, Volume 34, Issue 4, pp 917–933 | Cite as

Late-onset intermittent fasting dietary restriction as a potential intervention to retard age-associated brain function impairments in male rats

  • Rumani Singh
  • Dinesh Lakhanpal
  • Sushil Kumar
  • Sandeep Sharma
  • Hardeep Kataria
  • Manpreet Kaur
  • Gurcharan KaurEmail author


Lifelong dietary restriction (DR) is known to have many potential beneficial effects on brain function as well as delaying the onset of neurological diseases. In the present investigation, the effect of late-onset short-term intermittent fasting dietary restriction (IF-DR) regimen was studied on motor coordination and cognitive ability of ageing male rats. These animals were further used to estimate protein carbonyl content and mitochondrial complex I–IV activity in different regions of brain and peripheral organs, and the degree of age-related impairment and reversion by late-onset short-term IF-DR was compared with their levels in 3-month-old young rats. The results of improvement in motor coordination by rotarod test and cognitive skills by Morris water maze in IF-DR rats were found to be positively correlated with the decline in the oxidative molecular damage to proteins and enhanced mitochondrial complex IV activity in different regions of ageing brain as well as peripheral organs. The work was further extended to study the expression of synaptic plasticity-related proteins, such as synaptophysin, calcineurin and CaM kinase II to explore the molecular basis of IF-DR regimen to improve cognitive function. These results suggest that even late-onset short-term IF-DR regimen have the potential to retard age-associated detrimental effects, such as cognitive and motor performance as well as oxidative molecular damage to proteins.


Intermittent fasting–dietary restriction (IF-DR) Ageing Synaptic plasticity Mitochondrial electron transport chain (ETC) Morris water maze (MWM) Protein carbonyl content 



This grant was funded by Indian Council of Medical Research (ICMR) under the National Task Force Project—an initiative on ageing research. Rumani Singh and Sandeep Sharma are thankful to ICMR for the research fellowship grant during entire course of study.


