295

THE PIGMENTARY EFFECTOR SYSTEM V. THE

NERVOUS CONTROL OF EXCITEMENT
PALLOR IN REPTILES
BY LANCELOT T. HOGBEN AND LOUIS MIRVISH.
From the Department of Zoology, University of Cape Town.

(Received nth December 1927.)

(With Seven Text-figures.)

1. INTRODUCTION.
THE chromatic responses of the chameleon have excited curiosity from the earliest
times. They were known to Aristotle, who wrote concerning them: "the change in
the colour of its skin takes place when it is filled with air. It can acquire either
a black colour like that of a crocodile, or ochreous like that of a lizard, or spotted
with black like the panther; for the eyes also change like the rest of the body,
and so does the tail 1 " (Book 11, History of Animals).
Nevertheless, the phenomena of colour change have been studied less ex-
tensively in Reptiles than in Fishes and Amphibia. Among those who have in-
vestigated the physiology of pigmentary response in the chameleon may be men-
tioned Bert, Briicke, Krukenberg and Keller. The only recent investigations on
colour change in Reptiles worthy of note are embodied in the memoirs of Schmidt
(1912) on the histology of reptilian pigmentary effectors, and those of Parker (1906)
and Redfield (1918) on the behaviour of the Mexican horned "toad" Phrynosoma.
Though a variety of pigments occur in the skin of Reptiles, the predominant agents
of colour change in all cases are—as in Fishes and Amphibia—the melanophores.
The melanophores of Reptiles are unicellular branched cells, and according to the
unanimous testimony of Brucke, Keller, Thilenius, Carlton, Parker and Schmidt,
their activity depends on the migration of the pigment along the cell processes.
Three categories of external stimuli are known to be effective in the determination
of colour change—light, temperature and mechanical or other nocuous stimuli.
As in Amphibia and Fishes, Reptiles respond to warmth by pallor; but the reaction
to bright illumination is darkening of the skin, whereas generally speaking—
Necturus being a notable exception—the reverse is the case with Amphibia and
Fishes. Both these reactions are essentially local (vide infra) and it has been held
by many investigators that they depend on the direct excitation of the melano-
phores. In the case of nocuous stimulation however locally applied stimuli evoke
generalised pallor, so that a mechanism of co-ordination is evidently involved.
1
CresswelFs translation.
296 LANCELOT T. HOGBEN and Louis MIRVISH
The nature of this mechanism was regarded by earlier workers as nervous.
A somewhat different interpretation has been proffered on the other hand by
Redfield. The phenomenon of excitement pallor was extensively studied by
Redfield in Phrynosoma (1918). As the result of his investigations this worker
came to the conclusion that the main factor in distributing the stimulus to chromatic
response after nocuous stimulation is the liberation of adrenaline into the circu-
lation. It had long been known that adrenaline induces contraction of the melano-
phores of Vertebrates. And the new evidence adduced by Redfield seemed to
point very strongly to an endocrine control of excitement pallor. The chief points
which he records are: (a) failure of animals to respond to electrical stimulation of
the roof of the mouth after section of the cord at the point between the eighth and
thirteenth vertebra, (b) failure of animals to respond in most cases after epi-
nephrectomy, (c) failure of local section of nerves to selected areas to interfere with
the responses in any way.
Following as they did upon Cannan's researches into the role of the adrenals
in sympathomimetic accompaniments of excitement in the Mammal, these results
were fully consonant with what appeared to be the correct explanation of the
consequences of prolonged excitement in warm-blooded Vertebrates. The criti-
cisms that have been brought to bear upon the work of Caiman and his collaborators
by Stewart and his colleagues have re-opened the question; and it is now very
doubtful whether it is possible to define any specific conditions in which increased
liberation of adrenaline from the suprarenal medulla of intact mammals takes
place. Consequently the determination of excitement pallor in Reptiles acquires,
as Stewart himself has observed, a new interest in relation to the attempt to in-
terpret a physiological, as opposed to pharmacodynamic, role for adrenaline in
the animal body.
Phrynosoma, though displaying the chromatic reaction more noticeably than
most other common lizards of the North American continent, is by no means the
most felicitous type to select for the study of this phenomenon. As is proverbial,
the chameleons among Lacertilia afford the most striking display of colour change,
and this family is well represented in South Africa. The viviparous species
Chamaeleo pumilus is particularly abundant in the Cape Peninsula, and was the
form selected by the writers for investigation. There does not seem to have been
any investigations on the control of colour response in the chameleon, since the
intervention of endocrine agencies (cf. Hogben and Winton, 1922-23 and Hogben,
1924) in the co-ordination of pigmentary effector activity has been established.
Individual chameleons vary considerably in colour, a fact that had given rise
to the popular misconception that a single individual can change from any one
colour to any other. Needless to say any given individual can only change from a
definite light to a definite dark tint. Selected individuals which in the dark con-
dition were a deep olive, changing through green to yellow, were employed in the
