THE CORRELATION BETWEEN CHOLINE ACETYLTRANSFERASE AND ACETYLCHOLINESTERASE ACTIVITY IN DIFFERENT AREAS OF THE CEREBELLUM OF RAT AND GUINEA PIG

Abstract— Choline acetyltransferase (ChAc) and acetylcholinesterase (AChE) levels were measured quantitatively in samples from the archi- and paleocerebellar vermis (Larsell's Lobules IX c,d,-X, and Lobules VII-VIII, respectively) and from the cerebellar peduncles, nuclei and white matter of rat and guinea pig. Lesions to isolate archi- or paleocerebellar areas were made in some rats and the effect on enzyme levels and ultrastructure were studied. In the rat there was a striking correlation between the activity of ChAc and AChE in the different areas; thus in the archicerebellar cortex the levels of both enzymes were 3–4 times those in the paleocortex. Deafferentation caused a fall in ChAc and this practically paralleled the fall in AChE in the same area. The reduction in both enzymes was more pronounced in the archi- than in the paleocerebellar cortex. In the guinea pig the results were very different. The ChAc activity was much lower than in the rat and was equal in the archi- and paleocerebellum. The AChE activity was also uniform in the different areas but, in contrast to ChAc, was higher than in the rat.     

Regional and subcellular distribution of choline acetyltransferase in the brain of rats

—Homogenates of corpus striatum, cerebral cortex and hypothalamus excised from rat brain were fractionated on discontinuous Ficoll and sucrose density gradients, and the distribution of choline acetyltransferase (ChAc) in the mitochondrial and synaptosomal fractions was determined. In the hypothalamic and cortical regions the fractions enriched in synaptosomes showed much higher activity of ChAc than those containing mainly mitochondria. On the other hand, the corpus striatum showed an equal distribution of ChAc activity in those two fractions. The localization of ChAc was also studied in the postnuclear supernatants obtained from three brain regions, using continuous sucrose density gradients. The distribution of ChAc was compared to that of monoamine oxidase (MAO), potassium and protein. When the pellets obtained from the fractions collected from the gradient were suspended in sucrose, the peak of ChAc activity was close to that of MAO in all three brain regions. When 0.1 mm EDTA +1% butanol was used in order to liberate the occluded form of ChAc, the maximum liberation occurred in lighter fractions, resulting in a shift of the activity peak toward the top of the gradient. This was found with fractions from hypothalamic and cortical regions. In the striatum, the liberated ChAc remained in the same fractions as the occluded enzyme. The results indicate that ChAc is liberated only in those fractions where it is present in synaptosomes. In agreement with the results on the discontinuous gradients this occurs in particles of lower density than mitochondria in cortex and hypo-thalamus, but in particles of similar density to mitochondria in the corpus striatum, indicating regional differences in the distribution of ChAc in the brain. K+ containing particles centrifuged in less dense fractions than those containing ChAc, indicating that synaptosomes are heterogeneous with respect to these two marker substances.     

A histochemical study of the distribution of choline acetyltransferase and acetylcholinesterase activity in sensory ganglia and nerve roots of the bullfrog

Synopsis Histochemical techniques were employed for the localization of choline acetyltransferase (ChAc; EC 2.3.1.6.), acetylcholinesterase (AChE; EC 3.1.1.7) and cholinesterase (ChE; EC 3.1.1.8) activities in dorsal and ventral roots and dorsal root ganglia of the bullfrog. AChE activity was present in most of the neuronal elements of dorsal root ganglia, in some nerve fibres in the dorsal roots, and in all nerve fibres in ventral roots. ChE activity in dorsal root ganglia and in the dorsal roots was confined to non-neuronal elements. No ChE activity was demonstrable in the ventral roots. ChAc activity was localized in many neurons of the dorsal root ganglia and in some nerve fibres of the dorsal roots; however, none of the ventral root fibres were visibly reactive. Some supportive cells of the dorsal roots and ganglia contained small amounts of ChAc activity. Except for the ventral roots, the histochemical distribution of AChE and ChAc activity was similar. The results of solubility studies indicated that under the histochemical conditions, approximately 50% of the ChAc remained bound to the dorsal roots and ganglia, whereas more than 90% of the ChAc in the ventral roots was soluble. This would account for the lack of reactivity in ventral root fibres. Differences in ChAc solubility are discussed in relation to the interpretation of histochemical data and in relation to the concept of multiple forms of ChAc. The results of this study indicate that at least one-third of the neurons of the dorsal root ganglia contain significant levels of the enzymes involved in both the synthesis and hydrolysis of acetylcholine.     

