NO.
4741 September 10, 1960 NATURE
100
90
SO
70
60
50
~ antigonadotropic function of the pineal gland is in
40
30
20
10
.4 B second hypoactive. Investigations which are under
Fig.!. Oxytocic activity of dog para ventricular nucleus (mean
values of 6 animals), .4, normal activity (30±9 m!'); B, 5 hr,
after administration or pineal extract (90 ± 12 m!'). The means and
their standard errors are given as m!', in terms of U.S. Phal'ma·
copceia reference standard
If this hypothesis is correct, the antigonadotropic
function of the pineal gland ought to be exerted
through the blocking of the oxytocin as a releasing
factor in the nuclei of the neurosecretory hypothala-
mic system.
Our experiment was performed on 12 adult male
dogs weighing about 15 kgm. The animals were
divided into 2 groups: 6 animals served as controls
and another 6 were given intravenous injections of
10 ml. protein extract of bovine pineal gland contain-
ing a nitrogen equivalent of 0·03 mgm.jml. The
animals were killed 5 hr. after the administration of
the pineal extracts, with as little psychic disturbance
as possible, by the rapid intravenous injection of
magnesium as a 25 per cent wjv aqueous solution
of the sulphate. The whole brain and pituitary body
were removed in about 10 min. and placed in 20 vol.
of anhydrous acetone. The supra-optic nuclei, the
paraventricular nuclei, the tuber cinereum and the
neurohypophysis were isolated. Extraction was
performed according to the method of Van Dyke et al. 4.
Holton's methodS was used for the assay of oxytocin.
All determinations were performed in comparison
with U.S. Pharmacopceia posterior pituitary reference
standard.
The normal amounts of oxytocin that we found in
the paraventricular nucleus is in strong agreement
with those of Van Dyke et al.·.
Five hr. after the intravenous administration of
10 ml. protein extract of bovine pineal gland, the
oxytocic activity of the paraventricular nucleus
increased from 30 ± 9 to 90 ± 12 mfL (Fig. 1). We
consider this increased concentration of oxytocin to
be due to blocking of this polypeptide under the
action of the pineal extract.
In a recent paper6, a very low oxytocic activity was
described in the nuclei of the neurosecretory hypo-
thalamic system during the releasing of gonadotropic
hormones. In the present work, concomitantly with
the blocking of gonadotropic hormones, there is a
significant increase of the oxytocic activity in the
nuclei of the neurosecretory hypothalamic system.
This parallelism between the releasing or blocking of
gonadotropic hormones and releasing or blocking
© 1960 Nature Publishing Group
951
respect!vely of oxytocin in the neurosecretory hypo.
thalamIC system supports the hypothesis of Shibu-
saw:a and Martini according to which oxytocin or an
active. substance associated with it represents t.he
releasmg factor for the gonadotropic hormones. On
the other hand, this experiment shows that the anti·
gonadotropic function of the pineal gland is achieved
through blocking of the oxytocin or of an active
substance a~sociated with it in the neurosecretory
hypothalamic system, and not through the direct
interference with gonadotropic hormones. The
fact an antigonadotropic hormone function. This
hypothesis is strongly supported by Milin's histo-
physiological investigations 7 • He found a lowered
activity of the nuclei of the neurosecretory hypo-
thalamic system in the rabbit during the blocking
of the gonadotropic hormones by darkness and an
increased activity of these nuclei during the releasing
of gonadotropic hormones caused by light. In the
first case, the pineal gland is hyperactive, in t.he
way concerning the action of pineal extracts on vaso-
pressin in the neurosecretory hypothalamic systern
make us assume that the anticorticotropic function
of the pineal gland is exerted through the inter·
mediary of vasopressin as a releasing factor of
a substance associated with it, and not through
the direct interference with adrenocorticotropic
hormone.