experiments recorded below. All the individuals were females. It is a curious
fact that of about two hundred animals collected for these experiments, not more
than half a dozen were males.
 Excitement Pallor in Reptiles 297
2. THE NORMAL CHROMATIC REACTIONS OF THE CHAMELEON.
As indicated above, the main purpose of the investigation was to determine
whether the phenomenon of excitement pallor provides evidence of conditions
under which the adrenals discharge their active product into the blood stream in
increased amount. It was necessary however for this purpose to confirm earlier
observations on the normal sequence of pigmentary changes in the species in-
vestigated. As regards the influence of light, there is no doubt about the salient
facts which have been established with a few exceptions for reptiles in general,
viz. (1) that the exclusion of the organs of vision does not prevent the darkening
which accompanies exposure to bright illumination at ordinary temperatures
(10-200 C.) (Bert, Keller, Krukenberg); (2) that areas locally illuminated respond
locally by darkening (Bert, Briicke, Keller, Parker, Redfield). The conclusion
drawn from these facts by most investigators has been that reptilian melanophores
respond directly to light without the intervention of the nervous system, and
Redfield (1918) upholds this view with experiments in which peripheral sections
of the nerve supply of local areas was carried out. Briicke however regarded the
phenomenon as dependent on nervous co-ordination, a possibility not to be ex-
cluded in view of the evidence brought forward by Parker (1906) to show that
the skin of Amphibia possesses photo-receptors of a simple kind. Briicke based his
view on the effects of spinal transection. His results have been criticised by Fuchs
on the grounds that his preparations had not recovered effects of "shock." In the
course of the present investigations chameleons were kept alive after section of
the cord or cord and sympathetic chain, for more than a week, and in all cases
where the region posterior to the point of section failed to respond to stimulation
of the mouth, the exclusion of light only resulted in pallor anterior to the point of
section. Since these animals responded by pallor to electrical stimulation of the
cloaca on the side posterior to the point of section, the results of shock may be
said to have been adequately eliminated. Similarly with regard to temperature,
exposure to a source of warmth in chameleons in which the cord and chain had
been cut at say the fourteenth vertebra resulted in complete pallor anterior to the
point of section only. These data might be interpreted in the sense defined above,
as inferred by Briicke. His interpretation may be put explicitly as follows: (1) that
the melanophores of the chameleon are normally maintained in a state of "tone"
controlled by a centre in the anterior part of the c.N.s.; (2) that light acts reflexly
through this centre, by releasing them from this state of tone; (3) that warmth
reflexly augments the tonic control of the melanophores. On the other hand, the
data could equally well be harmonised with Redfield's view by different assump-
tions, namely, that the state of expansion of the melanophores at any given moment
is due partly to a more or less continuous state of tone, partly to the direct action
of light in releasing them from this extrinsic control, partly to the effect of warmth
in directly influencing them so as to augment cumulatively the tone effect. On
this view the pallor that normally occurs in darkness would result from the fact
that the tonic control is no longer antagonised by light while the failure when
298 LANCELOT T. HOGBEN and L o u i s MIRVISH
darkened to exhibit pallor in the region posterior to the point of section of the cord
would be due to the fact that the impulses tending to keep the melanophores in a
state of sustained contraction no longer reach the area in question. Without
cutting all dorsal roots of the spinal nerves, a well-nigh impossible operation, there
is no apparent method of distinguishing between the two alternatives experi-
mentally. The latter is however the more economical hypothesis.
All the experiments which follow deal with the analysis of excitement pallor.
Chameleons do not readily respond to rough handling by pallor; and even electrical
stimulation of the skin usually evokes only localised effects. On the other hand
the application of a faradic current from an ordinary shocking coil to the roof of
the mouth or merely mechanical stimulation of the cloaca calls forth a generalised
lightening of tint. Mechanical stimulation of the roof of the mouth failed to evoke
the response. The time relations of excitement pallor are worth noting in contrast
with the phenomena of colour change in Amphibia. They are illustrated in the
following protocol:
Time
h. m. s.
o 00 00 Stimulation of roof of mouth,
o 00 30 Stimulation ceased. Noticeable pallor,
o 00 50 Pallor complete,
o 02 00 Noticeably darker.
o 09 00 Darkening complete. Stimulation of roof on mouth,
o 09 30 Stimulation ceased,
o 09 50 Pallor complete,
o 11 00 Noticeably darker.
o 19 30 Stimulated roof of mouth. Darkening not quite complete,
o 20 00 Stimulation ceased,
o 20 05 Complete pallor.
In general it was found that pallor was complete within from half to two minutes
from the beginning of stimulation. All experiments in relation to this question
were performed on chameleons kept in bright light at room temperature (18-210 C).
The conditions were such as to ensure that the skin remained dark during the
course of th? operations except when stimulated.

