Psychopharmacologia (Berl.

) 19, 95--104 (1971)

9 by Springer-Verlag 1971
The Effects of Chronic Admi ni strati on of Ethanol
on Startle Thresholds i n Rats
R. J . ~IBBI1N-S, ~{. KALAI~T, A. E. L~BLA~c, a n d J . W. ~LARK
Addi ct i on Research Foundat i on of Ontario, and Depar t ment of
Pharmacology, Uni versi t y of Toronto, Toronto 5, Canada
Received J ul y 23, 1970
Abstract. The thresholds for st art l e responses to electric shock were measured
in adul t male Wi st ar st rai n r at s given ethanol dai l y in doses rising from 3 to 7 g/ kg
over a 30-day period, and in controls receiving equicaloric doses of sucrose. Tests
made 23, 36, or 47 h aft er ethanol (i.e., during par t i al or complete ethanol with-
drawal) gave t hreshol d values significantly lower t han those obt ai ned wi t h
sucrose-treated controls. The difference became great er aft er longer ethanol t r eat -
ment and larger doses. However, when t hreshol d measurements were made under
t he acut e influence of ethanol in t he experi ment al group, t he mean values were
vi r t ual l y equal t o those of t he sucrose controls. This normalization, by ethanol,
of a di st urbance produced by absence of ethanol in a chronically t r eat ed animal
is i ndi cat i ve of physi cal dependence. Following t ermi nat i on of ethanol t r eat ment
t here was a gradual r et ur n of st art l e thresholds almost to control values over a
rel at i vel y short period, i ndi cat i ng t hat t he changes underlying t he hyperexci t abi l i t y
are readi l y reversible.
Key-Words: Et hanol - - Tolerance - - Dependence - - Wi t hdrawal Re a c t i o n -
St art l e Threshold - - Rat .
The c hr oni c i n g e s t i o n o f l a r ge a mo u n t s of e t h a n o l i s wel l k n o wn t o
gi ve r i s e t o i n c r e a s e d t o l e r a n c e a n d t o p h y s i c a l d e p e n d e n c e whi c h i s
ma n i f e s t e d as wi t h d r a wa l s y mp t o ms r a n g i n g i n s e v e r i t y f r o m " s h a k e s "
a n d i n s o mn i a t o d e l i r i u m t r e me n s . Ac c o r d i n g t o one h y p o t h e s i s (Col l i er,
1965), i n c r e a s e d t o l e r a n c e a n d p h y s i c a l d e p e n d e n c e c oul d r e p r e s e n t
t wo a s p e c t s of t h e s a me c e l l ul a r r e s pons e t o c hr oni c e x p o s u r e t o a dr ug.
P h a r ma c o l o g i c a l i n v e s t i g a t i o n o f t h e me c h a n i s m of t he s e c ha nge s , a n d
of t h e r e l a t i o n s h i p b e t we e n t h e m, wo u l d be h e l p e d s u b s t a n t i a l l y b y t h e
d e v e l o p me n t o f s e ns i t i ve a n d r e l i a bl e me t h o d s f or me a s u r i n g t o l e r a n c e
a n d p h y s i c a l d e p e n d e n c e i n l a b o r a t o r y a ni ma l s . One me t h o d f or me a s u r -
i ng mi n o r de gr e e s of e t h a n o l i n t o x i c a t i o n wa s d e s c r i b e d r e c e n t l y ( Gi b-
bi ns et al . , 1968), a n d h a s be e n e mp l o y e d s uc c e s s f ul l y t o d e mo n s t r a t e t h e
t i me c h a r a c t e r i s t i c s of a c q u i s i t i o n a n d l oss o f t ol e r a nc e , d u r i n g a n d a f t e r
c hr oni c a d mi n i s t r a t i o n o f e t h a n o l t o r a t s ( Le Bl a nc et al . , 1969). The
p r e s e n t p a p e r de s c r i be s a me t h o d f or f ol l owi ng t h e cour s e o f d e v e l o p me n t
o f p h y s i c a l d e p e n d e n c e on e t h a n o l .
