• Biomolecules & Therapeutics
• /
• v.29 no.6
• /
• pp.630-636
• /
• 2021

Eupafolin, a constituent of the aerial parts of Phyla nodiflora, has neuroprotective property. Because reducing the synaptic release of glutamate is crucial to achieving pharmacotherapeutic effects of neuroprotectants, we investigated the effect of eupafolin on glutamate release in rat cerebrocortical synaptosomes and explored the possible mechanism. We discovered that eupafolin depressed 4-aminopyridine (4-AP)-induced glutamate release, and this phenomenon was prevented in the absence of extracellular calcium. Eupafolin inhibition of glutamate release from synaptic vesicles was confirmed through measurement of the release of the fluorescent dye FM 1-43. Eupafolin decreased 4-AP-induced [Ca²⁺]_i elevation and had no effect on synaptosomal membrane potential. The inhibition of P/Q-type Ca²⁺ channels reduced the decrease in glutamate release that was caused by eupafolin, and docking data revealed that eupafolin interacted with P/Q-type Ca²⁺ channels. Additionally, the inhibition of calcium/calmodulin-dependent protein kinase II (CaMKII) prevented the effect of eupafolin on evoked glutamate release. Eupafolin also reduced the 4-AP-induced activation of CaMK II and the subsequent phosphorylation of synapsin I, which is the main presynaptic target of CaMKII. Therefore, eupafolin suppresses P/Q-type Ca²⁺ channels and thereby inhibits CaMKII/synapsin I pathways and the release of glutamate from rat cerebrocortical synaptosomes.

• The Korean Journal of Physiology and Pharmacology
• /
• v.2 no.2
• /
• pp.225-232
• /
• 1998

To investigate the possible involvement of outward potassium ($K^+$) currents in nitric oxide-induced relaxation in intestinal smooth muscle, we used whole-cell patch clamp technique in freshly dispersed guinea-pig ileum longitudinal smooth muscle cells. When cells were held at -60 mV and depolarized from -40 mV to -50 mV in 10 mV increments, sustained outward $K^+$ currents were evoked. The outward $K^+$ currents were markedly increased by the addition of 10 ${\mu}M$ sodium nitroprusside (SNP). 10 ${\mu}M$ S-nitroso-N-acetylpenicillamine (SNAP) and 1 mM 8-Bromo-cyclic GMP (8-Br-cGMP) also showed a similar effect to that of SNP. 1 mM tetraethylammonium (TEA) significantly reduced depolarization-activated outward $K^+$ currents. SNP-enhanced outward $K^+$ currents were blocked by the application of TEA. High EGTA containing pipette solution (10 mM) reduced the control currents and also inhibited the SNP-enhanced outward $K^+$ currents. 5 mM 4-aminopyridine (4-AP) significantly reduced the control currents but showed no effect on SNP-enhanced outward $K^+$ currents. 0.3 ${\mu}M$ apamin and 10 ${\mu}M$ glibenclamide showed no effect on SNP-enhanced outward $K^+$ currents. 10 ${\mu}M$ 1H-[1,2,4]oxadiazolo [4,3-a]quinoxaline-1-one (ODQ), a specific inhibitor of soluble guanylate cyclase, significantly blocked SNP-enhanced $K^+$ currents. We conclude that NO donors activate the $Ca^{2+}-activated$ $K^+$ channels in guinea-pig ileal smooth muscle via activation of guanylate cyclase.

