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Sleep-inducing effect of piperidine
186
Brain Research, 88 (t975) 1,~6 I~!) Elsevier Scientific Publishing Company, Amsterdam - Printed il~ The Nethert~md~
Sleep-inducing effect of piperidine
RENI~ RAI]L DRUCKER-COL~N* AND EZIO GIACOB1NI Department of Psychobiology, University of California, [rvine, Calif. 92664 and Laboratory of Neuropsychopharmacology, Department of Biobehavioral Sciences, University of Connecticut, Storrs, Conn. 06268 (U.S.A.)
(Accepted December 3lst, 1974)
Piperidine, a volatile amine, has recently been identified by means of mass spectrometry in the CNS of the snail a and in the brain of the mouse as, rat and catL Kas~ et al. 1~ showed a pronounced sedative effect and a significant prolongation of barbiturate-induced sleep in the mice and rats after i.p. or i.v. injection of piperidine. Moreover, an accumulation of piperidine in the brain of dormant invertebrates (snails) and vertebrates (mice) has been recently demonstrated by the studies of Dolezalova et al.4, 5 and Stepita-Klauco et al. x8 in our laboratory. These findings are of particular interest since a selective and powerful synaptic action of piperidine has been found to occur on identified cholinoceptive neurons of the snail cerebral ganglia 19. Moreover, piperidine shows nicotine-like action in the peripheral and central nervous system of vertebratesla, 14. Using the technique of direct chemical stimulation 1° of the brain it is possible to selectively stimulate an anatomically circumscribed cholinergic hypnogenic pathway extending from the forebrain to the midbrain 9. This paper reports behavioral and electrophysiological manifestations following perfusion ofpiperidine with a push-pull cannula system into a defined area of the brain stem of the cat. Sixteen cats weighing between 2.5 and 3.5 kg were used. Under aseptic conditions and pentobarbital anestheSia a 17-gauge stainless steel guide tube was implanted stereotaxicall~. The guide tube extended 15 m m inside the brain with its tip lying 7 m m above the midbrain reticular formation ( M R F ) (A = 0; L == 2; V = --2). In addition stainless steel screw electrodes were inserted into the skull over the sensorimotor cortex and around the orbit in order to record neocortical activity and eye movements respectively. E M G was also recorded through electrodes introduced into the neck muscles. The electrodes were soldered to a 9-pin miniconnector and the entire assembly was fixed to the skull with dental acrylic. All animals were allowed 10 days of postoperative recovery. At the start of the experiments the animals were placed in a cage inside a * Permanent address: Dept. de Biologia Experimental, lnstituto de Biologia, UNAM, Apartado Postal 70-600, Mexico 20, D.F., Mexico; where reprint requests should be addressed.
187 TABLE I LATENCIES AND DURATIONS OF S W S AND
REM FOR EACH
RECIPIENT CAT AFTER 1NTRACEREBRAL PERFU-
SION OF R I N G E R OR PIPER1DINE
All values are in minutes. Note that the 6 piperidine subjects are independent from the l0 Ringer controls. Cat no.
Latency
Duration
Ringer control
Piperidine
Ringer control
Piperidine
SWS
REM
SWS
SWS
REM
SWS
REM
2 3 4 5 6 7 8 9 10
27 46 42 31 29 33 60 29 37 31
48 60 60 60 43 58 60 39 60 60
26 17 19 21 14 22
18 11 8 16 21 13
Mean S.E.
36.5 :~ 3.2
54.8 ~ 2,5
19.8 ~_ 1.7
14.5" L 1,9
1
REM
3
9
12
2
8 5 7 8 10
19 12 18 14 16
9 13 22 12 20 0 15 23 7
0 0 0 6 1 0 9 0 0
6.8* ± 1.0
14.6" ~ 1.5
13.7 ± 2.0
1.6 ± 0.91
* Significantly different from control, P < 0.001.
