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Effects of histamine on resting and action potentials of squid giant axons
Lüe Sciences Vol. 10, Part I, pp . 955-380, 1971 . Printed in Great Britain
Perga.mon Press
EFFECTS OF IìISTAMINE ON RESTING AND ACTION POTENTIALS OF SQUID GIANT AXONS M . Scuka Istituto di Fisiologia umana, dell~Università di Trieste, Trieste (Italy) (Received in final. form 10 February 1871) Sumnary The effects of histamine on the squid giant axon have been studied by means of intracellular electrode technique . Histamine depressed the magnitude of resting and action potentials and pro duced a repetitive excitation, which was associated with an incre ass in negative after-potential . Lowering the temperature decreased the amplitude of the negative after-potential thereby suppressing the repetitive after-discharge . These effects of histamine are discussed in relation to similar effects of other drugs . Introduction In a previous paper (11) it was reported that histamine decreases the amplitude and changes the time course of the falling phase of the end-plate potential (e .p .p .) in the sciatic nerve-sartoriua muscle preparation of the frog . The experiments did not exclude the possibility that the change in the e .p .p . was due, at least partially, to an action of histamine on the nerve fibers . In the present research this possibility was supported by the finding that histamine was effective on the squid giant axon in suppressing both resting and action potentials . Tdethods The giant axon (diameter 300-400~u) was isolated from the squid, Loligo pealei , available at the ldarine Biological Laboratory, ~~ooda Hole, Mass . (U .S .A .) . The preparation was careflzlly cleaned and placed in the nerve chamber continuously perf~zsed with ne.tural sea water. 355
358
Effect ad Histamine on Squid Axon
Vol. 10, No . 8
Intracellular recordings were made by means of a capillary microelectrode filled with 3 M-KC1, the resistance being in the range of 5-15 Mn. Action potentials were elicited by external stimulation . The resting and action potentials were fed to a d .c . amplifier and recorded on an oscilloscope and on a pen recorder . Histamine perfusion fluid was prepared by dissolving histamine dihydrochloride (Fisher)
in natural sea water. Hoth control
and test bathing solutions were perfused continuously through the nerve chamber. The experiments were carried out at room temperature (24-28°C) . When the effect of the temperature was tó be examined, the bathing fluid was passed through a cooling chamber before introducing into the nerve chamber, and the temperature was measured near the preparation . Results Histamine 0 .5-2 .5 mÀ2 depressed the magnitudes of both resting and action potentials . Examples of the changes in the magnitudes of the resting and action potentials caused by appli cation of histamine 0 .5, 1 and 2 .5 mM are shown in Table 1 . A few minutes after application of histamine the negative after-potential that followed the post-spike undershoot, gradually (Fig . 1b) increased to the point that a repetitive, damped after-discharge was elicited by a single stimulus (Fig . 1c, d) . Washing with natural sea water without containing histamine brought about complete recovery in tk~e magnitudes of resting and action potentials, and abolished the repetitive discharges, but the negative after-potential usually remained augmented . The duration of the repetitive excitation in the histamine-treated axon vas shortened with decreasing the temperature . When the temperature was decreased from 25°C to 10°C, the after-potential au ~nsnted by histamine decreased in height thereby decreasing ttie number of spikes in the repetitive after-discharg
Effect a~f Histamine on Squid Axon
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Effect a~f Histamine on Squid Axon
Vol . 10, No. ß
~o . .,v
d FIG. 1 Action potential before (a) and during (b, c, d) application of 0 .5 mM histamine elicited by a single stimulus . The axon eventually ceased to fire repetitively . It was also observed that lowering the temperature increased the amplitude and prolonged the duration of the action potential and slightly hyperpolarized the membrane both in the control and histamine-treated axons (Fig . 2) . The effect of temperature change was completely reversible .