• The Korean Journal of Physiology and Pharmacology
• /
• v.1 no.2
• /
• pp.169-183
• /
• 1997

We have investigated the two types of voltage-dependent outward potassium (K) currents, i.e. delayed rectifier K current ($I_{K(V)}$) and 'A-like' transient outward K current ($I_{to}$) with patch-clamp technique in single smooth muscle cells (SMCs) isolated from rabbit basilar artery, and investigated the characteristics of them. The time-courses of activation were well fitted by exponential function raised to second power ($n^2$) in $I_{K(V)}$ and fourth power ($n^4$) in $I_{to}$. The activation, inactivation and recovery time courses of $I_{to}$ were much faster than that of $I_{K(V)}$. The steady-state activation and inactivation of $I_{K(V)}$ was at the more hyperpolarized range than that of $I_{to}$ contrary to the reports in other vascular SMCs. Tetraethylammonium chloride (TEA; 10 mM) markedly inhibited $I_{K(V)}$ but little affected $I_{to}$. 4-Aminopyridine (4-AP) had similar inhibitory potency on both currents. While a low concentration of $Cd^{2+}$ (0.5 mM) shifted the current- voltage relationship of $I_{to}$ to the positive direction without change of maximum conductance, $Cd^{2+}$ did not cause any appreciable change for $I_{K(V)}$.

• Journal of Veterinary Clinics
• /
• v.7 no.2
• /
• pp.501-509
• /
• 1990

This study was designed to examine effects of $\alpha$$_2$-Adrenergic Antagonists on blood chemical values in xylazine-sedated dogs. Twenty-four crossbred dogs of both sexes were intramusculary injected with a standard dosage of xylazine(2.2mg/kg of body weight). Righting reflex was uniformly lost and considered to be the point of maximum sedation. When the dogs were maximally sedated, tested groups were in-travenously injected with yohimbine 0.125mg/kg, 4-aminopyridine(4-AP) 0.3mg/kg, and a combination of yohimbine with 4-AP. Control group was intravenously 1 $m\ell$ of physiological saline solution. Total protein(T.P), albumin, glucose, aspartate aminotransferase(AST), alanine aminotrnasferase(ALT), blood urea nitrogen(BUN) were analyzed in the conditions of 0-, 30-, 60- and 120-minute after the administration of drugs. The results obtained in the study were as follows. 1. Changes of T.P, albumin, AST, ALT and BUN values in the control group were not significant during or after xylazine administration for at least 120minutes. 2. No changes of T.P, albumin, AST, ALT and BUN values in the tested groups were observed during or after $\alpha$$_2$-Adrenergic Antagonists treatment. 3. Serum glucose values of control group were getting remarkably increased after xylazine injection. 4. The xylasine-induced hyperglycemia was reversed in the dogs administrated with $\alpha$$_2$-Adrenergic Antagonists. Therefore, the results of the study show that the combined treatment with antagonists may be useful for accidental overdoses of xylazine and rapid reversal of animals sedated with xylazine.

• The Korean Journal of Physiology
• /
• v.23 no.2
• /
• pp.301-312
• /
• 1989

It well known that the magnitude of contraction and the shape of action potential depend upon the stimulation frequency and the duration of resting period (positive and negative staircase). Although the underlying mechanism of the staircase phenomenon is not fully understood, it has been suggested that staircase could be related to the intracllular $Ca^{2+}$ concentration. In order to elucidate the role of intracellular $Ca^{2+}$ on the contraction and action potential staircases, we examined the effects of 1 mM 4-aminopyridine (4-AP), 0.5 uM verapamil, 1 uM ryanodine, or reduction of extracellular Na concentration to 30% $(substituted\;by\;equimolar\;Li^+)$ in small atrial strips of the rabbit $(3{\times}10\;mm)$. The results obitained were as follows; 1) When the stimulation frequency was increased from 0.1 Hz to 2 Hz, positive staircase of the contraction and elevation of plateau level in action potential were found in control and the conditions of Na reduction and treatments of 4-AP, verapamil and ryanodine. 2) When stimulation frequency returned to 0.1 Hz from 1 min rest just after 2 Hz stimulation fer 1 min, the magnitudes of initial few contractions were larger than that of steady state contraction (post-rest potentiation) except, ryanodine or Na-reduction groups. 3) Negative staircase of contraction was developed in control and 4-AP group at post-rest 0.1 Hz stimulation and the plateau level of the action potential was decreased at the same time. But the reduction of contraction or the plateau level was much smaller in 4-AP group and than in control. From the above results it can be concluded that contraction and action potential staircase is dependent upon transmembrane $Ca^{2+}-current\;and\;Ca^{2+}$release from the SR.

