PROGRESS I N BRAIN RESEARCH VOLUME 36 BIOCHEMICAL AND PHARMACOLOGICAL MECHANISMS UNDERLYING BEHAVIOUR doc

PROGRESS I N B R A I N RE SE ARCH VO L U ME 36 BIOCHEMICAL A N D PHARMACOLOGICAL MECHANISMS U N D E R L Y I N G BEHAVIOUR PROGRESS IN BRAIN RESEARCH ADVISORY B O A R D W Bargmann H T Chang E De Robertis J C Eccles J D French H HydCn J Arigns Kappers S A Sarkisov J P SchadC F Schmitt Kiel Shanghai Buenos Aires Canberra Los Angeles Goteborg Amsterdam Moscow Amsterdam Brookline (Mass.) T Tokizane Tokyo J Z Young London PROGRESS IN BRAIN RESEARCH V O L U M E 36 BIOCHEMICAL AND PHARMACOLOGICAL MECHANISMS UNDERLYING BEHAVIOUR EDITED B Y P B B R A D L E Y Department of Pharmacology (Preciinical), The Medical School, University of Birmingham, Birmingham (England) AND R W BRIMBLECOMBE Chemical Defence Establishment, Porton Down, Salisbury, Wiltshire (EngIand) ELSEVIER P U B L I S H I N G C O M P A N Y AMSTERDAM / LONDON / NEW YORK 1972 ELSEVIER P U B L I S H I N G C O M P A N Y P.O BOX 335 J A N VAN G A L E N S T R A A T 211, AMSTERDAM, T H E N E T H E R L A N D S AMERICAN ELSEVIER P U B L I S H I N G COMPANY, I N C V A N D E R B I L T AVENUE, N E W YORK, N.Y LIBRARY OF CONGRESS C A R D NUMBER ISBN WITH COPYRIGHT @ 1972 95 10017 72-190679 0-444-40992-0 ILLUSTRATIONS AND TABLES BY ELSEVIER PUBLISHINO COMPANY, AMSTERDAM A L L R I G H T S RESERVED NO P A R T OF T H I S P U B L I C A T I O N MAY BB R E P R O D U C E D , S T O R E D I N A R E T R I E V A L SYSTEM, OR T R A N S M I T T E D I N ANY FORM O R BY A N Y MEANS, E L E C T R O N I C , M E C H A N I C A L , P H O T O C O P Y I N G , R E C O R D I N G , O R O T H E R W I S E , W I T H O U T T H E P R I O R W R I T T E N PERMISSION OF T H E P U B L I S H E R , ELSEVIER P U B L I S H I N G COMPANY, J A N VAN G A L E N S T R A A T 335, AMSTERDAM PRINTED I N THE NETHERLANDS List of Participants ALDOUS, A B., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) F ANSELL, B., Department of Pharmacology (Prechical), University of Birmingham, Birmingham G (U.K.) BALLANTYNE,Chemical Defence Establishment, Porton Down, Salisbury (U.K.) B., BARSTAD, A B., Department of Toxicology, Norwegian Defence Research Establishment, Kjeller J (Norway) BEBBINGTON, Chemical Defence Establishment, Porton Down, Salisbury (U.K.) A., W BERRY, K., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) BESWICK, W., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) F BRADLEY, B., Department of Pharmacology (Precliinical), University of Birmingham, BirmingP ham (U.K.) BRIGGS, Department of Pharmacology (Precliicd), University of Birmingham, Birmingham I., (U.K.) BRIMBLECOMBE, Chemical Defence Establishment, Porton Down, Salisbury (U.K.) R W., BUXTON, A., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) D CALLAWAY,Chemical Defence Establishment, Porton Down, Salisbury (U.K.) S., COHEN, M., Medical Biological Laboratories, RVO-TNO & Department of Fundamental PharE macology, University of Leiden, Leiden (Netherlands) COOPER, H., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) G COULT, B., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) D Cox, T., School of Pharmacy, University of Nottingham, Nottingham (U.K.) CREASEY, H., Chemical Defence Establishment, Porton Down, Salisbury? (U.K.) N CROSSLAND, School of Pharmacy, University of Nottingham, Nottingham (U.K.) J., D a v i ~ s J., School of Pharmacy, University of Bath, Bath, (U.K.) , FISHER, B., Department of Pharmacology (Preclinical), University of Birmingham, Birmingham, R & CDE, Porton Down, Salisbury (U.K.) FONNUM, Department of Toxicology, Norwegian Defence Research Establishment, Kjeller F., (Norway) GORDON, J., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) J GREEN, M., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) D HANDS, H., Department of Pharmacology (Preclinical), University of Birmingham, Birmingham D U.K.) HEILBRONN, EDITH, Research Institute of National Defence, Sundbyberg (Sweden) HOLLAND, Chemical Defence Establishment, Porton Down, Salisbury (U.K.) P., HOLMES, Directorate of Biological & Chemical Defence, London (U.K.) R., HOWELLS, J., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) D HUGHES, ANNETTE, Chemical Defence Establishment, Porton Down, Salisbury (U.K.) INCH, D., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) T KEMP,K H., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) KERKUT, A., Department of Physiology & Biochemistry, University of Southampton, SouthampG ton (U.K.) KING,A R., Department of Pharmacology (Preclinical), University of Birmingham, Birmingham (U.K.) KNIGHT, JOSEPHINE, Department of Pharmacology (Preclinical), University of Birmingham, Birmingham (U.K.) LEADBEATER,Chemical Defence Establishment, Porton Down, Salisbury (U.K.) L., VI LIST OF PARTICIPANTS MEETER, Medical Biological Laboratories, RVO-TNO, Rijswijk (Netherlands) E., MOLENAAR,C., Department of Fundamental