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Learning behaviour and cerebral protein kinase C, antioxidant status, lipid composition in senescence-accelerated mouse: influence of a phosphatidylcholine–vitamin B12 diet

Published online by Cambridge University Press:  09 March 2007

Mei-Chu Hung
Laboratory of Nutrition Chemistry, Division of Bioresource and Bioenvironmental Sciences, Graduate School, Kyushu University, Fukuoka 812-8581, Japan
Koji Shibasaki
Laboratory of Nutrition Chemistry, Division of Bioresource and Bioenvironmental Sciences, Graduate School, Kyushu University, Fukuoka 812-8581, Japan
Riki Yoshida
Laboratory of Nutrition Chemistry, Division of Bioresource and Bioenvironmental Sciences, Graduate School, Kyushu University, Fukuoka 812-8581, Japan
Masao Sato
Laboratory of Nutrition Chemistry, Division of Bioresource and Bioenvironmental Sciences, Graduate School, Kyushu University, Fukuoka 812-8581, Japan
Katsumi Imaizumi*
Laboratory of Nutrition Chemistry, Division of Bioresource and Bioenvironmental Sciences, Graduate School, Kyushu University, Fukuoka 812-8581, Japan
*Corresponding author: Dr Katsumi Imaizumi, fax +81 092 642 3003, email
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Our objective was to determine whether dietary supplementation with phosphatidylcholine (PC) plus vitamin B12 could afford beneficial effects on biochemical and biophysical events in the brain of senescence-accelerated mouse (SAM) substrain SAMP8. We measured learning behaviour, hippocampal protein kinase C (PKC) activity, cerebral antioxidant status, phospholipid composition and fatty acid composition in 6-month-old SAMP8 and in age-matched controls (SAM substrain SAMR1). In comparison with SAMR1, SAMP8 showed a significant elevation in total grading score of senescence (P<0·05) and a significant decline in acquisition (P<0·05). SAMP8 had a lower hippocampal PKC activity and cerebral PKC-β mRNA abundance than SAMR1. SAMP8 had increased cerebral lipid peroxide levels and proportion of sphingomyelin, and a lower proportion of 20 : 4n-6 and 22 : 6n-3 in cerebral phosphtidylethanolamine than SAMR1. SAMP8 fed the PC combined with vitamin B12 diet had an increased PKC activity and a higher proportion of 22 : 6n-3 than SAMP8 fed the control diet. These results indicate the potential benefit of PC combined with vitamin B12 as a dietary supplement.

Research Article
Copyright © The Nutrition Society 2001


Armbrecht, HJ, Boltz, MA, Kumar, VB, Flood, JF & Morley, JE (1999) Effect of age on calcium-dependent proteins in hippocampus of senescence-accelerated mice. Brain Research 842, 287293.Google Scholar
Battaini, F, Del Vesco, R, Govoni, S & Trabucchi, M (1990) Regulation of phorbol ester binding and protein kinase C activity in aged rat brain. Neurobiology and Aging 11, 563566.Google Scholar
Bliss, TVP & Collingridge, GL (1993) A synaptic model of memory: long-term potentiation in the hippocampus. Nature 361, 3139.Google Scholar
Chomczynski, P & Sacchi, N (1987) Single-step method of RNA isolation by acid guanidium thiocyanate–phenol–chloroform extraction. Analytical Biochemistry 162, 156159.Google Scholar
Chung, SY, Moriyama, T, Uezu, E, Uezu, K, Hirata, R, Yohena, N, Masuda, Y, Kokubu, T & Yamamoto, S (1995) Administration of phosphatidylcholine increases brain acetylcholine concentration and improves memory in mice with dementia. Journal of Nutrition 125, 14841489.Google Scholar
Colley, PA & Routtenberg, A (1993) Long term potentiation as synaptic dialogue. Brain Research Review 18, 115122.Google Scholar
Delion, S, Chalon, S, Guilloteau, D, Lejeune, B, Besnard, J-C & Durand, G (1997) Age-related changes in phospholipid fatty acid composition and monoaminergic neurotransmission in the hippocampus of rats fed a balanced or an n-3 polyunsaturated fatty acid-deficient diet. Journal of Lipid Research 38, 680689.Google Scholar
Denisova, NA, Strain, JG & Joseph, JA (1997) Oxidant injury in PC12 cells – A possible model of calcium dysregulation in aging. 2. Interactions with membrane lipids. Journal of Neurochemistry 69, 12591266.CrossRefGoogle Scholar
Epand, RM & Lester, DS (1990) The role of membrane biophysical properties in the regulation of protein kinase C activity. Trends in Pharmaceutical Science 11, 317320.Google Scholar
