Rapid isolation of chloride cells from pinfish gill

Abstract

Enriched fractions of chloride cells with good ultrastructural integrity have been obtained from gill filaments of the euryhaline teleost, Lagodon rhomboides. The branchial epithelium from seawater‐adapted fish was dissociated by gentle mechanical means in a Ca⁺⁺, Mg⁺⁺ ‐free balanced salt solution. Density gradient centrifugation of the mixed cell suspensions through a Ficoll gradient yielded a fraction containing between 50 and 70% chloride cells. This fraction showed a 3‐ to 4‐fold enrichment over comparable gill homogenate values for sodium plus potassium‐activated adenosinetriphosphatase, (Na⁺, K⁺‐ATPase), an enzyme concentrated in chloride cells. Isolation of chloride cells from fish adapted to one‐third seawater was less successful, due to the smaller size and reduced number of these cells, although fractions with at least a 2‐fold enrichment of the enzyme were obtained. These results continue to support the belief that chloride cells are responsible for the osmoregulatory activity associated with the branchial epithelium of teleosts and that this vital function is mediated through the activity of the transport associated enzyme, Na⁺, K⁺ ‐ATPase, the specific activity of which increases with osmotic stress.