Pharmacology, University of Leiden, Leiden (NetherP lands) MOYLAN-JONES, J., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) R Mum, A W., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) OICONNOR, J., RAF Hospital, Wroughton (U.K.) P PATTLE, E., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) R PINDER, M., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) R POLAK, K., Medical Biological Laboratories, RVO-TNO, Rijswijk (Netherlands) R RAWLINS, S P., Department of Director General Medical Services, Royal Navy, London (U.K.) J REDFERN, School of Pharmacy, University of Bath, Bath (U.K.) P., RICK,J T., Department of Psychology, University of Birmingham, Birmingham (U.K.) J RUTLAND, P., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) SAINSBURY, Chemical Defence Establishment, Porton Down, Salisbury (U.K.) G., SAMUELS, GILLIAN R., Department of Pharmacology (Preclinical), University of Birmingham, M Birmingham (U.K.) SCHOCK, Chemical Defence Establishment, Porton Down, Salisbury (U.K.) C., D SINKINSON, V., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) SPANNER, SHEILA Department of Pharmacology (Preclinical), University of Birmingham, BirG., mingham (U.K.) STORM-MATHISEN, Department of Toxicology, Norwegian Defence Research Establishment, J., Kjeller (Norway) SWANSTON, W., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) D SZERB, C., Department of Physiology & Biophysics, Dalhousie University, Halifax (Canada) J UPSHALL, G., Chemical Defence Establishment, Porton Down, Salisbury (U.K.) D VANDER POEL,A M., Department of Fundamental Pharmacology, University of Leiden, Leiden (Netherlands) VINE,R S., Home Office, Romney House, London (U.K.) WALKER, Department of Physiology & Biochemistry, University of Southampton, Southampton R., (U.K.) WATTS, Chemical Defence Establishment, Porton Down, Salisbury (U.K.) P., WOODRUFFE, M., Department of Physiology & Biochemistry, University of Southampton, G Southampton (U.K.) List of Contributors G B ANSELL, Department of Pharmacology (Preclinical), Medical School, Birmingham B15 2TJ, England B C BARRASS, Chemical Defence Establishment, Porton Down, Salisbury, Wiltshire, England J A B BARSTAD, Norwegian Defence Research Establishment, Division of Toxicology, P.O Box 25, Kjeller, Norway P B BRADLEY, Department of Pharmacology (Preclinical), Medical School, Birmingham B15 2TJ, England R W BRIMBLECOMBE, Medical Division, Chemical Defence Establishment, Porton Down, Salisbury, Wiltshire, England D A BUXTON, Chemical Defence Establishment, Porton Down, Salisbury, Wiltshire, England J A DAVIES, School of Pharmacy, University of Bath, Bath, England F FONNUM, Norwegian Defence Research Establishment, Division for Toxicology, P.O Box 25, Kjeller, Norway D M GREEN, Medical Division, Chemical Defence Establishment, Porton Down, Salisbury, Wiltshire, England E HEILBRONN, Research Institute of the Swedish National Defence, Avdelning 1, Box 416, S-172 04 Sundbyberg 4, Sweden T D INCH, Chemical Defence Establishment, Porton Down, Salisbury, Wiltshire, England G A KERKUT, Department of Physiology and Biochemistry, University of Southampton, Southampton, England E MEETER, Medical Biological Laboratories, RVO-TNO, Lange Kleiweg 139, Rijswijk ZH, Netherlands J T RICK,Department of Psychology, University of Birmingham, Birmingham B15 2TT, England G M R SAMUELS, Tunstall Laboratory, Shell Research, Sittingbourne, Kent, England J STORM-MATHISEN, Norwegian Defence Research Establishment, Division for Toxicology, P.O Box 25, Kjeller, Norway J C SZERB, Department of Physiology and Biophysics, Dalhousie University, Sir Charles Tupper Medical Building, Halifax, Nova Scotia, Canada A M VAN DER POEL, Department of Fundamental Pharmacology, University of Leiden, Wassenaarseweg 62, Leiden, Netherlands This Page Intentionally Left Blank Preface The papers contained in this Volume were presented at a meeting held at the Chemical Defence Establishment, Porton Down, Salisbury, on March 22 and 23, 1971, and which was attended by government scientists from the U.K., Norway, Sweden and the Netherlands, together with a number of academic research workers While there exist in many countries brain research institutes where neurobiologists from various disciplines work side by side and have daily contact for the exchange of ideas and experimental findings, in the U.K such research is fragmented Thus, certain individuals working in departments of anatomy, physiology, pharmacology and psychology are engaged upon investigations into brain function in their own disciplines but there is no coordinated effort, nor are there adequate opportunities for liaison between different disciplines One way of attempting to overcome this isolation of brain research workers is by meetings or symposia and one of the purposes of the C.D.E Symposium was to achieve this end Additionally, the meeting served the purpose of bringing together