293

Bioelectrochemistry and Bioenergetics, 27 (1992) 2 9 3 - 3 0 4

A section o f J. Electroanal. Chem., and constituting Vol. 342 (1992)
Elsevier Sequoia S.A., L a u s a n n e
J E C BB 01485

Influence of weak non-thermic high-frequency

e l e c t r o m a g n e t i c fields o n t h e m e m b r a n e p o t e n t i a l o f n e r v e
cells *

U . Kullnick
Zoologisches Institut, Physiologic, Technische Unioersitiit Braunschweig, D-3300 Braunschweig (Germany)
( R e c e i v e d 9 O c t o b e r 1991)

Abstract

N e r v e cells o f the snail Helix pomatia w e r e subjected to high-frequency (159 M H z , 8.3 H z

m o d u l a t e d ) , n o n - t h e r m i e (maximum .qux density 124/~T) e l e c t r o m a g n e t i c fields. T h e e f f e c t o f the fields
on the m e m b r a n e potential o f various nerve cells was investigated. It was o b s e r v e d that short and
u n i q u e B e f e l d u n g e n t led to an alteration o f the m e m b r a n e potential o f the n e u r o n s examined. T h e
alteration almost always expressed itself as a long-term h y p e r p o l a r i z a t i o n o f the resting potential. A
clear c o n n e c t i o n b e t w e e n the negativity o f the m e m b r a n e p o t e n t i a l o f a nerve cell b e f o r e the B e f e l d u n g
and the strength o f the hyperpolarization c a u s e d by the B e f e l d u n g was seen. As well as this effect, an
a l t e r a t i o n in the threshold o f excitation o f b e f e l d e t cells could be m e a s u r e d .

INTRODUCTION

T h e influences o f so-called n o n - t h e r m i c h i g h - f r e q u e n c y ( H F ) e l e c t r o m a g n e t i c
fields ( E M F ) on biological systems a r e b e c o m i n g a g r e a t e r p a r t o f public interest.
T h e fields are b e i n g m a d e use o f m o r e a n d m o r e extensively for c o m m e r c i a l
t h e r a p y a p p a r a t u s . This explains t h e discussion a b o u t t h e i r m e d i c a l i m p o r t a n c e
a n d t h e i r possible effects on t h e e n v i r o n m e n t . It was against this b a c k g r o u n d t h a t
t h e effects of w e a k ac m a g n e t i c fields b e g a n to b e c o m e the object o f n u m e r o u s
r e s e a r c h projects [1]. In the m e a n t i m e , a n u m b e r o f clinical findings seem to

* P a p e r p r e s e n t e d at the syrn~osium " H i g h - F r e q u e n c y E l e c t r o m a g n e t i c ac Fields a n d their• Effects on

Biological Systems", B r a u n s c h w e i g , 9.-.1. .0. . .July
. . . . . . 1991.
....
.1 Definition: B e f e l d u n g is t r e a t m e n t with a w e a k (non-ionizing, n o n - t h e r m i c ) e l e c t r o m a g n e t i c field.

0302-4598/92/$05.00 © 1992 - Elsevier Sequoia S.A. All rights reserved

294

confirm t h a t t h e r e are effects which can be u s e d therapeutically. N o t only t h e