3. THE NERVOUS CONTROL OF EXCITEMENT PALLOR.

The generalised pallor which follows after stimulation of the roof of the mouth
or cloaca in the manner indicated implies the intervention of a co-ordinating
mechanism. To ascertain how far nervous and endocrine agencies enter into this
phenomenon, the first question to be investigated was the effect of section of the
c.N.s. at different levels; and as there was little difficulty in obtaining large
supplies of chameleons, it was possible to investigate this issue extensively by
experiments involving (1) section of the cord alone at different levels, (2) section
of the cord and both of the sympathetic chains simultaneously at different levels,
(3) section of the cord with unilateral section of the chain, (4) section of the chain
alone.
{a) Section of the cord alone. When the cord is cut at any level in the trunk
region, stimulation of the cloaca produces pallor only on the side posterior to the cut
 Excitement Pallor in Reptiles 299
(Text-fig. 3). The two regions as in all the segmental effects hereunder described
are very sharply defined (Text-fig. 2). When the cord is cut at any level anterior to a
point corresponding to the tenth or eleventh vertebra—in extreme cases the
critical point varied individually from behind the ninth to the twelfth in one
instance—stimulation of the roof of the mouth results in complete pallor on the
side anterior to the point of section, while the region behind remains completely
dark (see Text-fig. 1, a, b7 c). How striking is the localisation of the response may

a. 5th vertebra c. 10th vertebra

b. 8th vertebra d. 13th vertebra

Fig. i. Effects of stimulation of mouth after section of cord only at different levels.
(The shaded area remains dark.)

be inferred from an actual photograph here reproduced (p. 300). Behind this
region—from the eleventh or twelfth vertebra backwards that is to say—the effects
of section of the cord alone were quite different (Text-fig. 1, d). On stimulation
of the roof the mouth in animals so treated, the result obtained was a generalised
pallor affecting the whole body with the exception in nearly all cases of a short
region at the distal extremity of the tail. These experiences, obtained repeatedly
on a large series of animals, showed conclusively that a nervous agency of some
kind is the main factor in the co-ordinating mechanism; but they leave open the
possibility that adrenal secretion plays an adjuvant role in the phenomenon. For,
BJEB'Viv 2O
3<x> LANCELOT T. HOGBEN and Louis MIRVISH
if the nerve supply of the adrenals emerges just in front of the tenth or eleventh
vertebra, it is possible that the pallor of the posterior half of the animal when the
cord is cut behind this point results from liberation of adrenaline into the circula-
tion. This possibility did not seem at the outset a likely one in view of the conse-
quence obtained from cloacal stimulation after transection of the cord in the same

Fig. 2. Photograph of chameleon showing pallor anterior to point of section

at tenth vertebra after stimulation of roof of mouth.