scientists in government research laboratories and academic research workers who not often have such opportunities for exchange of ideas, etc It is to be hoped that further meetings will be held in the future along similar lines but on different topics As the Symposium had to be limited in the number of participants attendmg, it was decided to concentrate in this first meeting on two main aspects of brain research, namely the study of biochemical and pharmacological mechanisms In particular, since t-hese two approaches tend to be pursued independently, it was hoped that a greater degree of integration between biochemistry and pharmacology might ensue and that their relevance in terms of behaviour become more apparent The first day was therefore devoted to papers dealing in the main with biochemical mechanisms and on the second day papers on the actions of drugs producing changes in behaviour were presented The fact that more than haif the papers presented were concerned to a greater or lesser extent with cholinergic mechanisms in the central nervous system reflects, in our opinion, the relative importance of acetylcholine in brain function, although of late this transmitter has been somewhat neglected in favour of others We are grateful to the Director of C.D.E.,Mr G N Gadsby, for making facilities available for this Symposium and to the staff of the Establishment for their assistance with the organization Thanks are also due to Miss Sally Clements for helping to edit these proceedings and to Miss Josephine Knight for her excellent work during the meeting in keeping track of the discussion and discussants P B Bradley R W Brimblecombe 190 R A ANDERSEN et al P 40 20 L u V Fig In vivo reactivation of CNS ChE o guinea pigs Paraoxon (0.75 mg/kg) administered in f tracardially, was followed h later by P2S (60mg/kg) administered subcutaneously times at intervals of h The animals were killed and the ChE was determined 18 h after the last P2S dose (columns on the right) Controls (columns on the left) were treated similarly except that the administration of P2S was omitted Enzyme activities are expressed as percentages of normal activity, each column representing the mean value from animals, and the bars denoting f x S.E.M (Reproduced from Arch int Pharmacodyn (1968), AARSETH, AND BARSTAD, A B., 176,434442.) P J 1000 - 800 600 400 200 Fig In vivo ChE activity in various CNS areas and in whole blood after injection of a quarternary irreversible inhibitor (grey columns) compared with corresponding normal values (white columns) The inhibitor, 3-MPAM-ES, was administered subcutaneously in a dose of mg/kg 24 h before death The columns represent average enzyme activities @moles acetylthiocholine split per g wet weight per h) in male guinea pigs, except for the blood which is from animals only Bars show P f x S.E.M (Reproduced from Arch int Pharmacodyn (1968), AARSETH, AND BARSTAD, J A B., 176,434-442.) acetylcholinesterase (AChE), are situated inside the respective barriers This concept is supported by experiments in which quaternary AChE inhibitors failed to inhibit these enzymes (Koelle and Steiner 1956; Fredriksson, 1957, 1958; Andersen et al., 1970) By means of the technique used by Aarseth and Barstad (1968), the present authors have studied the permeability of the rat BBB and BRB to quaternary nitrogen compounds, viz an irreversible AChE inhibitor (2-MPAM-ES) (Fig 4A; see also Barstad et a/., 1969) and AChE reactivators (Fig 4B-D) The 2-MPAM-ES iodide was administered subcutaneously, 0.325 mg/kg, to male rats having received atropine sulphate (5 mg/kg) subcutaneously 30 in ACCESS OF QUATERNARY DRUGS TO THE 191 B A CH=W CNS CH-WH CH=NOH C CH=W D Fig The chemical constitution of the quaternary compounds employed A, The phosphorylated oxime, 2-MPAM-ES, a strong irreversible ChE inhibitor with the oxime moiety as the leaving group B-D, The reactivators P2S, Tx 10 and Toxogonin, respectively advance of 2-MPAM-ES administration Sixteen hours after the injection of 2MPAM-ES the animals were killed, and ChE-activities were determined in whole blood and homogenates from the liver, retina and cerebral cortex, the values being compared with those of untreated controls The method used for ChE activity determination was that described by Ellman et al (1961) The results (Fig ) indicate that the penetration of 2-MPAM-ES through both the barriers was slow To see whether active transport of the quaternary nitrogen compounds outwards through the BBB or BRB could play any role in these events, similar experiments were performed on two groups of animals pretreated with inhibitors of metabolismdependent transport, viz., 2,4-dinitrophenol and ouabain In some additional ex- '201 100 80 60 t 40 20 Fig AChE activities slightly more than 16 h after injection of the quaternary inhibitor Z-MPAMES iodide (0.325 mg/kg) administered subcutaneously in male rats treated with