorologists

© The Author(s), 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in EvoDevo 7 (2016): 19, doi:10.1186/s13227-016-0057-3Here we present a report on Ctenopalooza: A meeting of ctenophorologists held at the Whitney Laboratory for Marine Bioscience in St. Augustine, FL, USA, on March 14–15, 2016. In this report, we provide a summary of each of the sessions that occurred during this two-day meeting, which touched on most of the relevant areas of ctenophore biology. The report includes some major themes regarding the future of ctenophore research that emerged during Ctenopalooza. More information can be found at the meeting Web site: http://ctenopalooza.whitney.ufl.edu.Ctenopalooza was sponsored by a grant from the National Science Foundation’s Division of Integrative Organismal Systems to Joseph Ryan (#1619712). We also acknowledge funding was provided by a grant from the University of Florida’s Office of Research to Joseph Ryan (Project #00075235). Additional funding was provided by The Whitney Laboratory for Marine Bioscience

Incisional hernia repair after caesarean section: a population based study

BACKGROUND Incisional hernias occur at surgical abdominal incision sites but the association with caesarean section (CS) has not been examined. AIM: To determine whether CS is a risk factor for incisional hernia repair. MATERIAL and METHODS: Population-based cohort study in Australia using linked birth and hospital data for women who gave birth from 2000 to 2011. (n=642,578) Survival analysis was used to explore the association between CS and subsequent incisional hernia repair. Analyses were adjusted for confounding factors including other abdominal surgery. The main outcome measure was surgical repair of an incisional hernia. RESULTS: 217,555 women (33.9%) had at least one CS and 1,554 (0.2%) had an incisional hernia repair. The frequency of incisional hernia repair in women who had ever had a caesarean section was 0.47%, compared to 0.12% in women who never had a caesarean section. After controlling for different follow up lengths and known explanatory variables, the adjusted hazard ratio (aHR) was 2.73 (95%CI 2.45-3.06, P <0.001). Incisional hernia repair risk increased with number of caesarean sections: women with two CS had a threefold increased risk of incisional hernia repair, which increased to 6 fold after five CS (aHR=6.29, 95%CI 3.99-9.93, P<0.001) compared to women with no CS. Prior abdominal surgery including other hernia repair also increased the risk of incisional hernia repair (all p<0.001). CONCLUSIONS: There was a strong association between maternal CS and subsequent incisional hernia repair, which increased as the number of CSs increased, but the absolute risk of incisional hernia repair was low.We thank the New South Wales (NSW) Ministry of Health for access to the population health data and the NSW Centre for Health Record Linkage for linking the data sets. This work was supported by an Australian National Health and Medical Research Council (NHMRC) Centre for Research Excellence Grant (1001066). CLR is supported by a NHMRC Senior Research Fellowship (#APP1021025)

Cnidofest 2018: the future is bright for cnidarian research.

The 2018 Cnidarian Model Systems Meeting (Cnidofest) was held September 6-9th at the University of Florida Whitney Laboratory for Marine Bioscience in St. Augustine, FL. Cnidofest 2018, which built upon the momentum of Hydroidfest 2016, brought together research communities working on a broad spectrum of cnidarian organisms from North America and around the world. Meeting talks covered diverse aspects of cnidarian biology, with sessions focused on genomics, development, neurobiology, immunology, symbiosis, ecology, and evolution. In addition to interesting biology, Cnidofest also emphasized the advancement of modern research techniques. Invited technology speakers showcased the power of microfluidics and single-cell transcriptomics and demonstrated their application in cnidarian models. In this report, we provide an overview of the exciting research that was presented at the meeting and discuss opportunities for future research

Expression of multiple Sox genes through embryonic development in the ctenophore Mnemiopsis leidyi is spatially restricted to zones of cell proliferation

Background: The Sox genes, a family of transcription factors characterized by the presence of a high mobility group (HMG) box domain, are among the central groups of developmental regulators in the animal kingdom. They are indispensable in progenitor cell fate determination, and various Sox family members are involved in managing the critical balance between stem cells and differentiating cells. There are 20 mammalian Sox genes that are divided into five major groups (B, C, D, E, and F). True Sox genes have been identified in all animal lineages but not outside Metazoa, indicating that this gene family arose at the origin of the animals. Whole-genome sequencing of the lobate ctenophore Mnemiopsis leidyi allowed us to examine the full complement and expression of the Sox gene family in this early-branching animal lineage. Results: Our phylogenetic analyses of the Sox gene family were generally in agreement with previous studies and placed five of the six Mnemiopsis Sox genes into one of the major Sox groups: SoxB (MleSox1), SoxC (MleSox2), SoxE (MleSox3, MleSox4), and SoxF (MleSox5), with one unclassified gene (MleSox6). We investigated the expression of five out of six Mnemiopsis Sox genes during early development. Expression patterns determined through in situ hybridization generally revealed spatially restricted Sox expression patterns in somatic cells within zones of cell proliferation, as determined by EdU staining. These zones were located in the apical sense organ, upper tentacle bulbs, and developing comb rows in Mnemiopsis, and coincide with similar zones identified in the cydippid ctenophore Pleurobrachia. Conclusions: Our results are consistent with the established role of multiple Sox genes in the maintenance of stem cell pools. Both similarities and differences in juvenile cydippid stage expression patterns between Mnemiopsis Sox genes and their orthologs from Pleurobrachia highlight the importance of using multiple species to characterize the evolution of development within a given phylum. In light of recent phylogenetic evidence that Ctenophora is the earliest-branching animal lineage, our results are consistent with the hypothesis that the ancient primary function of Sox family genes was to regulate the maintenance of stem cells and function in cell fate determination