Thus, it seems that the antigonadotropic mechan-
ism consisting in the blocking of releasing factors
represents a general pattern according to which the
pineal gland exerts its antitropic functions. Investi.
gations now being carried out will, we hope, furnish
new data on the relationship between tropic hormones
and the releasing factors in the neurosecretory hypo-
thalamic system.
S.-M. MILCOl-
S. PAVEL
Institute of Endocrinology,
Bucharest.
1 :\Iilcou, S.-:\!., "Epitlza glanda endocrina", Congo Nat. Sci. )[0<1.,
Bucuresti (May, 1957).
, Shibusawa, K., Saito, S., Fukuda, M., Kawai, T., Yamado, K .. nnd
Tomizawa, K., Endocrinol. (Japan), 2, 3 (1955).
3 Martini, L., ~Iira, L., Pecile, A., and Saito, S., Acta Endocrinol., ~l1PP.
38 (1958).
, Van Dyke, H. B., Karlis, Adamson, jun., and Stanford, L. Engel, in
"Recent Progress in HOl'lnOne Research". 1 (Academic Pres~.
New York, 1955).
'Holton, P., Brit. J. Pharm. Ohem., 3, 328 (194R).
4\ l\iilcOll, S.-M., Pavel, S., and Pterescu, S., Rumanian JfNi. Rf'r.
(in the press).
':\Ii1in, R., Congo Nat. Sci. Med. Bucuresti (}Iay, 1957).
Autonomic Nerve - Smooth Muscle
Transmission
DESPITE extensive work on the morphology and
pharmacology of autonomic effector systems, very
little is known about the mechanism of transmission
of excitation from nerve to smooth muscle. In the
present work our approach has been to apply
modern electrophysiological methods to a suitable
autonomic nerve - smooth muscle preparation in a
manner similar to that described for the sl;;eletal
neuro·muscular junction'.
Smooth muscle cells from the longitudinal layer of
the vas deferens of the guinea pig were impaled with
capillary microelectrodes and membrane potential
 952 NATURE
Fig. 1. !.lembrane potential recorded from a single smooth muscle
fibre of the vas deferens of the guinea pig during repetitive stimu-
lation of the hypogastric nerve (stimuli 1 msec. duration). Suc-
cessive 'junction potentials' increase in amplitude aud rate of rise.
Summation occurs until at a critical depolarization a spike is iultl-
ated
changes recorded during repetitive stimulation of
the hypogastric nerve. The vas deferens was never
spontaneously active. In the experiment illustrated
in Fig. 1, the resting potential was 65 mY. Five
successive stimuli resulted in depolarizations of
progressively greater amplitude until, at a critical
membrane potential (37 mV.), a spike was initiated
and a vigorous contraction occurred. The membrane
potential at which the spike arose was remarkably
constant for the majority of cells (35-38 mV.), The
number of stimuli required to reach this threshold,
however, was variable.
The mean resting potential of the vas deferens was
57 mY. (SEM = 6 mV.). The spike was similal' in
configuration to those recorded from the taenia coli of
the guinea pig' and the uterus of the rata. Spike
amplitudes of up to 84 mY. were recorded (mean
68 mV., SEM = 8 mV.); overshoots were up to
20 mY. The positive phase of the spike (after-hyper-
polarization) generally brought the membrane poten-
tial back to, or beyond, its resting-level.
We have decided to call the successive depolariza-
tions in response to nerve stimulation 'junction
potentials'. We felt that the term 'end-plate poten-
tial' was best reserved for tissues where a definite end
plate region could be distinguished; the term 'post
synaptic potential' did not seem appropriate since,
at present, there is no histological evidence for any
structure resembling a neuronal synapse in smooth
muscle.
Both the amplitude and the rate of depolarization of
the junction potentials increased with successive
stimuli, this facilitation occurring even when the
interval between stimuli exceeded 500 msec. The
rate of depolarization of the junction potential was
slow compared with other post-synaptic potentials.
