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Cation ions modulate the ACh--sensitive current in type Ⅱ vestibular hair cells of guinea pigs

Guo Changkai, Zhang Song, Kong Weijia, Li Qingtian, Li Zhiwang

Department of Otolaryngology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology.Wuhan 430022,Hubei；China

Abstract

Molecular biological studies and electrophysiological data have demonstrated that acetylcholine （ACh） is the principal cochlear and vestibular efferent neurotransmitter among mammalians. However, the functional roles of ACh in type Ⅱ vestibular hair cells among mammalians are still unclear, with the exception of the well-known #alpha#9-containing nicotinic ACh receptor （#alpha#9-nAChR） in cochlear hair cells and frog saccular hair cells. In this study, the properties of the ACh-sensitive current were investigated by whole-cell patch clamp technique in isolated type Ⅱ vestibular hair cells of guinea pigs. The direct effect of extracellular ACh was to induce a hyperpolarization effect in type Ⅱ vestibular hair cells. Type Ⅱ vestibular hair cells displayed a sustained outward current in response to the perfusion of ACh. It took about 60 s for the ACh-sensitive current to get a complete re-activation. The reversal potential of the ACh-sensitive current was （-66±8） mV, which indicated that potassium ion was the main carrier of this current. The blocking effect by the submillimolar concentration of tetraethylammonium （TEA） further indicated that extracellular ACh stimulated the calcium-dependent potassium current. Following replacement of the compartment of NaCl in the normal external solution with TrisCl, LiCl or saccharose respectively, the amplitude of the ACh-sensitive current was not affected. Blocking of the release of intracellular Ca~(2+) stores by intracellular application of heparin failed to inhibit the ACh-sensitive current. Therefore, extracellular Na~(+) and the inositol 1,4,5-trisphosphate （IP_(3)）- dependent intracellular Ca~(2+) release were not involved in the activation of the ACh-sensitive current. However, the ACh-sensitive current was strongly affected by the concentration of the extracellular K~(+), extracellular Ca~(2+) and intracellular Mg~(2+). The amplitude of the ACh-sensitive current was strongly inhibited by high concentration of extracellular K~(+). In the Ca~(2+)-free external solution, ACh only activated a very small current; however, the ACh-sensitive current demonstrated a Ca~(2+)-dependent inhibition effect in high concentration of Ca~(2+) solution. In addition, the ACh-sensitive current was inhibited by increasing of the concentration of intracellular Mg~(2+). In conclusion, the present results demonstrate that ACh plays an important role in the vestibular efferent system. The fact that Na~(+) is not involved in the ACh-sensitive current will not favor the well-known profile of #alpha#9-nAChR, which is reported to display a small but important permeability to Na~(+). It is also suggested that, in vivo, the amplitude of the ACh-induced hyperpolarization may strongly depend on the concentration of extracellular Ca~(2+) and intracellular Mg~(2+).

Key words: vestibule;hair cells;Acetylcholine;Ca~(2+)-dependent K~(+) currents;Modulation

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Citing This Article：

Guo Changkai, Zhang Song, Kong Weijia, Li Qingtian, Li Zhiwang. Cation ions modulate the ACh--sensitive current in type Ⅱ vestibular hair cells of guinea pigs. Acta Physiol Sin 2006; 58 (2): (in Chinese with English abstract).