Stretch-Induced Increases in Intracellular Ca Stimulate Thick Ascending Limb O₂^- Production and Are Enhanced in Dahl Salt-Sensitive Rats.

ABSTRACT: Mechanical stretch raises intracellular Ca (Ca_i) in many cell types. Luminal flow-derived stretch stimulates O₂^- production by thick ascending limbs (THALs). Renal O₂^- is greater in Dahl salt-sensitive (SS) than salt-resistant (SR) rats. We hypothesized that mechanical stretch stimulates Ca influx via TRPV4 (transient receptor potential vanilloid type 4) which in turn raises Ca_i in THALs; these increases in Ca_i are necessary for stretch to augment O₂^- production; and stretch-stimulated, and therefore flow-induced, O₂^- production is enhanced in SS compared with SR THALs due to elevated Ca influx and increased Ca_i. Ca_i and O₂^- were measured in SS and SR THALs from rats on normal salt using Fura2-acetoxymethyl ester and dihydroethidium, respectively. Stretch raised Ca_i in SS by 270.4±48.9 nmol/L and by 123.6±27.0 nmol/L in SR THALs (P<0.02). Removing extracellular Ca eliminated the increases and differences in Ca_i between strains. Knocking down TRPV4 in SS THALs reduced stretch-induced Ca_i to SR levels (SS: 92.0±15.9 nmol/L; SR: 123.6±27.0 nmol/L). RN1734, a TRPV4 inhibitor, blunted stretch-elevated Ca_i by ?75% and ?66% in SS (P<0.03) and SR (P<0.04), respectively. Stretch augmented O₂^- production by 58.6±10.2 arbitrary fluorescent units/min in SS and by 24.4±2.6 arbitrary fluorescent units/min in SR THALs (P<0.05). Removal of extracellular Ca blunted stretch-induced increases in O₂^- and eliminated differences between strains. RN1734 reduced stretch-induced O₂^- by ?70% in SS (P<0.005) and ?60% in SR (P<0.01). Conclusions are as follows: (1) stretch activates TRPV4, which raises Ca_i in THALs; (2) the increase in Ca_i stimulates O₂^- production; and (3) stretch-induced O₂^- production is enhanced in SS THALs due to greater increases in Ca_i.

SUBMITTER: Saez F 

PROVIDER: S-EPMC7058965 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature