measurement-ion-concentration-unstirred-boundary-layer-with-open

Measurement of Ion Concentration in the Unstirred Boundary Layer with Open Patch-Clamp Pipette: Implications in Control of Ion Channels by Fluid Flow

Fluid flow is an important environmental stimulus that controls many physiological and pathological processes, such as fluid flow-induced vasodilation. ...

Konkuk University School of Medicine

MK801 and ketamine, which are phencyclidine (PCP) derivative N-methyl-d-aspartate receptor (NMDAr) blockers, reportedly enhance the function of 5-hydroxytryptamine (HT)-2A receptors (5-HTRs). Both are believed to directly affect the pathogenesis of schizophrenia, as well as hypertension. 5-HTR signaling involves the inhibition of Kv conductance. This study investigated the interaction of these drugs with Kv1.5, which plays important roles in 5-HTR signaling and in regulating the excitability of the cardiovascular and nervous system, and the potential role of this interaction in the enhancement of the 5-HTR-mediated response. Using isometric organ bath experiments with arterial rings and conventional whole-cell patch-clamp recording of Chinese hamster ovary (CHO) cells ectopically overexpressing Kv1.5, we examined the effect of ketamine and MK801 on 5-HTR-mediated vasocontraction and Kv1.5 channels. Both ketamine and MK801 potentiated 5-HTR-mediated vasocontraction. This potentiation of 5-HTR function occurred in a membrane potential-dependent manner, indicating the involvement of ion channel(s). Both ketamine and MK801 rapidly and directly inhibited Kv1.5 channels from the extracellular side independently of NMDArs. The potencies of MK801 in facilitating the 5-HTR-mediated response and blocking Kv1.5 were higher than those of ketamine. Our data demonstrated the direct inhibition of Kv1.5 channels by MK801/ketamine and indicated that this inhibition may potentiate the functions of 5-HTRs. We suggest that 5-HTR-Kv1.5 may serve as a receptor-effector module in response to 5-HT and is a promising target in the pathogenesis of MK801-/ketamine-induced disease states such as hypertension and schizophrenia.

Enhancement of 5-HT receptor function and blockade of Kv1.5 by MK801 and ketamine: implications for PCP derivative-induced disease models.

Oxidative stress is associated with many cardiovascular diseases, such as hypertension and arteriosclerosis. Oxidative stress reportedly activates the L-type voltage-gated calcium channel (VDCC) and elevates [Ca] in many cells. However, how oxidative stress activates VDCC under clinical setting and the consequence for arteries are unclear. Here, we examined the hypothesis that hydrogen peroxide (HO) regulates membrane potential (Em) by altering Na influx through cation channels, which consequently activates VDCC to induce vasoconstriction in rat mesenteric arteries. To measure the tone of the endothelium-denuded arteries, a conventional isometric organ chamber was used. Membrane currents and Em were recorded by the patch-clamp technique. [Ca] and [Na] were measured with microfluorometry using Fura2-AM and SBFI-AM, respectively. We found that HO (10 and 100 µM) increased arterial contraction, and nifedipine blocked the effects of HO on isometric contraction. HO increased [Ca] as well as [Na], and depolarized Em. Gd (1 µM) blocked all these HO-induced effects including Em depolarization and increases in [Ca] and [Na]. Although both nifedipine (30 nM) and low Na bath solution completely prevented the HO-induced increase in [Na], they only partly inhibited the HO-induced effects on [Ca] and Em. Taken together, the results suggested that HO constricts rat arteries by causing Em depolarization and VDCC activation through activating Gd-and nifedipine-sensitive, Na-permeable channels as well as Gd-sensitive Ca-permeable cation channels. We suggest that unidentified Na-permeable cation channels as well as Ca-permeable cation channels may function as important mediators for oxidative stress-induced vascular dysfunction.

Hydrogen peroxide constricts rat arteries by activating Na-permeable and Ca-permeable cation channels.

Estrogen has diverse effects on cardiovascular function, including regulation of the contractile response to vasoactive substances such as serotonin. The serotonin system recently emerged as an important player in the regulation of vascular tone in humans. However, hyperreactivity to serotonin appears to be a critical factor for the pathophysiology of hypertension. In this study, we examined the modulatory mechanisms of estrogen in serotonin-induced vasoconstriction by using a combinatory approach of isometric tension measurements, molecular biology, and patch-clamp techniques. 17β-Estradiol (E2) elicited a significant and concentration-dependent relaxation of serotonin-induced contraction in deendothelialized aortic strips isolated from male rats. E2 triggered a relaxation of serotonin-induced contraction even in the presence of tamoxifen, an estrogen receptor antagonist, suggesting that E2-induced changes are not mediated by estrogen receptor. Patch-clamp studies in rat arterial myocytes showed that E2 prevented Kv channel inhibition induced by serotonin. Serotonin increased Src activation in arterial smooth muscle required for contraction, which was significantly inhibited by E2. The estrogen receptor-independent inhibition of Src by E2 was confirmed in HEK293T cells that do not express estrogen receptor. Taken together, these results suggest that estrogen exerts vasodilatory effects on serotonin-precontracted arteries via Src, implying a critical role for estrogen in the prevention of vascular hyperreactivity to serotonin.

Article	Year
Enhancement of 5-HT receptor function and blockade of Kv1.5 by MK801 and ketamine: implications for PCP derivative-induced disease models. Experimental & molecular medicine\| PubMed ID: 29700292	2018
Hydrogen peroxide constricts rat arteries by activating Na-permeable and Ca-permeable cation channels. Free radical research\| PubMed ID: 30526150	2018
Estrogen modulates serotonin effects on vasoconstriction through Src inhibition. Experimental & molecular medicine\| PubMed ID: 30559345	2018

Jae Gon Kim

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