quiet l a b o r a t o r y r o o m , where they c o u l d move freely a n d be observed. T h e y were then c o n n e c t e d to a grass M o d e l 7 p o l y g r a p h , a n d a ' p u s h - p u l l ' c a n n u l a consisting o f a 27-gauge inner ' p u s h ' a n d a 20-gauge o u t e r ' p u l l ' p a i r o f needles was inserted t h r o u g h the guide tube to a p o s i t i o n o f 7 m m b e y o n d its tip. The c a n n u l a was c o n n e c t e d t h r o u g h p o l y e t h y l e n e tubes (PE-20 a n d PE-90) to a H a r v a r d infusion w i t h d r a w a l p u m p . T e n a n i m a l s were perfused with a c o m m e r c i a l R i n g e r s o l u t i o n ( A b b o t t , N a C I 8.6 g; KC10.3 g; CaCI 0.33 g; a n d distilled water to m a k e up 1 liter), a n d 6 a n i m a l s were perfused with piperidine chloride solution as follows : each solution was perfused d u r i n g 15 min at which time the p u m p was stopped. The E E G , however, c o n t i n u e d b e y o n d for a n o t h e r 45 rain. In o t h e r words, the p e r f u s i o n was d o n e for 15 rain b u t the recording o f the a n i m a l s ' s l e e p - w a k e cycle was d o n e for 60 min. The perfusion speed was 20 #l/rain so t h a t over 15 min 333 # g o f p i p e r i d i n e passed t h r o u g h the brain. The latencies a n d the d u r a t i o n o f the slow-wave sleep (SWS) a n d R E M sleep was det e r m i n e d in each cat by visual inspection o f the p o l y g r a p h i c recordings. A t the end o f the e x p e r i m e n t s the b r a i n o f each cat was perfused in situ with 10 ~ f o r m a l i n , with the a n i m a l u n d e r deep b a r b i t u r a t e anesthesia. F o l l o w i n g removal o f b r a i n a n d a 2 week fixation p e r i o d , frozen 40-50 # m sections, cut for histological verification o f electrode placements, i n d i c a t e d a d e q u a t e localization within the m i d brain t e g m e n t u m . T a b l e I s u m m a r i z e s o u r results. The a n i m a l s which were locally perfused with piperidine showed a highly significant decrease ( P < 0.001) in the latency to sleep
188 from 36.5 ~:i: 3.2 to 6.8 :k 1.0 for SWS and from 54.8 ::t: 2.5 to 14.6 1.5 ['or REM. Also, the duration of' REM sleep was significantly increased (P
1 13OBILLIER,P., FROMENT,J. L., SEGUIN,S., ET JOUVET,M., Effets de lap-chlorophCnylalanine et du 5-hydroxytryptophane sur le sommeil et le m6tabolisme central des monoamines et des prot~ines chez le chat, Biochem. Pharmacol., 22 (1973) 3077-3090. 2 BOHN, M., AND GIACOaINI, E., (1974) in preparation. 3 DOLEZALOVA,H., GIACOBINI,E., SELLER,N., AND SCHNEIDER,H. H., Determination of piperidine in snail brain (Helix pomatia), Brain Research, 55 (1973) 242-244. 4 DOLEZALOVA,H., STEPITA-KLAUCO,M., AND FAIRWEATHER,R., The accumulation of piperidine i,~ the central ganglia of dormant snails, Brain Research, 72 (1974) 115-122.
189 5 DOLEZALOVA, H., STEPITA-KLAUCO, M., AND GIACOBINI, E., Variation of endogenous and exogenous piperidine in the brain of active and dormant snails, Soe. Neurosei. (S. Diego), (1973) 349. 6 DOLEZALOVA, H., STEPITA-KLAUCO, M., AND GIACOBINI, E., The passage of tritiated piperidine into the central ganglia and other tissue of the snail, Int. J. Neurosci., 6 (1973) 89-95. 7 DRUCKER-COL]~N,R. R., SPANIS, C. W., COTMAN, C. W., AND McGAUGH, J. L., Proteins and REM sleep: studies on push-pull perfusates from the midbrain reticular formation. In Symposium on Endocrinology and Biochemistry of Sleep. 2nd European Congress on Sleep Research ( Rome), Karger, Basel, 1974, in press. 8 GIACOBINI, E., Seasonal and activity dependent variation of biogenic amine levels in the invertebrate CNS. In Symposium on Endocrinology and Biochemistry of Sleep, 2nd European Congress on Sleep Research (Rome), Karger, Basel, 1974, in press. 9 HERNANDEZ-PE6N, R., AND DRUCKER-COL[N, R. R., A neuronographic study of corticobu[bar hypnogenic pathways, Physiol. Behav., 5 (1970) 721-725. I 0 HERNANDEZ-PEON, R., CHAVEZ-IBARRA,G., MORGANE,P., AND TIMO-IARIA,C., Limbic cholinergic pathways involved in sleep and emotional behavior, Exp. Neurol., 3 (1963) 93 I 11. I 1 [-IERNANDEZ-PEON,R., O'FLAHERTY, J. J.~ AND MAZZUCHELLI-O'FLAHERTY, A. L., Sleep and other behavioral effects induced by acetylcholinic stimulation of boral temporal cortex and striate structure, Brain Research, 4 (1967) 243-267. 