RESUMEN
Pituitary cells were cultured as three-dimensional reaggregates in serum-free chemically defined medium supplemented with different concentrations of dexamethasone. Immunostaining of the cells using a polyclonal antiserum and three monoclonal antibodies raised against choline acetyl transferase (CAT), revealed the presence of CAT immunoreactivity in 4-10% of anterior pituitary cells depending on the antibody used. CAT immunoreactivity was also found in freshly dispersed anterior pituitary cells. CAT-immunoreactive cells could be enriched on BSA and Percoll gradients and codistributed with ACTH-immunoreactive cells in these gradients. Perifusion of the aggregates with the potent muscarinic receptor antagonist atropine (Atr) resulted in a dose-dependent (0.1-100 nM) increase in both basal PRL and GH secretion; the response was dependent on the dexamethasone concentration in the culture medium. A similar response to Atr was observed in organ-cultured pituitaries. The specificity of the Atr effect was supported by the findings that the potent and highly specific muscarinic receptor blocker dexetimide showed a similar action, whereas its inactive enantiomer levetimide and the nicotinic receptor blocker hexamethonium failed to do so. Two other muscarinic antagonists, benzatropine and pirenzepine, showed a dose-dependent hormone-releasing action similar to that of Atr, but were less potent than the latter. Pirenzepine was only effective at high molar concentrations, suggesting that an M2 muscarinic receptor subtype was mediating the present phenomenon. Atr also potentiated GH release stimulated by the beta-adrenergic agonist isoproterenol and PRL release stimulated by vasoactive intestinal peptide, but had no effect on GRF-stimulated GH release. The choline uptake blocker hemicholinium abolished the effect of Atr on GH and PRL release. These data suggest that certain pituitary cells can express CAT activity and that these cells exert a tonic inhibitory activity on GH and PRL release which is mediated by a cholinomimetic substance, possibly acetylcholine, through a muscarinic receptor.
Asunto(s)
Colina O-Acetiltransferasa/metabolismo , Hormona del Crecimiento/metabolismo , Adenohipófisis/metabolismo , Prolactina/metabolismo , Acetilcolina/fisiología , Animales , Atropina/farmacología , Benzotropina/farmacología , Carbacol/farmacología , Células Cultivadas , Dexametasona/farmacología , Femenino , Hemicolinio 3/farmacología , Histocitoquímica , Técnicas para Inmunoenzimas , Isoproterenol/farmacología , Masculino , Naftilvinilpiridina/farmacología , Técnicas de Cultivo de Órganos , Pirenzepina/farmacología , Ratas , Ratas Endogámicas , Receptores Muscarínicos/efectos de los fármacos , Receptores Muscarínicos/fisiología , Péptido Intestinal Vasoactivo/farmacologíaRESUMEN
This paper correlates the X-ray structures of two 4-(beta-1-naphthylvinyl)pyridine analogues (one cis and one trans) with chemical and biological activity data for this class of cholineacetylase inhibitors. Our results suggest that one of the two proposed mechanisms for inhibition by this class of compounds better describes their efficacy. Previous arguments about coplanarity of the aromatic rings and nucleophilicty across the vinyl linkage need to be modified. Quantum calculations are also included and substantiate previous suggestions about the charge distribution across the vinyl linkages. An alternate new mechanism of inhibition is proposed to encompass the published data and more recent results discussed in this paper.
Asunto(s)
Colina O-Acetiltransferasa/antagonistas & inhibidores , Naftilvinilpiridina/farmacología , Piridinas/farmacología , Fenómenos Químicos , Química , Cisteína , Modelos Moleculares , Naftilvinilpiridina/análogos & derivados , Teoría Cuántica , Relación Estructura-Actividad , Compuestos de Sulfhidrilo , Azufre , Difracción de Rayos XRESUMEN
Analogues of the potent inhibitor of choline acetyltransferase (CAT) (E)-4-(1-naphthylvinyl)pyridine methiodide were synthesized and evaluated for their ability to inhibit CAT and protect against nerve agent intoxication. Several compounds, notably (E)-1-(2-hydroxyethyl)-(1-naphthylvinyl)pyridinium bromide (3), (E)-1-methyl-4-(1-naphthylvinyl)-1,2,3,6-tetrahydropyridine hydrochloride (22), and (E)-1-methyl-4-(1-naphthylvinyl)piperidine hydrochloride (23), were found to afford significant protection against sarin in the mouse and against soman in the guinea pig. However, protection was apparently not related to CAT inhibition. Compound 23, our most effective compound in protecting against nerve agent, was without CAT inhibitory activity. Compound 22, which proved to be a potent CAT inhibitor, most likely owed this activity to being dehydrogenated back to the pyridinium quaternary salt by oxidative enzymes. Several of the (naphthylvinyl)pyridine quaternary salts, but not their tertiary amine analogues, were found to be effective in slowing the rate of aging of soman-inhibited acetylcholinesterase. Ability to slow the rate of aging was enhanced by introduction of methoxy substituents on the aryl moiety whereas the aging rate was actually accelerated by chloro substituents. To date, our most effective compound in slowing the rate of aging, (E)-4-[(4-methoxy-1-naphthyl)vinyl]pyridine methochloride (6), did not provide significant protection against soman in the mouse.