Folch, J, Lees, M & Sloane-Stanley, GH (1957) A simple method for the isolation and purification of total lipids from animal tissues. Journal of Biological Chemistry 226, 497509.Google Scholar
Freire-Moar, J, Cherwinski, H, Hwang, F, Ransom, J & Webb, D (1991) Expression of protein kinase C isoenzymes in thymocyte subpopulations and their differential regulation. Journal of Immunology 147, 405409.Google Scholar
Giorgione, J, Epand, RM, Buda, C & Farkas, T (1995) Role of phospholipids containing docosahexaenoyl chains in modulating the activity of protein kinase C. Proceedings of the National Academy of Sciences USA 92, 97679770.CrossRefGoogle ScholarPubMed
Harman, D (1981) The aging process. Proceedings of the National Academy of Sciences USA 78, 71247128.Google Scholar
Hirsch, MJ & Wurtman, RJ (1978) Lecithin consumption increases acetylcholine concentrations in rat brain and adrenal gland. Science 202, 223224.Google Scholar
Hissin, PJ & Hilf, R (1976) A fluorometric method for determination of oxidized and reduced glutathione in tissues. Analytical Biochemisry 74, 214226.Google Scholar
Hosokawa, M, Kasai, R, Higuchi, K, Takeshita, S, Shimizu, K, Hamamoto, H, Honoma, A & Irino, M (1984) Grading score system: a method for evaluation of the degree of senescence in senescence accelerated mouse (SAM). Mechanisms of Ageing and Development 26, 91102.CrossRefGoogle ScholarPubMed
Hung, MC, Shibasaki, K, Nishizono, S, Sato, M, Ikeda, I, Masuda, Y, Kunou, M, Kawamura, M, Yamashita, M, Inoue, S & Imaizumi, K (2000) Ibotenic acid-induced lesions of medial septum increase hippocampal membrane associated protein kinase C activity and reduce acetylcholine synthesis: prevention by phosphatidylcholine/vitamin B12 diet. Journal of Nutritional Biochemistry 11, 159164.CrossRefGoogle ScholarPubMed
Ikeda, I, Yoshida, H, Tomooka, M, Yosef, A, Imaizumi, K, Tsuji, H & Seto, A (1998) Effects of long-term feeding of marine oils with different positional distribution of eicosapentaenoic and docosahexaenoic acids on lipid metabolism, eicosanoid production, and platelet aggregation in hypercholesterolemic rats. Lipid 33, 897904.CrossRefGoogle ScholarPubMed
Jarvik, ME & Kopp, R (1967) An improved one-trial passive avoidance learning situation. Psychological Reports 21, 221224.Google Scholar
Jones, CR, Arai, T & Rapoport, SI (1997) Evidence for the involvement of docosahexaenoic acid in cholinergic stimulated signal transduction at the synapse. Neurochemical Research 22, 663670.Google Scholar
Jope, RS (1982) Effects of phosphatidylcholine administration to rats on choline in blood and choline and acetylcholine in brain. Journal of Pharmacology and Experimental Therapeutics 220, 322328.Google ScholarPubMed
Kessler, AR & Yehuda, S (1985) Learning-induced changes in brain membrane cholesterol and fluidity: implications for brain aging. International Journal of Neuroscience 28, 7382.Google Scholar
Kimura, S, Minami, M, Endo, T, Hirafuji, M, Monma, Y, Togashi, H, Saito, H & Parvez, SH (1998) Methylcobalamine (V-B-12) increases cerebral acetylcholine levels and improves passive avoidance response in stroke-prone spontaneously hypertensive rats. Biogenic Amines 14, 1524.Google Scholar
Lawrence, RA & Burk, RF (1976) Glutathione peroxidase activity in selenium-deficient rat liver. Biochemical and Biophysical Research Communications 71, 952958.Google Scholar
Leathwood, PD, Heck, E & Mauron, J (1982) Phosphatidylcholine and avoidance performance in 17 month-old SEC/1ReJ mice. Life Science 30, 10651071.Google Scholar
Loy, R, Heyer, D, Williams, CL & Meck, WH (1991) Choline-induced spatial memory facilitation correlates with altered distribution and morphology of septal neurons. Advances in Experimental Medicine and Biology 295, 373382.Google Scholar
Magil, SG, Zeisel, SH & Wurtman, RJ (1981) Effects of ingesting soy or egg lecithins on serum choline, brain choline and brain acetylcholine. Journal of Nutrition 111, 166170.Google Scholar
Masuda, Y, Kokubu, T, Yamashita, M, Ikeda, H & Inoue, S (1998) Egg phosphatidylcholine combined with vitamin B12 improved memory impairment following lesioning of nucleus basalis in rats. Life Sciences 62, 813822.CrossRefGoogle ScholarPubMed
Matsugo, S, Kitagawa, T, Minami, S, Esashi, Y, Oomura, Y, Tokumaru, S, Matsushima, K & Sasaki, K (2000) Age-dependent changes in lipid peroxide levels in peripheral organs, but not in brain, in senescence-accelerated mice. Neuroscience Letters 278, 105108.Google Scholar