12 JOUVET, M., The role of monoamines and acetylcholine containing neurons in the regulation of the sleep-waking cycle, Ergebn. Physiol., 64 (1972) 166-307. 13 KAS~, Y., MIYATA, T., AND YUIZONO, T., Pharmacological studies on alicyclic amines. I. Comparison of pharmacological activities of piperidine with those of other amines, Jap. J. Pharmaeol., 17 (1967) 475490. 14 KAS~, Y., MIYATA, T., KAMIKAWA, Y., AND KATAOKA, M., Pharmacological studies on alicyclic amines. 11. Central actions of piperidine, pyrrolidine and piperazine, Jap. J. Pharmaeol., 19 (1969) 300-314. 15 LEWIS, P. R., AND SHUETTE, C. C. A., The cholinergic limbic system: projection to hippocampal formation, medial cortex, nuclei of the ascending cholinergic reticular system and the subfornical organ and supraoptic crest, Brain, 90 (1967) 521 543. 16 ROJAS-RAMIREZ, J. A., AND DRUCKER-COLfN, R. R., Sleep induced by spinal cord cholinergic stimulation, hit. J. Neurosci., 5 (1973) 215-221. 17 SHUETTE, C. C. D., AND LEWIS, P. R., The ascending cholinergic reticular system: neocortical olfactory and subcortical projections, Brahl, 90 (1967) 497-520. 18 STEPITA-KLAUCO, M., DOLEZALOVA, H., AND FA1RWEATHER, R., Piperidine increase in the brain of dormant mice, Science, 183 (1974) 536-537. 19 STEP~TA-KLAUCO, M., DOLEZALOVA, H., AND GIACOBINI, E., The action of piperidine on cholinoceptive neurons of the snail, Brain Research, 63 (1973) 141 152.
Brain Research, 88 (t975) 1,~6 I~!) Elsevier Scientific Publishing Company, Amsterdam - Printed il~ The Nethert~md~
Sleep-inducing effect of piperidine
RENI~ RAI]L DRUCKER-COL~N* AND EZIO GIACOB1NI Department of Psychobiology, University of California, [rvine, Calif. 92664 and Laboratory of Neuropsychopharmacology, Department of Biobehavioral Sciences, University of Connecticut, Storrs, Conn. 06268 (U.S.A.)
(Accepted December 3lst, 1974)
Piperidine, a volatile amine, has recently been identified by means of mass spectrometry in the CNS of the snail a and in the brain of the mouse as, rat and catL Kas~ et al. 1~ showed a pronounced sedative effect and a significant prolongation of barbiturate-induced sleep in the mice and rats after i.p. or i.v. injection of piperidine. Moreover, an accumulation of piperidine in the brain of dormant invertebrates (snails) and vertebrates (mice) has been recently demonstrated by the studies of Dolezalova et al.4, 5 and Stepita-Klauco et al. x8 in our laboratory. These findings are of particular interest since a selective and powerful synaptic action of piperidine has been found to occur on identified cholinoceptive neurons of the snail cerebral ganglia 19. Moreover, piperidine shows nicotine-like action in the peripheral and central nervous system of vertebratesla, 14. Using the technique of direct chemical stimulation 1° of the brain it is possible to selectively stimulate an anatomically circumscribed cholinergic hypnogenic pathway extending from the forebrain to the midbrain 9. This paper reports behavioral and electrophysiological manifestations following perfusion ofpiperidine with a push-pull cannula system into a defined area of the brain stem of the cat. Sixteen cats weighing between 2.5 and 3.5 kg were used. Under aseptic conditions and pentobarbital anestheSia a 17-gauge stainless steel guide tube was implanted stereotaxicall~. The guide tube extended 15 m m inside the brain with its tip lying 7 m m above the midbrain reticular formation ( M R F ) (A = 0; L == 2; V = --2). In addition stainless steel screw electrodes were inserted into the skull over the sensorimotor cortex and around the orbit in order to record neocortical activity and eye movements respectively. E M G was also recorded through electrodes introduced into the neck muscles. The electrodes were soldered to a 9-pin miniconnector and the entire assembly was fixed to the skull with dental acrylic. All animals were allowed 10 days of postoperative recovery. At the start of the experiments the animals were placed in a cage inside a * Permanent address: Dept. de Biologia Experimental, lnstituto de Biologia, UNAM, Apartado Postal 70-600, Mexico 20, D.F., Mexico; where reprint requests should be addressed.