u n k n o w n effect mechanisms, but also t h e lack of knowledge about t h e best
t h e r a p e u t i c p a r a m e t e r s of t h e fields used (intensity, frequency, m o d u l a t i o n ) a r e
against sensible use. Also with regard to work a n d e n v i r o n m e n t a l risks, g r e a t e r
k n o w l e d g e of the interactions b e t w e e n E M F a n d biological systems is n e c e s s a r y at
all costs. This is of special i m p o r t a n c e b e c a u s e s t a n d a r d s for the p r o t e c t i o n of
m a n k i n d in the future could be f o r m u l a t e d not only from an e n e r g y p o i n t of view
[2]. It can be seen t h a t the o p i n i o n of m a n y r e s e a r c h e r s ( f r e q u e n t l y expressed up to
now) t h a t the effect of E M F can only be s u b s t a n t i a t e d e n e r g e t i c a l l y must now be
r e c o n s i d e r e d in the wake of more m o d e r n knowledge. F o r investigations of t h e
effect of w e a k e l e c t r o m a g n e t i c fields in this study, the central nervous system
(CNS) of the vineyard snail is used. T h e a d v a n t a g e s of using a snail as the object of
investigation can be found in the c o m p a r a t i v e l y small n u m b e r o f n e u r o n s (ap-
proximately 50 000), their easy accessibility, their specific size (up to 150 /xm) a n d
t h e i r robustness against i n t e r v e n t i o n s in t h e nervous system (e.g. p r e p a r a t i o n ) .
F u r t h e r , it is a well-known fact t h a t t h e f u n d a m e n t a l functions of nerve ceils are
practically identical in t h e various groups of animals, b o t h f l o m a m e t a b o l i c a n d an
electrophysical point of view (signal g e n e r a t i o n , c o n d u c t i o n a n d transmission). A
great deal of c o r r e s p o n d e n c e can be f o u n d especially in molluscs a n d t h e n e u r o n s
of v e r t e b r a t e s ( H o d g k i n , Huxley 1952). A p r i n c i p a l transferability of t h e effects of
e l e c t r o m a g n e t i c fields observed in the n e u r o n s of snails to h u m a n nerve cells can
t h u s be p r e s u p p o s e d . T h e n e u r o n s of molluscs m a k e it possible to recognize not
only short-term, brief effects, but also l o n g - t e r m or irreversible ones, as t h e y can
be m e a s u r e d intracellularly for a n u m b e r of hours a n d even days, as o p p o s e d to
the nerve cells of vertebrates. Helix pomatia is thus a suitable m o d e l o r g a n i s m for
t h e investigation of n e u r o n a l questions. In addition, e x p e r i m e n t s with v e r t e b r a t e
cell cultures a n d p r e p a r a t i o n s of brain slices will be carried out in our l a b o r a t o r y in
t h e n e a r future.
T h e first investigations of t h e nerve ceils of Helix pomatia a n d t h e i r bioelectric-
ity u n d e r t h e effects of w e a k H F m a g n e t i c fields (150 M H z , 8.3 Hz, L F pulsed)
gave i n t e r e s t i n g results [3]. T h e p a r a m e t e r which was p r e d o m i n a n t l y investigated,
t h e m e m b r a n e potential, showed clear h y p e r p o l a r i z a t i o n of the nerve cells. T h e s e
results w e r e confirmed, c o m p l e t e d a n d e x t e n d e d in f u r t h e r examinations. P e r h a p s
t h e y can form t h e basis for t h e solution to t h e p r o b l e m of cell-physiological
m e c h a n i s m s of t h e a t h e r m i c effect of H F - E M fields.

METHODS

F o r t h e experiments, adult v i n e y a r d snails ( 3 - 4 years old) of t h e species Helix

pomatia (t3astropoda, P u l m o n a t a ) w e r e used. T h e C N S of t h e animals was re-
moved w i t h o u t t h e use of medicines. T h e C N S consists of an o e s o p h a g e a I g a n g l i o n
w i t h cerebral a n d s u b o e s o p h a g e a l ganglia. F o r intracellular m e a s u r e m e n t s , only
giant nerve cells of the s u b o e s o p h a g e a l g a n g l i o n w e r e used (the a r e a m a r k e d by a
 295

Fig. 1. CNS of Helix pomatia with intact connective tissue. CG: cerebral ganglion; CPLK: cerebro-pleu-
ral connective; CPK: cerebro-pedal connective; P1-P10: pedal nerves; P A R G : parietal ganglion; PD:
pedal ganglion; PLG: pleural ganglion; ST: statoeyst; STN: stato nerve; VISC: visceral ganglion. T h e
neurons measured are those in the circle.

circle in Fig. 1). T h e cell d i a m e t e r was a p p r o x i m a t e l y 100 tzm. Selection of t h e

m e a s u r e d cells d e p e n d e d o n t h e i r position relative to the electrode.
M e a s u r e m e n t s of t h e m e m b r a n e p o t e n t i a l ( M P ) o f living cells, which w e r e not
obviously stimulated, by use o f suitable m e a s u r e m e n t systems always show charac-
teristic voltage values, called resting p o t e n t i a l s (RP), which can r e m a i n c o n s t a n t
for a long time as a rule. In all resting potentials, t h e m e m b r a n e o f t h e nerve cell
on t h e inside has a negative load c o m p a r e d with t h e outside. In this study, this
r e s t i n g p o t e n t i a l was t h e p r e d o m i n a n t l y investigated p a r a m e t e r , in o r d e r to estab-
lish t h e effects of h i g h - f r e q u e n c y e l e c t r o m a g n e t i c fields on nerve cells.
A l t e r a t i o n s of the m e m b r a n e p o t e n t i a l occur in physiological excitations or
artificial electrical stimulation of t h e cell. A n increase in the m e m b r a n e potential,
i.e. i n c r e a s e d n e g a t i v i z a t i o n o f t h e inside, is called h y p e r p o l a r i z a t i o n , a r e d u c t i o n
in t h e p o t e n t i a l b e i n g d e p o l a r i z a t i o n . T h e m e m b r a n e p o t e n t i a l of nerve cells
m a i n l y arises t h r o u g h :
(1) a s e m i - p e r m e a b l e cell m e m b r a n e ; or
( 2 ) an u n e v e n d i s t r i b u t i o n o f ions b e t w e e n t h e i n t r a c e l l u l a r a n d the extracellular
fluid, which is p r o d u c e d , i n t e r alia, by e n e r g y - c o n s u m i n g t r a n s p o r t processes.
296 •

