Abstract:
:We cloned 10 full-length variants of PIIH, the gene for I(h) from the spiny lobster, Panulirus interruptus, using reverse transcription-PCR (RT-PCR) and rapid amplification of cDNA ends (RACE). This gene shows a significant amount of alternative splicing in the S3-S4 and S4-S5 linkers, in the P-loop and the entire S6 transmembrane domain, in the cyclic nucleotide binding domain (CNBD), and near the 3' end of the gene. Functional expression of seven splice variants in Xenopus oocytes generated slowly activating hyperpolarization-activated inward currents, which were blocked by the I(h) channel blockers CsCl and ZD7288. The different splice variants had markedly varying activation kinetics and voltage dependence of activation. Bath application of 8-Br-cAMP shifted the V(1/2) to more positive potentials and accelerated the activation kinetics in an isoform-specific manner. Two variants containing a segment with an ER-retention motif in the S4-S5 loop did not produce currents in oocytes. Overexpression of one splice variant, PIIH AB(S)-I, in pyloric dilator (PD) neurons in the lobster stomatogastric ganglion produced an average threefold increase in I(h) without evoking a compensatory increase in I(A). The voltage for half-maximal activation of I(h) in PIIH AB(S)-I-expressing PDs was shifted in the depolarizing direction by 9 mV, whereas the slope factor decreased by 3.8 mV. Moreover, its activation kinetics were significantly faster than in control PDs. PIIH AB(S)-I overexpression enhanced PD neuron rhythmic firing in an amplitude-dependent manner above a minimal threshold two- to threefold increase in amplitude.
journal_name
J Neurophysioljournal_title
Journal of neurophysiologyauthors
Ouyang Q,Goeritz M,Harris-Warrick RMdoi
10.1152/jn.00246.2007subject
Has Abstractpub_date
2007-06-01 00:00:00pages
3880-92issue
6eissn
0022-3077issn
1522-1598pii
00246.2007journal_volume
97pub_type
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