Noradrenergic depression of neuronal excitability in the entorhinal cortex via activation of TREK-2 K+ channels.
ABSTRACT The entorhinal cortex is closely associated with the consolidation and recall of memories, Alzheimer disease, schizophrenia, and temporal lobe epilepsy. Norepinephrine is a neurotransmitter that plays a significant role in these physiological functions and neurological diseases. Whereas the entorhinal cortex receives profuse noradrenergic innervations from the locus coeruleus of the pons and expresses high densities of adrenergic receptors, the function of norepinephrine in the entorhinal cortex is still elusive. Accordingly, we examined the effects of norepinephrine on neuronal excitability in the entorhinal cortex and explored the underlying cellular and molecular mechanisms. Application of norepinephrine-generated hyperpolarization and decreased the excitability of the neurons in the superficial layers with no effects on neuronal excitability in the deep layers of the entorhinal cortex. Norepinephrine-induced hyperpolarization was mediated by alpha(2A) adrenergic receptors and required the functions of Galpha(i) proteins, adenylyl cyclase, and protein kinase A. Norepinephrine-mediated depression on neuronal excitability was mediated by activation of TREK-2, a type of two-pore domain K(+) channel, and mutation of the protein kinase A phosphorylation site on TREK-2 channels annulled the effects of norepinephrine. Our results indicate a novel action mode in which norepinephrine depresses neuronal excitability in the entorhinal cortex by disinhibiting protein kinase A-mediated tonic inhibition of TREK-2 channels.
- SourceAvailable from: Dina Simkin[show abstract] [hide abstract]
ABSTRACT: TREK-2 expressed in mammalian cells exhibits small ( approximately 52 pS) and large ( approximately 220 pS) unitary conductance levels. Here we tested the role of the N-terminus (69 amino acids long) in the control of the unitary conductance, and role of the alternative translation initiation as a mechanism that produces isoforms of TREK-2 that show different conductance levels. Deletion of the first half (Delta1-36) of the N-terminus had no effect. However, deletion of most of the N-terminus (Delta1-66) resulted in the appearance of only the large-conductance channel ( approximately 220 pS). In support of the critical function of the distal half of the N-terminus, the deletion mutants Delta1-44 and Delta1-54 produced approximately 90 pS and 188 pS channels, respectively. In Western blot analysis, TREK-2 antibody detected two immunoreactive bands at approximately 54 kDa and approximately 60 kDa from cells expressing wild-type TREK-2 that has three potential translation initiation sites (designated M(1)M(2)M(3)) within the N-terminus. Mutation of the second and third initiation sites from Met to Leu (M(1)L(2)L(3)) produced only the approximately 60 kDa isoform and the small-conductance channel ( approximately 52 pS). Mutants designed to produce translation from the second (M(2)L(3)) or third (M(3)) initiation site produced the approximately 54 kDa isoform, and the large conductance channel ( approximately 185-224 pS). M(1)L(2)L(3), M(2)L(3) and M(3) were relatively selectively permeable to K(+), as judged by the 51-55 mV shifts in reversal potential following a 10-fold change in [K(+)](o). P(Na)/P(K) values were also similar for M(1)L(2)L(3) ( approximately 0.02), M(2)L(3) ( approximately 0.02) and M(3) ( approximately 0.03). Arachidonic acid, proton and membrane stretch activated, whereas dibutyryl-cAMP inhibited all three isoforms of TREK-2, indicating that deletion of the N-terminus does not abolish modulation. These results show that the small and large conductance TREK-2 channels are produced as a result of alternative translation initiation, producing isoforms with long and short N-termini, and that the distal half of the N-terminus controls the unitary conductance.The Journal of Physiology 11/2008; 586(Pt 23):5651-63. · 4.38 Impact Factor
- [show abstract] [hide abstract]
ABSTRACT: Species differences in the distribution of beta-adrenergic receptors in the hippocampal and retrohippocampal regions of rats and guinea pigs were examined using in vitro autoradiographic techniques. beta 1-receptors were found in the hippocampal area CA1 of both species, although guinea pigs had significantly lower receptor densities in comparison to rats. In guinea pigs, beta 2-adrenergic receptors were predominant in hippocampal area CA1. Hippocampal area CA3 had very low levels of beta 1- and beta 2-receptors in both species. The retrohippocampal area was also found to have a distinct topographic distribution of beta-receptors. In rats, the subiculum and parasubiculum (layers II-III) were heavily labeled for beta 1-receptors; in contrast, guinea pigs had few receptors in these regions. beta 2-receptors were particularly prominent in the parasubicular region in rats. The entorhinal cortex laminae was found to contain beta-receptors in both rats and guinea pigs. Immunohistochemical techniques were used to compare the pattern of catecholaminergic innervation with the receptor distribution within each hippocampal subregion. Despite the general lack of beta-receptors in area CA3, abundant catecholamine immunoreactive fibers were observed in CA3 of rat and guinea pig hippocampus. Significant species differences were found in the distribution of hippocampal beta-adrenergic receptor subtypes, and moreover, in both species the distribution of beta-adrenergic receptors did not coincide with the pattern of hippocampal adrenergic innervation.Synapse 04/1993; 13(3):206-14. · 2.31 Impact Factor
- [show abstract] [hide abstract]
ABSTRACT: The adrenergic system is an essential regulator of neuronal, endocrine, cardiovascular, vegetative, and metabolic functions. The endogenous catecholamines epinephrine and norepinephrine activate G-protein-coupled receptors to transmit their signal across the plasma membrane. These adrenoceptors can be divided into three different groups: the alpha(1)-receptors (alpha(1A), alpha(1B), alpha(1D)), alpha(2)-receptors (alpha(2A), alpha(2B), alpha(2C)), and beta-receptors (beta(1), beta(2), beta(3)). This review summarizes recent findings in the field of adrenoceptor signaling in neurons and includes a discussion of receptor-associated proteins, receptor dimerization, subcellular trafficking, and fluorescence optical methods for studying the kinetics of adrenergic signaling. Spatio-temporal imaging may become an important future tool for identifying the physiological significance of these complex signaling mechanisms in vivo. Gene-targeted mouse models carrying deletions in alpha(2)-adrenoceptor have provided detailed insights into specific neuronal functions of the three alpha(2)-receptor subtypes.Cell and Tissue Research 12/2006; 326(2):541-51. · 3.68 Impact Factor