Role of sarcoplasmic reticulum in inhibitory junction potentials and hyperpolarizations by nitric oxide donors in opossum oesophagus.

1. Previous patch clamp studies of oesophageal circular muscle cells showed that nitric oxide (NO) modulated the opening of Ca2(+)-activated K+ channels involved in mediating the inhibitory junction potentials (i.j.ps). This study clarified the role of Ca2+ release from the superficial sarcoplasmic reticulum (SR) in the mechanism of i.j.ps or hyperpolarizing responses to NO-releasing compounds. Electrical and mechanical activities were simultaneously recorded by intracellular microelectrode or double sucrose gap techniques. 2. The NO-donors, sydnonimine (SIN-1) and sodium nitroprusside, each at 500 microM, hyperpolarized oesophageal circular muscle cells by 15-20 mV, like i.j.ps. 3. The selective inhibitors of SR Ca2(+)-ATPase (cyclopiazonic acid 10-30 microM and thapsigargin 5 microM) and the SR Ca2+ release channel activator (ryanodine 30 microM) caused depolarization and spontaneous contractions which were diminished after prolonged (> 30 min) incubation with these agents in Ca2(+)-containing medium. Moreover, these agents inhibited both the i.j.p. and NO-donor hyperpolarizations, suggesting that a functional SR Ca2+ uptake is necessary for the response to endogenous or exogenous NO. 4. These results, along with our previous findings of the dependence of i.j.ps and NO-donor hyperpolarizations on K+ channel activation and cyclic GMP elevation, support the hypothesis that subplasmalemmal (Ca2+)i elevation, via vectorial Ca2+ release from superficial SR toward the plasmalemma, may be an important mechanism by which NO, from NO-liberating compounds or released from inhibitory neurones induces relaxation and i.j.ps in opossum oesophagus.