  1. Adams MM, Shi L, Linville MC, Forbes ME, Long AB, Bennett C, Newton IG, Carter CS, Sonntag WE, Riddle D, Brunso-Bechtold JK (2008) Caloric restriction and age affect synaptic proteins in hippocampal CA3 and spatial learning ability. Exp Neurol 211:141–149PubMedCrossRefGoogle Scholar
  2. Ahmet I, Wan R, Mattson MP, Lakatta EG, Talan M (2005) Cardioprotection by intermittent fasting in rats. Circulation 112(20):3115–3121PubMedCrossRefGoogle Scholar
  3. Aksenov V, Long J, Lokuge S, Foster JA, Liu J, Rollo CD (2010) Dietary amelioration of locomotor, neurotransmitter and mitochondrial aging. Exp Biol Med 235:66–76CrossRefGoogle Scholar
  4. Albers DS, Beal MF (2000) Mitochondrial dysfunction and oxidative stress in aging and neurodegenerative disease. J Neural Transm Suppl 59:133–154PubMedGoogle Scholar
  5. Altun M, Bergman E, Edström E, Johnson H, Ulfhake B (2007) Behavioral impairments of the aging rat. Physiol Behav 92(5):911–923PubMedCrossRefGoogle Scholar
  6. Anson RM, Guo Z, de Cabo R, Iyun T, Rios M, Hagepanos A, Ingram DK, Lane MA, Mattson MP (2003) Intermittent fasting dissociates beneficial effects of dietary restriction on glucose metabolism and neuronal resistance to injury from calorie intake. Proc Natl Acad Sci U S A 100(10):6216–6220PubMedCrossRefGoogle Scholar
  7. Ayala V, Naudı A, Sanz A, Caro P, Portero-Otin M, Barja G, Pamplona R (2007) Dietary protein restriction decreases oxidative protein damage, peroxidizability index, and mitochondrial complex I content in rat liver. J Gerontol A Biol sci Med Sci 62:352–360PubMedCrossRefGoogle Scholar
  8. Calabrese EJ, Mattson MP (2011) Hormesis provides a generalized quantitative estimate of biological plasticity. J Cell Commun Signal 5(1):25–38PubMedCrossRefGoogle Scholar
  9. Cambon K, Hansen SM, Venero C, Herrero AI, Skibo G, Berezin V, Bock E, Sandi C (2004) A synthetic neural cell adhesion molecule mimetic peptide pomotes synaptogenesis, enhances presynaptic function and facilitates memory consolidation. J Neurosci 24:4197–4204PubMedCrossRefGoogle Scholar
  10. Carter SC, Leeuwenburgh C, Daniels M, Foster CT (2009) Influence of calorie restriction on measures of age related cognitive decline: role of increased physical activity. J Gerontol A Biol Sci Med Sci 64:850–859PubMedCrossRefGoogle Scholar
  11. Chaudhuri AR, Waal E, Pierce A, Remmen VA, Ward FW, Richardson F (2006) Detection of protein carbonyls in aging liver tissue: a fluorescence-based proteomic approach. Mech Ageing Dev 127:849–861PubMedCrossRefGoogle Scholar
  12. Chevion M, Berenshtein E, Stadtman ER (2000) Human studies related to protein oxidation: protein carbonyl content as a marker of damage. Free Radic Res 33:S99–S108PubMedGoogle Scholar
  13. Cremer H, Genevieve C, Carleton A, Gordis C, Vincent JD, Lledo PM (1998) Long term but not short term plasticity at mossy fiber synapses is impaired in neural cell adhesion molecule deficient mice. Proc Natl Acad Sci USA 95:13242–13247PubMedCrossRefGoogle Scholar
  14. Davies AH, Kelly A, Dhanrajan TM, Lynch MA, Rodrıguez JJ, Stewart GM (2003) Synaptophysin immunogold labelling of synapses decreases in dentate gyrus of the hippocampus of aged rats. Brain Res 986:191–195PubMedCrossRefGoogle Scholar
  15. Dineley KT, Hogan D, Zhang WR, Taglialatela G (2007) Acute inhibition of calcineurin restores associative learning and memory in Tg2576 APP transgenic mice. Neurobiol Learn Mem 88:217–224PubMedCrossRefGoogle Scholar
  16. Dityatev A, Dityateva G, Schachner M (2000) Synaptic strength as a function of post versus presynaptic expression of the neural cell adhesion molecule NCAM. Neuron 26:207–217PubMedCrossRefGoogle Scholar
  17. Djordjevic MA, Perovic M, Tesic V, Tanic N, Rakic L, Ruzdijic S, Kanazir S (2010) Long-term dietary restriction modulates the level of presynaptic proteins in the cortex and hippocampus of the aging rat. Neurochem Int 56(2):250–255CrossRefGoogle Scholar