I
!._ . . . . . . . . . . . . . . ,,J

Io -

tof--q ~ ol
Fig. 2. Experimental set-up for intracellular measurements. (1) Signal connection; (2) signal-ground
connection; (3) intercellular preamplifier; (4) oscilloscope; (5) D A T recorder; (6) Mega-Wave 150/1; (7)
E M F transmitter; (8) off-line computer.

• Resting p o t e n t i a l and o t h e r bioelectrical signals (AP, EPSP, IPSP) are r e c o r d e d

by m e a s u r e m e n t probes (electrodes). For intracellular m e a s u r e m e n t s , these are
fine glass capillaries with tip o p e n i n g s of a r o u n d 1 /xm, filled with a 3 M KC1
solution. W i t h the help o f such m i c r o - e l e c t r o d e s and their c o n d u c t i v e c o n n e c t i o n
( A g - A g C I wire) and a m e t a l s i g n a l - g r o u n d c o n n e c t i o n in the R i n g e r solution in
which the objects to be m e a s u r e d can be found, a n d also m e a s u r i n g i n s t r u m e n t s
with a high electrical resistance, the m e m b r a n e polarity o f a nerve cell can be
m e a s u r e d and portrayed. F o r reasons to be f o u n d in the physics o f h i g h - f r e q u e n c y
e l e c t r o m a g n e t i c fields, t h e r e should be no m e t a l parts in the a r e a o f the effects o f
the e l e c t r o m a g n e t i c fields used. In the a r e a of the m a g n e t i c field, we t h e r e f o r e
m a n a g e d w i t h o u t metal conductors, especially in the m e a s u r e m e n t area, in o r d e r
to avoid physical interactions which could falsify the m e a s u r e m e n t or possibly even
m a k e it useless. T h e s e d e m a n d s were fulfilled by the d e v e l o p m e n t o f a new,
m o d i f i e d intracellular m e a s u r i n g m e t h o d .
M e a s u r e m e n t s o f the m e m b r a n e p o t e n t i a l in the m a g n e t i c field were m a d e
possible by replacing the c h l o r i n a t e d silver wire (sig,ial a n d s i g n a l - g r o u n d connec-
tion) inside the field with a 3 M H C l - a g a r - a g a r bridge in thin plastic hoses. T h e
agar bridges only pass into the silver wires, which c o n d u c t the m e a s u r e d signals to
t h e amplifier (Fig. 2), at a distance which basically rules out an i n f l u e n c e of the
a p p l i e d m a g n e t i c field. This m e t h o d m a k e s intracellular m e a s u r e m e n t o f nerve
Cells in high-frequency e l e c t r o m a g n e t i c fields possible. In o r d e r to avoid f u r t h e r
possible m e a s u r e m e n t artefacts caused by an u n f a v o u r a b l e m e a s u r e m e n t geome-
try, t h e signal a n d s i g n a l - g r o u n d c o n n e c t i o n s were a r r a n g e d m o r e or less in the
d i r e c t i o n o f the spread of the e l e c t r o m a g n e t i c field (Fig. 3).
 297

15 c m
I
r2 c m-,-)

Fig, 3. Modified intracellular measurement. (1) Glass signal connection (3 M KCI); (2) KCl-agar
bridges (3 M KCI); (3) connection of agar bridges and Ag-AgCI wife; (4) EMF transmitter; (5) ganglion.