12th vertebra 15 th vertebra

Fig. 3. Effects of stimulation of cloaca after section of cord only in posterior
region (12th and 15th vertebra).

region of the trunk, and in view of the fact that the generalised pallor resulting
from stimulation of the mouth after transection at the level of e.g. the thirteenth
or fourteenth vertebra was perfectly uniform both as regards its extent and rate
of development. As will be seen, the effects of section of the cord and chain simul-
taneously provide conclusive evidence for rejecting the view that adrenal secretion
has any significant influence in contributing to excitement pallor. One fact however
is worthy of mention here. In order to exclude the possibility that adrenaline
secretion may reinforce the effects of continued stimulation, stimulation of the
 12th vertebra 14th vertebra
Fig. 4. Effects of stimulation of mouth after section of cord and sympathetic
chain of both sides in posterior region.

la

lla lib
Fig. 5. Effects of stimulation of mouth after section of cord and unilateral
section of sympathetic chain:
I a. Left side: chain cut at same level as cord (13th vertebra).
I b. Right side: chain intact.
II a. Left side: chain intact.
II b. Right side: chain cut at same level as cord (12th vertebra).
20-2
302 LANCELOT T. HOGBEN and Louis MIRVISH
mouth or cloaca in cases, where purely segmental responses occurred, was in some
cases protracted for a period of ten minutes or even a quarter of an hour.
(b) Section of the cord and chain. When in addition to cutting the cord, the
sympathetic chain is cut on both sides at the same level, the result of stimulating
the mouth is a pallor confined to the region anterior to the point of section, even
when the cut is made in the region behind the eleventh vertebra (Text-fig. 4). In
two experiments in which the cord was cut at the level of the twelfth and thirteenth
vertebra, and the chain was cut on both sides at the level of the sixteenth vertebra,
pallor extended after stimulation of the roof of the mouth beyond the point of
transection of the cord as far as the level, where the chain was cut. In this case
the chain was cut behind the region where the adrenals are located.
(c) Section of the cord with unilateral section of the chain. The most conclusive
evidence in favour of a purely nervous
interpretation of the determination of ex-
citement pallor in the chameleon lies in the
effects of spinal transection with unilateral
section of the sympathetic chain (see Text-
fig. 5). In this case the region posterior to
the point of section remained dark on the
side on which the chain was cut, while the
side on which the chain was intact exhibited
generalised pallor after stimulation of the
roof of the mouth.
(d) Section of the chain only. Section of F i g 6 E f f e c t o f s t i m u i a t i o n o f t h e m o u t h
the chain only does not interfere with the after section of chain involving partial de-
generalised pallor which follows stimulation
of the roof of the mouth. But in one ex-
periment (Text-fig. 6) in which the cut went through a large ganglion in the
posterior region of the trunk a narrow band of skin remained dark in the
segment of the cut.

4. THE NATURE OF NERVOUS CONTROL OF PIGMENTARY EFFECTOR

ACTIVITY IN THE CHAMELEON.
The evidence brought forward in section 3 clearly indicates that the pallor
resulting from nocuous stimulation is predominantly, if not wholly, nervous. Two
alternatives now present themselves: is this control exercised through direct inner-
vation of the pigmentary effectors, or is the production of pallor due to some
chemical conditions such as oxygen-want, etc., resulting from extreme constriction
(or dilation) of the peripheral arterioles? As has been pointed out by the senior
author, the latter possibility has too frequently been overlooked. In this case it
can be rejected on experimental grounds by cutting out the influence of the circu-
lation. If the body is divided into strips by section at right angles to the caudal
cephalad axis, pallor can always be induced in isolated segments of the trunk by
 Excitement Pallor in Reptiles 303
electrical stimulation of the caudal end of the c.N.s. The experience can more-
over be repeated again and again on the same strip. It is very difficult to believe
that a reversible effect of this kind could be repeated after the circulation had
been stopped, unless the pigmentary effector organs were in direct connection
with the central nervous system. And it thus seems legitimate to draw the con-
clusion that the production of excitement pallor depends on the direct innervation
of the melanophores in Reptiles. It should be borne in mind that there is no
available histological evidence for the direct innervation of the pigmentary effectors
in Reptiles and Amphibia, though the nervous connections of the Melanophores
of Fishes have been observed by more than one investigator.

STinULftT/onr OF
r-JOVTH

TAIL

HIND LIMBS
FORE Unas
Fig. 7. Diagrammatic representation of the nerve paths involved in the control of the pigmentary
effector system of the chameleon on the basis of the experiments recorded in the text.
For the purpose of diagrammatisation the number of ganglia is reduced, and the ascending and
descending afferent paths from cloaca and mouth respectively are represented in each case by
a single neurone. Section of the cord alone anterior to A restricts the pallor following stimulation
of the mouth to the region in front of the cut. After section of the cord alone at the level
indicated by B, generalised pallor involving the hind extremities with the exception of the tip of the
tail follows stimulation of the roof of the mouth.

