Rosenthal, J. J. | Search bioRxiv

bioRxiv2025

Squid primary cell culture as a model system and experimental tool

Squid primary cells from various tissues and ages are isolated, maintained in culture, and express exogenous genes. This protocol opens up numerous opportunities in molecular biology, neuroscience, and marine biology, enabling molecular and cellular-level investigations into processes specific to squids, including their complex behaviors, rapid color change mechanisms, RNA editing capabilities, with broader implications in basic cell physiology. We also describe procedures that harness life-stage tractability in Euprymna berryi squids towards future studies of aging in this model marine organism. Highlights- Squid primary cells are isolated from optic lobes, gills, eyes, and the skin - Cells from these tissues can be dissociated from any life stage of the squid - Isolated cells can express exogenous genes through mRNA transfections and are subject to live and/or fixed cell imaging - Specific details for optimized media conditions, trypsinization, plating, and passaging are included

cell biology↗

bioRxiv2022

The FMRF-NH2 Gated Sodium Channel of Biomphalaria glabrata: Localization and Expression Following Infection by Schistosoma mansoni

The neglected tropical disease schistosomiasis impacts the lives of over 700 million people globally. Schistosoma mansoni, the trematode parasite that causes the most common type of schistosomiasis, requires planorbid pond snails of the genus Biomphalaria to support its larval development and transformation to the form that can infect humans. A greater understanding of neural signaling systems that are specific to the Biomphalaria intermediate host could lead to novel strategies for parasite or snail control. This study characterized a Biomphalaria glabrata neural receptor that is gated by the molluscan neuropeptide FMRF-NH2. The Biomphalaria glabrata FMRF-NH2 gated sodium channel (Bgl-FaNaC) amino acid sequence was highly conserved with FaNaCs found in related gastropods, especially the planorbid Planorbella trivolvis (91% sequence identity). In common with the P. trivolvis FaNaC, the B. glabrata receptor exhibited a low affinity (EC50: 3 x 10-4 M) and high specificity for the FMRF-NH2 agonist. Its expression in the central nervous system, detected by immunohistochemistry and in situ hybridization, was widespread, with the protein localized mainly to neuronal fibers and the mRNA confined to cell bodies. Colocalization was observed with the FMRF-NH2 tetrapeptide precursor in some neurons associated with male mating behavior. At the mRNA level, Bgl-FaNaC expression in the visceral and left parietal ganglia decreased at 20 days post infection by S. mansoni and in the shedding phase. Altered FMRF-NH2 signaling could be vital for parasite survival and proliferation in its snail intermediate host.

neuroscience↗