Asunto(s)
Antídotos/síntesis química , Colina O-Acetiltransferasa/antagonistas & inhibidores , Naftilvinilpiridina/análogos & derivados , Piridinas , Acetilcolinesterasa/metabolismo , Envejecimiento/efectos de los fármacos , Animales , Inhibidores de la Colinesterasa/toxicidad , Cobayas , Dosificación Letal Mediana , Ratones , Naftalenos/síntesis química , Naftalenos/farmacología , Naftilvinilpiridina/farmacología , Piridinas/síntesis química , Piridinas/farmacología , Sarín/toxicidad , Soman/toxicidad , Relación Estructura-ActividadRESUMEN
Corneal epithelium is known to have a high acetylcholine (ACh) concentration, but its role remains uncertain. Furthermore, rabbit corneal epithelium is devoid of cholinergic receptors. ACh concentration in calf, rabbit, and frog corneal epithelium was determined with Fellman's fluorometric assay to be 16.9, 11.1, and 21.8 micrograms of ACh per gram of epithelium, respectively. The isolated frog cornea was used to examine a possible role of the cholinergic system on active ionic transport. A 2 mM concentration of exogenous ACh has a moderate inhibitory effect on Na transport but no effect on Cl transport. A 10(-4)M concentration of 4-(1-naphthylvinyl) pyridine (NVP), a choline acetyltransferase inhibitor, reversibly inhibited both Na and Cl transport by about 70%. NVP also reduced ACh content of frog corneal epithelium by 51%. Thus endogenous ACh, but not exogenous ACh, seems to be stimulatory of active ionic transport. Of several muscarinic or nicotinic blockers tested, 10(-3)M atropine inhibited Na transport by 55% and Cl transport by 83%; 10(-3)M nicotine inhibited Na transport by 33% and Cl transport by 17%. If frog cornea (like rabbit cornea) contains no cholinergic receptors, the effects of ACh, nicotine, and atropine on ionic transport may be mediated through a nonspecific pathway.
Asunto(s)
Acetilcolina/fisiología , Córnea/fisiología , Acetilcolina/metabolismo , Acetilcolina/farmacología , Animales , Transporte Biológico Activo/efectos de los fármacos , Bovinos , Cloruros/metabolismo , Córnea/metabolismo , Epitelio/metabolismo , Epitelio/fisiología , Naftilvinilpiridina/farmacología , Parasimpaticomiméticos/farmacología , Conejos , Rana catesbeiana , Sodio/metabolismoRESUMEN
Rat hippocampal minces were loaded with N-methyl-[3H]acetylcholine ([3H]ACh) in the presence of the 'poorly penetrating' acetylcholinesterase (EC 3.1.1.7, AChE) inhibitor echothiophate and the effect of the depolarizing agent veratridine determined on the subcellular storage and release of [3H]ACh and [3H]choline. Results indicated that veratridine stimulated the release of [3H]ACh from a crude vesicular fraction (P3) by a Ca2+-dependent process, while simultaneously accelerating the breakdown of cytosolic (S3) [3H]ACh. A portion of the [3H]choline derived from the hydrolyzed S3 [3H]ACh was donated to the P3 fraction for [3H]ACh formation and release. When the identical experiment was done using hippocampal minces from septal lesioned rats, veratridine did not stimulate either the Ca2+-dependent release of [3H]ACh or the hydrolysis of cytosolic [3H]ACh. Incubation of control hippocampal minces with paraoxon, an AChE inhibitor which can penetrate cholinergic nerve terminals more rapidly than echothiophate, prevented veratridine from stimulating the Ca2+-dependent release of [3H]ACh from the P3 fraction. Instead, it then stimulated the Ca2+-independent release of [3H]ACh from the S3 fraction. When minces were incubated with the choline O-acetyltransferase (EC 2.3.1.6, ChAT) inhibitor 4-(1-naphthyl)vinyl pyridine (NVP), veratridine was no longer able to stimulate the Ca2+-dependent release of labelled ACh either. Instead, veratridine stimulated the Ca2+-independent release of labelled ACh from the S3 fraction. NVP also abolished the veratridine-induced, Ca2+-dependent release of total ACh. Both paraoxon and NVP inhibited the reversible reaction