W
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0
"Histamine 2,5rr~INh
-20 ~-40
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FI G . 2 The effects of temperature on the magnitudes of resting (RP) and action (AP) potentials
Vol. 10, No. 8
Effect od Histamine oa Squid Axon
359
Discussion The experiments described above demonstrate that histamine decreases both the resting and action potentials in the squid giant axon . Several other drugs, which are chemically unrelated to histamine, have been shown to produce repetitive discharges in nerve fibers . They include veratrine alkaloids (3, 12, 16),
pyrethrins (1, 2, 17), allethrin (4, 7), DDT (8, 9, 10, 16, 17,
18), Condylactis toxin (14), and p-xylene (15) . In many of these
cases, repetitive discharges have been shown to be associated with an increase in negative after-potential, i .e . allethrin (4, 5, 7), DDT (8, 9), Condylactia toxin (14), and veratrine alkaloids (13) . The effect of temperature on the negative aftexLpotential and repetitive after-discharge in the histamine-treated squid axon is similar to that in the allethrin-treated cockroach axon (4) . In both cases lowering the temperature decreased the amplitude of the negative after-potential thereby suppressing the repetitive after-discharge . However, these two agents are different in their temperature dependency of the suppressive action on the action potential. Although the suppressive action of histamine is not affected by temperature change, the suppressive action of allethrin is potentiated by lowering the temperature (6) . Histamine is different from DDT in the temperature dependency . Unlike histamine, the repetitive discharge caused by DDT is facilitated by lowering the temperature (19) . In the squid axon treated with p-xylene, lowering the temperature also causes repetitive responses (15) . Acknowledgments The author expresses her thanks hospitality at the Marine Biological (Massachusetts, U .S .A .), and for his encouragement during the development
to Dr . T . Narahashi for his Laboratory, Woods Hole helpfla.l criticism and of this work .
380
Vol. 10, No . 8
Eüect od Histamine on Squid Aaon
References 1 .
I . HAYASHI,
Shokubutsu Oyobi Dobutsu 7, 2001
2.
0 . LÖWENSTEIN, Nature 150, 760 (1842) .
3.
H . MEVES, Pflug . Arch . Res . Physiol . 290, 211 (1966) .
4.
T . NARAHASHI, J . Cell . Comp . Physiol . 59, 61
(1962) .
5.
T . NARAHASHI, J . Cell . Comp . Physiol . 59, 67
(1962) .
6.
T . NARAHASHI, Effects of insecticides on excitable tissues, Advances of Insect Physiol . , Academic Press, London and New York, in presa .
7.
T . NARAHASHI and N .C . ANDERSON, Toxicol . Appl . Pharmacol . 10, 529 (1967) .
(1939) .
8.
T . NAR.AHASHI and H. G . HAAS, J . Gen . Physiol . 51, 177 (1968) .
9.
T . NARAHASHI and T . YAMASAKI, J . Physiol . 152,
122 (1960) .
10 .
g.D . ROEDER and E .A . WEIANT, J . Cell . Comp . Physiol . 3?, 175 (1948) .
11 .
M . SCUgA, Boll . Soc . It . Biol . Sper . , in press .
12 .
A .M . SHANES, J . Gen. Physiol . 33, 57 (1949) .
13 .
A .M . SHANES, J . Pharmacol . Exp . Ther . 105, 216 (1952) .
14 .
B .I . SHAPIRO and G. LILLEHEIL, Comp . Biochem . Physiol . 1225 (1969) .
15 .
R .A . SJODIN and L .J . MULLINS, J . Gen. Physiol . 42, 39 (1958) .
16 .
J .H . WELSH and H.T . GORDON, J . Cell . Comp . Physiol . _30, 147 (1947) .
17 .
8,
T . YAMASAgI and T . ISHII, J. Nippon . Soc . Appl . Entom . 7, 157 (1952) .
18 .
T . YAMASAKI and T . ISHII, Botyu-Kagaku (Scientific Insect Control) 19, 1 (1954) .
19 .
T . YAMASAKI and T . ISHII, Botyu-Ka.gaku (Scientific Insect Control) 19, 39 (1954) .