PURIFICATION AND SOME PROPERTIES OF CHOLINE ACETYLTRANSFERASE (EC 2.3.1.6) FROM BOVINE BRAIN

Abstract— Choline acetyltransferase (ChAc) has been isolated and highly purified from the caudate nuclei of bovine brains. The procedure involved: (1) making an acetone- and chloroform-insoluble powder from the tissue; (2) treating the powder with aqueous buffer and chromatographing the extractable soluble proteins on an organomercurial-sepharose column; (3) removing impurities by passage through columns of DEAE-cellulose and hydroxyapatite; and (4) separation of the heme-containing protein from ChAc by denaturing the former with a mixture of chloroform and n -butanol. The purified ChAc was essentially homogeneous as judged by polyacrylamide gel electrophoresis and exhibited a pH optimum at about pH 7. The partially purified ChAc dissociated into two non-identical subunits when chromatographed with a dilute buffer on Bio-gel A. It did not dissociate when a more concentrated buffer was used. The purified ChAc dissociated on the Bio-gel A even in the presence of a high salt concentration. The dissociation was accompanied by a great loss of enzymatic activity, and we concluded that the presence of other proteins tends to prevent the dissociation of ChAc on gel filtration.     

Androgen receptors in cranial nerve motor nuclei of male and female rats

This study compared AR proteins in four cranial nerve motor nuclei among male and female rats that were intact, gonadectomized, or gonadectomized and given TP by immunohistochemistry. AR-immunoreactive (ir) neurons were found, in descending order of abundance, in the nucleus ambiguus, hypoglossal nucleus, and the facial and trigeminal motor nuclei of both males and females of intact and gonadectomized plus TP rats. Virtually every neuron of the nucleus ambiguus was AR-ir. In contrast, AR-ir neurons were either restricted to a specific area of the hypoglossal nucleus, or randomly distributed in the facial and trigeminal motor nuclei. The predominant AR-ir site shifted from cell nuclei to the cytoplasm, depending upon the presence or absence of ligand. Sex differences in the amount and staining intensity of AR-ir neurons were discernable in all four motor nuclei of intact rats, and these differences were maintained in gonadectomized plus TP rats, with the exception of the nucleus ambiguus. The immunostaining results were complemented by results from AR binding studies. Cytosolic AR binding values for the hypoglossal and facial motor nuclei of females were only approximately 50% of those of males despite the absence of a sex difference in neuron number. These results indicate that intrinsic sex differences in AR levels and androgenic regulation of AR exist in cranial nerve motor nuclei, and that there are differences in the abundance and distribution pattern of AR responsive neurons in cranial nerve motor nuclei. These results are consistent with the idea that sex differences in AR could account for sex differences observed in nerve regeneration and neuron loss following cranial nerve injury.     

DISTRIBUTION OF CHOLINE ACETYLTRANSFERASE AND GLUTAMATE DECARBOXYLASE WITHIN THE SUBSTANTIA NIGRA AND IN OTHER BRAIN REGIONS FROM CONTROL AND PARKINSONIAN PATIENTS

—The distribution of choline acetyltransferase (ChAc, EC 2.3.1.6) and l -glutamate 1-carboxylyase (glutamate decarboxylase, GAD, EC 4.1.1.15) was studied in serial frontal slices of the substantia nigra (SN) (pars compacta, PC; pars reticulata, PR; an intermediate region, IR) as well as in other brain areas from post mortem tissue of control and Parkinsonian patients. Within the SN from control brain ChAc and GAD activities showed a distinctive distribution: ChAc activity in PC was higher than in PR and IR by 427% and 253% respectively and within PC the enzyme activity in the rostral part exceeded that in the control part by 353%. The GAD activity in PC was higher by 41% than that in PR and within PC seemed to be higher in the caudal than in the rostral part. For both enzyme activities there were no significant differences between PR and IR or within these regions. In Parkinsonian brain both ChAc and GAD activities were reduced to 15-25% of controls in all 3 regions of the SN. The distinctive distribution of ChAc and GAD activity found in the SN of control brain was abolished: no difference was observed between the 3 regions. However, within PC the ChAc activity was lower in the medial than in the rostral part. Since nigral ChAc is possibly located in interneurons, the decrease in enzyme activity may be connected with the cell loss observed in the SN of Parkinsonian brain. By contrast, nigral GAD is probably contained in terminals of strio-nigral neurons and the decrease in enzyme activity in Parkinson's disease in the absence of striatal cell loss, may reflect a change in the functional state of these GABA neurons. Among various areas of control brains ChAc activity was highest in caudate nucleus and putamen while GAD was highest in SN. caudate nucleus, putamen and cerebral cortex. In Parkinsonian brain the most severe reduction in ChAc and GAD activities was found in the SN.     

