Hyperhomocysteinemia induced by methionine dietary nutritional overload modulates acetylcholinesterase activity in the rat brain
Methionine is the only endogenous precursor of homocysteine, sulfur—containing amino acid and well known as risk factor for various brain disorders. Acetylcholinesterase is a serine protease that rapidly hydrolyzes neurotransmitter acetylcholine. It is widely distributed in different brain regions. The aim of this study was to elucidate the effects of methionine nutritional overload on acetylcholinesterase activity in the rat brain. Males of Wistar rats were randomly divided into control and experimental group, fed from 30th to 60th postnatal day with standard or methionine-enriched diet (double content comparing to standard, 7.7 g/kg), respectively. On the 61st postnatal day, total homocysteine concentration was determined and showed that animals fed with methionine-enriched diet had significantly higher serum total homocysteine concentrations comparing to control rats (p < 0.01). Acetylcholinesterase activity has been determined spectrophotometrically in homogenates of the cerebral cortex, hippocampus, thalamus, and nc. caudatus. Acetylcholinesterase activity showed tendency to decrease in all examined brain structures in experimental comparing to control rats, while statistical significance of this reduction was achieved in the cerebral cortex (p < 0.05). Brain slices were stained with haematoxylin and eosin (H&E) and observed under light microscopy. Histological analysis of H&E-stained brain slices showed that there were no changes in the brain tissue of rats which were on methionine-enriched diet compared to control rats. Results of this study showed selective vulnerability of different brain regions on reduction of acetylcholinesterase activity induced by methionine-enriched diet and consecutive hyperhomocysteinemia.
KeywordsMethionine Homocysteine Acetylcholinesterase Brain Rats
This work was supported by the Ministry of Education, Science and Technological Development of Serbia (Grant #175032). We are grateful to reviewers for their constructive suggestions.
- 5.Djurić D, Jakovljević V, Rašić-Marković A, Đurić A, Stanojlović O (2008) Homocysteine, folic acid and coronary artery disease: possible impact on prognosis and therapy. Indian J Chest Di Allied Sci 50:39–48Google Scholar
- 15.Rasić-Marković A, Stanojlović O, Hrnčić D, Krstić D, Čolović M, Šusić V, Radosavljević T, Djuric D (2009) The activity of erythrocyte and brain Na+/K+ and Mg2+-ATPases in rats subjected to acute homocysteine and homocysteine thiolactone administration. Mol Cell Biochem 327:39–45PubMedCrossRefGoogle Scholar
- 20.Scherer EB, da Cunha AA, Kolling J, da Cunha MJ, Schmitz F, Sitta A, Lima DD, Delwing D, Vargas CR, Wyse AT (2011) Development of an animal model for chronic mild hyperhomocysteinemia and its response to oxidative damage. Int J Dev Neurosci 29(7):693–699. doi: 10.1016/j.ijdevneu.2011.06.004 PubMedCrossRefGoogle Scholar
- 21.Petrović M, Fufanović I, Elezović I, Čolović M, Krstić D, Jakovljević V, Đurić D (2010) The effect of homocysteine thiolactone on acetylcholinesterase activity in rat brain, blood and brain. Ser J Exp Clin Res 11(1):19–22Google Scholar
- 23.Whittaker VP, Barker LA (1972) The subcellular fractionation of brain tissue with special reference to the preparation of synaptosomes and their component organelles. Method Neurochem 2:1–52Google Scholar
- 31.Matté C, Mackedanz V, Stefanello FM, Scherer EB, Andreazza AC, Zanotto C, Moro AM, Garcia SC, Gonçalves CA, Erdtmann B, Salvador M, Wyse AT (2009) Chronic hyperhomocysteinemia alters antioxidant defenses and increases DNA damage in brain and blood of rats: protective effect of folic acid. Neurochem Int 54(1):7–13PubMedCrossRefGoogle Scholar
- 32.Schweinberger BM, Schwieder L, Scherer E, Sitta A, Vargas CR, Wyse AT (2014) Development of an animal model for gestational hypermethioninemia in rat and its effect on brain Na+, K+-ATPase/Mg2+-ATPase activity and oxidative status of the offspring. Metab Brain Dis 29(1):153–160. doi: 10.1007/s11011-013-9451-x PubMedCrossRefGoogle Scholar
- 37.Shi QS, Savage JE, Hufeisen SJ, Rauser L, Grajkowska E, Ernsberger P (2003) l-Homocysteine sulfinic acid and other acidic homocysteine derivates are potent and selective metabotropic glutamate receptor agonists. J Pharmacol Exp Ther 305:131–142. doi: 10.1124/jpet.102.047092 PubMedCrossRefGoogle Scholar
- 42.Vučević D, Petronijević N, Radonjić N, Rašić-Marković A, Mladenović D, Radosavljević T, Hrnčić D, Djuric D, Sušić V, Dj Macut, Stanojlović O (2009) Acetylcholinesterase as a potential target of acute neurotoxic effects of lindane in rats. Gen Physiol Biophys 8:18–24Google Scholar
- 43.Mikroulis AV, Psarropoulou C (2012) Endogenous ACh effects on NMDA-induced interictal-like discharges along the septotemporal hippocampal axis of adult rats and their modulation by an early life generalized seizure. Epilepsia 53(5):879–887. doi: 10.1111/j.1528-1167.2012.03440.x PubMedCrossRefGoogle Scholar
- 45.Scherer EB, Loureiro SO, Vuaden FC, da Cunha AA, Schmitz F, Kolling J, Savio LE, Bogo MR, Bonan CD, Netto CA, Wyse AT (2014) Mild hyperhomocysteinemia increases brain acetylcholinesterase and proinflammatory cytokine levels in different tissues. Mol Neurobiol. (in press)Google Scholar