atropine Enzyme activities are expressed as percentages of the normal Bars denote scatter at 95% fiducial probability ( k P = 0.05) , References PP 194 192 R A ANDERSEN et al, periments the animals instead were pretreated with high X-ray doses through the head In the first series, animals were injected subcutaneously with 2,4-dinitrophenol (10 mg/kg) Two hours later, atropine sulphate (5 mg/kg) was given subcutaneously and then 2-MPAM-ES, etc., as described above The second series was similar except that ouabain (1 mg/kg) was used instead of 2,4-dinitrophenol In the third series, only animals were employed These were irradiated with a dose of 5,000 rad uniformly over the head 24 h prior to administration of 2-MPAM-ES The results (Fig 6A) indicate that none of these pretreatments had any significant effect on the permeability of either the BBB or the BRB by 2-MPAM-ES In another T A B Fig A, ChE activity of hornogenates from the cerebrum (white columns) and retina (hatched columns) of atropinized male rats 16 h after subcutaneous administration of 2-MPAM-ES iodide (0.325 mg/kg) Two hours prior to the administration of the inhibitor the animals were pretreated with subcutaneous injections of 2,Cdinitrophenol (10 mg/kg), ouabain (1 mg/kg), and X-ray irradiation, 5,000 rads, through the head Enzyme activities are expressed as percentages of the normal Erythrocyte ChE activity (not shown on the figure) was reduced to 24% (see also Fig 3) B, Columns denoting ChE in the cerebrum and retina as in A, but from animals pretreated with paraoxon (0.3 mg/kg) intravenously The left pair of columns represents control animals which were given no subsequent reactivator therapy, whereas the following pairs of columns show the effect of the enzyme reactivators indicated on figure The ChE reactivation is seen to be more pronounced in the retina than in the cerebrum Reactivation of erythrocyte ChE (not shown) was materially complete in these experiments For further details (reactivator dosage, number and time-spacing of doses, etc.) see text series of experiments, intravenous doses of paraoxon (0.3 mg/kg) were administered intravenously to male rats Starting 2.5 h later, doses of pralidoxime methanesulphonate (P2S) (60 mg/kg each) were given subcutaneously at intervals of h to animals The animals were killed 18 h later and the ChE in the blood and homogenates of the brain and retina determined as stated above Similar experiments were performed on 11 animals with Toxogonin (1,l’-oxydimethylenebis (4-hydroxyiminomethyl pyridinium chloride)) as enzyme reactivator, each dose being 1/5 the P2S dose In yet another series Tx 10 (Fig 4C) (3/5 the P2S dose) was used for reactivation The results from the reactivation experiments were compared with controls ACCESS OF QUATERNARY DRUGS TO THE CNS 193 similarly pretreated, but having received no P2S The doses of P2S, Tx 10 and Toxogonin were roughly equiactive with regard to reactivation of diethyl-phosphorylAChE in rat erythrocytes in vitro and in vivo The reactivation of AChE in the brain or retina by the different reactivators should therefore reveal any gross differences between the reactivators with regard to penetration of the BBB or the BRB The results of these experiments are shown in Fig 6B The BRB is seen to be significantly more permeable to the reactivators than is the BBB These results were supported by experiments with P2S, in which ChE determinations in brain and retina were performed histochemically according to the method described by Koelle and Friedenwald (1949) No significant differences between the reactivators with regard to penetration of either of the two barriers were demonstrated More detailed investigations, on discrete areas of the barriers concerned, as we11 as under more varied conditions, are desirable With this reservation the following general conclusions seem justified: (1) Whereas the BBB seems to vary very little with gross location, and to be highly impermeable to quaternary nitrogen compounds, the BRB - ontogenetically a part of the BBB - seems to be somewhat more leaky to quaternary nitrogen compounds than the BBB proper To the phosphorylated oxime employed, which contained a quaternary nitrogen group, even the BRB was, however, highly impermeable (2) The penetration of quaternary nitrogen compounds through the BBB and the BRB does not seem to be influenced by high doses of ionizing radiation, nor by 2,4-dinitrophenol or ouabain, drugs known to interfere with active transport through biological membranes (3) There is apparently no marked difference between the AChE reactivators P2S, Toxogonin or Tx 10 with regard to penetration of either barrier concerned SUMMARY Four quaternary nitrogen compounds, one of them an irreversible cholinesterase inhibitor (2-MPAM-ES), the remaining cholinesterase reactivators (P2S, Tx 10 and Toxogonin), were used in a study of the permeability of the