  18. Drapeau E, Mayo W, Aurousseau C, Le Moal M, Piazza PV, Abrous DN (2003) Spatial memory performances of aged rats in the water maze predict levels of hippocampal neurogenesis. Proc Natl Acad Sci U S A 100:14385–14390PubMedCrossRefGoogle Scholar
  19. Feuers RJ (1998) The effects of dietary restriction on mitochondrial dysfunction in aging. Ann N Y Acad Sci 854:192–201PubMedCrossRefGoogle Scholar
  20. Filburn CR, Edris W, Tamatani M, Hogue B, Kudryashova I, Hansford RG (1996) Mitochondrial electron transport chain activities and DNA deletions in regions of the rat brain. Mech Ageing Dev 87(1):35–46PubMedCrossRefGoogle Scholar
  21. Forster MJ, Dubey A, Dawson KM, Stutts WA, Lal H, Sohal RS (1996) Age-related losses of cognitive function and motor skills in mice are associated with oxidative protein damage in the brain. Proc Natl Acad Sci USA 93:4765–4769PubMedCrossRefGoogle Scholar
  22. Forster MJ, Sohal BH, Sohal RS (2000) Reversible effects of long-term caloric restriction on protein oxidative damage. J Gerontol A Biol Sci Med Sci 55:B522–B529PubMedCrossRefGoogle Scholar
  23. Giese KP, Fedorov NB, Filipkowski RK, Silva AJ (1998) Autophosphorylation at Thr286 of the alpha calcium-calmodulin kinase II in LTP and learning. Science 279:870–873PubMedCrossRefGoogle Scholar
  24. Goto S (2006) Health span extension by later-life caloric or dietary restriction: a view based on rodent studies. Biogerontology 7:135–138PubMedCrossRefGoogle Scholar
  25. Goto S, Takahashi R, Radak Z, Sharma R (2007) Beneficial biochemical outcomes of late-onset dietary restriction in rodents. Ann N Y Acad Sci 1100:431–441PubMedCrossRefGoogle Scholar
  26. Gredilla R, Barja G, Lopez-Torres M (2001a) Effect of short-term caloric restriction on H2O2 production and oxidative DNA damage in rat liver mitochondria and location of the free radical source. J Bioenerg Biomembr 33:279–287PubMedCrossRefGoogle Scholar
  27. Gredilla R, Sanz A, Lopez-Torres M, Barja G (2001b) Calorie restriction decreases mitochondrial free radical generation at complex I and lowers oxidative damage to mitochondrial DNA in the rat heart. FASEB J 15:U481–U496Google Scholar
  28. Griffths D (1989) Clarification and extraction. In: Harris ELV, Angal S (eds) Protein purification methods a practical approach. IRL, Oxford, pp 91–97Google Scholar
  29. Grune T, Shringarpure R, Sitte N, Davies K (2001) Age-related changes in protein oxidation and proteolysis in mammalian cells. J Gerontol A Biol Sci Med Sci 56A:B459–B467CrossRefGoogle Scholar
  30. Hamadeh MJ, Rodriguez MC, Kaczor JJ, Tarnopolsky MA (2005) Caloric restriction transiently improves motor performance but hastens clinical onset of disease in the Cu/Zn-superoxide dismutase mutant G93A mouse. Muscle Nerve 31:214–220PubMedCrossRefGoogle Scholar
  31. Harvie MN, Pegington M, Mattson MP, Frystyk J, Dillon B, Evans G, Cuzick J, Jebb SA, Martin B, Cutler RG, Son TG, Maudsley S, Carlson OD, Egan JM, Flyvbjerg A, Howell A (2011) The effects of intermittent or continuous energy restriction on weight loss and metabolic disease risk markers: a randomized trial in young overweight women. Int J Obes 35(5):714–727CrossRefGoogle Scholar
  32. Hashimoto T, Watanabe S (2005) Chronic food restriction enhances memory in mice—analysis with matched drive levels. Neuroreport 16:1129–1133PubMedCrossRefGoogle Scholar
  33. Hatefi Y, Rieske JS (1967) The preparation and properties of DPNH-cytochrome C reductase (Complex I of respiratory chain). In: Estabrook RW, Pullman ME (eds) Methods in enzymology, vol 10. Academic, New York, pp 235–239Google Scholar
  34. Hsu KS, Huang CC, Liang YC, Wu HM, Chen YL, Lo SW, Ho WC (2002) Alterations in the balance of protein kinase and phosphatase activities and age-related impairments of synaptic transmission and long-term potentiation. Hippocampus 12:787–802PubMedCrossRefGoogle Scholar
