Voltage-dependent blockade of connexin40 gap junctions by spermine.
The effects of spermine and spermidine, endogenous polyamines that block many forms of ion channels, were investigated in homotypic connexin (Cx)-40 gap junctions expressed in N2A cells. Spermine blocked up to 95% of I(j) through homotypic Cx40 gap junctions in a concentration- and transjunctional voltage (V(j))-dependent manner. V(j) was varied from 5 to 50 mV in 5-mV steps and the dissociation constants (K(m)) were determined from spermine concentrations ranging from 10 micro M to 2 mM. The K(m) values ranged from 4.9 mM to 107 micro M for 8.6 < or = V(j) < or = 37.7 mV, within the physiological range of intracellular spermine for V(j) > or = 20 mV. The K(m) values for spermidine were > or = 5 mM. Estimates of the electrical distance (delta) for spermine (z = +4) and spermidine (z = +3) were 0.96 and 0.76 respectively. Cx40 single channel conductance was 129 pS in the presence of 2-mM spermine and channel open probability was significantly reduced in a V(j)-dependent manner. Similar concentrations of spermine did not block I(j) through homotypic Cx43 gap junctions, indicating that spermine selectively blocks Cx40 gap junctions. This is contrary to our previous findings that large tetraalkylammonium ions, also known to block several forms of ion channels, block junctional currents (I(j)) through homotypic connexin Cx40 and Cx43 gap junctions.
Musa
Hassan
H
Department of Pharmacology, SUNY Upstate Medical University, Syracuse, New York 13210, USA.
Veenstra
Richard D
RD
D000818:Animals; D018031:Connexin 43 / Q000187:drug effects / Q000502:physiology; D017630:Connexins / Q000037:antagonists & inhibitors* / Q000502:physiology*; D004305:Dose-Response Relationship, Drug; D004553:Electric Conductivity; D017629:Gap Junctions / Q000187:drug effects / Q000502:physiology; D018014:Gene Transfer Techniques; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology; D051379:Mice; D009447:Neuroblastoma / Q000503:physiopathology; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000187:drug effects / Q000502:physiology; D012680:Sensitivity and Specificity; D013095:Spermidine / Q000494:pharmacology*; D013096:Spermine / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D000097003:Gap Junction alpha-5 Protein
D018031:Connexin 43; D017630:Connexins; D011993:Recombinant Fusion Proteins; D013095:Spermidine; D013096:Spermine; D000097003:Gap Junction alpha-5 Protein