CL- CHANNELS IN BASOLATERAL RENAL MEDULLARY VESICLES .5. COMPARISON OF BASOLATERAL MTALH CL- CHANNELS WITH APICAL CL- CHANNELS FROM JEJUNUM AND TRACHEA

Cl- channels from basolaterally-enriched rabbit outer renal medullary membranes are activated either by increases in intracellular Cl- activity or by intracellular protein kinase A (PKA). Phosphorylation by PKA, however, is not obligatory for channel activity since channels can be activated by intracellular Cl- in the absence of PKA. The PKA requirement for activation of Cl- channels in certain secretory epithelia is. in contrast, obligatory. In the present studies, we examined the effects of PKA and intracellular Cl- concentrations on the properties of Cl- channels obtained either from basolaterally-enriched vesicles derived from highly purified suspensions of mouse medullary thick ascending limb (mTALH) segments, or from apical membrane vesicles obtained from two secretory epithelia. bovine trachea and rabbit small intestine. Our results indicate that the Cl- channels from mTALH suspensions were virtually identical to those previously described from rabbit outer renal medulla. In particular, an increase in intracellular (trans) Cl- concentration from 2 to 50 mm increased both channel activity (P0) and channel conductance (g(Cl), pS). Likewise, trans PKA increased mTALH Cl- channel activity by increasing the activity of individual channels when the trans solutions were 2 mm Cl. Under the latter circumstance, PKA did not activate quiescent channels, nor did it affect g(Cl). Moreover, when mTALH Cl- channels were inactivated by reducing cis Cl- concentrations to 50 mM, cis PKA addition did not affect P0. These results are consistent with the view that these Cl- channels originated from basolateral membranes of the mTALH. Cl- channels from apical vesicles from trachea and small intestine were completely insensitive to alterations in trans Cl- concentrations and demonstrated markedly different responses to PKA. In the absence of PKA, tracheal Cl- channels inactivated spontaneously after a mean time of 8 min, addition of PKA to trans solutions reactivated these channels. The intestinal Cl- channels did not inactivate with time. Trans PKA addition activated new channels with no effect on basal channel activity. Thus the regulation of Cl- channel activity by both intracellular Cl- and by PKA differ in basolateral mTALH Cl- channels compared to apical Cl- channels from either the tracheal or small intestine.