• The Korean Journal of Pharmacology
• /
• v.32 no.3
• /
• pp.301-309
• /
• 1996

Since it has been reported that the depolarization-induced NE release is inhibited by activation of presynaptic $A_1-adenosine$ heteroreceptor in hippocampus, a large body of experimental data on the post-receptor mechanism of this process has been accumulated. But, the post-receptor mechanism of presynaptic $A_1-adenosine$ receptor on the NE release has not been clearly elucidated yet. Therefore, it was attempted to clarify the participation of $K^+-channel$ in the post-receptor mechanisms of the $A_1-adenosine$ receptor-mediated control of NE release in this study. Slices from rat hippocampus were equilibrated with $^3H-norepinephrine$ and the release of the labelled products was evoked by electrical stimulation (3 Hz, 5 $VCm^{-1}$, 2 ms, rectangular pulses), and the influence of various agents on the evoked tritium-outflow was investigated. Adenosine, in concentrations ranging from $1{\sim}30\;{\mu}M$, decreased the NE release in a dose-dependent manner, without affecting the basal rate of release. 4AP $(1{\sim}30{\mu}M)$, a specific A-type $K^+-channel$ blocker, increased the evoked NE release in a dose-related fashion, and the basal rate of release is increased by 10 and $30{\mu}M$. TEA $(1{\sim}10{\mu}\;M)$, a nonspecific $K^+-channel$ blocker, increased the evoked NE release in a dose-dependent manner without affecting basal release. The adenosine effects were significantly inhibites by 3 ${\mu}M$ 4AP and 10 mM TEA treatment. 4AP $(30{\mu}M)-$ and TEA (10 mM)-induced increments of evoked NE release were completely abolished in $Ca^{++} free, but these were recoverd in low $Ca^{++} medium. And the effects of $K^+-channel$ blockers in low $Ca^{++} medium were inhibites and abolishes by $Mg^{++} (4 mM) adding and TTX $(0.3{\mu}M)$ adding medium, respectively. These results suggest that the decrement of the evoked NE-release by $A_1-adenosine$ receptor is mediated by 4AP and TEA sensitive $K^+-channel$.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.24 no.4
• /
• pp.592-597
• /
• 2010

Cinnamomi Ramulus is one of the medicinal plants that have been used to improve various diseases caused by insufficient blood circulation. This study was performed for the investigation of vasodilation efficacy ethanol extract of Cinnamomi Ramulus (CR). CR exhibited vascular relaxation against phenylephrine (PE, $10^{-6}M$)-, KCl- and NaF-induced contraction in rat thoracic aorta. In addition, its relaxation was endothelium-independent. Treatment of potassium channel blockers such as gilbenclamide (Gli, $10^{-5}M$), tetraethylammonium (TEA, 1 mM) and 4-aminopyridine (4-AP, 0.2 mM) did not effect on the relaxation of CR. The relaxant effects were also not inhibited by pre-treatment of rat aorta with L-NAME ($10^{-4}M$), methylene blue ($10^{-5}M$), indomethacin ($10^{-5}M$), and atropine ($10^{-6}M$). However, nifedipine ($10^{-5}M$), L-type $Ca^{2+}$ channel blocker, in part attenuated the relaxation of CR ($0.2\;mg/m{\ell}$), but SK&F96365 ($3{\times}10^{-5}M$), receptor activated $Ca^{2+}$ channel blocker and 2-APB ($10^{-4}M$), store operated $Ca^{2+}$ channel blocker did not affact dilation of CR. These findings suggest that the endothelium-independent relaxation effect of CR is partly related with inhibition of $Ca^{2+}$ influx via voltage dependent $Ca^{2+}$ channel.

