Abstract
Homocystinuria is due to enzymatic deficiencies resulting in elevated blood levels of homocysteine (Hcy), homocystine (Hci), and/or methionine (Met) and the clinical presentation of mental retardation, seizures, and cardiovascular disease. Since these symptoms may be closely implicated with acetylcholinesterase (AChE) activity, we aimed to investigate whether this metabolic disorder affects the hippocampal AChE activity in 21 days suckling Wistar rat hippocampus. Various concentrations of Hcy, Hci (0.05–0.5 mM), or Met (0.05–2 mM) as well as Mixture A (Mix A) (0.3 mM (Hcy)+0.2 mM (Hci)+1.0 mM (Met) = in vitro cystathionine β-synthase deficiency homocystinuria), Mix B1 (Hcy 0.3 mM + Hci 0.2 mM=in vitro severe methylenetetrahydrofolate reductase deficiency homocystinuria) or Mix B2 (Hcy 0.1 mM+Hci 0.05 mM=in vitro mild methylenetetrahydrofolate reductase deficiency homocystinuria) were preincubated with homogenized hippocampii or with eel Electrophorus electricus pure AChE. AChE was evaluated spectrophotometrically. Hcy or Met stimulated hippocampal AChE by 50% (p < 0.001) at low concentrations of the amino acids (up to 0.3–0.5 mM), whereas Hci inhibited the enzyme by 40% (p < 0.001). Mix A, Mix B1, or Mix B2 activated hippocampal AChE by 40, 30, (p < 0.001), and 12% (p < 0.01), respectively. In contrast, the S-containing amino acids, Mix A, Mix B1, Mix B2 failed to affect the pure AChE activity. Conclusions: a) The presence of −SH group in Hcy and Met may result in hippocampal AChE stimulation and the redox isomer Hci in the inhibition of the enzyme, probably by producing free radicals, and b) The SH-amino acids seem to affect the hippocampal enzyme indirectly, possibly by lipid(s)-protein modifications(s) and Hci by inducing oxidative stress, since no effect was observed on pure AChE activity.
Similar content being viewed by others
References
Allen IC, Grieve A, Griffiths R (1986) Differential changes in the content of amino acid neurotransmitters in discrete regions of the rat brain prior to the onset and during the course of homocysteine-induced seizures. J Neurochem 46:1582–1592
Ballantyne BM, Marro TC (1992) In: Clinical and experimental toxicology of organophosphates and carbamates. Butterworth-Heinemann, Oxford, pp 641–650
Charlton CG, Crowell B Jr (1995) Striatal dopamine, depletion, tremors, and hypokinesia following the intracranial injection of S-adenosylmethionine. A possible role of hypermethylation in Parkinsonism. Mol Chem Neuropathol 26:269–284
Clarke R, Daly L, Robinson K, Naughten E, Cahalane S, Fowler B, Graham I (1991) Hyperhomocysteinemia: an independent risk factor for vascular disease. N Engl J Med 324:1149–1155
Committee on Care and Use of Laboratory Animals (1985) Guide for the care and use of laboratory animals. Institute of Laboratory Animal Resources, National Research Council, Washington, DC, pp 83
Dewhurst IC, Hagan JJ, Morris RG, Griffiths R (1983) Hippocampal electrical activity and gamma-aminobutyrate metabolism in brain tissue following administration of homocysteine. J Neurochem 40:752–757
Ellman GL, Courtney D, Andres V, Featherstone RM (1961) A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–95
Finkelstein JD, Martin JJ, Harris BJ (1988) Methionine metabolism in mammals. The methionine-sparing effect of cystine. J Biol Chem 263:11750–11754
Kouniniotou-Krontiri P, Tsakiris S, Hadjigeorgiou G (1994) Involvement of sulfhydryl groups in time dependent changes of diaphragm acetylcholinesterase activity by monovalent (Na+, Li+) cations. Biochem Mol Bio Int 33:485–496
Lehmann A, Isacsson H, Hamberger A (1983) Effects of in vivo administration of kainic acid on the extracellular amino acid pool in the rabbit hippocampus. J Neurochem 40:1314–1320
Lotti M (1995) Cholinesterase inhibition: complexities in interpretation. Clin Chem 41:1814–1818
Lou MF, Poulsen LL, Ziegler DM (1987) Cellular protein mixed disulfides In: Methods in enzymology, JacobyWB, Griffin OWE(eds), vol 143, Academic, Orlando FL, pp 124–129
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275
McCully KS (1993) Chemical pathology of homocysteine I Atherogenesis. Ann Clin Lab Sci 23:447–493
McCully KS (1997) Pathology of homocystinuria. In: Homocysteine metabolism.: From Basic Science to Clinical Medicine, Kluwer Academic,NY pp 251–257
Meyer EM, Cooper JR (1981) Correlations between Na+,K+-ATPase activity and acetylcholine release in rat cortical synaptosomes. J Neurochem 36:467–475
Mudd SH, Levy H, Skovby F (2000) In: ScriverCR, Beadet AL, Sly WS, Valle D (eds), Disorders of transsulfation: In the metabolic basis of inherited diseases, pp 488–499. McGraw-Hill, New York
Olszewski AJ, McCully KS (1993) Homocysteine metabolism and the oxidative modification of proteins and lipids. Free Radic Biol Med 14:683–693
Pinardi G, Pelissier T, Kramer V, Paeile C, Miranda HF (1994) Effects of CDP-choline on acetylcholine-induced relaxation of the perfused carotid vascular beds of the rat. Gen Pharmacol 25:635–638
Plataras C, Angelogianni P, Tsakiris S (2003) Effect of CDP-choline on hippocampal acetylcholinesterase and Na+,K+-ATPase in adult and aged rats. Z Naturforsch 58c:277–281
Stipanuk MH (1986) Metabolism of sulfur-containing amino acids. Ann Rev Nutr 6:179–209
Streck EL, Zugno AI, Tagliari B, Sarkis JJ, Wajner M, Wannmacher CM, Wyse AT (2002) On the mechanism of the inhibition of Na+, K+-ATPase activity caused by homocysteine. Int J Dev Neurosci 20:77–81
Sussman JL, Harel M, Frolow F, Oefner C, Goldman A, Toker L, Silman I (1991) Atomic structure of acetylcholinesterase from Torpedo californica: A prototypic acetylcholine-binding protein. Science 253:872–879
Tsakiris S, Angelogianni P, Schulpis KH, Stavridis J (2000) Protective effect of L-phenylalanine on rat brain acetylcholinesterase inhibition induced by free radicals. Clin Biochem 33:103–106
Ungvari Z, Csiszar A, Edwards JG, Kaminski PM, Wollin MS, Kaley G, Koller A (2003) Increased superoxide production in coronary arteries in hyperhomocysteinemia: role of tumor necrosis factor-alpha, NAD(P)H oxidase and inducible nitric oxide synthase. Arterioscler Thromb Vasc Biol 23:418–424
Acknowledgments
This study was funded by the University of Athens. We thank Dr. Filia Stratigea (veterinary surgeon) and Mrs. Anna Stamatis for their significant assistance.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Schulpis, K.H., Kalimeris, K., Bakogiannis, C. et al. The Effect of in Vitro Homocystinuria on the Suckling Rat Hippocampal Acetylcholinesterase. Metab Brain Dis 21, 20–27 (2006). https://doi.org/10.1007/s11011-006-9001-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11011-006-9001-x