Cells with high s p o n t a n e o u s activity w e r e not used in the e x p e r i m e n t . C e l l s

whose m e m b r a n e p o t e n t i a l showed many E P S P (excitatory postsynaptical p o t e n -
tials) a n d IPSP (inhibitory postsynaptical p o t e n t i a l s ) were also n o t u s e d f o r t h e
evaluation. Basically, only nerve cells with a resting p o t e n t i a l which r e m a i n e d
u n c h a n g e d (2 rnV) for at least 40 min ( p r e l i m i n a r y stage) w e r e used in the
evaluation. Following the experiment, no i d e n t i f i c a t i o n of the p e n e t r a t e d "cell was
carried o u t by n e u r o - a n a t o m i c staining or f u n c t i o n a l checks.
T h e m e a s u r e m e n t w a s always carried out in t h e following sequence: 40 rain
p r e l i m i n a r y phase, 40 rain run-up, 10 min Befeldung, 40 min run-down, 40 rain
s u b s e q u e n t phase. T h e fields used h a d t h e following characteristics: c a r r i e r fre-
quency: 150 MHz; m o d u l a t i o n frequency; 8.3 Hz; form o f m o d u l a t i o n : n e e d l e
impulse; m a g n e t i c flux density; 124 tzT 50% (2 cm from the t r a n s m i t t e r , I n s t i t u t e
for E x p e r i m e n t a l Physics, F r e e University o f Berlin); time of application: 10 min+
T h e d a t a r e c o r d e d by a D A T r e c o r d e r (48 kHz scan frequency) were r e a d i n t o
an off-line c o m p u t e r . T h e c o m p u t e r e v a l u a t i o n was c a r r i e d o u t o n a C o m m o d o r e
PC by m e a n s of an analysis p r o g r a m d e v e l o p e d in o u r d e p a r t m e n t . This p r o g r a m
works with a scan f r e q u e n c y o f 32 kHz. T h e m e a n v a l u e s a r e t a k e n from t e n
m e a s u r e d values. This averaging is carried o u t until a m e a n value for 1 min of
m e a s u r i n g is p r o d u c e d . T h e p r o g r a m recognizes technical i n t e r f e r e n c e , action
p o t e n t i a l s a n d s u d d e n slight a l t e r a t i o n s o f t h e m e m b r a n e potential. T h e s e are
f a d e d o u t with slight r u n - u p or r u n - d o w n times (1 s) a n d are n o t used in t h e
c o m p u t a t i o n of the m e a n value. T h e times f a d e d out are displayed a n d should n o t
be longer t h a n 1 rain in an e x p e r i m e n t a l p e r i o d of 9 0 - 1 4 0 min.

Measurement o f the threshold o f excitation

T h e n e r v e cells were excited intracellulary just above t h e t h r e s h o l d by signal
c o n n e c t i o n s w i t h r e c t a n g u l a r impulses in a c c o r d a n c e with the s t a n d a r d m e t h o d .
E a c h cell was excited ten times at intervals of 10 s. This t o o k p l a c e 10 rain b e f o r e
the Befeldung, during t h e Befeldung, 10 rain after this and, for a final time, 1 h
after t h e end o f t h e Befeldung.
298

RESULTS

Measurement of the resting potential of the so-called quiet cell

T h e single Befeldung of a nerve cell almost always resulted in the resting
p o t e n t i a l being more or less strongly h y p e r p o l a r i z e d . T h e h y p e r p o l a r i z a t i o n some-
times started shortly after the start of the Befeldung. However, h y p e r p o l a r i z a t i o n
n o r m a l l y occurred only in the last m i n u t e of t h e B e f e l d u n g or, quite frequently,
only 1 - 1 0 rain after the e n d o f the B e f e l d u n g (Fig. 4). T h e nerve cell s o m a t a
m e a s u r e d had m e m b r a n e p o t e n t i a l s b e t w e e n - 2 3 and - 6 8 mV. T h e h y p e r p o l a r -
izatmn in these cases was - 2 and - 1 3 mV (cf. T a b l e 1).
T h e strength o f the evoked h y p e r p o l a r i z a t i o n s is closely c o n n e c t e d with the
m e m b r a n e p o t e n t i a l s o f the cells, Nerve cells with highly negative R P (e.g. --68
mV) were only slightly h y p e r p o l a r i z e d by the B e f e l d u n g (approximately - 2 mV).
On the o t h e r hand, cells with a weak R P (e.g. - 2 3 mV) were strongly h y p e r p o l a r -
ized (e.g. 13 mV) (Fig. 5). Thus, t h e r e is a c o n n e c t i o n b e t w e e n the value o f a
resting p o t e n t i a l set by the cell and its h y p e r p o l a r i z a t i o n caused by the field.
T h e h y p e r p o l a r i z a t i o n also lasted for a n u m b e r of hours. As the p r e p a r a t i o n , as
described in the M e t h o d s section, was not in a n u t r i t i o n solution b u t in a b l o o d
r e p l a c e m e n t solution, it was not possible to establish u n d e r these exper.lmental
c o n d i t i o n s w h e t h e r the d e p o l a r i z a t i o n of t h e m e m b r a n e p o t e n t i a l , which some-
times only occurred after several hours, was d u e to a d r o p in the effect of the
B e f e l d u n g or to physiological reactions of the cell to a lack of supply (Fig. 6).
Measurement of the hyperpolarized resting potential after a second Befeldung
W h e n the h y p e r p o l a r i z a t i o n of a b e f e l d e t ceil had c o m e to a standstill, i.e. the
m e m b r a n e p o t e n t i a l s h a d t a k e n on a stable value, a s e c o n d B e f e l d u n g was carried

nu . . . . I . . . . I . . . . I . . . .

