Takayuki Tokimasa

Synaptic plasticity preserved with arachidonic acid diet in aged rats

We examined whether synaptic plasticity was preserved in aged rats administered an arachidonic acid (AA) containing diet. Young male Fischer-344 rats (2 mo of age), and two groups of aged rats of the same strain (2 y of age) who consumed either a control diet or an AA ethyl ester-containing diet for at least 3 mo were used. In the Morris water maze task, aged rats on the AA diet had tendency to show better performance than aged rats on the control diet. Long-term potentiation induced by tetanic stimulation was recorded from a 300 microm thick hippocampal slice with a 36 multi-electrode-array positioned at the dendrites of CA1 pyramidal neurons. The degree of potentiation after 1 h in aged rats on the AA diet was comparable as that of young controls. Phospholipid analysis revealed that AA and docosahexaenoic acid were the major fatty acids in the hippocampus in aged rats. There was a correlation between the behavioral measure and the changes in excitatory postsynaptic potential slope and between the physiologic measure and the total amount of AA in hippocampus.

Effects of quinine on three different types of potassium currents in bullfrog sympathetic neurons.

Whole-cell/voltage-clamp recordings were made from dissociated bullfrog sympathetic neurons to examine the sensitivity of potassium currents to a potassium channel blocker quinine (1-500 microM). Among three currents tested, a rapidly inactivating A-type current (I(A)) was the most sensitive to the block by quinine (IC50 approximately 22 microM). A non-inactivating M-type current (I(M)) was the least sensitive (IC50 approximately 445 microM), and the sensitivity of a slowly inactivating delayed rectifier-type current (I(K)) was in between (IC50 approximately 115 microM). Results suggest that the ability of quinine to block different types of potassium currents such as I(A) and I(M) with significantly different IC50 values would be of help for the potassium channel pharmacology in amphibian autonomic ganglion cells.

Evidence for the calcium-dependent potentiation of M-current obtained by the ratiometric measurement of the fura-2 fluorescence in bullfrog sympathetic neurons.

Intracellular Ca2+ concentration ([Ca]i) was measured following the activation of an inward Ca2+ current and subsequent potentiation of an M-type K+ current (IM) in bullfrog sympathetic neurons. Fura-2 was used as an indicator for [Ca]i. The fluorescence ratio at 340 and 380 nm (F340/F380) was elevated from 0.36 to 1.22 when IM was potentiated by 68% following the Ca2+ current. Based on the in vivo calibration curve obtained from cells permeabilized with digitonin (20 microM), the F340/F380 value of 1.22 was equivalent to a [Ca]i of 0.97 microM. We therefore propose that a rise in [Ca]i into the micromolar range can lead to the potentiation of IM in amphibian autonomic neurons.

Purinergic cation channels in neurons of rabbit vesical parasympathetic ganglia.

Membrane current was recorded from neurons in rabbit vesical parasympathetic ganglia, utilizing single electrode voltage clamp techniques. ATP (0.1-1 mM) caused an inward current (IATP) associated with an increased conductance at a holding potential of -50 mV. ADP (0.1-1 mM) and 5-O-3-thiotriphosphate (0.1-0.6 mM) but not AMP (0.3-2 mM) and adenosine (0.1-2 mM) mimicked the actions of ATP. The IATP reversed its polarity at -12.1 +/- 1.4 mV. The amplitude of the IATP was depressed in low sodium solutions and in nominally calcium-free solutions but not in low chloride solutions. Suramin (10-100 microM) and reactive blue 2 (10-100 microM), P2-antagonists, reversibly depressed the IATP. In contrast, hexamethonium (100 microM) did not affect the IATP. These data suggest that ATP activates cation channels through P2X receptor subtypes in parasympathetic neurons.

Calcium-dependent potentiation of M-current in bullfrog sympathetic neurons ☆

Whole-cell voltage-clamp recordings were made from cultured bullfrog sympathetic neurons to measure the steady-state activation curve of M-type potassium current. When measured with a calcium-deficient (10 nM) pipette solution M-conductance was 4.8 nS at -35 mV having the 50%-activation voltage at-20 mV. Respective values were 17.2 nS at -35 mV with the 50%-activation voltage at -42 mV when measured with a calcium-rich (1 microM) solution, indicating the hyperpolarizing displacement of the activation curve with high internal calcium. It is suggested that intracellular calcium ions can modulate kinetics of M-current which thereby regulate the number of M-channels being open at given membrane potentials.

