Journal of Neural Transmission

, Volume 115, Issue 7, pp 1027–1035 | Cite as

MKC-231, a choline uptake enhancer: (2) Effect on synthesis and release of acetylcholine in AF64A-treated rats

  • Ken TakashinaEmail author
  • Tomoko Bessho
  • Reiko Mori
  • Junichi Eguchi
  • Ken-Ichi Saito
Alzheimer's Disease and Related Disorders - Original Article


The effect of MKC-231 on acetylcholine (ACh) synthesis and release was studied in the hippocampus of normal and AF64A-treated rats. AF64A (3 nmol/brain, i.c.v.) produced significant reduction of high-affinity choline uptake (HACU) and high K+-induced ACh release in hippocampal synaptosomes. Treatments with MKC-231 (10−8 and 10−7 M) showed significant reverse of the decrease in both HACU and ACh release. In hippocampal slices superfused with choline-containing artificial cerebro-spinal fluid (ACSF), high K+-induced ACh release was gradually decreased by repeated alteration of resting and high K+ stimulations in AF64A-treated rats. However, addition of MKC-231 (10−8 to 10−7 M) in the superfusate reduces this decrease. In vivo microdialysis studies indicate MKC-231 (10 mg/kg, p.o.) significantly reversed reduction of basal ACh concentrations in AF64A-treated rats, measured by radioimmunoassay without a cholinesterase inhibitor in the perfusate. These results indicate MKC-231 improves AF64A-induced cholinergic hypofunction by enhancing HACU, subsequently facilitating ACh synthesis and release in vitro and in vivo.


MKC-231 AF64A High-affinity choline uptake (HACU) Acetyl choline (ACh) Microdialysis Radioimmunoassay 



The authors thank Prof. Koichiro Kawashima and Dr. Kazuko Fujimoto (Department of Pharmacology, Kyoritsu College of Pharmacy, Tokyo) for their invaluable technical advices and support for the microdialysis study and RIA assays for ACh.