of ionically bound ChAT prepared from rat brain when tested in vitro, yet paraoxon was much less potent than NVP, and was unable to inhibit this reaction at the low concentration which prevented the veratridine induced breakdown of S3 [3H]ACh during mince incubation. Veratridine depolarization of hippocampal minces stimulated the activity of a membrane-bound fraction of ChAT associated with the P3 fraction, but this fraction of ChAT did not become more sensitive to inhibition by paraoxon during tissue incubation. Veratridine depolarization of minces also increased the activity of membrane-bound AChE, but this enzyme was not inhibited by the low NVP concentration which prevented the veratridine-induced breakdown of S3 [3H]ACh. The veratridine-induced increase in membrane-bound ChAT activity was dependent on the presence of extracellular Ca2+ in the incubation medium.(ABSTRACT TRUNCATED AT 400 WORDS)
Asunto(s)
Acetilcolina/metabolismo , Acetilcolinesterasa/metabolismo , Colina O-Acetiltransferasa/metabolismo , Hipocampo/efectos de los fármacos , Veratridina/farmacología , Veratrina/análogos & derivados , Animales , Yoduro de Ecotiofato/farmacología , Hipocampo/metabolismo , Técnicas In Vitro , Masculino , Naftilvinilpiridina/farmacología , Paraoxon/farmacología , Ratas , Tabique Pelúcido/fisiología , Fracciones Subcelulares/metabolismoRESUMEN
The effects of hemicholinium-3 (HC-3) or 4-(l-naphthylvinyl)pyridine (4-NVP) alone and together with cholinolytics and/or cholinesterase inhibitors on brain acetylcholine (ACh) levels and survival were studied. Intracerebroventricular (ICVT) injection of 10 micrograms HC-3 280 min before euthanasia by microwave irradiation reduced rat cerebral ACh levels from 28.4 to 5.4 nmoles ACh/g wet tissue. In rats pretreated with HC-3 alone or with other pretreatment drugs prior to giving up to 2.7 LD50 of soman, iv, cerebral ACh levels increased very little, but in animals not receiving HC-3, brain ACh levels increased to 67.1 nmoles. Treatment of unpoisoned rats with 4-NVP resulted in a significant (26%) reduction in ACh. The inclusion of atropine with 4-NVP caused sign-free doses of physostigmine to produce toxic signs in rabbits and did not enhance the efficacy of carbamate pretreatment against soman. Pretreatment of rabbits with pyridostigmine and atropine methyl nitrate (AMN) failed to provide any protection against soman, but when HC-3, ICVT, was included with those drugs, the protective ratio (PR), against soman was increased excess ACh is a primary lesion in organophosphorus anticholinesterase intoxication and that the central nervous system is quite sensitive to excesses of ACh.
Asunto(s)
Acetilcolina/metabolismo , Atropina/farmacología , Encéfalo/metabolismo , Hemicolinio 3/farmacología , Naftilvinilpiridina/farmacología , Compuestos Organofosforados/farmacología , Piridinas/farmacología , Soman/farmacología , Animales , Encéfalo/efectos de los fármacos , Hemicolinio 3/administración & dosificación , Inyecciones Intraventriculares , Masculino , Bromuro de Piridostigmina/farmacología , Conejos , Ratas , Ratas EndogámicasRESUMEN
Cholinergic neuronal influences on the function of male rat's hypothalamo-hypophyseal-adrenocortical (HHA) system both during basal and stressful situations (hypoxia and hypercapnia) were investigated using a cholinergic agonist (eserine) and antagonists (atropine, methyl atropine, mecamylamine and 4-(1-naphthylvinyl) pyridine). The results indicate that the transmitter, acetylcholine (ACh), plays a partial role in the regulation of the HHA system. The muscarinic (m) effects of ACh were stimulatory peripherally and inhibitory centrally. The nicotinic (n) effects were stimulatory and possibly affected the HHA system by inhibiting the central m-inputs. The cholinergic regulation of the HHA system for both non-stressed and hypercapnic animals is probably mediated via a common nm-cholinergic pathway.
