Functional characterisation and subcellular localisation of HCN1 channels in rabbit retinal rod photoreceptors.
Gating of voltage-dependent conductances in retinal photoreceptors is the first step of a process leading to the enhancement of the temporal performance of the visual system. The molecular components underlying voltage-dependent gating in rods are presently poorly defined. In the present work we have investigated the isoform composition and the functional characteristics of hyperpolarisation-activated cyclic nucleotide-gated channels (HCN) in rabbit rods. Using immunocytochemistry we show the expression in the inner segment and cell body of the isoform 1 (HCN1). Electrophysiological investigations show that hyperpolarisation-activated currents (I(h)) can be measured only from the cell regions where HCN1 is expressed. Half-activation voltage (-75.0 +/- 0.3 mV) and kinetics (t(1/2) of 101 +/- 8 ms at -110 mV and 20 degrees C) of the I(h) in rods are similar to those of the macroscopic current carried by homomeric rabbit HCN1 channels expressed in HEK 293 cells. The homomeric nature of HCN1 channels in rods is compatible with the observation that cAMP induces a small shift (2.3 +/- 0.8 mV) in the half-activation voltage of I(h). In addition, the observation that within the physiological range of membrane potentials, cAMP does not significantly affect the gain of the current-to-voltage conversion, may reflect the need to protect the first step in the processing of visual signals from changes in cAMP turnover.
Demontis
Gian Carlo
GC
Dipartimento di Psichiatria, Neurobiologia, Farmacologia e Biotecnologie, Università di Pisa, Via Bonanno, 6-56126 Pisa, Italy. demontis@farm.unipi.it
Moroni
Anna
A
Gravante
Biagio
B
Altomare
Claudia
C
Longoni
Biancamaria
B
Cervetto
Luigi
L
DiFrancesco
Dario
D
D000465:Algorithms; D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D000242:Cyclic AMP / Q000494:pharmacology; D054815:Cyclic Nucleotide-Gated Cation Channels; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007473:Ion Channels / Q000378:metabolism* / Q000648:ultrastructure; D007700:Kinetics; D008564:Membrane Potentials / Q000502:physiology; D018613:Microscopy, Confocal; D018408:Patch-Clamp Techniques; D020419:Photoreceptor Cells, Vertebrate / Q000378:metabolism* / Q000648:ultrastructure; D015221:Potassium Channels; D012333:RNA, Messenger / Q000096:biosynthesis; D011817:Rabbits; D017948:Retinal Rod Photoreceptor Cells / Q000378:metabolism* / Q000648:ultrastructure; D013347:Subcellular Fractions / Q000378:metabolism
D054815:Cyclic Nucleotide-Gated Cation Channels; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; D007473:Ion Channels; D015221:Potassium Channels; D012333:RNA, Messenger; D000242:Cyclic AMP