blood-brain barrier (BBB) and the blood-retinal barrier (BRB), the latter being, from an ontogenetic point of view, a part of the former In contrast to other parts of the BBB, which have been found in earlier work to show slight mutual differences with regard to permeability, the BRB was found to be more permeable to the reactivators than was the BBB proper Both the BBB and the BRB were highly impermeable to 2-MPAM-ES This impermeability was not significantly influenced by high X-ray doses (5,000 rad), nor by toxic doses of 2,4-dinitrophenol or ouabain No marked differences were found between the cholinesterase reactivators with regard to penetration of either barrier References p 194 194 R A ANDERSEN et al REFERENCES J AARSETH, AND BARSTAD, A B (1968) Blood-brain barrier permeability in various parts of the P central nervous system Arch inf Pharmacodyn., 176, 434442 ANDERSEN, A., BARSTAD, A B AND LAAKE, (1970) Permeability of the blood-retina barrier R J K to quaternary nitrogen compounds Acra Pharmacol Toxicoi., Suppl 1,28, 31 BAKAY, (1957) Dynamic aspects of the blood-brain barrier In Mefabolisrn ofrhe Nervous System, L D RICHTER (Ed.), Pergamon Press, London, pp 136152 BARSTAD, A B., LILLEHEIL.,G AND SKOBBA, J (1969) Phosphorylated oximes Some pharmacoJ T toxicological and biochemical features Arch inf.Pharmacodyn., 179, 352-363 ELLMANN, L., COURTNEY, D., ANDRES, V JR AND FEATHERSTONE, M (1961) A new and G K R rapid colorimetric determination of acetylcholinesterase activity Biochem Pharmacol., , 88-95 FREDRIKSSON,T (1957) Pharmacological properties of methyl-fluorophosphorylcholines.Two synthetic cholinergic drugs Arch int Pharmacodyn., 113, 101-1 13 FREDRIKSSON, T (1958) Further studies on fluorophosphorylcholines Pharmacological properties of two new analogues Arch inf Pharmacodyn., 115, 474-482 KOELLE, B AND STENR, E C (1956) The cerebral distributions of a tertiary and a quaternary G anticholinesterase agent following intravenous and intraventricular injection J Pharmacol., 118,42&434 KOELLE, B AND FRIEDENWALD, (1949) A histochemical method for localising cholinesterase G J S activity Proc SOC exp Biol (N.Y.), 617-622 70, REESE,T s AND KARNOVSKY, J (1967) Fine structural localisation of a blood-brain barrier to M exogenous peroxidase J Cell Biol., 34,204-217 ROSENBERG, (1960) In vivo reactivation by PAM of brain cholinesterase inhibited by paraoxon P Biochem Pharmacoi., 3, 212-219 DISCUSSION I BERRY: was unhappy about your phosphorylated oxime since it has been found that the toxic and anticholinesterase effects of ethyl-sarin were more reversible than those of sarin However, your control experiments with liver show that the brain results cannot be attributed to rapid spontaneous reactivation BARSTAD: are right, the inhibitor is slightly reversible However, with the liver and blood controls You and the timing taken into account, the reversibility would probably not be an important source of error MEETER:only want to confirm your findings on the relative abilities of the various oximes to pass the I blood-brain barrier With the anticholinesterase hypothermia as a criterion we tested oxime and found no evidence whatsoever for differences in penetrating abilities HEILBRONN: When you compared the effects of the oximes in the brain you used totally different concentrations What were these related to? BARSTAD: They were approximately equiactive with regard to AChE reactivation in vitro &HEN: Are you sure that in the experiments in which you measured central ChE activity after inhibition with an anticholinesterase and treatment with an oxime there was no “reinhibition” by anticholinesterase which was still present in the tissues? BARSTAD: time course of the experiments makes it unlikely that a residue of inhibitor (or reactiThe vator) high enough to be a serious source of error should persist in the tissues at the time of homogenization We injected paraoxon intravenously to keep the dose low, thereby avoiding a depot effect The oximes were administered so as to outlast the inhibitor, and the animals were given 18 h more to get rid of the oximes With regard to the experiments with 2-MPAM-ES, a residue would have caused inhibition of AChE ACCES OF QUATERNARY DRUGS TO THE CNS 195 during homogenization of the brair, and retina This can be ruled out since no enzyme inhibition was demonstrated in these tissues BERRY: the point of the alleged presence of free organo-phosphate in tissues there is no reliable On evidence that there is any detectable amount after barely lethal doses This was shown in experiments which Rutland and I did a few years ago, and Creasey even earlier BARSTAD: I agree; but just to play safe we still took this into account This Page Intentionally Left Blank Author Index Aldous, F A B., 143-157 Ancill, R J., 79-95 Andersen, R A., 189-194 Ansell, G B., 3-10 Inch, T D., 59-64 Kerkut, G A., 65-77 Laake, K., 189-194 Barrass, B