-45

-551

!iiiiiiiii!iiiii!iiiiiiii
i!ili i i i i i i i !i i i!ilili!i i i i !i!i !i !ili i !i
mmmw mm
-551 ~[l~H~fii ,,.~,,
,,,I,,,,,,,,,1.......~".... "~' t~,i~!

0 1o 20 zo 4o SO ~0 70 80 90
e'eAn ~

Fig. 4. Single Befeldung after one Befeldung. T h e resting potential o f the measured hyperpolarizations
was approximately 7 mV.
 299

TABLE 1
Dependence of the hyperpolarization values on the membrane potential of some nerve cells. Neurons
with highly negative membrane potentials hyperpolarize slightly after one Befeldungl Neurons with
weakly negative membrane potentials hyperpolarize strongly after one Befeldung
Resting potential Resting potential Hyperpolarization/mV
~tart v a l u e / m V after 1st B e f e l d u n g / m V
- 68 --70 2
- 67 --68 I
--67 --70 3
-- 66 -- 69 3
-- 63 -- 68 5
- 62 --65 3
-58 -62 4
- 57 -61 4
-56 -62 5
-46 -55 9
--40 -47 7
--39 --45 6
--39 --47 8
--38 -47 9
--28 -38 10
-- 27 -37 10
-- 25 -36 11
-- 24 -37 13

out on a n u m b e r of occasions. T h e newly set m e m b r a n e potential did not

hyperpolarize further. A third B e f e l d u n g also h a d no m e a s u r a b l e effect on the
d e v e l o p m e n t of potential (Fig. 7).

Befeldung of non-quiet cells

(a) A l o n g s i d e quiet cells, o t h e r nerve cells were also included in the experiment.
N e u r o n s w e r e befeldet, the resting potential of which was not stable but h a d a
depolarizing tendency. This t e n d e n c y was s t o p p e d by t h e B e f e l d u n g and, in some
cases, even reversed (Fig. 8).
(b) S o m e ceils, which w e r e possibly excessively d a m a g e d in the p e n e t r a t i o n of
the signal connection, showed strong depolarization. T h e Befeldung did not, as in
case (a), achieve a d r o p in the depolarization, b u t the exact opposite (Fig. 9). T h e
d e p o l a r i z a t i o n was clearly increased a n d practically led to potential c o m p e n s a t i o n
after a short time (approximately 1 h).

Measurement o f the threshold of excitation

N a t u r a l l y , i t w a s interesting to see how the t h r e s h o l d of excitation of a nerve

cell would b e h a v e if its m e m b r a n e potential was hyperpolarized by Befeldung.
300

MV - 7 o

-GO

-40

- 30

-20

--tO

. v la

13

Fig. 5. H y p e r p o l a r i z a t i o n s after one B e f e l d u n g as a function o f the initial m e m b r a n e potential. U p p e r

line: m e m b r a n e potentials of various cells b e f o r e the Befeldung; lower line: hyperpolarizations o f these
cells after c n e Befeldung.

U n f o r t u n a t e l y , t h e r e are few r e s u l t s c o n c e r n i n g this q u e s t i o n . Y e t t h e s e a r e v e r y

i n t e r e s t i n g , for w h i c h r e a s o n t h e y a r e p o r t r a y e d h e r e ( T a b l e 2). U n f o r t u n a t e l y , it
h a s only b e e n possible to c a r r y o u t t h e e x p e r i m e n t t h r e e t i m e s u p to now, t h e
r e s u l t b e i n g t h e s a m e e a c h time. T h e e l e c t r i c a l i n t r a c e l l u l a r s t i m u l a t i o n s , to w h i c h
a r e a c t i o n c a m e w i t h a c t i o n p o t e n t i a l s ( A P ) in a b o u t 8 3 % o f t h e c a s e s in t h e
r u n - u p , w h i c h b r o u g h t a b o u t a r e a c t i o n of 3 6 . 6 % d u r i n g t h e B e f e l d u n g . T e n

TABLE 2
M e a s u r e m e n t of the threshold o f excitation before, during and after a Befeldung (three different nerve
cells). Evoked action potentials ( c A P ) c a u s e d by electrical r e c t a n g u l a r stimuli before, during, 10 min
and 1 h after a Befeldung
RP/ Before During 10 min after 1 h after Hyperpolarization
mV Befeldung Befeldung Befeldung Befeldung mV
cAP cAP eAP cAP
--58 28 12 17 22 --5
--40 27 10 15 20 - 7
-56 28 11 16 22 --6

c A P total = 83% 36.6% 53.3% 71%

 301

ely .,.l .... i. , ;', i .... . . . . I . . . . I . . . . I . . . . I . . . . l . . . . I . . . . I . . . .