7 Psychopharmacologia (Berl.), Vol. 19
96 R. J. Gibbins, H. Kalant, A. E. LeBlanc, and J. W. Clark:
I t has been r epor t ed t ha t et hanol , admi ni st ered by gavagc i n doses
of 1- - 4 g/kg, produces a significant el evat i on of t he t hr eshol d stimulus
for el ect roshock seizures i n bot h r at s (Allan and Swi nyard, 1949) and
mice (McQuarrie and Fingl, 1958), I n t he l at t er s t udy i t was not ed t ha t
t he el evat i on of t hreshol d was followed by a decline t o subnor mal val ues
f or a vari abl e peri od of t i me following t er mi nat i on of t he alcohol i nt oxi ca-
t i on. The dur at i on and i nt ensi t y of t hi s r eact i ve hyper exci t abi l i t y were
f ound t o be pr opor t i onal t o t he i nt ensi t y and l engt h of t he precedi ng
peri od of i nt oxi cat i on, rangi ng f r om a few hours af t er a single dose
of et hanol , t o several days following a t wo week pr ogr am of adminis-
t r at i on of 2 g/ kg ever y 8 h. McQuarric and Fi ngl suggest ed t ha t t he post -
wi t hdrawal hyper exci t abi l i t y was an easily measurabl e mani f est at i on of
physi cal dependence.
However , el ect roshock seizures are compl ex responses whi ch are
difficult t o correl at e wi t h spont aneous wi t hdr awal phenomena. Spont an-
eous convulsions have been observed in severe wi t hdr awal react i ons i n
man (Victor, 1966), dogs (Essig and Lam, 1968), monkeys (Ellis and
Pi ck, 1970) and mice (Freund, 1969), but not i n rat s. Moreover, et hanol
was shown t o have di fferent effects upon di fferent par amet er s of electro-
shock seizures (MeQuarrie and Fingl, 1958). Ther ef or e i t seemed @sir-
able t o expl ore anot her measure of hyper exci t abi l i t y, mor e closely r el at ed
t o nor mal sensory pat hways and mot or responses, whi ch mi ght pr ovi de
an i ndi cat i on of lesser degrees of wi t hdrawal react i on. The " f l i n c h j ump"
pr ocedur e (Kimble, 1955; Evans, 1961, 1962; Hof f man et al . , 1964) was
selected for t hi s purpose. Essent i al l y, t hi s consists of measur ement of t he
t hreshol d i nt ensi t i es of electric shock, del i vered t hr ough a grid on whi ch
t he ani mal is st andi ng, r equi r ed t o make i t flinch (i.e., t o lift one paw)
or t o j ump off t he grid. Compari son of t he t hreshol ds for t he ani mal s
before, duri ng, and af t er chroni c et hanol t r eat ment was f ound t o pr ovi de
a useful i ndex of t he degree of alcohol dependence produced.
Methods
The appar at us consists of 4 Luci t e t est chambers, each about 20 cm
long, 13 cm wide, and 25 cm high, wi t h a grid floor t hr ough which br i ef
shocks of vari ous intensities can be del i vered at vari abl e t i me i nt erval s.
The gri d is composed of 16 stainless steel rods, 3.2 mm in di amet er set
1.25 cm apar t . The back and bot h ends of each compar t ment are pai nt ed
black. The grid is electrified (al t ernat e bars connect ed, shock not scrambl-
ed) by a st ep-up t r ansf or mer wi t h a cent er - t apped secondary t hr ough a
small aut ot r ansf or mer f r om an A.C. power source. The secondar y vol t age
r at i ng of t h e t r ansf or mer is 240-0-240, and t he out put 42 vol t amps.
A double-pole 4-position switch serves t he dual f unct i on of changing t he
out put vol t age connect i ons f r om a gi ven val ue t o doubl e t ha t value, and
Measurement of Ethanol Dependence in Rats by Startle Thresholds 97
swi t chi ng a vol t met er t o sui t t he connect i ons. The shock voltage is
del i vered t o each of t he i ndi vi dual gri d floors t hr ough a 450,000 ohm
resi st or (non-i nduct i ve) so t ha t each ani mal has his own Hmi t i ng resi st or.
