RNA isolation from corals and other cnidarian species using urea-LiCl as a denaturant.

A method of RNA isolation using a solution of urea-LiCl as a denaturing agent was tested on stony coral. As the method does not require homogenization of tissues prior to their incubation in the denaturant, specimen collected in the field can be immediately transferred to the urea-LiCl solution. The method was also tested on tissues of other cnidarian species.

A SLC6 transporter cloned from the lion's mane jellyfish (Cnidaria, Scyphozoa) is expressed in neurons.

In the course of recent comparative genomic studies conducted on nervous systems across the phylogeny, current thinking is leaning in favor of more heterogeneity among nervous systems than what was initially expected. The isolation and characterization of molecular components that constitute the cnidarian neuron is not only of interest to the physiologist but also, on a larger scale, to those who study the evolution of nervous systems. Understanding the function of those ancient neurons involves the identification of neurotransmitters and their precursors, the description of nutrients used by neurons for metabolic purposes and the identification of integral membrane proteins that bind to those compounds.

Semantic Assembly and Annotation of Draft RNAseq Transcripts without a Reference Genome.

Transcriptomes are one of the first sources of high-throughput genomic data that have benefitted from the introduction of Next-Gen Sequencing. As sequencing technology becomes more accessible, transcriptome sequencing is applicable to multiple organisms for which genome sequences are unavailable. Currently all methods for de novo assembly are based on the concept of matching the nucleotide context overlapping between short fragments-reads.

Immunolocalization of a voltage-gated calcium channel β subunit in the tentacles and cnidocytes of the Portuguese man-of-war, Physalia physalis.

This study investigated the localization of a voltage-gated calcium channel (VGCC) β subunit in the tentacles and cnidocytes of the Portuguese man-of-war using confocal immunocytochemistry. An antibody specific to the Ca(2+) channel β subunit of the Portuguese-man-of-war (PpCaVβ) was generated, and characterized by Western immunoblotting. The antibody labeling was widespread in the ectoderm of cnidosacs of the tentacles.

Escape variants of the XPR1 gammaretrovirus receptor are rare due to reliance on a splice donor site and a short hypervariable loop.

Entry determinants in the XPR1 receptor for the xenotropic/polytropic mouse leukemia viruses (XP-MLVs) lie in its third and fourth putative extracellular loops (ECLs). The critical ECL3 receptor determinant overlies a splice donor and is evolutionarily conserved in vertebrate XPR1 genes; 2 of the 3 rare replacement mutations at this site destroy this receptor determinant. The 13 residue ECL4 is hypervariable, and replacement mutations carrying an intact ECL3 site alter but do not abolish receptor activity, including replacement of the entire loop with that of a jellyfish (Cnidaria) XPR1.

The regulation of cnidocyte discharge.

Because cnidocytes are exceedingly complex cells which can only be used once, their discharge is highly regulated by way of a variety of chemosensory, mechanosensory and endogenous pathways. The integration of these various inputs ultimately results in exocytosis and then discharge of the cnidocyte's diagnostic organelle, the cnidocyst. Here we review what is known about the sensory pathways that regulate cnidocytes, the electrical events that manifest in cnidocytes following sensory stimulation and the ionic mechanisms that underlie discharge.

Identification of a novel aminergic-like G protein-coupled receptor in the cnidarian Renilla koellikeri.

Biogenic amines exert various physiological effects in cnidarians, but the receptors involved in these responses are not known. We have cloned a novel G protein-coupled receptor cDNA from an anthozoan, the sea pansy Renilla koellikeri, that shows homology to mammalian catecholamine receptors and, to a lesser extent, to peptidergic receptors. This putative receptor, named Ren2, has a DRC pattern that replaces the well-conserved DRY motif on the cytoplasmic side of the transmembrane III and lacks the cysteine residues usually found in the second extracellular loop and C-terminus tail.

A new G protein-coupled receptor from a primitive metazoan shows homology with vertebrate aminergic receptors and displays constitutive activity in mammalian cells.

Biogenic amine receptors mediate wide-ranging hormonal and modulatory functions in vertebrates, but are largely unknown in primitive invertebrates. In a representative of the most basal multicellular animals possessing a nervous system, the cnidarian Renilla koellikeri, aminergic-like receptors were previously characterized pharmacologically and found to engender control of the animal's bioluminescent and peristaltic reactions. Using degenerate oligonucleotides in a RT-PCR strategy, we obtained a full-length cDNA encoding a polypeptide with typical G protein-coupled receptor (GPCR) characteristics and which displayed a significant degree of sequence similarity (up to 45%) to biogenic amine receptors, particularly dopamine and adrenergic receptors.