Expression of multiple Sox genes through embryonic development in the ctenophore Mnemiopsis leidyi is spatially restricted to zones of cell proliferation

Background: The Sox genes, a family of transcription factors characterized by the presence of a high mobility group (HMG) box domain, are among the central groups of developmental regulators in the animal kingdom. They are indispensable in progenitor cell fate determination, and various Sox family members are involved in managing the critical balance between stem cells and differentiating cells. There are 20 mammalian Sox genes that are divided into five major groups (B, C, D, E, and F). True Sox genes have been identified in all animal lineages but not outside Metazoa, indicating that this gene family arose at the origin of the animals. Whole-genome sequencing of the lobate ctenophore Mnemiopsis leidyi allowed us to examine the full complement and expression of the Sox gene family in this early-branching animal lineage. Results: Our phylogenetic analyses of the Sox gene family were generally in agreement with previous studies and placed five of the six Mnemiopsis Sox genes into one of the major Sox groups: SoxB (MleSox1), SoxC (MleSox2), SoxE (MleSox3, MleSox4), and SoxF (MleSox5), with one unclassified gene (MleSox6). We investigated the expression of five out of six Mnemiopsis Sox genes during early development. Expression patterns determined through in situ hybridization generally revealed spatially restricted Sox expression patterns in somatic cells within zones of cell proliferation, as determined by EdU staining. These zones were located in the apical sense organ, upper tentacle bulbs, and developing comb rows in Mnemiopsis, and coincide with similar zones identified in the cydippid ctenophore Pleurobrachia. Conclusions: Our results are consistent with the established role of multiple Sox genes in the maintenance of stem cell pools. Both similarities and differences in juvenile cydippid stage expression patterns between Mnemiopsis Sox genes and their orthologs from Pleurobrachia highlight the importance of using multiple species to characterize the evolution of development within a given phylum. In light of recent phylogenetic evidence that Ctenophora is the earliest-branching animal lineage, our results are consistent with the hypothesis that the ancient primary function of Sox family genes was to regulate the maintenance of stem cells and function in cell fate determination.publishedVersionPeer Reviewe

Non-excitable fluorescent protein orthologs found in ctenophores

Background: Fluorescent proteins are optically active proteins found across many clades in metazoans. A fluorescent protein was recently identified in a ctenophore, but this has been suggested to derive from a cnidarian, raising again the question of origins of this group of proteins. Results: Through analysis of transcriptome data from 30 ctenophores, we identified a member of an orthologous group of proteins similar to fluorescent proteins in each of them, as well as in the genome of Mnemiopsis leidyi. These orthologs lack canonical residues involved in chromophore formation, suggesting another function. Conclusions: The phylogenetic position of the ctenophore protein family among fluorescent proteins suggests that this gene was present in the common ancestor of all ctenophores and that the fluorescent protein previously found in a ctenophore actually derives from a siphonophore

Effect of plant chemical variation and mutualistic ants on the local population genetic structure of an aphid herbivore

Plants exhibit impressive genetic and chemical diversity, not just between species but also within species, and the importance of plant intraspecific variation for structuring ecological communities is well known. When there is variation at the local population level, this can create a spatially heterogeneous habitat for specialised herbivores potentially leading to non-random distribution of individuals across host plants. Plant variation can affect herbivores directly and indirectly via a third species, resulting in variable herbivore growth rates across different host plants. Herbivores also exhibit within-species variation, with some genotypes better adapted to some plant variants than others. We genotyped aphids collected across 2 years from a field site containing ~200 patchily distributed host plants that exhibit high chemical diversity. The distribution of aphid genotypes, their ant mutualists, and other predators was assessed across the plants. We present evidence that the local distribution of aphid (Metopeurum fuscoviride) genotypes across host-plant individuals is associated with variation in the plant volatiles (chemotypes) and non-volatile metabolites (metabotypes) of their host plant tansy (Tanacetum vulgare). Furthermore, these interactions in the field were influenced by plant-host preferences of aphid-mutualist ants. Our results emphasise that plant intraspecific variation can structure ecological communities not only at the species level but also at the genetic level within species and that this effect can be enhanced through indirect interactions with a third species