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Biophys J, January 2000, p. 1-12, Vol. 78, No. 1

Evidence for Cooperativity Between Nicotinic Acetylcholine Receptors in Patch Clamp Records

Asbed M. Keleshian,* Robert O. Edeson,dagger Guo-Jun Liu,Dagger and Barry W. MadsenDagger

 *Department of Physiology and Biophysics, State University of New York at Buffalo, Buffalo, New York 14214 USA;  dagger Department of Anaesthesia, Sir Charles Gairdner Hospital, Nedlands 6009, Western Australia; and  Dagger Department of Pharmacology, University of Western Australia, Nedlands 6907, Western Australia

It is often assumed that ion channels in cell membrane patches gate independently. However, in the present study nicotinic receptor patch clamp data obtained in cell-attached mode from embryonic chick myotubes suggest that the distribution of steady-state probabilities for conductance multiples arising from concurrent channel openings may not be binomial. In patches where up to four active channels were observed, the probabilities of two or more concurrent openings were greater than expected, suggesting positive cooperativity. For the case of two active channels, we extended the analysis by assuming that 1) individual receptors (not necessarily identical) could be modeled by a five-state (three closed and two open) continuous-time Markov process with equal agonist binding affinity at two recognition sites, and 2) cooperativity between channels could occur through instantaneous changes in specific transition rates in one channel following a change in conductance state of the neighboring channel. This allowed calculation of open and closed sojourn time density functions for either channel conditional on the neighboring channel being open or closed. Simulation studies of two channel systems, with channels being either independent or cooperative, nonidentical or identical, supported the discriminatory power of the optimization algorithm. The experimental results suggested that individual acetylcholine receptors were kinetically identical and that the open state of one channel increased the probability of opening of its neighbor.

Biophys J, January 2000, p. 1-12, Vol. 78, No. 1
© 2000 by the Biophysical Society   0006-3495/00/01/01/12  $2.00


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