  1. Aarsland D, Mosimann UP, McKeith IG (2004) Role of cholinesterase inhibitors in Parkinson’s disease and dementia with Lewy bodies. J Geriatr Psychiatry Neurol 17:164–171PubMedCrossRefGoogle Scholar
  2. Bessho T, Takashina K, Tabata R, Oshima C, Chaki H, Yamabe H, Egawa M, Tobe A, Saito K-I (1996) Effects of the novel high affinity choline uptake enhancer 2-(2-oxopyrrolidin-1-yl)-N-(2, 3-dimethyl-5, 6, 7, 8-tetrahydrofuro[2, 3-b]quinolin-4-yl) acetoamide on deficits of water maze learning in rats. Arzneimittelforschung 46:369–373PubMedGoogle Scholar
  3. Buyukuysal RL, Holmes T, Wurtman RJ (1991) Interactions of 3, 4-diaminopyridine and choline in stimulating acetylcholine release and protecting membrane phospholipids. Brain Research 541:1–6PubMedCrossRefGoogle Scholar
  4. Chrobak JJ, Hanin I, Schmechel TJ, Walsh TJ (1988) AF64A-induced working memory impairment: behavioral, neurochemical and histological correlates. Brain Res 463:107–117PubMedCrossRefGoogle Scholar
  5. Coleman PD, Yao PJ (2003) Synaptic slaughter in Alzheimer’s disease. Neurobiol Aging 24:1023–1027PubMedCrossRefGoogle Scholar
  6. Collerton D (1986) Cholinergic function and intellectual decline in Alzheimer’s disease. Neuroscience 19(1):1–28PubMedCrossRefGoogle Scholar
  7. De Bore P, Westerink BHC, Horn AS (1990) The effects of acetylcholine from the striatum in vivo: interaction with autoreceptor responses. Neurosci Lett 116:357–360CrossRefGoogle Scholar
  8. Fisher A, Mantione CR, Abraham DJ, Hanin I (1982) Long-term central cholinergic hypofunction induced in mice by ethylcholine aziridinium ion (AF64A) in vivo. J Pharmacol Exp Ther 222:140–145PubMedGoogle Scholar
  9. Gower AJ, Rousseau D, Jamsin P, Gobert J, Hanin I, Wulfert E (1989) Behavioral and histological effects of low concentrations of intraventricular AF64A. Eur J Pharmacol 166:271–281PubMedCrossRefGoogle Scholar
  10. Hortnagl H (1994) AF64A-induced brain damage and its relation to dementia. J Neural Transm Suppl 44:245–257PubMedGoogle Scholar
  11. Jarrard LE, Kant GJ, Meyerhoff JL, Levy A (1984) Behavioral and neurochemical effects of intraventricular AF64A administration in rats. Pharmacol Biochem Behav 21:273–80PubMedCrossRefGoogle Scholar
  12. Kasa P, Rakonczay Z, Gulya K (1997) The cholinergic system in Alzheimer’s disease. Prog Neurobiol 52:511–535PubMedCrossRefGoogle Scholar
  13. Kawashima K, Sato A, Yoshizawa M, Fujii T, Fujimoto K and Suzuki T (1994) Effects of the centrally acting cholinesterase inhibitors tetrahydroaminoacridine and E2020 on the basal concentration of extracellular acetylcholine in the hippocampus of freely moving rats. Naunyn-Schmiedeberg’s Arch Pharmacol 350: 523-528Google Scholar
  14. Kawashima K, Hayakawa T, Kashima Y, Suzuki T, Fujimoto K, Oohata H (1991) Determination of acetylcholine release in the striatum of anesthetized rats using in vivo microdialysis and radioimmunoassay. J Neurochem 57:882–887PubMedCrossRefGoogle Scholar
  15. Krištofiková Z, Fales E, Majer E, Klaschka J (1995) (3H) Hemicholinium-3 binding sites in postmortem brains of human patients with Alzheimer’s disease and multi-infarct dementia. Exp Gerontol 30:125–136PubMedCrossRefGoogle Scholar
  16. Leventer S, McKeag D, Clancy M, Wulfert E, Hanin I (1985) Intracerebroventricular administration of ethylcholine mustard aziridinium ion (AF64) reduces release of acetylcholine from rat hippocampal slices. Neuropharmacology 24:453–459PubMedCrossRefGoogle Scholar
  17. Murai S, Saito H, Abe E, Masuda Y, Odashima J, Itoh T (1994) MKC-231, a choline uptake enhancer, ameliorates working memory deficits and decreased hippocampal acetylcholine induced by ethylcholine aziridinium ion in mice. J Neural Trasm 98:1–13CrossRefGoogle Scholar
  18. Paxinos G, Watson C (1986) The rat brain in stereotaxic coordinates, 2nd ed. Academic Press, New YorkGoogle Scholar
  19. Potter PE, Nitta S (1993) Alterations in modulation of acetylcholine release following lesion of hippocampal cholinergic neurons with the neurotoxin AF64A. Neuropharmacology 32:519–526PubMedCrossRefGoogle Scholar
  20. Rodriguez-Puertas R, Pazos A, Zarranz JJ, Pascual J (1994) Selective cortical decrease of high-affinity choline uptake carrier in Alzheimer’s disease: autoradiographic study using 3H-hemicholinium-3. J Neural Transm 8:161–169CrossRefGoogle Scholar
  21. Rylett RJ, Ball MJ, Colhoun EH (1983) Evidence for high affinity choline transport in synaptosomes prepared from hippocampus and neocortex of patients with Alzheimer’s disease. Brain Res 289:169–175PubMedCrossRefGoogle Scholar
  22. Scheff SW, Price DA (2003) Synaptic pathology in Alzheimer’s disease: a review of ultrastructural studies. Neurobiol Aging 24(8):1029–1046PubMedCrossRefGoogle Scholar
  23. Schliebs R, Arendt T (2006) The significance of the cholinergic system in the brain during aging and in Alzheimer’s disease. J Neural Transm 113(1):1625–1644PubMedCrossRefGoogle Scholar
  24. Sims NR, Bowen DM, Allen SJ, Smith CCT, Neary D, Thomas DJ, Davison AN (1983) Presynaptic cholinergic dysfunction in patients with dementia. J Neurochem 40(2):503–509PubMedCrossRefGoogle Scholar
  25. Smith G (1988) Animal models of Alzheimer’s disease: experimental cholinergic denervation. Brain Research Review 13:103–118CrossRefGoogle Scholar
  26. Stip E, Chouinard S, Boulay LJ (2005) On the trail of a cognitive enhancer for the treatment of schizophrenia. Prog Neuropsychopharmacol Biol Psychiatry 29:219–232PubMedCrossRefGoogle Scholar
  27. Terry AV Jr., Buccafusco JJ (2003) The cholinergic hypothesis of age and Alzheimer’s disease-related cognitive deficits: Recent challenges and their implications for novel drug development. J Pharmacol Exp Ther 306:821–827PubMedCrossRefGoogle Scholar
  28. Ulus IH, Wurtman RJ, Mauron C, Blusztajn JK (1989) Choline increases acetylcholine release and protect against the stimulation-induced decrease in phosphatide levels within membrane of rat corpus striatum. Brain Research 484:217–227PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Ken Takashina
    • 1
    Email author
  • Tomoko Bessho
    • 1
  • Reiko Mori
    • 1
  • Junichi Eguchi
    • 1
  • Ken-Ichi Saito
    • 2
  1. 1.Pharmacology Department IV, Pharmacology Laboratory, Research Division 1000Mitsubishi Tanabe Pharma CorporationYokohamaJapan
  2. 2.Global Product Strategy DepartmentMitsubishi Tanabe Pharma CorporationTokyoJapan

Personalised recommendations