Assumi ng t h a t t he ani mal cont r i but es an addi t i onal 50,000 ohms, t he
t ot al resi st ance i n each ci rcui t is 500,000 ohms. Thi s pr ovi des a conve-
ni ent basi s f or det er mi ni ng t he i nt ens i t y of t he shock f r om t he vol t age
r egi st er ed on t he vol t met er . A t a pe - pr ogr a mme r and i nt er val t i mer are
used t o cont r ol t he pr es ent at i on of shock st i mul i . A si gnal l i ght is
act i vat ed each t i me t he equi pment del i vers a st i mul us, even when t he
vol t age of t he l at t er is set a t zero.
Thr eshol ds were det er mi ned b y t he me t hod of cons t ant st i mul i
(Guilford, i 954; Hof f ma n e~ a l . , i964). Each ani mal was gi ven a 2 rai n
per i od of a da pt a t i on i n t he c ompa r t me nt af t er whi ch 6 di fferent shock
i nt ensi t i es (0.0, 0. i , 0.2, 0.3, 0.4 and 0.5 mA) were pr esent ed l 0 t i mes
each i n a pr e- dct er mi ned r a ndom order. The dur at i on of each shock
was 0.5 see and t he i nt er - shock i nt er val var i ed r a ndoml y about a mean
of 12 sec. At t he end of each bl ock of 12 t ri al s t he ani mal was r emoved
f r om t he c ompa r t me nt and t he gri d floor wi ped clean.
The subj ect s' responses t o t he shock st i mul i were classified as "fl i nch",
" j u mp " , or " no r esponse". " Fl i nch" was r ecor ded when t he ani mal made
an a br upt st art l e-l i ke move me nt wi t hout r emovi ng mor e t ha n one paw
f r om t he grid. " J u mp " was r ecor ded i f t wo or mor e paws were r emoved
f r om t he gri d i n response t o t he shock st i mul us (Ki mbl e, 1955; Evans ,
1961). Thr oughout t he exper i ment t he ani mal s were t es t ed b y t he same
t hr ee exper i ment er s. One set t he shock i nt ensi t i es while each of t he
ot her t wo obser ved and r ecor ded t he responses of t wo ani mal s at a
t i me, as di ct at ed b y t he si gnal lights. The l at t er t wo obser ver s di d not
know whi ch ani mal s had recei ved whi ch t r eat ment . I n a pr el i mi nar y
exper i ment i n whi ch t he same t wo obser ver s i ndependent l y classified
t he responses of t he same pai r s of l 0 ani mal s duri ng 8 t es t sessions of
i 20 t ri al s each, t he i nt er - obser ver r el i abi l i t y coefficients were all f ound
t o exceed r ~ 0.93. These resul t s agree ve r y well wi t h t hose r epor t ed b y
Eva ns (196i) and s uppor t his concl usi on t h a t t he vi sual l y det ect ed
response pa t t e r ns ar e sat i sf act or i l y di scri mi nabl e.
Twent y- f our nai ve, mal e Wi s t ar st r ai n r at s (initial wei ght s 288 t o
4i 6 g) were r a ndoml y assi gned t o 2 gr oups of equal size. For t he first
6 days of t he exper i ment bot h gr oups were i nt uba t e d wi t h a 500/0 (w/v)
sol ut i on of sucrose i n t a p wat er, i n an a mount cal ori cal l y equi val ent t o
a 3 g/ kg dose of et hanol , and t es t ed on days 2 , 4 and 6, 23 h af t er i nt uba-
t i on. Ther eaf t er , t he exper i ment al ani mal s were gi ven et hanol (30~ w/ v
i n t a p wat er ) in r egul ar l y i ncreasi ng doses and t he cont rol s equal vol umes
of t he equi cal ori c sol ut i on of sucrose. Begi nni ng on da y 6 wi t h 3 g/ kg
t he et hanol dose was i ncreased b y 1 g/ kg ever y 6 days unt i l a l evel of
7.
98 R. J. Gibbins, H. Kalant, A. E. LeBlanc, and J. W. Clark:
7 g/kg was r eached on day 31. Thi s dose was mai nt ai ned unt i l day 37,
af t er which bot h et hanol and sucrose t r eat ment s were di scont i nued.
