Project description:CRISPR/Cas9 mediated gene knockout show that nanos2+ progenitor cells are indispensable for male and female germline maintenance in Nematostella. This suggests nanos and piwi genes have a conserved role in somatic and germline stem cells in cnidarians.
Project description:MicroRNAs of bilaterian animals undergo posttranscriptional modifications such as methylation, tailing and trimming that regulate miRNA stability and function. To gain insight on the evolution of miRNA posttranscriptional modification, we studied regulation of miRNA stability by methylation in the sea anemone Nematostella vectensis, a representative of Cnidaria, the sister group of Bilateria.
Project description:NvNcol3::mOrange2 is a stable transgenic line that labels cnidocytes(stinging cells) of the sea anemone Nematostella vectensis (Nakanishi et al., Development 2012). Two week old primary polyps were dissociated and the NvNcol3::mOrange2 positive and negative cells were enriched by FACS.
Project description:Bilaterian animals have evolved complex sensory organs comprised of distinct cell types that function coordinately to sense the environment. Each sensory unit has a defined architecture built from component cell types, including sensory cells, non-sensory support cells, and dedicated sensory neurons. Whether this characteristic cellular composition is present in the sensory organs of non-bilaterian animals is unknown. Here, we interrogate the cell type composition and gene regulatory networks controlling development of the larval apical sensory organ in the sea anemone Nematostella vectensis. Using single cell RNA sequencing and imaging approaches, we reveal two unique cell types in the Nematostella apical sensory organ, GABAergic sensory cells and a putative non-sensory support cell population. Further, we identify the paired-like (PRD) homeodomain gene prd146 as a specific sensory cell marker and show that Prd146+ sensory cells become post-mitotic after gastrulation. Genetic loss of function approaches show that Prd146 is essential for apical sensory organ development. Using a candidate gene knockdown approach, we place prd146 downstream of FGF signaling in the apical sensory organ gene regulatory network. Further, we demonstrate that an aboral FGF activity gradient coordinately regulates the specification of both sensory and support cells. Collectively, these experiments define the genetic basis for apical sensory organ development in a non-bilaterian animal and reveal an unanticipated degree of complexity in a prototypic sensory structure.
Project description:NvElav1::mOrange is a stable transgenic line that labels a large fraction of the nervous system of the sea anemone Nematostella vectensis (Nakanishi et al., Development 2012). Two week old primary polyps were dissociated and the NvElav1::mOrange positive cells were enriched by FACS.
Project description:The aim of this part of the wider project is to identify neuropeptide precursors, investigate cleavage sites on neuropeptide precursors and predict mature peptides in the sea anemone Nematostella vectensis. This was done to create a synthetic library of N. vectensis neuropeptides which were then used to test neuropeptide receptor candidates for activation by the different peptides.
Project description:We assessed genome-wide temporal transcript expression patterns in the sea anemone, Nematostella vectensis, in Great Sippewissett Marsh in Massachusetts, where anemones experienced a natural light cycle with intensity varying from 0-200 lum/ft2, daily temperature fluctuations of ~9C. We measured ‘in situ’ gene expression from recaptured anemones every hour from 0800 to 1700 and identified six time-dependent gene clusters, represented by several genes involved in metabolism, stress, and transcription-translation related functions.