  35. Hursting SD, Smith SM, Lashinger LM, Harvey AE, Perkins SN (2009) Calories and carcinogenesis: lessons learned from 30 years of calorie restriction research. Carcinogenesis 31:83–89PubMedCrossRefGoogle Scholar
  36. Jabr RI, Wilson JA, Riddervold M, Jenkins AH, Perrino BA, Clapp LH (2007) Nuclear translocation of calcineurin Aα but not calcineurin Aβ by platelet-derived growth factor in rat aortic smooth muscle. Am J Physiol Cell Physiol 292:C2213–C2225PubMedCrossRefGoogle Scholar
  37. Jouvenceau A, Dutar P (2006) A role for the protein phosphatase 2B in altered hippocampal synaptic plasticity in the aged rat. J Physiol Paris 99:154–161PubMedCrossRefGoogle Scholar
  38. Kaur M, Sharma S, Kaur G (2008) Age-related impairments in neuronal plasticity markers and astrocytic GFAP and their reversal by late-onset short term dietary restriction. Biogerontology 9:441–454PubMedCrossRefGoogle Scholar
  39. Keenan KP, Coleman JB, McCoy CL, Hoe CM, Soper KA, Laroque P (2000) Chronic nephropathy in ad libitum overfed Sprague–Dawley rats and its early attenuation by increasing degrees of dietary (caloric) restriction to control growth. Toxicol Pathol 28:788–798PubMedCrossRefGoogle Scholar
  40. Keller JN, Schmitt FA, Scheff SW, Ding Q, Chen Q, Butterfield DA, Markesbery WR (2005) Evidence of increased oxidative damage in subjects with mild cognitive impairment. Neurology 64:1152–1156PubMedCrossRefGoogle Scholar
  41. King DL, Arendash GW (2002) Maintained synaptophysin immunoreactivity in Tg2576 transgenic mice during aging: correlations with cognitive impairment. Brain Res 926:58–68PubMedCrossRefGoogle Scholar
  42. Klee CB, Ren H, Wang X (1998) Regulation of the calmodulin-stimulated protein phosphatase, calcineurin. J Biol Chem 273:13367–13370PubMedCrossRefGoogle Scholar
  43. Kumar S, Parkash J, Kataria H, Kaur G (2010) Interactive effect of excitotoxic injury and dietary restriction on neurogenesis and neurotrophic factors in adult male rat brain. Neurosci Res 65:367–374CrossRefGoogle Scholar
  44. Lemaire V, Koehl M, Le Moal M, Abrous DN (2000) Prenatal stress produces learning deficits associated with an inhibition of neurogenesis in the hippocampus. Proc Natl Acad Sci U S A 97:11032–11037PubMedCrossRefGoogle Scholar
  45. Lesne S, Koh MT, Kotilinek L, Kayed R (2006) A specific amyloid-beta protein assembly in the brain impairs memory. Nature 440:352–357PubMedCrossRefGoogle Scholar
  46. Levine RL, Garland D, Oliver CN et al (1990) Determination of carbonyl content in oxidatively modified proteins. Methods Enzymol 186:464–478PubMedCrossRefGoogle Scholar
  47. Lin SJ, Defossez PA, Guarente L (2000) Requirement of NAD and SIR2 for life-span extension by calorie restriction in Saccharomyces cerevisiae. Science 289:2126–2128PubMedCrossRefGoogle Scholar
  48. Liu Y, Fiskum G, Schubert D (2002) Generation of reactive oxygen species by the mitochondrial electron transport chain. J Neurochem 80:780–787PubMedCrossRefGoogle Scholar
  49. Liu HX, Zhang JJ, Zheng P, Zhang Y (2005) Altered expression of MAP-2, GAP-43, and synaptophysin in the hippocampus of rats with chronic cerebral hypoperfusion correlates with cognitive impairment. Brain Res Mol Brain Res 139:169–177PubMedCrossRefGoogle Scholar
  50. Lopez-Torres M, Gredilla R, Sanz A, Barja G (2002) Influence of aging and long-term caloric restriction on oxygen radical generation and oxidative DNA damage in rat liver mitochondria. Free Rad Biol and Med 32:882–889CrossRefGoogle Scholar
  51. Love R (2005) Calorie restriction may be neuroprotective in AD and PD. Lancet Neurol 4:84PubMedCrossRefGoogle Scholar
  52. Mansuy MI (2003) Calcineurin in memory and bidirectional plasticity. Biochem Biophys Res Commun 311:1195–1208PubMedCrossRefGoogle Scholar