Thr eshol d det er mi nat i ons were made ever y second day duri ng t he
chroni c t r eat ment peri od and dai l y duri ng t he wi t hdrawal period. Wi t h
t he except i ons not ed below, t he ani mal s were t est ed on t he second and
f our t h days of a gi ven dose level, 23 h af t er i nt ubat i on, at whi ch t i me
previ ous experi ence (Hawki ns e t a l . , 1966; Khanna e t a l . , 1967) had
i ndi cat ed t ha t t her e woul d be l i t t l e or no residual alcohol i n t he blood.
On t he si xt h day at each dose level t he t est was done 30- - 35 rain af t er
i nt ubat i on, at t he probabl e t i me of peak bl ood level. Thi s t i me was
chosen because earlier wor k ( Kal ant and Czaja, 1962) had shown t ha t
doses of as much as 4 g/kg by gar age pr oduced t hei r maxi mum i nt oxi cat -
i ng effect i n 30- - 60 min. The t est s on days 32 and 37 were gi ven 36 h
af t er i nt ubat i on, and t he t est on day 34 at 47 h aft er. The reason is
expl ai ned i n t he section on "Resul t s". Bl ood samples (0.05 ml f r om t he
t i p of t he taft) were t aken i mmedi at el y af t er t he t est sessions on days 12,
18, 24 and 30, and i mmedi at el y before t est i ng on day 28, and l at er
anal ysed f or alcohol concent r at i on by t he depr ot ci ni zat i on and gas-
chr omat ogr aphi c met hods descri bed by LeBl anc (1968).
Results
The mean val ues for t he st art l e t hreshol ds of t he t wo groups were
qui t e st abl e and not i mpor t ant l y di fferent duri ng t he first six days of
t he exper i ment . Fig.1 shows t ha t begi nni ng on day 8 and cont i nui ng
t hr oughout t he chroni c t r eat ment peri od t he t r end lines di verge con-
spicuously, t he et hanol group showing lower mean t hreshol ds t han t he
cont rol groups. The overal l mean flinch t hreshol ds for t he et hanol and
sucrose groups duri ng t he chroni c t r eat ment peri od (but excl udi ng t he
val ues on days 12, 18, 24 and 30) were 0.21 mA and 0.24 mA respect i vel y;
t he j ump t hreshol ds were 0.34 mA and 0.36 mA respect i vel y. Duri ng
t he wi t hdr awal peri od (from da y 38 on) t her e is a gradual increase of
bot h flinch and j ump t hreshol ds i n t he et hanol group al most t o cont r ol
values.
Tabl es 1 and 2 summari ze t he analysis of t he t hr eshol d dat a. I t can
be seen t ha t drugs, doses, and drugs doses were all significant for bot h
measures, al t hough t o a lesser degree for t he flinch t han for t he j ump
t hreshol ds. Thi s means t ha t t he flinch and j ump t hreshol ds of t he et hanol
animals were lower t ha n t hose of t he cont rol s; t ha t t he t hreshol ds var i ed
wi t h t he amount of sucrose or et hanol admi ni st ered, and t ha t t he
effects of change i n dose were di fferent for t he t wo t r eat ment s.
I n addi t i on, days, and days drugs are significant for j ump t hreshol d.
Thi s means t ha t t hreshol ds on t he second t est day at a gi ven dose l evel
were di fferent f r om t hose on t he first, and t ha t t he t wo t r eat ment s differ
Measur ement of Et hanol Dependence in Rat s by St ar t l e Thresholds 99
~ o . I t . I , T ~
" ~ Y r " - '
I '
" ~ / ~ , , t . t , , ' 0 ; , : , , t , : . . t , , ' 0 ~ ,: , , t , ~ 4 , ' 0 ~ 4 , t , t . t , ~ ,I ~ : , ~ ,: ,: o o r : 4 ~
6 8 10 12 14 16 18 20 22 24 26 28 ~0 32 3 4 36 38 4 0 42 4 4 46
DAY
Fig. i . Effects of chronic admi ni st rat i on of ethanol , and of its wi t hdrawal , on j ump
and flinch t hr eshol ds in r at s : ~ j ump t hreshol d, sucrose gr oup; o j ump t hreshol d,
et hanol gr oup; 9 flinch t hreshol d, sucrose gr oup; 9 flinch t hreshol d, et hanol group.