-JLO

-20

-3(]

-40

_ ~ ' ' ; ' ' , ' ' : ' , ' . , '

-60
' ' . * * . ' ' . ' O , ' ' , ' ' . ' . 0.°'. "
: ;::::: ;:: ;:: ;:: ,." .,.......
: :-.: :.:/.: :..: :.; :.: :,j ~.: :." _.-£-.
-70

zo 60 ~0 120 xse

Fig. 6. A l t e r a t i o n s o f the m e m b r a n e p o t e n t i a l o f a nerve cell a f t e r o n e h y p e r p o l a r i z a t i o n . A p p r o x i -

m a t e l y 90 rain a f t e r the e n d of the B e f e l d u n g , t h e h y p e r p o l a r i z a t i o n o f t h e m e m b r a n e p o t e n t i a l due to
the field has finished in this ease. A f t e r a p l a t e a u phase, slow d e p o l a r i z a t i o n starts a b o u t 3 h a f t e r the
e n d o f the B e f e l d u n g .

m i n u t e s after the e n d of the B e f e l d u n g , this figure rose to 53.3% again, e v e n rising

to 71% after 1 h. H o w e v e r , it m u s t be m e n t i o n e d that t h e cells u s e d in t h e s e
e x p e r i m e n t s had a relatively strong negative resting potential, w h i c h m e a n s that
the h y p e r p o l a r i z a t i o n w a s not very strong.

Measurement of artefacts

T h e p r o b l e m with e l e c t r o p h y s i o l o g i c a i m e a s u r e m e n t in e l e c t r o m a g n e t i c fields is
to cover all possible artefacts w h i c h may i n f l u e n c e the nerve cell wire o f the

i . . . . i . . . . ] . . . . . . . . | . . . . | ~'-'- .

-LO

li !
--20
E

-30

-40

-~0

,,-,--,---~.~.....;..~-.:.....;

; : :- 2".. ~'- :'_. : '. :'.. :', 3", :', ;"

60 cJO 188 2411

Fig. 7. A l t e r a t i o n o f the m e m b r a n e p o t e n t i a l o f a nerve cell with t h r e e B e f e l d u n g e n . O n l y the first

B e f e l d u n g leads to h y p e r p o l a r i z a t i o n o f the m e m b r a n e potential. B e f e l d u n g 2 a n d 3 have n o effect.
 302

mY

-10

-20

-~O

-40

-50

iiiiiiiiiiiiiiiiiiiiiiii!iii
-50

-70iii0 ~6 50 90 120 159

nln
Fig, 8. A l t e r a t i o n o f the m e m b r a n e p o t e n t i a l o f a n o n - q u i e t cell a f t e r a single B e f e l d u n g . T h e low
d e p o l a r i z a t i o n of the m e m b r a n e p o t e n t i a l o f a nerve cell is s t o p p e d a f t e r o n e B e f e l d u n g a n d t h e n even
b r o u g h t to hyperpolarization.

r e c o r d i n g electrode. T h e r e are some physical p h e n o m e n a ( m a g n e t i c induction,

r a d i o aerial effect, influention, skin effect, etc.) which have p r o p e r t i e s m a s k i n g t h e
ability to influence the m e a s u r e m e n t s . In o r d e r to escape from m a g n e t i c voltage
i n d u c t i o n we i n t e r r u p t e d the m e a s u r i n g loop d u r i n g the e l e c t r o m a g n e t i c Befel-
dung. T h e d i s a d v a n t a g e of this type of m e a s u r e m e n t , however, is t h a t t h e n e u r o n s
( R P ) c a n n o t actually be t e s t e d d u r i n g E M F stimulation. T h e electrical i n f l u e n t i o n
was tested e x p e r i m e n t a l l y a n d we could not see any artefact. O t h e r effects w e r e
calculated (Institute of E x p e r i m e n t a l Physics, F U , Berlin) but t h e r e was no
indication of any electrical or o t h e r influence on the m e a s u r i n g system.