187 TABLE I LATENCIES AND DURATIONS OF S W S AND
REM FOR EACH
RECIPIENT CAT AFTER 1NTRACEREBRAL PERFU-
SION OF R I N G E R OR PIPER1DINE
All values are in minutes. Note that the 6 piperidine subjects are independent from the l0 Ringer controls. Cat no.
Latency
Duration
Ringer control
Piperidine
Ringer control
Piperidine
SWS
REM
SWS
SWS
REM
SWS
REM
2 3 4 5 6 7 8 9 10
27 46 42 31 29 33 60 29 37 31
48 60 60 60 43 58 60 39 60 60
26 17 19 21 14 22
18 11 8 16 21 13
Mean S.E.
36.5 :~ 3.2
54.8 ~ 2,5
19.8 ~_ 1.7
14.5" L 1,9
1
REM
3
9
12
2
8 5 7 8 10
19 12 18 14 16
9 13 22 12 20 0 15 23 7
0 0 0 6 1 0 9 0 0
6.8* ± 1.0
14.6" ~ 1.5
13.7 ± 2.0
1.6 ± 0.91
* Significantly different from control, P < 0.001.
quiet l a b o r a t o r y r o o m , where they c o u l d move freely a n d be observed. T h e y were then c o n n e c t e d to a grass M o d e l 7 p o l y g r a p h , a n d a ' p u s h - p u l l ' c a n n u l a consisting o f a 27-gauge inner ' p u s h ' a n d a 20-gauge o u t e r ' p u l l ' p a i r o f needles was inserted t h r o u g h the guide tube to a p o s i t i o n o f 7 m m b e y o n d its tip. The c a n n u l a was c o n n e c t e d t h r o u g h p o l y e t h y l e n e tubes (PE-20 a n d PE-90) to a H a r v a r d infusion w i t h d r a w a l p u m p . T e n a n i m a l s were perfused with a c o m m e r c i a l R i n g e r s o l u t i o n ( A b b o t t , N a C I 8.6 g; KC10.3 g; CaCI 0.33 g; a n d distilled water to m a k e up 1 liter), a n d 6 a n i m a l s were perfused with piperidine chloride solution as follows : each solution was perfused d u r i n g 15 min at which time the p u m p was stopped. The E E G , however, c o n t i n u e d b e y o n d for a n o t h e r 45 rain. In o t h e r words, the p e r f u s i o n was d o n e for 15 rain b u t the recording o f the a n i m a l s ' s l e e p - w a k e cycle was d o n e for 60 min. The perfusion speed was 20 #l/rain so t h a t over 15 min 333 # g o f p i p e r i d i n e passed t h r o u g h the brain. The latencies a n d the d u r a t i o n o f the slow-wave sleep (SWS) a n d R E M sleep was det e r m i n e d in each cat by visual inspection o f the p o l y g r a p h i c recordings. A t the end o f the e x p e r i m e n t s the b r a i n o f each cat was perfused in situ with 10 ~ f o r m a l i n , with the a n i m a l u n d e r deep b a r b i t u r a t e anesthesia. F o l l o w i n g removal o f b r a i n a n d a 2 week fixation p e r i o d , frozen 40-50 # m sections, cut for histological verification o f electrode placements, i n d i c a t e d a d e q u a t e localization within the m i d brain t e g m e n t u m . T a b l e I s u m m a r i z e s o u r results. The a n i m a l s which were locally perfused with piperidine showed a highly significant decrease ( P < 0.001) in the latency to sleep
188 from 36.5 ~:i: 3.2 to 6.8 :k 1.0 for SWS and from 54.8 ::t: 2.5 to 14.6 1.5 ['or REM. Also, the duration of' REM sleep was significantly increased (P
1 13OBILLIER,P., FROMENT,J. L., SEGUIN,S., ET JOUVET,M., Effets de lap-chlorophCnylalanine et du 5-hydroxytryptophane sur le sommeil et le m6tabolisme central des monoamines et des prot~ines chez le chat, Biochem. Pharmacol., 22 (1973) 3077-3090. 2 BOHN, M., AND GIACOaINI, E., (1974) in preparation. 3 DOLEZALOVA,H., GIACOBINI,E., SELLER,N., AND SCHNEIDER,H. H., Determination of piperidine in snail brain (Helix pomatia), Brain Research, 55 (1973) 242-244. 4 DOLEZALOVA,H., STEPITA-KLAUCO,M., AND FAIRWEATHER,R., The accumulation of piperidine i,~ the central ganglia of dormant snails, Brain Research, 72 (1974) 115-122.
