Hi,

We are currently performing in-vitro tissue bath experiments, but we recently had a problem with our experiment. In our lab, we performed a series of in-vitro organ bath experiments in the fundal and antral parts of the rats to detect the pathways by which electrical field stimulation (EFS) activates non-adrenergic non-cholinergic (NANC) pathway, in turn, causes muscle relaxation. For this purpose, we added 1 uM atropine and 1 uM guanethidine to the organ bath in the 15 min. preincubation phase. We then applied EFS (60 V, 16 Hz, 2ms, 30 sec.), but we observed muscle contraction instead of relaxation. When we increased the concentrations of atropine and guanethidine ten-fold and tried to apply EFS in different voltages and frequencies, we did not observe any significant change.

I will very appreciate  it if you give us any suggestion to solve this problem.