Each poi nt is t he mean of 12 ani mal s; S. E. i ndi cat ed by ver t i cal bars. The boxes
i ndi cat e days on whi ch specified doses of et hanol or equi cal ori e sucrose were gi ven.
Arrows i ndi cat e t i mes of i nt ubat i on, ei t her 30 mi n before (days 12, 18, 24 and 30)
or i mmedi at el y af t er t he t hr eshol d t est s. Threshol d measur ement s made on days
12, 18, 24 and 30 are not r epr esent ed (see Fi gs. 2 and 3)
Tabl e 1. Anal ysi s el variance o] /linch response
Source d_~ Mean F P
square r at i o
Dr ugs 1 0. i 7658 11.23 < 0. 005
Subj ect s (drugs) 22 0.01573
Doses 4 0.02477 29.14 < 0.001
Dr ugs x doses 4 0.00456 5.36 < 0.001
Subj ect s x doses (drugs) 88
Days 1 0.00187 3.74 hT.S.
Dr ugs x days 1 0.00077 1.54 N. S.
Subj ect s X days (drugs) 22
Doses X days 4 0.00234 2.41 /q.S.
Dr ugs X doses x days 4 0.00040 0.41 N. S.
Subj ect s X doses X days (drugs) 88 0.00097
100 R. d. Gibbins, H. Kalant, A. E. LeBlanc, andd. W. Clark:
Table 2. Anal ysi s o/ variance o / j u mp response
Source dF Mean F P
square ratio
Drugs 1 0.16068 8.68 < 0.01
Subjects (drugs) 22 0.01851
Doses 4 0.02987 23.90 < 0.001
Drugs doses 4 0.00370 2.96 < 0.025
Subjects doses (drugs) 88 0.00125
Days 1 0.00392 5.76 < 0.05
Drugs days 1 0.00345 5.07 < 0.05
Subjects days (drugs) 22 0.00068
Doses days 4 0.00321 3.91 N.S.
Drugs doses days 4 0.00006 0.07 N.S.
Subjects doses days (drugs) 88 0.00082
in this respect. Specifically, in the ethanol group the threshold tends to
be lower on the second day t han on the first, while the reverse is true in
the sucrose group.
To illustrate further the differences between groups, the ratios of
alcohol:control group mean threshold values were calculated for each
test day. The results are shown in Figs. 2 and 3. A fairly steady decline
in threshold values for the ethanol group relative to the controls is seen
between days 6 and 22. Following cessation of alcohol t reat ment there
was a return of the relative thresholds for bot h flinch and jump t oward
the control values, which was almost complete by day 45.
On days 12, 18, 24 and 30, when the experimental animals were
tested under the acute influence of ethanol, the mean threshold values
were virtually equal t o those of the control group. The mean blood
alcohol levels of the experimental group immediately after testing on
days 12, 18, 24, and 30 were 104, 144, 175 and 200 mg/100 ml respectively.
On day 26 an unexpected rise in relative thresholds for bot h flinch
and jump was not ed in the alcohol group. Blood samples t aken immedi-
ately before the test on day 28 revealed t hat 5 of the alcohol-treated
animals had blood ethanol levels ranging between 50 and 150 rag/100 ml,
indicating t hat the preceding daily dose had not been completely meta-
bolised in all eases. The results on days 26 and 28 therefore cannot be
considered true measures of maximal post-alcohol hyperexeitability.
The same consideration presumably applied to the tests on days 32--38,
especially since the dose had been increased further. To test this inter-
pretation, no alcohol was given on day 33, so t hat the test on day 34
was made 47 h rather t han 23 h after the last preceding dose of ethanol.