Asunto(s)
Corteza Suprarrenal/fisiología , Glándulas Suprarrenales/fisiología , Hipercapnia/fisiopatología , Hipoxia/fisiopatología , Parasimpatolíticos/farmacología , Parasimpaticomiméticos/farmacología , Corteza Suprarrenal/efectos de los fármacos , Hormona Adrenocorticotrópica/farmacología , Animales , Atropina/farmacología , Derivados de Atropina/farmacología , Corticosterona/sangre , Dexametasona/farmacología , Masculino , Mecamilamina/farmacología , Naftilvinilpiridina/farmacología , Fisostigmina/farmacología , Ratas , Factores de TiempoAsunto(s)
Colina O-Acetiltransferasa/metabolismo , Músculos/enzimología , Sistema Nervioso/enzimología , Células de Schwann/enzimología , Potenciales de Acción , Animales , Anuros , Carnitina O-Acetiltransferasa/metabolismo , Desnervación Muscular , Músculos/fisiología , Músculos/ultraestructura , Naftilvinilpiridina/farmacología , Degeneración Nerviosa , Sistema Nervioso/ultraestructura , Fenómenos Fisiológicos del Sistema Nervioso , Rana esculenta , Factores de TiempoAsunto(s)
Acetilcolina/metabolismo , Acetiltransferasas/antagonistas & inhibidores , Encéfalo/metabolismo , Colina O-Acetiltransferasa/antagonistas & inhibidores , Colina/metabolismo , Inhibidores de la Colinesterasa/farmacología , Naftilvinilpiridina/farmacología , Piridinas/farmacología , Animales , Compuestos de Bencilideno/farmacología , Encéfalo/ultraestructura , Cobayas , Masculino , Ratones , Ratones Endogámicos , Estrés Fisiológico/metabolismo , Natación , Factores de TiempoAsunto(s)
Acetilcolina/biosíntesis , Acetiltransferasas/antagonistas & inhibidores , Colina O-Acetiltransferasa/antagonistas & inhibidores , Naftoquinonas/farmacología , Naftilvinilpiridina/farmacología , Piridinas/farmacología , Acetilcolina/metabolismo , Animales , Encéfalo/enzimología , Encéfalo/metabolismo , Colina/farmacología , Técnicas In Vitro , Cinética , Masculino , Ratones , ConejosAsunto(s)
Colina/metabolismo , Hemicolinio 3/farmacología , Hipocampo/efectos de los fármacos , Naftilvinilpiridina/farmacología , Piridinas/farmacología , Acetilación , Acetilcolina/metabolismo , Animales , Transporte Biológico Activo/efectos de los fármacos , Colina/farmacología , Hipocampo/metabolismo , Técnicas In Vitro , RatasAsunto(s)
Encéfalo/enzimología , Colina O-Acetiltransferasa/metabolismo , Membranas Sinápticas/enzimología , Acetilcolina/biosíntesis , Acetilcolina/farmacología , Acetilcolinesterasa/farmacología , Animales , Colina/metabolismo , Colina O-Acetiltransferasa/antagonistas & inhibidores , Cinética , Masculino , Ratones , Naftilvinilpiridina/farmacología , Sodio/farmacología , Solubilidad , Sinaptosomas/enzimologíaAsunto(s)
Acetilcolina/metabolismo , Consumo de Bebidas Alcohólicas , Encéfalo/metabolismo , Colina/metabolismo , Acetilcolinesterasa/metabolismo , Animales , Encéfalo/efectos de los fármacos , Colinesterasas/metabolismo , Litio/farmacología , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Naftilvinilpiridina/farmacología , Especificidad de la EspecieAsunto(s)
Acetilcolina/fisiología , Cuerpo Carotídeo/fisiología , Hipoxia/fisiopatología , Proteínas de Transporte de Membrana , Transducción de Señal/fisiología , Acetilcoenzima A/metabolismo , Animales , Azepinas/farmacología , Cuerpo Carotídeo/citología , Proteínas Portadoras/antagonistas & inhibidores , Gatos , Hipoxia de la Célula , Colina/metabolismo , Colina O-Acetiltransferasa/antagonistas & inhibidores , Inhibidores Enzimáticos/farmacología , Nervio Glosofaríngeo/fisiología , Hemicolinio 3/farmacología , Modelos Neurológicos , Naftilvinilpiridina/farmacología , Proteínas del Tejido Nervioso/antagonistas & inhibidores , Proteínas del Tejido Nervioso/metabolismo , Piperidinas/farmacología , Piridinas/farmacología , Receptores Presinapticos/fisiología , Transducción de Señal/efectos de los fármacos , Vesículas Sinápticas/efectos de los fármacos , Vesículas Sinápticas/fisiologíaRESUMEN