Perga.mon Press
EFFECTS OF IìISTAMINE ON RESTING AND ACTION POTENTIALS OF SQUID GIANT AXONS M . Scuka Istituto di Fisiologia umana, dell~Università di Trieste, Trieste (Italy) (Received in final. form 10 February 1871) Sumnary The effects of histamine on the squid giant axon have been studied by means of intracellular electrode technique . Histamine depressed the magnitude of resting and action potentials and pro duced a repetitive excitation, which was associated with an incre ass in negative after-potential . Lowering the temperature decreased the amplitude of the negative after-potential thereby suppressing the repetitive after-discharge . These effects of histamine are discussed in relation to similar effects of other drugs . Introduction In a previous paper (11) it was reported that histamine decreases the amplitude and changes the time course of the falling phase of the end-plate potential (e .p .p .) in the sciatic nerve-sartoriua muscle preparation of the frog . The experiments did not exclude the possibility that the change in the e .p .p . was due, at least partially, to an action of histamine on the nerve fibers . In the present research this possibility was supported by the finding that histamine was effective on the squid giant axon in suppressing both resting and action potentials . Tdethods The giant axon (diameter 300-400~u) was isolated from the squid, Loligo pealei , available at the ldarine Biological Laboratory, ~~ooda Hole, Mass . (U .S .A .) . The preparation was careflzlly cleaned and placed in the nerve chamber continuously perf~zsed with ne.tural sea water. 355
358
Effect ad Histamine on Squid Axon
Vol. 10, No . 8
Intracellular recordings were made by means of a capillary microelectrode filled with 3 M-KC1, the resistance being in the range of 5-15 Mn. Action potentials were elicited by external stimulation . The resting and action potentials were fed to a d .c . amplifier and recorded on an oscilloscope and on a pen recorder . Histamine perfusion fluid was prepared by dissolving histamine dihydrochloride (Fisher)
in natural sea water. Hoth control
and test bathing solutions were perfused continuously through the nerve chamber. The experiments were carried out at room temperature (24-28°C) . When the effect of the temperature was tó be examined, the bathing fluid was passed through a cooling chamber before introducing into the nerve chamber, and the temperature was measured near the preparation . Results Histamine 0 .5-2 .5 mÀ2 depressed the magnitudes of both resting and action potentials . Examples of the changes in the magnitudes of the resting and action potentials caused by appli cation of histamine 0 .5, 1 and 2 .5 mM are shown in Table 1 . A few minutes after application of histamine the negative after-potential that followed the post-spike undershoot, gradually (Fig . 1b) increased to the point that a repetitive, damped after-discharge was elicited by a single stimulus (Fig . 1c, d) . Washing with natural sea water without containing histamine brought about complete recovery in tk~e magnitudes of resting and action potentials, and abolished the repetitive discharges, but the negative after-potential usually remained augmented . The duration of the repetitive excitation in the histamine-treated axon vas shortened with decreasing the temperature . When the temperature was decreased from 25°C to 10°C, the after-potential au ~nsnted by histamine decreased in height thereby decreasing ttie number of spikes in the repetitive after-discharg
Effect a~f Histamine on Squid Axon
Vol. 10, No . ß
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358
Effect a~f Histamine on Squid Axon
Vol . 10, No. ß
~o . .,v
d FIG. 1 Action potential before (a) and during (b, c, d) application of 0 .5 mM histamine elicited by a single stimulus . The axon eventually ceased to fire repetitively . It was also observed that lowering the temperature increased the amplitude and prolonged the duration of the action potential and slightly hyperpolarized the membrane both in the control and histamine-treated axons (Fig . 2) . The effect of temperature change was completely reversible .