Measur ement of Et hanol Dependence in Rat s by St ar t l e Threshol ds 10l
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DAY
~"ig.2. ]~ffects of chroni c admi ni st r at i on of et hanol ~nd of i t s wi t hdr awal on t hresh-
ol d r at i o for flinch responses. Each poi nt represent s t he mean t hr eshol d for t he
et hanol group di vi ded by t he mean t hr eshol d for t he cont rol group. The boxes
i ndi cat e t he days upon whi ch t he specified et hanol doses were gi ven. The arrows
i ndi cat e whet her t he ani mal s were i nt uba~ed 30 rain before (days 12, 18, 24 and 30
mar ked by ver t i cal br oken lines), or i mmedi abel y af t er t he t hreshol d t est s
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B A Y
Fi g. 3. Effect s of chroni c admi ni s t r at i on of et hanol ~nd of its wi t hdr awal on t hresh-
ol d r at i o for j ump responses. Ea c h poi nt r epr esent s t he mean t hr eshol d ~'or t he
et hanol group di vi ded by ~he r aean t hr eshol d for t he cont rol group. The boxes
i ndi cat e t he days upon whi ch t he specified et hanol doses were given. The arrows
i ndi cat e whet her t he ani mal s were i ncubat ed 30 rai n before (days 12, 18, 24 and 30
shown by ver t i cal broken lines), or i mmedi at el y af t er t he t hreshol d bests
102 R. J. Gibbins, H. Kalant, A. E. LeBlanc, and J. W. Clark:
Ther e appear ed t o be a subst ant i al decline i n t he rel at i ve t hr eshol d for
flinch but not for j ump. Despi t e cont i nued hi gh dosage, t her e di d not
appear t o be any f ur t her lowering of t hr eshol d on subsequent t est s.
Following cessation of et hanol t r eat ment , f r om da y 38 on t her e was
a gradual r et ur n of bot h t hreshol ds al most t o cont rol val ues over an
8- day peri od.
Di scussi on
Measurement s of st i mul us t hreshol ds for flinch and j ump responses
i n t he pr esent s t udy were made under several di fferent condi t i ons wi t h
respect t o et hanol t r eat ment and t o envi r onment al factors. The mean
val ues for bot h responses by t he cont r ol group f l uct uat ed duri ng t he
l at er par t of t he exper i ment , appar ent l y because of var i at i on i n measur ed
at mospheri c humi di t y. However , since t he analysis of t he act ual t est
val ues (Tables 1 and 2) showed significant differences bet ween t he t wo
t r eat ment groups for compari sons on t he same day, i t is permissible t o
use t he i nt er-group t hr eshol d r at i o (alcohol : controls) as an i l l ust rat i ve
simplification.
I t is evi dent f r om Figs. 2 and 3 t ha t t he flinch and j ump t hreshol ds
changed i n paral l el i n response t o t r eat ment . Despi t e some views t o t he
cont r ar y, t he t wo responses are general l y consi dered t o r epr esent mer el y
di fferent degrees of t he same st art l e reflex (Landi s and Hunt , 1939;
Hof f man et al . , 1964). Ther ef or e t hei r paral l el course reinforces t he
conclusions dr awn f r om t he exper i ment . As t r e a t me nt was cont i nued
t he et hanol ani mal s became progressi vel y mor e i rri t abl e and difficult t o
handl e, al t hough gross t r emor or convulsions were not observed. Ther e
was a cl earl y evi dent fall in t hr eshol d r at i o for bot h responses f r om t he
beginning of t he alcohol t r eat ment period. Thi s is consi st ent wi t h t he
finding t ha t alcohol wi t hdrawal signs, i ncl udi ng t r emor and convulsions,
coul d be pr oduced af t er as l i t t l e as 5 days of alcohol t r eat ment i n mice
(Freund, 1969) and 10- - 18 days i n monkeys (Ellis and Pi ck, 1970). The
decrease was gr eat er wi t h increasing alcohol dosage f r om da y 6 t hr ough
t o day 22, wi t h t he sole except i on of t he flinch t hr eshol d on day 16.
The rise in r at i o on days 26 and 28 refl ect ed an ar t i f act of experi-
ment al procedure. The precedi ng dosage i ncr ement had evi dent l y
rai sed t he dai l y dose t o more t ha n t he ani mal s coul d met abol i ze in 23 h.