Acetylcholine synthesis in homogenates of frog sartorius muscle was measured by a radiometric method with a low blank. Choline acetyltransferase activity was very low (Vmax, 2nmol . g-1 . h-1, Km for choline, approx. 50 microM). The enzyme was found only in the endplate area and disappeared after denervation; it was inactivated by 4-(1-naphthylvinyl)pyridine. At high substrate concentrations its activity was overshadowed by the acetylcholine-synthesizing activity of a different enzyme not saturated by 10 mM-choline. The non-specific enzyme was present at and away from the endplate area, and it was not affected by denervation.
Asunto(s)
Acetilcolina/biosíntesis , Colina O-Acetiltransferasa/metabolismo , Músculos/enzimología , Animales , Colina/farmacología , Femenino , Cinética , Masculino , Placa Motora/enzimología , Desnervación Muscular , Naftilvinilpiridina/farmacología , Rana temporaria , Distribución TisularRESUMEN
The choline analog homocholine is not acetylated in vitro by choline-O-acetyltransferase (ChAT, EC 2.3.1.6), which is solubilized by 100 mM-sodium phosphate buffer washes of a crude vesicular fraction of mouse forebrain. However, both homocholine and choline are acetylated by a form of ChAT which is nonionically associated with a subcellular fraction of mouse forebrain containing membrane-associated organelles and occluded acetylcholine (P4). Acetylation of homocholine by membrane-associated ChAT is saturable. 4-(1-Naphthylvinyl)pyridine (NVP) inhibits the acetylation of both choline (60%) and homocholine (40%) by membrane-associated ChAT but reduces the acetylation of choline alone by soluble ChAT (76%). Choline and homocholine serve as competitive alternative substrates for the same membrane-associated ChAT, whereas homocholine acts only as a competitive inhibitor of choline acetylation by soluble ChAT. Acetylhomocholine competitively inhibits the acetylation of choline by both soluble and membrane-associated ChAT more dramatically than does the natural end product, acetylcholine.
Asunto(s)
Encéfalo/enzimología , Colina O-Acetiltransferasa/metabolismo , Colina/análogos & derivados , Colina/metabolismo , Neuronas/enzimología , Acetilación , Animales , Cinética , Masculino , Ratones , Naftilvinilpiridina/farmacología , Especificidad por SustratoRESUMEN
A new type of extended pi-system aza-analogue of (E)-4-[2-(1-naphthylvinyl)]-1-substituted pyridinium salts (NVP+) has been designed and its inhibitory activity towards choline acetyltransferase (ChAT) has been evaluated in vitro. Among the several examples of the title quaternary salts synthesized 2 and 3, the indolylvinylpyridinium salt 2e is the only one to show a very low ChAT inhibition. The molecular modeling study is highly illustrative of their behavior towards ChAT and interaction with the recognition site. Thus, several selected cations together with the reference NVP+ compound 1a were studied at the PM3 and AM1 levels respectively. At the global minima, all the compounds are planar, which, from the electron charge distribution, shows a degree of polarization similar to the NVP+ model compound 1a. However, the fitting of all optimized structures indicated that only the indole derivative 2e showed the same aromatic fragment orientation as 1a, which allows us to define a volume that is not accessible to ligands in the enzyme and consequently to a refined model of the choline acetyltransferase recognition site.
