We have developed a method for the primary culture of gill epithelial cells from freshwater rainbow trout on permeable supports, polyethylene terephthalate membranes ('filter inserts'). Primary cultures of gill cells (6-9 days in Leibowitz L-15 culture medium plus foetal bovine serum and glutamine) are trypsinized and the cells seeded onto the inserts. After 6 days of growth with L-15 medium on both surfaces (approximately isotonic to trout plasma), the cells form a tight epithelium as judged from a progressive rise in transepithelial resistance which reaches a stable plateau for a further 6 days, as long as L-15 exposure is continued on both surfaces. The cultured epithelium (approximately 8 µm thick) typically consists of 2-4 overlapping cell layers organized as in the lamellae in vivo, with large intercellular spaces, multiple desmosomes and putative tight junctions. The cells appear to be exclusively pavement-type cells with an apical surface glycocalyx, an abundance of rough endoplasmic reticulum, no selective DASPEI staining and relatively few mitochondria. Transepithelial resistance (approximately 3.5 k cm2), permeability to a paracellular marker (polyethylene glycol-4000; 0.17x10(-6) cm s-1) and unidirectional flux of Na+ and Cl- (approximately 300 nmol cm-2 h-1) all appear realistic because they compare well with in vivo values; net fluxes of Na+ and Cl- are zero. The preparation acidifies the apical medium, which accumulates a greater concentration of ammonia. Upon exposure to apical freshwater, resistance increases six- to elevenfold and a basolateral-negative transepithelial potential (TEP) develops as in vivo. These responses occur even when mannitol is used to prevent changes in apical osmotic pressure. Net Na+ and Cl- loss rates are low over the first 12 h (-125 nmol cm-2 h-1) but increase substantially by 48 h. The elevated resistance and negative TEP gradually attenuate but remain significantly higher than pre-exposure values after 48 h of apical freshwater exposure. The preparation may provide a valuable new tool for characterizing some of the mechanisms of active and passive ion transport in the pavement cells of the freshwater gill.
In the field: an interview with Harald Wolf
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In our new Conversation, Harald Wolf talks about his fieldwork experiences working with desert ants in Tunisia to understand their navigation.
Propose a new Workshop
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Our Workshops bring together leading experts and early-career researchers from a range of scientific backgrounds. Applications are now open to propose Workshops for 2024, one of which will be held in a Global South country.
Julian Dow steps down and John Terblanche joins the JEB team
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After 15 years with the journal, Julian Dow from University of Glasgow, UK, is stepping down as a Monitoring Editor. We wish Julian all the best for the future and welcome John Terblanche, Stellenbosch University, South Africa, who is joining the team. Julian talks about his long association with The Company of Biologists and the journal and John tells us about his life and career in this News article.
The capture of crude oil droplets by filter feeders at high and low Reynolds numbers
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Researchers from the University of Montreal, Canada, reveal how tiny filter feeding barnacles and Daphnia entrap and consume minute droplets of crude oil, introducing the pollutant at the bottom of the food chain.
Patterns and processes in amphibious fish
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In their Review, Keegan Lutek, Cassandra Donatelli and Emily Standen discuss the biomechanics and neural control of terrestrial locomotion in amphibious fish. They explore how locomotor mode depends on body shape, physical constraints and phylogeny.
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