C., 97-104 Barstad, J A B., 189-194 Beesley, P., 65-77 Bradley, P B., 183-186 Brimblecombe, R W., 65-77, 115-125 Buxton, D A., 115-125, 171-180 Malthe-Ssrenssen, D., 13-26 Meter, E., 139-140 Oliver, G W., 65-77 van der Poel, A M., 127-137 Coult, D B., 97, 104 Davies, J A., 79-95 Emson, P C., 65-77 Redfern, P H., 79-95 Rick, J T., 105-112 Fonnum, F., 13-26,41-57 Fulker, D W., 105-112 Samuels, G M R., 167-168 Spanner, S., 3-10 Storm-Mathisen, J., 41-57 Szerb, J C., 159-165 Green, D M., 143-157 Walker, R J., 65-77 Heilbronn, E., 29-39 This Page Intentionally Left Blank 199 Subject Index Absorption of ChAc to membranes 18 AC (see Auditory cortex) Acetylcholine applied iontophoretically to single brain neurones, 183 from hypo-osmotically treated synaptosomes, 22 metabolism in cerebral cortex, atropine on, 159 release from cholinergic vesicles after phospholipase A, 30, 36 synthesis from labelled precursors, 22 role of ChAc, 13 uptake into synaptic vesicles, 23 Acetylcholinesterase activity in cockroach CNS, 67 distribution in hippocampus, 42 in PZfraction of brain tissue, membrane-bound in synaptosomes, 14 reactivators, P2S, T x 10 and Toxogonin, penetration of BBB and BRB, 190 role in ACh synthesis 27 ACh (see Acetylcholine) AChE (see Acetylcholinesterase) Acridine orange binding by AChE in cockroach, 73 Ambulation in rat after anticholinergic drugs, 120 after substituted amphetamines, 175 Amitryptyline on oxidation of NA and 5-HT by caeruloplasmin, 103 (&)-Amphetamine effect on locomotor activity in rat, 80 electrocortical desynchronisation, increased prostaglandin release, 167 enantiomeric comparisons, 63 facilitation of learning in cockroach, 66 on 24-hour rhythm of locomotor activity, 85, 89 (+)-Amphetamine behavioural effects in rats compared to DOM and DOET, 172 iontophoretically applied to single brain neurones, 185 Anticholinergic drugs behavioural actions of, 115 on evoked potentials, EEG and behaviour, 143 potency of glycollate enantiomers, 59 Area dentata ChAc and AChE activities, 43 Atropine electrocortica1 synchronisation, decreased prostaglandin release, 167 on evoked potentials, EEG and behaviour, 143 on metabolism of ACh in cerebral cortex, 159 Atropine sulphate on behaviour in mice and rats, 119 Auditory cortex evoked potentials and anticholinergic drugs, 146 BBB (see Blood-brain-barrier) Blood ChE activity after quaternary nitrogen compounds, 19 Blood-brain- barrier permeability to quaternary nitrogen compounds, 189 Blood-retinal-bamer permeability to quaternary nitrogen compounds, 189 Brain subcellular fractions by zonal centrifugation, BRB (see Blood-retinal-barrier) Brom-LSD (BOL-148) on oxidation of NA and 5-HT by caeruloplasmin, 99 on single brain neurones, actions of ACh, NA and 5-HT, 185 Caeruloplasmin and centrally acting drugs, 97 catalysis of oxidation of NA and 5-HT, 99 CAR (see Conditioned avoidance response) Carbachol -induced hypothermia in rat, 139 contractions in guinea pig ileum, anticholinergic drugs on, 60 Central Nervous System of cockroach, AChE activity, 65 access by quaternary compounds, 189 Cerebral cortex homogenates, ChE activity after quaternary compounds, 191 200 SUBJECT INDEX ChAc (see Choline acetyltransferase) ChE (see Cholinesterase) Chlorpromazine electrocortical synchronisation, decreased prostaglandin release, 167 on evoked potentials, EEG and behaviour, 143 on single brain neurones, actions of ACh, NA and 5-HT, 183 (3H)-Choline uptake by rat brain, 31 (Me-l4C)-Choline in vivo uptake into rat brain tissue, Choline acetyltransferase in hippocampal region of brain, 42 molecular properties of, 13 Cholinergic vesicles effect of phospholipase A, 31 from electric organ of Torpedo, 30 Cholinesterase -activities after quaternary compounds, 191 activity and relationship with GABA production, 106 levels in cockroach; relationship with learning, 69 Cholinolytic drugs and choice behaviour in rats, 127 Circadian rhythms; hallucinogenic drug effects, 79 CNS (see Central Nervous System) Cockroach changes in chemistry of NS following behavioural changes, 65 Conditioned avoidance response after anticholinergic drugs, 117, 148 after substituted amphetamines, 173 Cycloheximide effect on learning in cockroach, 66 Defaecation in rats after anticholinergic drugs, 120 due to mild stress using different strains, 106 after substituted amphetamines, 175 2,5-Dimethoxy-4-ethylamphetamine behavioural effect in rats compared to LSD, mescaline and amphetamine, 171 2,5-Dimethoxy-4-methylamphetamine behavioural effect compared to LSD, mescaline and amphetamine, 171 on action of caeruloplasmin, 0 3,4-Dimethoxyphenylethylamine on action of caeruloplasmin, 100 2,CDinitrophenol pretreatment, permeability BBB and BRB to quaternary compounds, 191 DOET (see 2,5-Dimethoxy-4-ethylamphetamine) DOM (see 2,5-Dimethoxy-4-methylamphetamine) Dopamine level in Drosophila after GHB, 108 Drosophila rnelanogaster GHB on spontaneous