m~ . . . . i . . . . I . . . . I . . . .
. . . . | . . . . I . . . . I ' • " ~ - ' i • ",T,_,.J

-10
r. . : "*'~ ~.'~t ~:i" =~" ;~:,~ :,

-Z6

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-40

-51] !:!:~ :!: !~ L:!i:~" ! :i!~" !:,:: :~:!!.!!!.:~ :,,:!': ! ~! ::,:!'! :'~: !~
0 IO 20 ~O 40 50 50 70 80 BO min,
Fig. 9. A l t e r a t i o n o f the m e m b r a n e p o t e n t i a l o f a nerve cell d e p o l a r i z i n g d u e to injury, a f t e r o n e
Befeldung. T h e d e p o l a r i z a t i o n of t h e cells is strongly a c c e l e r a t e d a f t e r the E;efeldung.
 303

N a t u r a l l y , B e f e l d u n g w a s also c a r r i e d o u t o n cells w h i c h h a d c o m p l e t e l y
c o l l a p s e d p h y s i o l o g i c a l l y ( a l m o s t c o m p l e t e c o m p e n s a t i o n of p o t e n t i a l , n o E P S P ,
I P S P o r A P , a n d n o electrical excitability). I n t h e B e f e l d u n g of t h e s e ceils, t h e r e
w a s n o a l t e r a t i o n of t h e p o t e n t i a l m e a s u r e d . I n t h e e x p e r i m e n t s w i t h q u i e t cells, a
r e l a t i v e l y u n i f o r m t e n d e n c y t o w a r d s h y p e r p o l a r i z a t i o n w a s seen. T h e r e l a t i o n s h i p
b e t w e e n t h e r e s t i n g p o t e n t i a l a n d t h e s t r e n g t h o f t h e h y p e r p o l a r i z a t i o n to b e
a t t r i b u t e d to t h e B e f e l d u n g ( h i g h r e s t i n g p o t e n t i a l - - w e a k h y p e r p o l a r i z a t i o n ; low
r e s t i n g p o t e n t i a l - - s t r o n g h y p e r p o l a r i z a t i o n ) c o r r e s p o n d s to t h e q u a n t i t a t i v e rela-
t i o n s h i p b e t w e e n t h e c o n c e n t r a t i o n r a t i o a n d t h e e q u i l i b r i u m p o t e n t i a l on n e r v e
cell m e m b r a n e s ( N e r n s t ' s e q u a t i o n ) . T h i s is also w h y r e p e a t e d B e f e l d u n g in t h e
e x p e r i m e n t did n o t l e a d to a n y f u r t h e r h y p e r p o l a r i z a t i o n s .