189 5 DOLEZALOVA, H., STEPITA-KLAUCO, M., AND GIACOBINI, E., Variation of endogenous and exogenous piperidine in the brain of active and dormant snails, Soe. Neurosei. (S. Diego), (1973) 349. 6 DOLEZALOVA, H., STEPITA-KLAUCO, M., AND GIACOBINI, E., The passage of tritiated piperidine into the central ganglia and other tissue of the snail, Int. J. Neurosci., 6 (1973) 89-95. 7 DRUCKER-COL]~N,R. R., SPANIS, C. W., COTMAN, C. W., AND McGAUGH, J. L., Proteins and REM sleep: studies on push-pull perfusates from the midbrain reticular formation. In Symposium on Endocrinology and Biochemistry of Sleep. 2nd European Congress on Sleep Research ( Rome), Karger, Basel, 1974, in press. 8 GIACOBINI, E., Seasonal and activity dependent variation of biogenic amine levels in the invertebrate CNS. In Symposium on Endocrinology and Biochemistry of Sleep, 2nd European Congress on Sleep Research (Rome), Karger, Basel, 1974, in press. 9 HERNANDEZ-PE6N, R., AND DRUCKER-COL[N, R. R., A neuronographic study of corticobu[bar hypnogenic pathways, Physiol. Behav., 5 (1970) 721-725. I 0 HERNANDEZ-PEON, R., CHAVEZ-IBARRA,G., MORGANE,P., AND TIMO-IARIA,C., Limbic cholinergic pathways involved in sleep and emotional behavior, Exp. Neurol., 3 (1963) 93 I 11. I 1 [-IERNANDEZ-PEON,R., O'FLAHERTY, J. J.~ AND MAZZUCHELLI-O'FLAHERTY, A. L., Sleep and other behavioral effects induced by acetylcholinic stimulation of boral temporal cortex and striate structure, Brain Research, 4 (1967) 243-267. 12 JOUVET, M., The role of monoamines and acetylcholine containing neurons in the regulation of the sleep-waking cycle, Ergebn. Physiol., 64 (1972) 166-307. 13 KAS~, Y., MIYATA, T., AND YUIZONO, T., Pharmacological studies on alicyclic amines. I. Comparison of pharmacological activities of piperidine with those of other amines, Jap. J. Pharmaeol., 17 (1967) 475490. 14 KAS~, Y., MIYATA, T., KAMIKAWA, Y., AND KATAOKA, M., Pharmacological studies on alicyclic amines. 11. Central actions of piperidine, pyrrolidine and piperazine, Jap. J. Pharmaeol., 19 (1969) 300-314. 15 LEWIS, P. R., AND SHUETTE, C. C. A., The cholinergic limbic system: projection to hippocampal formation, medial cortex, nuclei of the ascending cholinergic reticular system and the subfornical organ and supraoptic crest, Brain, 90 (1967) 521 543. 16 ROJAS-RAMIREZ, J. A., AND DRUCKER-COLfN, R. R., Sleep induced by spinal cord cholinergic stimulation, hit. J. Neurosci., 5 (1973) 215-221. 17 SHUETTE, C. C. D., AND LEWIS, P. R., The ascending cholinergic reticular system: neocortical olfactory and subcortical projections, Brahl, 90 (1967) 497-520. 18 STEPITA-KLAUCO, M., DOLEZALOVA, H., AND FA1RWEATHER, R., Piperidine increase in the brain of dormant mice, Science, 183 (1974) 536-537. 19 STEP~TA-KLAUCO, M., DOLEZALOVA, H., AND GIACOBINI, E., The action of piperidine on cholinoceptive neurons of the snail, Brain Research, 63 (1973) 141 152.