W
C
0
"Histamine 2,5rr~INh
-20 ~-40
f0
20 30 40 Time(mln)
50
FI G . 2 The effects of temperature on the magnitudes of resting (RP) and action (AP) potentials
Vol. 10, No. 8
Effect od Histamine oa Squid Axon
359
Discussion The experiments described above demonstrate that histamine decreases both the resting and action potentials in the squid giant axon . Several other drugs, which are chemically unrelated to histamine, have been shown to produce repetitive discharges in nerve fibers . They include veratrine alkaloids (3, 12, 16),
pyrethrins (1, 2, 17), allethrin (4, 7), DDT (8, 9, 10, 16, 17,
18), Condylactis toxin (14), and p-xylene (15) . In many of these
cases, repetitive discharges have been shown to be associated with an increase in negative after-potential, i .e . allethrin (4, 5, 7), DDT (8, 9), Condylactia toxin (14), and veratrine alkaloids (13) . The effect of temperature on the negative aftexLpotential and repetitive after-discharge in the histamine-treated squid axon is similar to that in the allethrin-treated cockroach axon (4) . In both cases lowering the temperature decreased the amplitude of the negative after-potential thereby suppressing the repetitive after-discharge . However, these two agents are different in their temperature dependency of the suppressive action on the action potential. Although the suppressive action of histamine is not affected by temperature change, the suppressive action of allethrin is potentiated by lowering the temperature (6) . Histamine is different from DDT in the temperature dependency . Unlike histamine, the repetitive discharge caused by DDT is facilitated by lowering the temperature (19) . In the squid axon treated with p-xylene, lowering the temperature also causes repetitive responses (15) . Acknowledgments The author expresses her thanks hospitality at the Marine Biological (Massachusetts, U .S .A .), and for his encouragement during the development
to Dr . T . Narahashi for his Laboratory, Woods Hole helpfla.l criticism and of this work .
380
Vol. 10, No . 8
Eüect od Histamine on Squid Aaon
References 1 .
I . HAYASHI,
Shokubutsu Oyobi Dobutsu 7, 2001
2.
0 . LÖWENSTEIN, Nature 150, 760 (1842) .
3.
H . MEVES, Pflug . Arch . Res . Physiol . 290, 211 (1966) .
4.
T . NARAHASHI, J . Cell . Comp . Physiol . 59, 61
(1962) .
5.
T . NARAHASHI, J . Cell . Comp . Physiol . 59, 67
(1962) .
6.
T . NARAHASHI, Effects of insecticides on excitable tissues, Advances of Insect Physiol . , Academic Press, London and New York, in presa .
7.
T . NARAHASHI and N .C . ANDERSON, Toxicol . Appl . Pharmacol . 10, 529 (1967) .
(1939) .
8.
T . NAR.AHASHI and H. G . HAAS, J . Gen . Physiol . 51, 177 (1968) .
9.
T . NARAHASHI and T . YAMASAKI, J . Physiol . 152,
122 (1960) .
10 .
g.D . ROEDER and E .A . WEIANT, J . Cell . Comp . Physiol . 3?, 175 (1948) .
11 .
M . SCUgA, Boll . Soc . It . Biol . Sper . , in press .
12 .
A .M . SHANES, J . Gen. Physiol . 33, 57 (1949) .
13 .
A .M . SHANES, J . Pharmacol . Exp . Ther . 105, 216 (1952) .
14 .
B .I . SHAPIRO and G. LILLEHEIL, Comp . Biochem . Physiol . 1225 (1969) .
15 .
R .A . SJODIN and L .J . MULLINS, J . Gen. Physiol . 42, 39 (1958) .
16 .
J .H . WELSH and H.T . GORDON, J . Cell . Comp . Physiol . _30, 147 (1947) .
17 .
8,
T . YAMASAgI and T . ISHII, J. Nippon . Soc . Appl . Entom . 7, 157 (1952) .
18 .
T . YAMASAKI and T . ISHII, Botyu-Kagaku (Scientific Insect Control) 19, 1 (1954) .
19 .
T . YAMASAKI and T . ISHII, Botyu-Ka.gaku (Scientific Insect Control) 19, 39 (1954) .