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Fig. 1. Hypertonicity-induced cell volume change and the expression of
Na+/K+-ATPase,
Na+/K+/2Cl– cotransporter (NKCC) and
Na+/H+ exchanger-1 (NHE-1) in freshwater gill pavement
cells (PVCs). (A) Percoll-purified PVCs were incubated in either normal (317
mOsmol l–1) or hypertonic (500 mOsmol l–1)
medium. Blockers for three ion transporters (ouabain, bumetanide or EIPA) were
added to the cells before the application of hyperosmotic stress. The change
in cell volume was monitored using a Multisizer for a period of 60 min. Note
that the three blockers reduced the RVI response in the cells. (B) A primary
freshwater PVC culture was established. Over 80% of gill epithelial cells
attached after overnight incubation (Bi); the cell suspension was obtained
from trypsin digestion. (Bii) Gill epithelial cells cultured for 1 day. (C)
Cell lysates were obtained from 6 h hypertonic treatment (HT) of the PVC
culture and analysed by western blot (right). The amounts of the respective
ion transporter protein were quantified and tabulated in graphical form
(left). Significant induction of 
and ß subunits of
Na+/K+-ATPase, NKCC and NHE-1 were observed. (D) The
gene expression levels for Ostf1 in the control and hypertonic-treated cells.
*P<0.05 compared with the control. The results were obtained from
more than five independent experiments.
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