The rate of decay of the junction potential varied
greatly from preparation to preparation. We think
it is unlikely that this slow decay is dependent on the
time-constant of the membrane, but that it may
represent slow destruction of the transmitter. Brown
and Gillespie' suggested that at least 100 msec. were
required for destruction of the noradrenaline released
from cat spleen by splenic nerve stimulation.
Junction potentials could be recorded in cells
along the entire surface of the vas deferens. Theil'
amplitude and latency varied somewhat from cell to
cell, but there was no indication of a defined end-plate
region. The functional distribution of nerve endings
t,h erefore appears to resemble that of crustacean
© 1960 Nature Publishing Group
September 10, 1960 VOL. 187
muscle' . The amplitude of the junction potentials
was dependent on the number of n erve fibres stimu-
lated. If the hypogastric nerve was stimulated sub-
maximally, the amplitude of junction potentials
all over the muscle was reduced.
In the a bsence of n erve stimulation small 'miniature
junction potentials' were recorded which presum-
ably indicated the spontaneous re lease of transmitter .
Their frequency was lower than that observed at the
skeletal neuromuscular junctions. Their amplitude
was larger (up to 18 mV.); but this may be accounted
for in terms of the small diameter of the muscl E'
fibres' .
The response of the vas deferens to hypogastric
nerve stimulation was unaffected by hexamethonium
(5 x 10- ·) and atropine (10- 6 ), but was blocked by the
anti-adrenergic drug darenthin (5 x 10- 5 ). Thus,
stimulation appears to have been post-ganglionic
and the transmitter is most likely to b e noradrenaline.
These results will be published in detail elsewhere.
We wish to thank Dr. Michael Rand for intl'o-
ducing us to this preparation and for UAsist ing us
with the pharmacology.
G. BURNSTOCK
MOLLIE E. HOLMAN
Departments of Zoology and Physiology,
University of Melbourne,
Victoria.
I del CastUlo, J., and Katz, n., "Progress in Biophys ics", 6, 12~ (Per-
gamon Press , Londou and New York, 1956).
, Holman, Mollie E., J. Physiol., 141, 464 (1958).
, Marshall, Jean M., Amer. J. Physio/ .. 197, 935 (1959).
, Brown , G. L., and Gillespie, J . Soo J. Physiol., 138, 81 (1957).
• Fatt, P. , a nd Katz, B., J. Exp. Bioi., 30, 433 (1 953).
• Boyd, 1. A .. and Martin, A. R. , J . Physiol., 132, 61 (1956).
'Katz. Boo a nd The"leff, S., J . Phy.iol., 137, 267 ( 1957).
PSYCHIATRY
Paradoxical Blocking and Arousal in the
Drowsy State
THERE have been many attempts to investigate
the hypnotic state making use of the electroencephalo-
graph. Though clinically the subject might have the
appearance of sleep, the electroencephalograph was
not found to be different from that found in the
wide-awake hypnotized state. Darrow et al.' believed
that the electroencephalograph in hypnosis had char'-
acteristics that distinguished it from both tme sleep
and full consciousness. Our experience does not con-
firm either of these views, but we have not found the
electroencephalograph of a hypnotized person to hav E'
any distinctive characteristics. Rather the electro-
encephalograph of the hypnotized person can bt'
characteristic of the waking or drowsing electro-
encephalogram without hypnosis, depending upon
the situation and the suggestions given. This finding
is therefore in accord with those of older writers on
the subject, including Paul Schilder'.
In the caRe we are describing, the pat ien t had b een
hypnotized several times. Suggestions were given to
" Allow yourself to go deeper and deepe,· aslee p . . .'.
with explanations to the effect that the word 'sleep'
was h ere being used in a special sense, because the
subject was told that he could still h eal' and compre-
hend the experimenter. On other occasions he waf;
urged to go to sleep and b ecome "oblivious to evet'y ·
thing and be completely relaxed". On one such
occasion when t.he record gave evidence of drowsiness