DISCUSSION

T h e r e s u l t s o f t h e B e f e l d u n g o f n o n - q u i e t cells m a t c h t h e a b o v e - m e n t i o n e d
a t t e m p t s at e x p l a n a t i o n . T h e e n d of d e p o l a r i z a t i o n t e n d e n c i e s c o r r e s p o n d s to t h a t
of t h e h y p e r p o l a r i z a t i o n a n d could b e s u b j e c t to t h e s a m e e f f e c t m e c h a n i s m s . T h e
m e a s u r e m e n t o f h i g h l y d a m a g e d n e r v e cells is d i f f e r e n t . T h e a c c e l e r a t i o n of t h e
potential c o m p e n s a t i o n could be substantiated by the fact that the mechanisms
w h i c h led to t h e h y p e r p o l a r i z a t i o n of q u i e t cells c a n n o l o n g e r w o r k effectively.
N e v e r t h e l e s s , t h e y c o u l d possibly r e p r e s e n t a p r o c e s s w h i c h c o n s u m e s a l a r g e
a m o u n t of e n e r g y . T h i s e n e r g y c o n s u m p t i o n c o u l d l e a d to a q u i c k c o l l a p s e o f the
b i o e t e c t r i c i t y of t h e n e r v e cell.
T h e m e a s u r e m e n t o f t h e t h r e s h o l d o f e x c i t a t i o n o f a n e r v e ceil s h o w e d q u i t e
s p e c i a l results. A n i n t e r p r e t a t i o n c a n o n l y b e m a d e very r e t i c e n t l y here. Obviously,
t h e s t r o n g e s t effect o f t h e e l e c t r o m a g n e t i c fields d u r i n g t h e B e f e l d u n g is t h a t it
c a u s e s a d r o p in t h e e v o k e d p o t e n t i a l s to 3 6 % . T h e r e a c t i o n to electrical stimuli
i n c r e a s e s a g a i n a f t e r t h e B e f e l d u n g . T h i s a p p e a r s to c o n t r a d i c t the results o b t a i n e d
u p to now, as t h e excitability is at its lowest at t h e m o m e n t w h e n t h e h y p e r p o l a r -
i z a t i o n c a u s e d b y t h e field is also at its lowest, s o m e t i m e s h a r d l y m e a s u r a b l e . T h e
excitability i n c r e a s e s a g a i n w i t h t h e a m o u n t o f h y p e r p o l a r i z a t i o n o r w i t h t h e d r o p
in t h e B e f e l d u n g . All t h e f i n d i n g s m e a s u r e d i n d i c a t e t h a t v a r i o u s effect m e c h a -
n i s m s exist in t h e field e f f e c t o n n e r v e cells.
M o r e p r e c i s e a n d m o r e n u m e r o u s m e a s u r e m e n t s in o u r i n s t i t u t e , i n t e r alia o n
b r a i n sclices a n d n e r v e cell cultures, a n d e x a m i n a t i o n s o n ion c h a n n e l s will
p r o d u c e f u r t h e r r e s u l t s in t h e n e a r f u t u r e a n d t h u s possibly c o n t r i b u t e to t h e
f o r m u l a t i o n of a g e n e r a l t h e o r y of effects. A s t h e r e h a s b e e n f r e q u e n t s p e c u l a t i o n
in t h e m e d i c a l l i t e r a t u r e in t h e r e c e n t p a s t c o n c e r n i n g t h e effects ~f w e a k
e l e c t r o m a g n e t i c fields in t h e i m m u n e s y s t e m o f m a n , i n v e s t i g a t i o n s a r e c u r r e n t l y
b e i n g c a r r i e d o u t in o u r l a b o r a t o r y o n n e u r o s e c r e t o r y n e r v e cells o n m o l l u s c s
(Lymnaea stagnalis), t h e a i m b e i n g to e x a m i n e t h e field effects r e f e r r e d to in this
paper.
O w i n g to t h e c h a n g e in m e t h o d s in t h e i n t r a c e l l u l a r m e a s u r e m e n t of n e r v e cells,
it w a s p o s s i b l e for t h e first t i m e to m e a s u r e t h e m e m b r a n e p o t e n t i a l f r e e o f
304

artefacts during a n d after the B e f e l d u n g with w e a k H F e l e c t r o m a g n e t i c fields.

Thus, the effects which the fields used had on the bioelectricity of the nerve cells
(clear h y p e r p o l a r i z a t i o n ) were proven. T h e s e results were e s p e c i a l l y i m p o r t a n t
because it was possible to show clear influences o n the electrical propt~rties of
nerve cells, despite the use of field strengths below the m e a n energy of m o l e c u l a r
t h e r m a l motion.
As t h e r m i c effects t h r o u g h t h e fields used are ruled out [4], o n e or m o r e
different effects must cause the a l t e r a t i o n of the bioelectricity of the cells investi-
gated.
As the m e m b r a n e p o t e n t i a l is basically a biophysical p h e n o m e n o n , which can be
explained by the specific p r o p e r t i e s of the cell m e m b r a n e , it would be possible to
imagine effect m e c h a n i s m s which have an effect o n e l e m e n t s of the m e m b r a n e ,
e.g. various p r o t e i n structures, on the basic structure o f the m e m b r a n e (phos-
pholipid d o u b l e layer) in its i n t e r p r e t a t i o n as a liquid crystal or m e m b r a n e
c o n d e n s e r or on the energetics forming the basis of the active t r a n s p o r t m e c h a -
nisms. T h e a l t e r a t i o n s of the ion c h a n n e l s which result from this, a n d t h e r e f o r e
also of the p e r m e a b i l i t y and the resistance of the cell m e m b r a n e , could explain the
m e a s u r e d a l t e r a t i o n s of the bioelectricity o f ~he iielwe cell.

REFERENCES

1 R. Glaser, Bioelectrochem. Bioenerg., 27 (1992) 255.

2 "Nichtionisierende Strahlung", Klausurtagung der Strahlenschutzkommission 7.-9. Dez. 1988,
VerSffentlichung der Strahlensehutzkommission, Bd. 16, Bundesminister t-fir Umwelt, Naturschutz u.
Reaktorsicherheit, Fischer, 1990.
3 LI. Kullniek and H.G. Wolff, in N. Eisner and H. Penzlin (Eds.), Synapse-Transmission-Modulation,
Proceedings of the 19th GSttingen Neurobiology Conference, Thieme, Stuttgart, NY, 1991.
4 K.D. Kramer, personal communication.