  53. Markowska AL, Savonenko A (2002) Retardation of cognitive aging by life-long diet restriction: implications for genetic variance. Neurobiol Aging 23:75–86PubMedCrossRefGoogle Scholar
  54. Martin B, Mattson MP, Maudsley S (2006) Caloric restriction and intermittent fasting: two potential diets for successful brain aging. Ageing Res Rev 5(3):332–353PubMedCrossRefGoogle Scholar
  55. Maswood N, Young J, Tilmont E, Zhang Z, Gash DM, Gerhardt GA, Grondin R, Roth GS, Mattison J, Lane MA, Carson RE, Cohen RM, Mouton PR, Quigley C, Mattson MP, Ingram DK (2004) Caloric restriction increases neurotrophic factor levels and attenuates neurochemical and behavioral deficits in a primate model of Parkinson's disease. Proc Natl Acad Sci USA 101:17887–17888CrossRefGoogle Scholar
  56. Mattson MP (2003) Gene–diet interactions in brain aging and neurodegenerative disorders. Ann Intern Med 139:441–444PubMedGoogle Scholar
  57. Mattson MP (2008) Dietary factors, hormesis and health. Ageing Res Rev 7(1):43–48PubMedCrossRefGoogle Scholar
  58. Mattson MP (2010) The impact of dietary energy intake on cognitive aging. Front Aging Neurosci 2:1–12Google Scholar
  59. Mattson MP, Pedersen WA, Duan W, Culmsee C, Camandola S (1999) Cellular and molecular mechanisms underlying perturbed energy metabolism and neuronal degeneration in Alzheimer's and Parkinson's diseases. Ann N Y Acad Sci 893:154–175PubMedCrossRefGoogle Scholar
  60. Monville C, Torres ME, Dunnett BS (2006) Comparison of incremental and accelerating protocols of the rotarod test for the assessment of motor deficits in the 6-OHDA model. J Neurosci Methods 158:219–223PubMedCrossRefGoogle Scholar
  61. Navarro A, Gomez C, Lopez-Cepero MJ, Boveris A (2004) Beneficial effects of moderate exercise on mice aging: survival, behavior, oxidative stress and mitochondrial electron transfer. Am J Physiol Regul Integr Comp Physiol 286:R505–R511PubMedCrossRefGoogle Scholar
  62. Navarro A, Gomez C, Sanchez-Pino MJ, Gonzalez H, Bandez MJ, Boveris AD, Boveris A (2005) Vitamin E at high doses improves survival, neurological performance, and brain mitochondrial function in aging male mice. Am J Physiol Regul Integr Comp Physiol 289:R1329–R1399CrossRefGoogle Scholar
  63. Navarro A, Lopez-Cepero JM, Bandez MJ, Sanchez-Pino MJ, Gomez C, Cadenas E, Boveris A (2008) Hippocampal mitochondrial dysfunction in rat aging. Am J Physiol Regul Integr Comp Physiol 294:R501–R509PubMedCrossRefGoogle Scholar
  64. Okada M, Nakanishi H, Amamoto T, Urae R, Ando S, Yazawa K, Fujiwara M (2003) How does prolonged caloric restriction ameliorate age-related impairment of long-term potentiation in the hippocampus? Brain Res Mol Brain Res 111:175–181PubMedCrossRefGoogle Scholar
  65. Olgun A, Akman S, Serdar AM, Kutluay T (2002) Oxidative phosphorylation enzyme complexes in caloric restriction. Exp Gerontol 37:639–645PubMedCrossRefGoogle Scholar
  66. Pandya JD, Pauly JR, Nukala VN, Sebastian AH, Day KM, Korde AS, Maragos WF, Hall ED, Sullivan PG (2007) Post injury administration of mitochondrial uncouplers increases tissue sparing and improves behavioral outcome following traumatic brain injury in rodents. J Neurotrauma 24:798–811PubMedCrossRefGoogle Scholar
  67. Pathan AR, Viswanad B, Sonkusare SK, Ramarao P (2006) Chronic administration of pioglitazone attenuates intracerebroventricular streptozotocin induced-memory impairment in rats. Life Sci 79(23):2209–2216PubMedCrossRefGoogle Scholar
  68. Rieske JS (1967) Preparation and properties of reduced coenzyme Q cytochrome C reductase (complex III of the respiratory brain). In: Estabrook RW, Pullman ME (eds) Methods in Enzymology, vol 10. Academic Press, New York, pp 239–245Google Scholar
  69. Ripple MJ, Verweij M, Brand K, van de Ven M, Goemaere N, van den Engel S, Chu T, Forrer F, Müller C, de Jong M, van IJcken W, IJzermans JN, Hoeijmakers JH, de Bruin RW (2009) Short-term dietary restriction and fasting precondition against ischemia reperfusion injury in mice. Aging Cell 9:40–53Google Scholar
