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Citrulline diet supplementation improves specific age-related raft changes in wild-type rodent hippocampus

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The levels of molecules crucial for signal transduction processing change in the brain with aging. Lipid rafts are membrane microdomains involved in cell signaling. We describe here substantial biophysical and biochemical changes occurring within the rafts in hippocampus neurons from aging wild-type rats and mice. Using continuous sucrose density gradients, we observed light-, medium-, and heavy raft subpopulations in young adult rodent hippocampus neurons containing very low levels of amyloid precursor protein (APP) and almost no caveolin-1 (CAV-1). By contrast, old rodents had a homogeneous age-specific high-density caveolar raft subpopulation containing significantly more cholesterol (CHOL), CAV-1, and APP. C99-APP-Cter fragment detection demonstrates that the first step of amyloidogenic APP processing takes place in this caveolar structure during physiological aging of the rat brain. In this age-specific caveolar raft subpopulation, levels of the C99-APP-Cter fragment are exponentially correlated with those of APP, suggesting that high APP concentrations may be associated with a risk of large increases in beta-amyloid peptide levels. Citrulline (an intermediate amino acid of the urea cycle) supplementation in the diet of aged rats for 3 months reduced these age-related hippocampus raft changes, resulting in raft patterns tightly close to those in young animals: CHOL, CAV-1, and APP concentrations were significantly lower and the C99-APP-Cter fragment was less abundant in the heavy raft subpopulation than in controls. Thus, we report substantial changes in raft structures during the aging of rodent hippocampus and describe new and promising areas of investigation concerning the possible protective effect of citrulline on brain function during aging.

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This work was supported by INSERM (ATC Vieillissement 2002), Université Paris Descartes (ATP aging). We thank Dr. Kenneth L. Moya for his helpful comments and his expert editing of the manuscript. We are grateful to Servane Le Plenier for her help with animal care.

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Correspondence to Bernadette Allinquant.

Additional information

Perrine Marquet-de Rougé and Christine Clamagirand contributed equally to this work.

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Fig. 1S

Detection of hippocampal rodent raft subpopulations. (A) Representative immunoblots showing the co-detection of F3 and Flotillin-1 in YNG, NEAA, and CIT rats. A 12–30 % continuous sucrose gradient was used. Similar patterns were obtained for the two proteins. (B) Representative co-detection of GM1 (dot blots) and FLOT-1 in mouse rafts. A 5–30 % continuous sucrose gradient was used. GM1 and FLOT-1 presented similar patterns. These three raft-specific markers displayed maximal levels of expression in the same fractions (JPEG 38 kb)

Fig. 2S

Hippocampal raft subpopulations of 20-month-old rats. (A) Representative immunoblots of hippocampal rafts from 20-month-old rats probed for CAV-1, FLOT-1, and APP. A 12–30 % continuous sucrose gradient was used. (B) CAV-1 and APP levels in total rafts and in the heavy raft subpopulation in the hippocampi of 20-CTRL rats (n = 3), compared with YNG (n = 6), NEAA (n = 7), and CIT rats (n = 8). Data are expressed in arbitrary units (AU) and are presented as mean values ± SEMs (JPEG 41 kb)

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Marquet-de Rougé, P., Clamagirand, C., Facchinetti, P. et al. Citrulline diet supplementation improves specific age-related raft changes in wild-type rodent hippocampus. AGE 35, 1589–1606 (2013). https://doi.org/10.1007/s11357-012-9462-2

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  • Aging
  • Amyloid precursor protein
  • Brain
  • Cholesterol
  • Lipid rafts
  • Caveolin-1
  • Citrulline diet
  • Hippocampus
  • Rodent