With this direct evidence available, it is possible to map the innervation of the
pigmentary effector system in the chameleon on the basis of the experiments re-
corded in the preceding section. All the results therein recorded are represented
diagrammatically in Text-fig. 7. The experiments show that the efferent impulses
are distributed by the sympathetic nervous system, and from the fact that stimu-
lation of the cloaca after transection of the cord at any level results in pallor
posterior to the cut only, it may be concluded that post-ganglionic neurones
supplying each segment of the trunk are connected by pre-ganglionic neurones to
the cord in the same segment; and further, that there are no ascending nerve paths
in the chain. The same assumption explains why the section of the chain alone
does not prevent the production of pallor behind the cut, when the mouth is
stimulated. The segmental character of the response to stimulation of the mouth
after section of the cord in front of the tenth vertebra indicates that up to this
304 LANCELOT T. HOGBEN and Louis MIRVISH
level there are no descending pre-ganglionic neurones in the sympathetic chain;
while the fact that generalised pallor follows after the stimulation of the mouth
when the cord alone is cut behind the nth-i2th vertebra, indicates that there
pass into the chain about the level of the ioth-i2th vertebra some pre-ganglionic
neurones which have a descending course, distributing impulses leaving the cord
at this point to segments posterior to it. The fact that the tip of the tail remains
dark after section of the cord in the region behind the nth-i2th vertebra is
explicable on the assumption that these descending neurones terminate in front of
the region where the post-ganglionic neurones supplying the pigmentary effector
organs at the extremity of the tail arise. The same assumptions account for the
effects of unilateral section of the chain at the same level as the cord and of section
of the chain at a lower level than that at which the cord is cut in the same region.

5. ACTION OF ADRENALINE.
Some further light is thrown on the possibility that adrenaline secretion enters
significantly into the phenomenon of excitement pallor in the chameleon by a
study of the minimal effective dose. In the first two experiments synthetic adrenaline
(suprarenin hydrochloride synth. Hoechst) was employed. In all cases the injection
was intraperitoneal to ensure rapid absorption, and the dosage indicates the
equivalent amount in 1 c.c. of saline solution.
Experiment I. Two medium sized chameleons were injected with 1 : 10,000.
General pallor resulted within five minutes. The pallor was more intense, and the
hue more yellowish than in any case of pallor which supervened after nocuous
stimulation. The condition persisted for about 36 hours.
A chameleon of the same size (as nearly as possible) became completely pale
within ten minutes after an injection of 1 : 50,000. The condition persisted for
the period of three hours during which it was under observation. The pallor in
this case was accompanied by the same intensely yellow tint. Three other chame-
leons of about the same size received respectively 1 : 150,000, 1 : 300,000 and
1 : 500,000. An incomplete and somewhat patchy pallor in no case extending over
the whole body ensued in all three cases, and subsided within an hour. Four
chameleons which received a dosage of 1 : 1,000,000, 1 : 5,000,000, 1 : 10,000,000,
1 : 50,000,000 displayed no response whatever.
Experiment II. A second series of chameleons were injected via the peritoneum
with 1 c.c. in each case of synthetic adrenaline in the following dilutions 1 : 25,000,
1 : 50,000, 1 : 100,000, 1 :200,000, 1 : 400,000,1 : 800,000,1 : 1,000,000. Those
that received 1 : 100,000 or the stronger solutions displayed complete pallor after
15 minutes. The pallor was still maximal after two hours. When examined ten