Matsukawa, N, Tooyama, I, Kimura, H, Yamamoto, T, Tsugu, Y, Oomura, Y & Ojika, K (1999) Increased expression of hippocampal cholinergic neurostimulating peptide-related components and their messenger RNAs in the hippocampus of aged senescence-accelerated mice. Neuroscience 88, 7992.Google Scholar
Matsumoto, Y, Kaneyuki, T, Moriyama, T, Uezu, K, Chung, SY, Uezu, E, Masuda, N, Kokubu, T & Yamamoto, S (1994) Effect of phoshatidylcholine administration on learning and memory and brain acetylcholine level in SAM. In The SAM Model of Senescence. International Congress Series 1062, pp. 423426 [Takeda, T, editor]. Amsterdam, London, New York and Tokyo: Excerpta Medica.Google Scholar
Misra, HP & Fridovich, I (1977) Superoxide dismutase: a photochemical augmentation assay. Archives of Biochemistry and Biophysics 181, 308312.Google Scholar
Nadeau, A & Roberge, AG (1988) Effects of Vitamin B12 supplementation on choline acetyltransferase activity in cat brain. International Journal for Vitamin and Nutrition Research 58, 402406.Google Scholar
Nishizaki, T, Nomura, T, Matsuoka, T, Enikolopov, G & Sumikawa, K (1999) Arachidonic acid induces a long-lasting facilitation of hippocampal synaptic transmission by modulating PKC activity and nicotinic Ach receptors. Brain Research 69, 263272.Google Scholar
Nourhashemi, F, Gillette-Guyonnet, S, Andrieu, S, Ghisolfi, A, Ousset, PJ, Grandjean, H, Grand, A, Pous, J, Vellas, B & Albarede, JL (2000) Alzheimer disease: protective factors. American Journal of Clinical Nutrition 71, 643S649S.Google Scholar
Ohkawa, H, Ohishi, N & Yagi, K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Analytical Biochemistry 95, 351358.Google Scholar
Reeves, PG, Nielsen, FH & Fahey, GC (1993) AIN-93 purified diets for laboratory rodents: final report of the American Institute of Nutrition Ad Hoc Writing Committee on the reformulation of the AIN-76A rodent diet. Journal of Nutrition 123, 19391951.Google Scholar
Rouser, G, Siakotos, AN & Fleischer, S (1966) Quantitative analysis of phospholipids by thin-layer chromatography and phosphorus analysis of spots. Lipids 1, 8586.Google Scholar
Sacktor, TC, Osten, P, Valsamis, H, Jiang, X, Naik, MU & Sublette, E (1993) Persistent activation of the δ isoform of protein kinase C in the maintenance of long-term potentiation. Proceedings of the National Academy of Sciences USA 90, 83428346.CrossRefGoogle Scholar
Sasaki, H, Matsuzaki, Y, Meguro, K, Igarashi, Y, Maruyama, Y, Yamaguchi, S & Sekizawa, K (1992) Vitamin B12 improves cognitive disturbance in rodents fed a choline-deficient diet. Pharmacology Biochemistry and Behavior 43, 635639.Google Scholar
Sato, E, Oda, N, Ozaki, N, Hashimoto, SI, Kurokawa, T & Ishibashi, S (1996) Early and transient increase in oxidative stress in the cerebral cortex of senescence-accelerated mouse. Mechanisms of Aging and Development 86, 105114.Google Scholar
Shearman, MS, Shinomura, T, Oda, T & Nishizuka, Y (1991) Synaptosomal protein kinase C subspecies: A dynamic change in the hippocampus and cerebral cortex concomitant with synaptogenesis. Journal of Neurochemistry 56, 12551262.Google Scholar
Sokolov, BP & Prockop, DJ (1994) A rapid and simple PCR-based method for isolation of cDNAs from differentially expressed genes. Nucleic Acid Research 22, 40094015.Google Scholar
Takeda, T, Hosokawa, M, Takeshita, S, Irino, M, Higuchi, K, Matsushita, T, Tomita, Y, Yasuhira, K, Hamamoto, H, Shimizu, K, Ishii, M & Yamamuro, T (1981) A new murine model of accelerated senescence. Mechanisms of Aging and Development 17, 183194.Google Scholar
Tokunaga, K, Taniguchi, H, Yoda, M, Shimizu, M & Sakiyama, S (1986) Nucleotide sequence of a full-length cDNA for mouse cytoskeletal β-actin mRNA. Nucleic Acids Research 14, 28292831.Google Scholar
Vanderzee, EA, Luiten, PGM & Disterhoft, JF (1997) Learning-induced alterations in hippocampal PKC-immunoreactivity and hypothesis of its functional significance. Progress in Neuro-Psychopharmacology and Biological Psychiatry 21, 531572.Google Scholar
Wree, A, Erselius, R, Tønder, N & Beck, T (1993) Time course of hippocampal glucose utilization and persistence of parvalbumin immunoreactive neurons after ibotenic acid-induced lesions of the rat dental area. Journal of Cerebral Blood Flow and Metabolism 13, 9981005.CrossRefGoogle Scholar
Zeisel, SH & Blusztajn, JK (1994) Choline and human nutrition. Annual Review of Nutrition 14, 269296.Google Scholar