Asunto(s)
Inhibidores de la Colinesterasa/química , Inhibidores de la Colinesterasa/síntesis química , Naftilvinilpiridina/análogos & derivados , Animales , Pollos , Inhibidores de la Colinesterasa/farmacología , Diseño de Fármacos , Evaluación Preclínica de Medicamentos , Técnicas In Vitro , Modelos Moleculares , Naftilvinilpiridina/química , Naftilvinilpiridina/farmacología , Relación Estructura-ActividadRESUMEN
It has been postulated that the developing sympathetic innervation of rat eccrine sweat glands changes from adrenergic to cholinergic under the influence of its target. In agreement with previous evidence that the sympathetic innervation of adult rat sweat glands is cholinergic, we found that choline acetyltransferase (CAT)-immunoreactive nerve fibers are present in adult glands, and that gland-rich chunks of adult footpads contain CAT enzyme activity. We were therefore interested in determining when CAT activity is first expressed in the developing gland innervation. Low levels of acetylating activity were observed in rat footpads as early as postnatal day 4, when sympathetic fibers first contact the glands. A greater than fourfold increase in CAT specific activity occurred between postnatal days 11 and 21. Neonatal treatment of rats with the adrenergic neurotoxin 6-hydroxydopamine (6-OHDA) eliminated most of the CAT activity in 14 and 19 d footpads. In contrast, the acetylating activity observed prior to day 11 was unaffected by neonatal 6-OHDA treatment, and only slightly reduced by the selective CAT inhibitor, naphthylvinylpyridine. These results indicate that the sympathetic fibers that innervate rat sweat glands do not acquire detectable levels of CAT activity until a full week after they contact the glands.
Asunto(s)
Colina O-Acetiltransferasa/metabolismo , Glándulas Sudoríparas/inervación , Sistema Nervioso Simpático/enzimología , Acetilcolina/metabolismo , Animales , Animales Recién Nacidos , Colina/metabolismo , Hemicolinio 3/farmacología , Histocitoquímica , Hidroxidopaminas/farmacología , Naftilvinilpiridina/farmacología , Oxidopamina , Ratas , Sistema Nervioso Simpático/efectos de los fármacosRESUMEN
Following amphetamine administration both NVP and SKF-525A cause elevations in whole brain levels of amphetamine compared with saline treated controls. NVP was found to be about 3 times more active than SKF-525A; significant effects were obtained after doses of 0.625mg/kg and above. NVP also inhibited brain choline acetyltransferase in vitro (I50 of 4.0 x 10-5 M). Following the administration of NVP (20-200 MG/KG, I.P), there was a dose-dependent inhibition of this enzyme in brain. These results suggest that potentiation of the behavioral effects of amphetamine by NVP, which occur after doses of 5 mg/kg of NVP, are probably related to its inhibitory effects on amphetamine metabolism.
Asunto(s)
Química Encefálica/efectos de los fármacos , Dextroanfetamina/metabolismo , Naftilvinilpiridina/farmacología , Piridinas/farmacología , Animales , Encéfalo/enzimología , Encéfalo/metabolismo , Colina O-Acetiltransferasa/antagonistas & inhibidores , Dextroanfetamina/análisis , Masculino , Proadifeno/farmacología , RatasRESUMEN
Three forms of acetyl coenzyme A: choline-O-acetyltransferase (EC 2.3.1.6, ChAT) have been isolated from mouse and rat forebrain synaptosomes with a 100 mM sodium phosphate (NaP) buffer of pH 7.4, a high-salt solution (500 mM NaCl), and a 2% Triton DN-65 solution, respectively. The Triton-solubilized form of ChAT differed from the other two forms in its capacity to acetylate homocholine, its pH profile, and its sensitivity to denaturation. NaCl-solubilized ChAT could be distinguished from the other two forms with respect to pH profile, sensitivity to inhibition by 4-(1-naphthylvinyl) pyridine (in the presence of Triton), and apparent Km value for choline acetylation. The caudate and putamen of rat brain contained the highest amount of ChAT activity, based on tissue wet weight, and the cerebellum contained the least of the brain regions examined; only the cerebellum had more membrane-bound than soluble ChAT. Septal lesion reduced ChAT activity in the NaP- and Triton-solubilized fractions prepared from hippocampus by 68% and 64%, respectively, whereas it reduced the activity of the NaCl-solubilized fraction by only 21%. These results suggest that three different forms of ChAT may exist in both mouse and rat brain.