• The Korean Journal of Physiology and Pharmacology
• /
• v.12 no.4
• /
• pp.131-135
• /
• 2008

The profile of membrane currents was investigated in differentiated neuronal cells derived from human neural stem cells (hNSCs) that were obtained from aborted fetal cortex. Whole-cell voltage clamp recording revealed at least 4 different currents: a tetrodotoxin (TTX)-sensitive $Na^+$ current, a hyperpolarization-activated inward current, and A-type and delayed rectifier-type $K^+$ outward currents. Both types of $K^+$ outward currents were blocked by either 5 mM tetraethylammonium (TEA) or 5 mM 4-aminopyridine (4-AP). The hyperpolarization-activated current resembled the classical $K^+$ inward current in that it exhibited a voltage-dependent block in the presence of external $Ba^{2+}$ (30 ${\mu}$M) or $Cs^+$ (3${\mu}$M). However, the reversal potentials did not match well with the predicted $K^+$ equilibrium potentials, suggesting that it was not a classical $K^+$ inward rectifier current. The other $Na^+$ inward current resembled the classical $Na^+$ current observed in pharmacological studies. The expression of these channels may contribute to generation and repolarization of action potential and might be regarded as functional markers for hNSCs-derived neurons.

• YAKHAK HOEJI
• /
• v.41 no.4
• /
• pp.399-406
• /
• 1997

The effects of different $K^+$ channel blockers were investigated on the non-adrenergic non-cholinergic (NANC) relaxations in the circular muscle of the rabbit proximal stomach. Non-selective blockers of $K^+$ channels, 4-aminopyridine (4-AP, 3~30${\mu}M$) and tetraethylammonium (TEA, 100~1000${\mu}M$) significantly enhanced the NANC relaxations in a concentration-dependent manner. The enhancement was more prominent for the NANC relaxations induced by the electric field stimulation (EFS) with lower frequencies. Blockers of large conductance $Ca^{2+}$-activated $K^+$ channels, charybdotoxin and iberiotoxin, a blocker of small conduntance $Ca^{2+}$-activated $K^+$ channels, apamin and a blocker of ATP-sensitive $K^+$ channels, glibenclamide had no effect on the NANC relaxations, respectively. Exogeneous administration of nitric oxide (NO, 1~30${\mu}M$) caused concentration-dependent relaxations which showed a similarity to those obtained with EFS. None of the $K^+$ channel blockers had an effect on the concentration-dependent relaxation in response to NO. These results suggest that prejunctional $K^+$ channels regulate the release of NO from the NANC nerve in the rabbit proximal stomach as the inhibition of prejunctional $K^+$ channels increases the NANC relaxation induced by the EFS.

• The Korean Journal of Physiology and Pharmacology
• /
• v.16 no.5
• /
• pp.297-303
• /
• 2012

This study was designed to elucidate high $K^+$-induced relaxation in the human gastric fundus. Circular smooth muscle from the human gastric fundus greater curvature showed stretch-dependent high $K^+$ (50 mM)-induced contractions. However, longitudinal smooth muscle produced stretch-dependent high $K^+$-induced relaxation. We investigated several relaxation mechanisms to understand the reason for the discrepancy. Protein kinase inhibitors such as KT 5823 (1 ${\mu}M$) and KT 5720 (1 ${\mu}M$) which block protein kinases (PKG and PKA) had no effect on high $K^+$-induced relaxation. $K^+$ channel blockers except 4-aminopyridine (4-AP), a voltage-dependent $K^+$ channel ($K_V$) blocker, did not affect high $K^+$ -induced relaxation. However, N(G)-nitro-L-arginine and 1H-(1,2,4)oxadiazolo (4,3-A)quinoxalin-1-one, an inhibitors of soluble guanylate cyclase (sGC) and 4-AP inhibited relaxation and reversed relaxation to contraction. High $K^+$-induced relaxation of the human gastric fundus was observed only in the longitudinal muscles from the greater curvature. These data suggest that the longitudinal muscle of the human gastric fundus greater curvature produced high $K^+$-induced relaxation that was activated by the nitric oxide/sGC pathway through a $K_V$ channel-dependent mechanism.