Novel Excitatory Effects of Adenosine Triphosphate on Contractile and Pacemaker Activity in Rabbit Urethral Smooth Muscle

Purpose

Adenosine triphosphate is thought to be an important neurotransmitter in urethral smooth muscle but its physiological role is still unclear. We characterized the effects of adenosine triphosphate on contractile and pacemaker activity in rabbit urethral smooth muscle.

Materials and Methods

Tension recordings were made from strips of rabbit proximal urethral smooth muscle. Membrane currents from freshly isolated smooth muscle cells and interstitial cells of Cajal were recorded using the patch clamp technique. Intracellular Ca²⁺ was measured using confocal microscopy.

Results

Exogenous application of adenosine triphosphate (10 μM) evoked robust contractions that were inhibited by the type 2 purinergic receptor blocker suramin (100 μM) and the selective type 2 purinergic Y1 receptor antagonist MRS2500 (Tocris Bioscience, Ellisville, Missouri) (100 nM). Application of the type 2 purinergic Y receptor agonist 2-MeSADP (1 μM) mimicked the effects of adenosine triphosphate. When smooth muscle cells were studied under voltage clamp at −60 mV, adenosine triphosphate evoked a large single inward current (greater than 1.2 nA) but 2-MeSADP produced a small current (about 16 pA). In contrast, when interstitial cells of Cajal were held at −60 mV, they showed spontaneous transient inward currents that were increased in frequency by adenosine triphosphate and 2-MeSADP. These excitatory effects were inhibited by suramin and MRS2500. Interstitial cells of Cajal showed spontaneous Ca²⁺ waves that were increased in frequency by adenosine triphosphate and 2-MeSADP. These effects were also inhibited by suramin and MRS2500.

Conclusions

Contractile effects of adenosine triphosphate in urethral smooth muscle are mediated by the activation of type 2 purinergic Y receptors on interstitial cells of Cajal.

Key Words: urethra, muscle, smooth, urinary incontinence, muscle contraction, adenosine triphosphate

Abbreviations and Acronyms: 2-MeSADP, 2 methylthio adenosine diphosphate, ATP, adenosine triphosphate, AVP, arginine vasopressin, EFS, electrical field stimulation, ICC, interstitial cells of Cajal, NO-ARG, N^G-nitro-L-arginine, P2, type 2 purinergic, SMC, smooth muscle cell, STIC, spontaneous transient inward current