  70. Roth GS, Lane MA, Ingram DK, Mattison JA, Elahi D, Tobin JD, Muller D, Metter EJ (2002) Biomarkers of caloric restriction may predict longevity in humans. Science 297:811PubMedCrossRefGoogle Scholar
  71. Rutten BP, Vander Kolk NM, Zandvoort V, Bayer MA, Steinbusch TA, Schmitz C (2005) Age-related loss of synaptophysin immunoreactive presynaptic boutons within the hippocampus of APP751SL, PS1M146L, and APP751SL/PS1M146L transgenic mice. Am J Pathol 167:161–173PubMedCrossRefGoogle Scholar
  72. Sandhu SK, Kaur G (2002) Alterations in oxidative stress scavenger system in aging rat brain and lymphocytes. Biogeronotology 3(3):161–173CrossRefGoogle Scholar
  73. Sandhu SK, Kaur G (2003) Mitochondrial electron transport chain complexes in aging rat brain and lymphocytes. Biogerontology 4:19–29PubMedCrossRefGoogle Scholar
  74. Scheff SW, Price DA, Hicks RR, Baldwin SA, Robinson S, Brackney C (2005) Synaptogenesis in the hippocampal CA1 field following traumatic brain injury. J Neurotrauma 22:719–732PubMedCrossRefGoogle Scholar
  75. Sharma S, Singh R, Kaur M, Kaur G (2010) Late-onset dietary restriction compensates for age-related increase in oxidative stress and alterations of HSP 70 and synapsin1 protein levels in male Wistar rats. Biogerontology 11:197–209PubMedCrossRefGoogle Scholar
  76. Soderling TR (1993) Calcium/calmodulin-dependent protein kinase II: role in learning and memory. Mol Cell Biochem 127–128:93–101PubMedCrossRefGoogle Scholar
  77. Toescu EC, Verkhratsky A, Landfield PW (2004) Ca2+ regulation and gene expression in normal brain aging. Trends Neurosci 27:614–620PubMedCrossRefGoogle Scholar
  78. Wang P, Wang WP, Sun-Zhang WHX, Yan-Lou FYH (2008) Impaired spatial learning related with decreased expression of calcium/calmodulin-dependent protein kinase IIα and cAMP-response element binding protein in the pentylenetetrazol-kindled rats. Brain Res 1238:108–117PubMedCrossRefGoogle Scholar
  79. Weindruch R (1996) The retardation of aging by caloric restriction: studies in rodents and primates. Toxicol Pathol 24:742–745PubMedCrossRefGoogle Scholar
  80. Wharton DC, Tzagoloff A (1967) Cytochrome oxidase from beef heart mitochondria. Methods Enzymol 10:245–250CrossRefGoogle Scholar
  81. Witte AV, Fobker M, Gellner R, Knecht S, Floela A (2009) Caloric restriction improves memory in elderly humans. Proc Natl Acad Sci USA 106:1255–1260PubMedCrossRefGoogle Scholar
  82. Wu A, Ying Z, Gomez-Pinnilla F (2006) Dietary curcumin counteracts the outcome of tramatic brain injury on oxidative stress, synaptic plasticity and cognition. Exp Neurol 197:309–317PubMedCrossRefGoogle Scholar
  83. Yanai S, Okaichi Y, Okaichi H (2004) Long-term dietary restriction causes negative effects on cognitive functions in rats. Neurobiol Aging 25(3):325–332PubMedCrossRefGoogle Scholar
  84. Yonetani T (1967) Cytochrome oxidase: beef heart. In: Estabrook RW, Pullman ME (eds) Methods in enzymology, vol 10. Academic, New York, pp 332–335Google Scholar
  85. Zainal TA, Oberley TD, Allison DB, Szweda LI, Weindruch R (2000) Caloric restriction of rhesus monkeys lowers oxidative damage in skeletal muscle. FASEB J 14:1825–1836PubMedCrossRefGoogle Scholar
  86. Ziegler D, Rieske JS (1967) Preparation and properties of succinate dehydrogenase coenzyme Q reductase (complex II). In: Estabrook RW, Pullman ME (eds) Methods in enzymology, vol 10. Academic, New York, pp 231–235Google Scholar

Copyright information

© American Aging Association 2011

Authors and Affiliations

  • Rumani Singh
    • 1
  • Dinesh Lakhanpal
    • 1
  • Sushil Kumar
    • 1
  • Sandeep Sharma
    • 1
  • Hardeep Kataria
    • 1
  • Manpreet Kaur
    • 1
  • Gurcharan Kaur
    • 1
    Email author
  1. 1.Department of BiotechnologyGuru Nanak Dev UniversityAmritsarIndia

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