activity and dopamine level, 107 Echothiophate iodide pretreatment in cats, effect on ACh output, 162 ECoG (see Electrocorticogram) Edrophonium facilitation of learning in cockroach, 66 EEG (see Electroencephalogram) Electrocorticogram synchronising and desynchronising drugs; prostaglandin release, 167 Electroencephalogram anticholinergic drugs, LSD and chlorpromazine on, 145 Electron microscopy cholinergic vesicles with phospholipase A, 33 histochemistry of ChAc, 22 Enantiomers of glycollates, anticholinergic potency, 60 EncEphallp &olP correlation between prostaglandin release and neuronal activity, 167 N-Ethyl-3-piperidyl benzilate (HCI) on behaviour in mice and rats, 119 N-Ethyl-3-piperidyl phenylcyclopentyl glycollate (HCI) on behaviour in mice and rats, 119 N-Ethyl-2-pyrrolidylmethylphenylcyclopent yl glycollate (HCI) on behaviour in mice and rats, 119 Evoked potentials effects of anticholinergic drugs, LSD and chlorpromazine, 145 Fimbria transection; effect on ChAc and AChE of hippocampal region, 44 GABA (see Gamma-aminobutyric acid) GAD (see L-Glutamate l-carboxylase) Gamma-aminobutyric acid in cerebral cortex, 49 production, ChE activity in sensori-motor cortex of rat, 106 Gamma-hydroxybutyric acid on spontaneous activity and dopamine level in Drosophilia, 108 GHB (see Gamma-hydroxybutyric acid) &Glucuronidase distribution in PZfraction of brain tissue, L-Glutamate 1-carboxylase activity in hippocampal region, 48 SUBJECT INDEX Glycollates behavioural effects of stereo-isomers, 59 behavioural effects in mice and rats, 119 alternation behaviour in trained and untrained rats, 128 Hallucinogenic compounds on oxidation NA and 5-HT by caeruloplasmin, 99 drugs and circadian rhythms, 79 Halothane -NzO anaesthesia; atropine and ACh output in cat, 160 Harmine on oxidation of NA and 5-HT by caeruloplasmin, 99 Harmol on oxidation of NA and 5-HT by caeruloplasmin, 99 Helix aspersa AChE activity in CNS, 73 Hemicholinium-3 on ACh output and content with or without atropine, 162 Hippocampal region localisation of transmitter substances, Hippocampus regio superior ChAc and AChE activities, 43 Histochemical staining of hippocampus regio superior and area dentata, 42 5-HT (see 5-Hydroxytryptamine) 3-Hydroxy-4-methoxyphenylethylamine acceleration of oxidation NA and 5-HT by caeruloplasmin, 100 5-Hydroxytryptamine iontophoretically applied to single brain neurones, 183 loss from hippocampus after transection of fimbria, 53 Hyoscine on evoked potentials, EEG and behaviour, 143 Hyoscine hydrobromide on behaviour in mice and rats, 119 Hypothermia due to organo-phosphorous ChE inhibitors or carbachol, 139 Ibogaine on oxidation of NA and 5-HT by caeruloplasmin, 99 Imipramine on oxidation of NA and 5-HT by caeruloplasmin, 103 Inherited behavioural differences, some biochemical correlates of, 105 20 Intertrial interval lengthening of on alternation behaviour in rats, 128 Ion tophoretically applied drugs on single brain neurones, 183 Irradiation of rats prior to administration of 2-MPAMES, 192 Isoelectric focusing of ChAc from brain, 18 Isoenzymes of ChAc from rat brain synaptosomes, Lactic dehydrogenase occluded; distribution in PZ fraction of brain tissue, Lateral lemniscus evoked potentials and anticholinergic drugs, 15 (3H)-Lecithin preparation of; test for phospholipase A activity, 35 Leech muscle-test, release of ACh from cholinergic vesicles, 31 Liver homogenates, ChE activity after quaternary compounds, 191 LL (see Lateral lemniscus) Locomotor activity of rat “sympathomimetic psychotomimetic” drugs on, 80 LSD (see Lysergic acid diethylamide) Lysergic acid diethylamide behavioural effects in rat compared to DOM and DOET, 172 on evoked potentials, EEG and behaviour, 143 on locomotor activity of rat, 87 on oxidation of NA and 5-HT by caeruloplasmin, 99 on single brain neurones, actions of ACh, NA and 5-HT, 184 stereochemical and electronic comparisons with tryptamine and amphetamine, 63 (3H)-Lysolecithin formed from (3H)-lecithin with phosphoIipase A, 35 Lysosomal fraction of brain tissue by zonal centrifugation, Mescaline behavioural effects in rats compared to DOM and DOET, 172 on action of caeruloplasmin, 100 Mescaline hydrochloride effect on locomotor activity in rat, 83, 88 202 SUBJECT INDEX N-Methyl-3-piperidyl benzilate on evoked potentials, EEG and behaviour, 143 N-Methyl-3-piperidyl benzilate (HCl) on behaviour in mice and rats, 119 N-Methyl-Cpiperidyl cyclopentyl methylethynyl glycollate on alteration behaviour in trained and untrained rats, 128 N-Methyl-3-piperidyl phenylcyclopentyl glycollate (HCI) on behaviour in mice and rats, 119 Methysergide on single brain neurones, actions of ACh, Na and 5-HT, 185 Mitochondria fraction of brain tissue by zonal centrifugation, 2-MPAM-ES penetration of BBB and BRB, 190 Mydriasis in mice, anticholinergic potencies of glycollate enantiomers, 60 Myelin removal from brain tissue, NA (see Noradrenaline) Naja