The finding i ndi cat es t ha t observat i ons of alcohol wi t hdrawal phenomena
made at a single fi xed t i me af t er admi ni st r at i on of t he l ast precedi ng
dose ma y be misleading, and t ha t i t is pr obabl y desirable in such cases
t o ver i f y by bl ood analysis t ha t t he subj ect s are act ual l y in a st at e of
wi t hdrawal . The i deal pr ocedur e for compar at i ve studies woul d be t o use
a fixed t i me af t er r et ur n of t he bl ood alcohol level t o zero. Since no bl ood
et hanol measur ement s were made f r om day 28 on, i t is impossible t o
Measurement of Ethanol Dependence in Rats by Startle Thresholds 103
know whet her t he absence of cont i nued decrease i n t hr eshol ds dur i ng
t he l ast 9 days of t he t r e a t me nt per i od was due t o resi dual et hanol or t o
t he a t t a i nme nt of ma x i mu m degree of physi cal dependence. I t is i nt er est -
i ng t o not e t h a t in pr evi ous wor k (LeBl anc e t a l . , 1969) t he same t ype of
dosage schedul e r esul t ed i n t he pr oduct i on of ma x i mu m at t ai nabl e
t ol er ance t o et hanol in 19- - 21 days, and f ur t her dose i ncr ement s di d not
i ncrease it.
On days 12, 18, 24 and 30 t he t hr eshol d meas ur ement s were made
dur i ng t he peri od 30- - 35 mi n af t er t he pr ecedi ng dose of et hanol . Under
t hese condi t i ons, af t er doses of 3, 4, 5 and 6 g/ kg t he t hr eshol ds in t he
al cohol - t r eat ed ani mal s, whi ch were ma r ke dl y r educed when meas ur ed
23 h post - et hanol , r et ur ned t o a ppr oxi ma t e l y t he same levels as in t he
cont rol s. Thi s is b y defi ni t i on an evi dence of physi cal dependence,
i nasmuch as i t is a cor r ect i on or nor mal i zat i on b y et hanol of a di st ur b-
ance pr oduced b y absence of et hanol in a chroni cal l y t r e a t e d ani mal .
Fur t her , t he pr esent met hod pr ovi des a means of quant i f yi ng t he degree
of dependence a t earlier st ages t ha n t he ful l y devel oped pi ct ur e descri bed
b y ot her i nvest i gat or s. The changes under l yi ng t he hyper exci t abi l i t y are
r eadi l y reversi bl e. The t i me of r et ur n t o nor mal in t he pr esent wor k is
of t he s ame order as t h a t r equi r ed for nor mal i zat i on of t he el ect ro-
convul si ve seizure t hr eshol d i n ani mal s whi ch had recei ved et hanol
f or t wo weeks ( ~eQuar r i e and Fi ngl , 1958), and for r ever sal of t ol er ance
t o et hanol (LeBl anc e t a l . , 1969) and of cross-t ol erance t o amobar bi t al
i n et hanol - t r eat ed r at s (Ratcliffe, 1969).
The pr esent wor k shows t ha t chroni c t r e a t me nt wi t h i ncreasi ng doses
of et hanol gi ves rise t o an i ncreasi ng degree of post - al cohol hyper -
exci t abi l i t y, as demons t r at ed b y r educt i on of t he t hr eshol d for response
t o st i mul at i on vi a nor mal sensor y pat hways . Thi s schedul e of admi ni s-
t r at i on was chosen specifically because i t had been used in earlier st udi es
of t he devel opment of t ol er ance (LeBl ane e t a l . , 1969) and because i t
causes mi ni mal mor t a l i t y among t he exper i ment al ani mal s. Unf or t un-
at el y i t cont ai ns t wo si mul t aneous vari abl es, i.e. t he dur at i on of t r eat -
me nt and t he dose level. The r el at i ve effects of t hese t wo mus t be exami ned
s epar at el y i n f ut ur e work. The pr esent findings do not per mi t a con-
clusion a bout t he maxi mal degree of dependence whi ch can be pr oduced,
but t he y are compat i bl e wi t h t he hypot hesi s t ha t t ol er ance t o, and
physi cal dependence on, et hanol are i nt i mat el y r el at ed processes, or
t wo mani f est at i ons of t he same process.
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Dr. 1~. J. Gibbins
Addiction Research Foundation
33 Russell Street
Toronto 149, Ontario, Canada