TRANSPORT, TURNOVER AND DISTRIBUTION OF CHOLINE ACETYLTRANSFERASE AND ACETYLCHOLIN-ESTERASE IN THE VAGUS AND HYPOGLOSSAL NERVES OF THE RABBIT

Abstract— The transport, distribution and turnover of choline O -acetyltransferase (ChAc, EC 2.3.1.6) and acetylcholinesterase (AChE, EC 3.1.1.7) in the vagus and hypoglossal nerves were studied in adult rabbits. The enzymes accumulated proximally and distally to single and double ligatures on both nerves and thus indicated both a proximo-distal and retrograde flow of the enzymes. Double ligature experiments indicated that only 5–20 per cent of the enzymes were mobile in the axon. The rate of accumulation of both enzymes above a single ligature corresponded to the slow rate of axonal flow provided that all the enzymes were mobile, but to an intermediate or fast flow if only a small part of the enzymes was transported. The distribution of ChAc along the hypoglossal neurons was studied and only 2 per cent of ChAc was confined to cell bodies, 42 per cent was localized to the main hypoglossal nerve trunks and 56 per cent to the preterminal axons and axon terminals in the tongue. The ratio of AChE to ChAc was about 3 in the hypoglossal nerve and 32 in the vagus nerve.
Transection of the hypoglossal nerve was followed by a decrease in the activity of ChAc in the hypoglossal nucleus and nerve and in the axons and their terminals in the tongue. The activity of AChE decreased in the hypoglossal nucleus and nerve but not in the tongue. The half-life of ChAc in preterminal axons and terminals of the hypoglossal nerve was estimated to be 16-21 days from the results obtained on transport, axotomy and distribution of the enzyme. Intracisternal injection of colchicine inhibited the cellulifugal transport of both enzymes and led to an increase in enzyme activity in the hypoglossal nucleus.     

REGIONAL AND SUBCELLULAR DISTRIBUTION AND KINETIC PROPERTIES OF RAT BRAIN CHOLINE ACETYLTRANSFERASE–SOME FUNCTIONAL CONSIDERATIONS

The kinetic properties of soluble and membrane-bound choline acetyltransferase (ChAc) were determined as a function of homogenization media and solubilization procedure in various regions of rat brain. Treatment of homogenate and/or subcellular fractions with KCl, Triton X-100, or ether dramatically altered the apparent V_max and the degree of solubilization of the enzyme, but no fraction exhibited K_m values substantially different from 12 μM for acetyl-CoA and 200 μM for choline. On the other hand, increasing the ionic strength of the assay medium for a given fraction from 0-02 M to 0-5 M increased both V_max and K_m values for both substrates. The absolute levels and subcellular distribution of ChAc were determined in 11 brain regions to localize cholinergic cell bodies and nerve endings. Levels of ChAc varied from 139 m-units/g tissue in caudate-putamen to 5-7 m-units/g tissue in cerebellum. The fraction of ChAc activity associated with synaptosomes varied from near 75 per cent in caudate-putamen, hippocampus and cortical regions to near 20 per cent in septum, locus coeruleus area and substantia nigra area. The apparent parallel distribution of cholinergic and catecholaminergic nerve endings is discussed in terms of a hypothetical model for the pathophysiology and treatment of Parkinson's syndrome.     

