NDLI: Cultured trout gill epithelia enriched in pavement cells or in mitochondria-rich cells provides insights into Na$^{+}$ and Ca$^{2+}$ transport

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Cultured trout gill epithelia enriched in pavement cells or in mitochondria-rich cells provides insights into Na$^{+}$ and Ca$^{2+}$ transport

Content Provider	Springer Nature Link
Author	Galvez, Fernando Tsui, Tommy Wood, Chris M.
Copyright Year	2008
Abstract	The lack of a suitable flat epithelial preparation isolated directly from the freshwater fish gill has led, in recent years, to the development of cultured gill epithelia on semipermeable supports. To date, their minimal capacity to actively transport ions has limited their utility as ionoregulatory models. The current study describes a new method of culturing gill epithelia consisting either of an enriched population of pavement (PV) cells or a mixed population of PV cells and mitochondria-rich (MR) cells from the gills of adult rainbow trout. Although the cell culture approach is similar to the double-seeded insert (DSI) technique described previously, it makes use of Percoll density centrifugation to first separate populations of PV and MR cells, which are then seeded on cell culture supports in varying proportions on successive days so as to produce preparations enriched in one or the other cell types. Based on rhodamine staining, the MR cell-rich epithelia exhibited a threefold higher enrichment of MR cells compared to traditional DSI preparations. In general, MR cell-rich epithelia developed extremely high transepithelial resistances (TER; >30 kΩ cm$^{2}$) and positive transepithelial potentials (TEP) under symmetrical conditions (i.e., L15 medium on both apical and basolateral sides). Apical exposure of cell cultures to freshwater reduced TER and produced a negative TEP in all the epithelial preparations, although MR cell-rich epithelia maintained relatively high TER and negative TEP for over 2 d under these asymmetrical conditions. Measurement of unidirectional Na$^{+}$ fluxes and application of the Ussing flux ratio criterion demonstrated active Na$^{+}$ uptake in PV cell-rich and MR cell-rich epithelia under both symmetrical and asymmetrical conditions. In comparison, Ca$^{2+}$ uptake and Na$^{+}$/K$^{+}$-ATPase activity were significantly elevated in MR cell-rich preparations relative to the traditional DSI or PV cell-rich cultures under symmetrical conditions. This new methodology enhances our ability to tailor cultured gill epithelia on semipermeable supports with different proportions of PV cells and MR cells, thereby illuminating the ionoregulatory functions of the two cell types.
Starting Page	415
Ending Page	425
Page Count	11
File Format	PDF
ISSN	10712690
Journal	In Vitro Cellular & Developmental Biology - Animal
Volume Number	44
Issue Number	10
e-ISSN	1543706X
Language	English
Publisher	Springer-Verlag
Publisher Date	2008-09-23
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Mitochondria-rich cells Pavement cells Ussing flux criterion Transepithelial resistance Transepithelial potential Animal Genetics and Genomics Cell Culture Stem Cells Developmental Biology Cell Biology
Content Type	Text
Resource Type	Article
Subject	Cell Biology Developmental Biology Medicine

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Cultured trout gill epithelia enriched in pavement cells or in mitochondria-rich cells provides insights into Na$^{+}$ and Ca$^{2+}$ transport

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Cultured trout gill epithelia enriched in pavement cells or in mitochondria-rich cells provides insights into Na$^{+}$ and Ca$^{2+}$ transport