Actions of zinc on rapidly inactivating A-type and non-inactivating M-type potassium currents in bullfrog sympathetic neurons

The actions of zinc on A-type potassium current (I(A)) were studied in dissociated bullfrog sympathetic neurons. Zinc (1-300 microM) caused a parallel shift in the activation and inactivation curves to a depolarizing direction, thereby enhancing I(A) around physiological resting potential. An EC50 value was 70-100 microM for these actions. The zinc actions were non-selective in a sense that zinc inhibited M-type potassium current (I(M)) with an IC50 value of 300 microM. Zinc was without effect on the maximum conductance for I(A) and the kinetic behavior for I(M). The ability of low concentrations of zinc to modulate separate set of potassium currents such as I(A) and I(M) in conceptually distinct manner may therefore assume pathophysiological importance for autonomic neurons.

Pyridoxalphosphate-6-azophenyl-2′,4′-disulfonic acid can antagonize the purinoceptor-mediated inhibition of M-current in bullfrog sympathetic neurons

Whole-cell recordings of an M-type potassium current (I(M)) were made from dissociated bullfrog sympathetic neurons. Purinoceptor agonists inhibited I(M) with UTP>ADP>adenosine triphosphate=UDP>ATPgammaS=guanosine triphosphate (GTP)>>amyloid precursor protein (APP)(NH)P as the rank order of potency. The IC(50) was 35 nM for UTP, and 2.6 microM for GTP. Under conditions in which I(M) was abolished by UTP (1 microM), a sulfonic acid derivative, pyridoxalphosphate-6-azophenyl-2,4-disulfonic acid (PPADS) (30-300 microM) recruited I(M) to 15 to 90% of its control in a reversible and concentration-dependent manner. These results indicate that PPADS can be useful as an antagonist for the purinoceptors presumably P2Y subtypes in amphibian autonomic neurons.

Effects of lanthanides on voltage-dependent potassium currents in bullfrog sympathetic neurons

The effects of lanthanides (La(3+), Gd(3+), Lu(3+) and Sm(3+)) on voltage-dependent potassium currents were studied in dissociated bullfrog sympathetic neurons. A-type current (I(A)) and M-type current (I(M)) were blocked by lanthanides (0.1-30 microM) with I(M) being much less sensitive to these ions than I(A). The order of potency was Gd(3+)>/=Lu(3+) approximately La(3+) approximately Sm(3+) for I(A) and Gd(3+)&z.Gt;Lu(3+) approximately La(3+)>Sm(3+) for I(M). The I(M) block occurred independently of its activation kinetics while the I(A) block was associated with a positive shift of the activation and inactivation curves. Gd(3+) (100 microM) blocked the delayed rectifier-type current (I(K)) by less than 20%; Lu(3+), La(3+) and Sm(3+) (100 microM for each) were without effect on I(K). It is concluded that I(A) was the most sensitive to lanthanides, and Gd(3+) was the most potent for all the currents in amphibian autonomic neurons.

Actions of barium on rapidly inactivating potassium current in bullfrog sympathetic neurons.

Whole-cell/voltage-clamp recordings were made from dissociated bullfrog sympathetic neurons to examine the inhibitory actions of barium (0.01-3 mM) on a rapidly inactivating A-type potassium current (IA). The IC50 value was about 0.9 mM. Barium (1 mM) approximately halved the maximum amplitude of IA (approximately 1.7 nA near 0 mV) without significantly affecting a voltage for the 50%-activation (approximately -40 mV) and that for the 50%-inactivation (approximately -90 mV), nor did it affected the time course of IA. The results suggest that the barium block is independent of the kinetics of the A-channels in bullfrog sympathetic neurons.

Mechanisms underlying the M-current block by barium in bullfrog sympathetic neurons

Whole-cell/voltage-clamp recordings were made from dissociated bullfrog sympathetic neurons to examine the channel blocking actions of barium (3-2000 microM) on an M-type potassium current (I(M)). Barium (IC(50) approximately 105 microM) blocked I(M) without affecting the 50%-activation voltage ( approximately -35 mV) and the slope factor ( approximately 11 mV) of the activation curve. The results indicate that the barium block is independent of the kinetics of I(M).