CHOLINE ACETYLTRANSFERASE ACTIVITY OF HUMAN BRAIN TISSUE DURING DEVELOPMENT AND AT MATURITY

Abstract— (1) On analysis of human brain tissue to determine its choline acetyltransferase (ChAc) content the recovery of enzyme from many regions is very poor when the tissue is acetone-dried and then extracted in the standard manner; for this reason the method is unsuitable when quantitative recoveries are required; it is preferable to prepare sucrose homogenates and activate these with ether before incubation.
(2) From measurements made on homogenates of one adult brain the highest concentration of ChAc was found in the putamen and the lowest in the corpus callosum. The caudate nucleus also had a high activity. As in other mammals, the concentration of enzyme in the cerebellum was found to be low. Analogous results were obtained on a nine-year-old brain but the level of ChAc activity was generally higher than in the older brain.
(3) During foetal development up to thirty-two weeks, ChAc is higher in the cerebellum than in the caudate, the thalamus, corpora quadrigemina, medulla and spinal cord. In all regions the concentration and total amount of enzyme rise fairly steadily up to this time; between 24 and 32 weeks, however, its concentration in the cerebellum and corpora quadrigemina falls slightly although the total increases considerably.
(4) Comparison of the results with the data of other authors indicates general agreement between the distribution of the enzyme in the human brain and its distribution in other mammals, especially the rhesus monkey. The corpus callosum may be an exception since in man it contains little ChAc while in lower mammals it seems to have relatively high concentrations of both ACh and ChAc.
(5) In comparing the values for ChAc reported here with the values for AChE reported by others, three tissues, the globus pallidus, substantia nigra and cerebellum are found to be exceptional in that relative to their concentration in the caudate the activity of ChAc is only about one-tenth that of AChE.     

TRANSPORT OF CHOLINE ACETYLTRANSFERASE AND ACETYLCHOLINESTERASE IN THE RAT SCIATIC NERVE: A BIOCHEMICAL AND ELECTRON HISTOCHEMICAL STUDY

The transport of acetylcholinesterase (AChE) and choline acetyltransferase (ChAc) were investigated by biochemical and histochemical methods. After ligature of one of the sciatic nerves of the rat for varying times—4, 14, 20 and 44 h—the normal levels and the accumulation of AChE and ChAc activities were investigated. It can be inferred from the results that there is a rapid accumulation of AChE activity just proximal to the ligature, while the increase in ChAc activity is less pronounced. Distal to the ligature the level of AChE is above the control value whereas, in contrast to this, the ChAc activity is significantly decreased. Histochemical demonstration of the two enzymes indicates that they are present in the cholinergic axons. The reaction end-product produced by AChE occurs within vesicles and neurotubules, while the endproduct due to ChAc appears to be free in the axoplasm, bound to neurofilaments and on the outer surface of vesicles and tubules.     

Immunological properties of rat brain choline acetyltransferase.

Abstract— Four antisera active against choline acetyltransferase (ChAc) were obtained by injecting 22 rabbits with rat brain ChAc. The ChAc preparations used for immunization (specific activity from 015 to 2 μmol/min/mg of protein) were not pure and the antisera produced were not monospecific. The antisera inhibited and precipitated ChAc, but the precipitated enzyme-antibody complexes still retain ChAc activity. One millilitre of the most active serum precipitates 0–5 μmol/min of rat brain ChAc at the equivalence point. Its titre expressed in mg/ml of immunoglobulins precipitated with the antigen and the equivalence point was calculated at about 0.08 mg/ml of serum. This relatively low titre explains the lack of any visible ChAc immunoprecipitate in an immunodiffusion test. Cross-reactivity studies revealed that ChAc has undergone few changes during evolution, since antisera produced against rat brain ChAc still precipitate ChAc from fish (Torpedo).     

THE SYNTHESIS OF ACETYLCHOLINE BY THE OLIVOCOCHLEAR BUNDLE

Abstract— Choline acetyltransferase (ChAc; EC 2.3.1.6) was assayed in the membranous cochlea and in the eighth cranial nerve (both the vestibular and cochlear components) from the point where it leaves the brain stem to the internal auditory meatus of the cat. To determine the contribution of the efferent innervation of the cochlea to this enzymatic activity both eighth nerves and both membranous cochleae were assayed at 17–29 days following section of the right, crossed and uncrossed olivo-cochlear bundles (OCB) in the cat. The lesion was produced along the right sulcus limitans on the floor of the fourth ventricle. The left eighth nerves and cochleae served as controls in the ChAc assay. There was a significant decrease in ChAc activity in the right cochlea and eighth nerve after OCB section and degeneration. The mean activity of ChAc in the right cochleae of the 6 operated cats was 15 ± 7 μg of ACh formed. h⁻¹ (g wet wt. of tissue) ⁻¹ in comparison to the rate of all the intact cochleae of 156 ± 38 μg of ACh formed. h⁻¹. (g of tissue)⁻¹, a statistically significant difference ( P < 0005). The mean activity of ChAc in the right eighth nerves of the cats with OCB lesions was 30 ± 8 n-g of ACh formed. h⁻¹. (g of tissue)⁻¹in comparison to 91 ± 19 fig of ACh formed . h⁻¹. (g of tissue)⁻¹ found for intact eighth nerves. This difference was also significant ( P < 0005). Thus, section and degeneration of the crossed and uncrossed OCB reduce the activity of ChAc in the eighth nerve and membranous cochlea. This finding provides support for the hypothesis suggesting the cholinergic nature of olivo-cochlear transmission.     