hours later, the pallor had not completely disappeared. Those that received doses
of 1 : 200,000 and 1 : 400,000 did not display complete pallor, but gave a noticeable
reaction persisting at least two hours. With solutions of 1 : 800,000 and 1 :1,000,000
the results were entirely negative.
Experiment III. Seven chameleons were injected (intraperitoneal) each with
 Excitement Pallor in Reptiles 305
1 c.c. of the following solutions of Parke Davis' adrenaline 1 :100,000,1 :150,000,
1 : 200,000, 1 : 300,000, 1 : 400,000, 1 : 500,000, 1 : 600,000, After 15 minutes
the first three were incompletely pale. The last showed no reaction. A slight
pallor was displayed by the remainder. After 30 minutes, Nos. 1, 2, 3 were
completely pale, Nos. 4, 5, 6 still displayed a slight degree of pallor, and No. 7 a
doubtful reaction. After 1 \ hours, Nos. 1,2,3 showed complete pallor, Nos. 4,5,6
slight pallor. After ten hours, Nos. 1,2,3 were still noticeably paler than the controls
and No. 4 showed a slight pallor. Nos. 5, 6, 7 were all completely dark.
From these observations it is seen that there is a fairly definite threshold
effective dose for the adrenaline pallor reaction, that the minimal quantity is from
a physiological standpoint very considerable, and that this quantity produces a
response which persists for a period which is very protracted in comparison with
the effects observed in connection with the phenomenon of excitement pallor. It
is clear that neither of these conclusions reinforce the likelihood that adrenal
secretion plays any significant part in the production of excitement pallor.
The persistence of the response to adrenaline secretion is in keeping with the
effects of adrenaline on other effector systems in cold-blooded animals. Thus the
pressor effect which in mammals is so characteristically evanescent may be pro-
longed over a period of over a quarter of an hour in the case of the tortoise (Hogben
and Schlapp, 1924). Whatever explanation may be given to this persistence, it
implies that if adrenal secretion entered into the phenomenon of excitement pallor,
prolonged nocuous stimulation should evoke a more lasting pallor than stimulation
for shorter periods. Whether such is actually the case may be inferred from the
following protocol:
Time
m. s.
00 00 Stimulated i minute. Complete pallor at end.
04 00 Dark again.
05 00 Stimulated 5 minutes. Complete pallor.
12 30 Dark again.
13 00 Stimulated 15 minutes: darkening began about 8 minutes after stimulation
began and pallor complete by time current was cut off.
Thus in the case of a stimulus applied for one minute recovery took three minutes.
With a stimulus of five minutes duration, recovery took place in two and a half
minutes, and with more prolonged stimulation recovery supervened before the
cessation of stimulation.
Effects of Epinephrectomy. The effects of removal of the adrenals were investi-
gated on a few acute preparations only. Had the results been otherwise the con-
clusion drawn from them would be of doubtful value unless confirmed by experi-
ment on chronic preparations. Unfortunately, although the adrenals are in an
eminently accessible situation for operative procedure, the chameleon did not prove
to be a very viable animal for laboratory purposes, and preparations that apparently
recovered well immediately after the operation did not survive till the wound was
healed. However the application of nocuous stimuli to the mouth in animals in
which the cord was intact or cut at the level of the thirteenth to fourteenth vertebra
3o6 LANCELOT T. HOGBEN and L o u i s MIRVISH
produced generalised pallor, except in cases where the chain which lies very close
to the adrenals was cut or damaged in the course of removing the latter. It was
thus possible to find no evidence of participation of increased adrenal secretion
in the phenomenon of excitement pallor in the chameleon.