naja siamensis venom, isolation of phospholipase A, 29, 35 Neostigmine competition with acridine orange to bind onto ChE, 73 on learning in cockroach, 66 Noradrenaline concentration in rat brain; variation with clock-hour, 82 iontophoretically applied to single brain neurones, 183 level in rat hippocampus after transection of fimbria, 53 Open-field test after anticholinergic drugs, 117 after substituted amphetamines, 173 Organo-phosphorous ChE inhibitors and resulting hypothermia in rats, 139 Ouabain pretreatment; permeability BBB and BRB to quaternary compounds, 191 Oxotremorine in mice, anticholinergic potencies of glycollate enantiomers, 60 in mice, anticholinergic drugs thereafter, 116 1,l'-Oxydimethylenebis (4hydroxyiminomethyl pyridinium chloride) effect on ChE activity in atropinised rats, 192 Oxygen consumption after carbachol in rats, 139 Pargyline differential uptake by synaptosomes, PCMG (see N-Methyl-Cpiperidyl cyclopentyl methylethylnyl glycollate) Pemoline facilitation of learning by cockroach, 66 Pentobarbitone electrocortical synchronisation, decreased prostaglandin release, 167 Permeability of BBB and BRB to quaternary nitrogen compounds, 189 P fraction of brain, use of sucrose gradient, Phospholipase A action on synaptic transmission, 29 Physostigmine ECoG desynchronisation, increased prostaglandin release, 167 Plasma membrane fraction of brain tissue by zonal centrifugation, Polygenic pleiotropy, interrelation of biochemistry and behaviour, 109 Pralidoxime methanesulphonate effect on ChE activity in rat, 192 Pretrigeminally sectioned non-anaesthetised cats; atropine on ACh, 161 Prostaglandins factors influencing release from cerebral cortex, 167 Prostigmine facilitation of learning in cockroach, 66 P2S (see Pralidoxime methanesulphonate) Psychotomimetic drugs on circadian rhythms with lighting variations, 79 Quaternary nitrogen compounds, access to CNS, 189 Rabbit cortex, subcellular fractions, Rat brain subcellular fractions, Reticular formation stimulation, ACh output and atropine, 160 Retina homogenates, ChE activity after quaternary compounds, 191 Salivation anticholinergic potencies of glycollate enantiomers, 60 SUBJECT INDEX oxotremorine-induced, anticholinergic drugs on, 116 Scopolamine on running speeds of rats in T-maze, 130 Sensori-motor cortex of rats, GABA production and ChE activity, 107 Serotonin (see 5-Hydroxytryptamine) Sodium dodecyl sulphate separation of AChE in snail brain, 74 Spontaneous locomotor activity in rats after anticholinergic drugs, 117 in rats after substituted amphetamines, 172 in Drosophila, GHB on, 108 State-dependent learning in rats treated with cholinolytic drugs, 128 Stimulation of surface and reticular formation, atropine on ACh output, 160 STP(see DOM) Subcellular fractions of brain by zonal centrifugation, fractionation of hippocampal region, ChAc in synaptosomes, 43 Substituted amphetamines, behavioural effects in rats, 112 Succinic dehydrogenase distribution in PZ fraction of brain tissue, Suxose gradient preparation of PZfraction from brain tissue, Synaptic transmission, phospholipase A on, 29 vesicles from rat brain cortex, phospholipase A on, 31 uptake of ACh, 23 Synaptosomal fractions of brain tissue after zonal centrifugation, fractions of hippocampal region, ChAc distribution, 43 203 Synaptosome disruption, release ChAc at different ionic strength and pH, 15 species difference with regard to particulate ChAc, 14 Thermoregulation of rat, central cholinergic mechanisms, 139 T-maze alternation behaviour, cholinolytic drugs on, 128 Torpedo nobiliana cholinergic vesicles from electric organ, 30 Toxogonin (see 1,l '-Oxydimethylenebis(4hydroxyiminomethyl pyridinium chloride)) Tranquillizers on oxidation of NA and 5-HT by caeruloplasmin, 103 Tremors oxotremorine-induced anticholinergic drugs on, 116 anticholinergic potencies of glycollate enantiomers, 60 Tryptamine stereochemical and electronic comparisons with LSD and amphetamine, 63 UML (see Methysergide) Vasodilation -free hypothermia in rats after carbachol, 139 X-radiation of rats prior to administration of 2-MPAMES, 192 Zinc concentrations in mossy fibres of hippocampal region, 54 Zonal centrifugation of brain subcellular fractions, isolation of cholinergic vesiclesfrom Torpedo, 30 This Page Intentionally Left Blank ... investigations into brain function in their own disciplines but there is no coordinated effort, nor are there adequate opportunities for liaison between different disciplines One way of attempting to... rat and cat brains have higher isoelectric points than those from pigeon and guinea-pig brains In addition, ChAc from rat and cat brains consists of two or three different isoenzymes with different... The subcellular distribution of endogenous E S and exogenous serotonin in brain tissue: comparison of synaptosomes storing serotonin, norepinephrine and y-aminobutyric acid J Neurochem., 18 333-343