HISTOCHEMICAL MAPPING OF MONOAMINE OXIDASE AND LACTIC DEHYDROGENASE IN THE PONS AND MESENCEPHALON OF SQUIRREL MONKEY (SAIMIRI SCIUREUS)

The detailed distribution of lactate dehydrogenase and monoamine oxide has been described at various levels through the pons and mesencephalon of the brain of the squirrel monkey. A comparison of the two enzymes has given an interesting picture of their selective localization in the different nuclei. Particular attention has been paid to select sections at comparative levels. Marked monoamine oxidase activity has been observed in the nucleus interpeduncularis, the nucleus centralis superior, the nucleus annularis, and in the neuropil and in the cytoplasm of neurons of some other nuclei. Particularly strong lactate dehydrogenase activity has, however, been observed in the corpus geniculatum laterale and mediale, the nucleus pulvinaris, the superior and inferior colliculi, the substantia grisea centralis and some of the nuclei of cranial nerves.     

Huntington's disease: regional alteration in muscarinic cholinergic receptor binding in human brain.

Huntington's Disease, an autosomal dominant neurological disorder, is characterized by diffuse neuronal degeneration particularly in the basal ganglia and cerebral cortex. The purpose of this study was to examine various discrete regions of choreic and control brains for alterations in muscarinic cholinergic receptor binding and choline acetyltransferase (ChAc) activity. Nine postmortem brains, three from patients with Huntington's Disease and six controls, were dissected into 17 discrete regions. Each regional homogenate was assayed for muscarinic receptor concentration by measuring specific membrane binding of [³H]-QNB, a potent muscarinic antagonist which selectively labels brain muscarinic receptors. Aliquots from each brain region were also assayed for ChAc activity. Of significance was the marked reduction in specific [³H]-QNB receptor binding in the caudate nucleus, putamen and globus pallidus of choreic brain while no significant alterations were detected in other brain regions. Significant decreases in ChAc activity were found in the caudate nucleus, putamen, and globus pallidus with no alterations in ChAc activity in the rest of the brain regions examined. The tissues were chosen such that protein levels were similar in both choreic and normal brain samples. The apparent reduction in the number of muscarinic cholinergic receptors in the choreic brains suggests that treatment with cholinomimetic drugs might be beneficial in Huntington's Disease.     

CHOLINE ACETYLTRANSFERASE (ChAc) ACTIVITY IN DEVELOPING CHICK OPTIC CENTRES AND THE EFFECTS OF MONOLATERAL REMOVAL OF RETINA AT AN EARLY EMBRYONIC STAGE AND AT HATCHING

The choline acetyltransferase (ChAc) activity was measured in the optic centres of chick embryos after early removal of the optic cup and of young chicks after monolateral extirpation of the right eyeball after hatching. The contralateral optic lobes were thus deprived of their complement of retinal fibres. The following results were obtained: in chick embryos the ChAc was slightly lower in the deafferented lobe between the 10th and the 14th day of incubation; between the 14th and the 17th day a critical fall in activity was observed leading to a significant ChAc loss of 71 per cent. In eye deprived chicks no significant change in total ChAc activity occurred during the first postoperative month; significant changes were found only in the second month. The results reached so far suggest that removal of retinal fibres does not cause short term changes in optic centre ChAc in either the embryo or the chick. ChAc contained in nerve cell bodies seems independent of synapses and its behaviour is interpreted as a reflection of metabolic disturbance of the centre.     