6. THE ACTION OF CERTAIN DRUGS.

Far too much importance has been attached by comparative physiologists to
the action of drugs, as indicative of nervous control of effector structures, appa-
rently under the misapprehension in some cases that drug action is a good deal
more specific than is actually the case. It cannot be too strongly emphasised that
most of the well-known drugs have a general protoplasmic action, and that their
selective action is generally speaking quantitative rather than qualitative. Con-
clusions based on this line of evidence as brought forward in a recent paper by
Carter (1925) on the supposed innervation of the cilia of veliger larvae are of little
significance, if unsupported by more direct forms of experimental procedure. In
the experiments which have been conducted by us an attempt was therefore made
to discriminate between peripheral and central action. The results are essentially
what would be expected from the direct evidence that the melanophores of the
chameleon are under c.N.s. control, tending to keep them in the contracted state.
The following experiments show that strychnine and caffeine act through the
C.N.S. by augmentation of the activity of the chromatic centre, and that the equally
characteristic action of curare and atropine in the opposite sense is due to paralysis
of nerve endings. Taken in conjunction with the foregoing experiments, the new
data reinforce conclusions already inferred from an independent source.
A few data regarding the action of drugs on the pigmentary effector system of
Reptiles are available from the work of previous authors, who did not attempt to
identify the seat of action, except in the case of curare, where the darkening pre-
viously observed by Bert to follow injection was shown conclusively by Krukenberg
to be due to peripheral action on the nerve endings. Pallor following the injection
of strychnine is recorded by Briicke, Krukenberg and Keller, and an identical
response to caffeine is recorded by Krukenberg, who also observed darkening after
administration of atropine.
Action of Strychnine. Conclusive proof that the pallor which accompanies the
excited state following strychnine injection acts through the c.N.s. is afforded by
experiments of which the following protocol is typical:
Time
m. s.
00 00 Injection of 0-0003 &&- strychnine, dark animal.
00 10 Excitable but still completely dark.
00 15 Pleurothotonos: pallor commences to develop.
00 20 Pallor complete.
00 25 Cord and chain cut at 13th vertebra.
00 27 Pallor in front of cut complete: region behind noticeably darker.
00 35 Pallor in front of cut complete: region behind completely dark.
00 50 Ditto. Still breathing.
 Excitement Pallor in Reptiles 307
Action of Cocaine. The action of cocaine at first raised the hope of discriminating
between the alternative hypotheses to account for the light reaction adumbrated
in § 2. A dose of o-ooi gm. injected into the cord in the tail region produced
pallor which in a few seconds manifested itself near the seat of injection and
gradually crept forwards till maximal pallor of that intense yellowish tint so charac-
teristic of the response to adrenaline was general within 3—5 minutes later. Since
cocaine acts specially on the sensory nerve endings, and since the propagation of
the effect suggested a nervous route, the inference at first appeared to justify the
view that light inhibits the chromatic centre reflexly through photo-receptors in
the skin (cf. § 2 above). This conclusion did not prove to be sustained by further
examination, as is shown by the following facts:
(1) Injection of cocaine into a chronic preparation with section of cord and
chain at about the level of the thirteenth vertebra produced generalised pallor.
(2) In chameleons injected with cocaine subsequent section of the cord and
chain did not affect the generalised character of the response, nor did complete
destruction of the cord in the posterior part of the trunk region.
It is thus clear that the cocaine pallor is not due simply to interference with
afferent paths, since it still occurs when all efferent routes are destroyed.
Action of Histamine. Histamine has a very similar action to that of adrenaline
and cocaine. An injection of o*ooi gm. (intraperitoneal) produces complete general-
ised pallor within ten minutes accompanied by twitchings with complete recovery
within 12 hours. The influence of histamine is not upon the chromatic centre
since section of the cord and chain does not affect the character of the response as
is the case with strychnine.
Action of Caffeine. Injection of 1 c.c. 1 per cent, solution of caffeine produced
pallor within ten minutes as would be expected. After section of the cord and
chain at about the thirteenth vertebra injection of the same quality produced pallor
anterior to the point of section. The following experiment illustrates both the
well-known action of caffeine on the higher centres of the c.N.s. and the de-
cussation of fibres below the chromatic centre. The right side of the brain of a
chameleon was destroyed after section of the cord about the level of the ninth
vertebra. Injection of the minimal effective dose of caffeine then produced pallor
on the right side of the body anterior to the point of section of the cord.
Action of Curare and Atropine. An injection of 1 c.c. 1 per cent, curare in pale
chameleons kept in darkness produces melanophore expansion with general mortu-
paralysis. The same quantity of atropine produced darkening without paralysis.
Stimulation of the cord after darkening produced by atropine and curare did not
result in pallor over the region innervated by the part of the c.N.s. stimulated.

7. CONCLUSIONS.
The main outcome of this investigation may be stated as follows:
(1) The pigmentary effector system of the chameleon is directly innervated
through the C.N.S., and the production of excitement pallor is determined through
this agency.
308 LANCELOT T. HOGBEN and Louis MIRVISH
(2) The light and heat reactions are not wholly independent of the C.N.S.
A chromatic centre in the brain at least plays a static role in tending to keep the
melanophores in a state of contraction, except when released from that condition
by external stimuli.
(3) While a number of circumstances make it unlikely that adrenaline plays
any part in the production of excitement pallor, no evidence could be found
to support the view that excitement pallor is conditioned or reinforced by active
liberation of adrenaline into the circulation.
The authors are unable to offer any explanation of the difference between their
own findings and those of Redfield. It seems unlikely, in view of the striking
uniformity of the characteristic features of Reptilian pigmentary effector activity,
that different mechanisms are involved in the production of excitement pallor in
the chameleon and in Phrynosoma. Until the influence of adrenal secretion has
been demonstrated in material more favourable for the study of colour response
than in Phrynosoma, it is legitimate to express the doubt that the phenomena of
colour response in Reptiles provides conclusive evidence of the possibility of
defining conditions in which the liberation of adrenaline into the vertebrate circu-
lation in increased quantity takes place.

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