PURIFICATION OF RAT BRAIN CHOLINE ACETYLTRANSFERASE1

Abstract— The purification of choline acetyltransferase (ChAc) has been hampered by the increasing instability of the enzyme in the course of purification. By working with a high concentration of protein and by adding glycerol to the enzyme, the stability was increased. The purification was performed by centrifuging twice, at low and high salt concentrations, precipitation by ammonium sulphate and chromatography on carboxymethyl–Sephadex, hydroxylapatite and Sephadex G 100. The final steps were performed by using chromatography on an immunoabsorbent; this consists of agarose-coupled gammaglobulins of antisera devoid of any activity against ChAc itself and directed against other proteins still present in the purest ChAc preparation achieved by conventional biochemical techniques. The purest rat brain ChAc preparation had a specific activity of 20 μmol/min/mg of protein after a 30,000-fold purification. The enzyme was not homogeneous in polyacrylamide gel electrophoresis performed either at pH 4.5 or with sodium dodecyl sulphate. Pure ChAc from rat brain would have a specific activity of approximately 100 μmol/min/mg of protein.     

SURFACE CHARGE OF CHOLINE ACETYLTRANSFERASE FROM DIFFERENT SPECIES

—The adsorption of partially purified choline acetyltransferase (ChAc) from cat, rat, guinea-pig and pigeon brains by the cation exchange resins, CM-Sephadex (C-50) and Amberlite CG-50 II, was studied at various pH values and ionic strengths. ChAc from cat and rat were more strongly adsorbed by cation exchangers and therefore have a stronger net positive surface charge than those from guinea pig and pigeon. Experiments showed that the difference in adsorption between these two groups of enzymes could not be explained by overloading of the resin, by competitive effect of other proteins present in the enzyme preparations or by the presence of any component suppressing the adsorption of ChAc in any of the enzyme preparations. The adsorption of ChAc by a cation exchanger is very similar to its binding to synaptosome membranes. The significance of the positive surface charge of ChAc in studies on the compartmentation of ChAc in synaptosomes is discussed.     

Distribution of simple esterase in the nuclei and fiber tracts of the medulla oblongata of squirrel (Funambulus palmarum)

The distribution of simple esterase has been studied in the nuclei and fiber tracts of the medulla oblongata. The simple esterase activity has mainly been observed in the grey matter. The white matter did not reveal enzymatic activity. In the grey matter also the cranial nerve nuclei are intensely stained as compared to intrinsic and reticular nuclei. The distributive pattern of simple esterase in the nuclei has been discussed in relation to its metabolic involvement in brain.     

Developmental changes of choline acetyltransferase (ChAc) activity in chick embryo spinal and sympathetic ganglia

The ChAc activity of spinal and sympathetic ganglia was measured throughout the embryonic life of the chick. In spinal ganglia, the ChAc activity reached a first peak when the maximal proliferation of neuroblasts occurred. Then, the relative ChAc activity decreased. After the 12th day of incubation, the enzyme activity increased again and reached a second peak on the 16th day. In sympathetic ganglia, the general course of the development of ChAc activity was similar to spinal ganglia. However, higher enzymic activity was found. Furthermore, the earlier peak of ChAc activity occurred 48 hr later than the corresponding peak in spinal ganglia. The behaviour of ChAc activity in these two areas of the developing nervous system is interpreted as a function of their histogenesis.     

Chorein,the protein responsible for chorea-acanthocytosis,interacts with β-adducin and β-actin

Chorea-acanthocytosis (ChAc) is an autosomal, recessive hereditary disease characterized by striatal neurodegeneration and acanthocytosis, and caused by loss of function mutations in the vacuolar protein sorting 13 homolog A (VPS13A) gene. VPS13A encodes chorein whose physiological function at the molecular level is poorly understood. In this study, we show that chorein interacts with β-adducin and β-actin. We first compare protein expression in human erythrocyte membranes using proteomic analysis. Protein levels of β-adducin isoform 1 and β-actin are markedly decreased in erythrocyte membranes from a ChAc patient. Subsequent co-immunoprecipitation (co-IP) and reverse co-IP assays using extracts from chorein-overexpressing human embryonic kidney 293 (HEK293) cells, shows that β-adducin (isoforms 1 and 2) and β-actin interact with chorein. Immunocytochemical analysis using chorein-overexpressing HEK293 cells demonstrates co-localization of chorein with β-adducin and β-actin. In addition, immunoreactivity of β-adducin isoform 1 is significantly decreased in the striatum of gene-targeted ChAc-model mice. Adducin and actin are membrane cytoskeletal proteins, involved in synaptic function. Expression of β-adducin is restricted to the brain and hematopoietic tissues, corresponding to the main pathological lesions of ChAc, and thereby implicating β-adducin and β-actin in ChAc pathogenesis.