a-faster-high-resolution-mtpa-gfp-based-mitochondrial-fusion-assay

A Faster, High Resolution, mtPA-GFP-based Mitochondrial Fusion Assay Acquiring Kinetic Data of Multiple Cells in Parallel Using Confocal Microscopy

Mitochondrial fusion plays an essential role in mitochondrial calcium homeostasis, bioenergetics, autophagy and quality control. Fusion is quantified in ...

Boston University Medical Center

eNOS activation resulting in mitochondrial biogenesis is believed to play a central role in life span extension promoted by calorie restriction (CR). We investigated the mechanism of this activation by treating vascular cells with serum from CR rats and found increased Akt and eNOS phosphorylation, in addition to enhanced nitrite release. Inhibiting Akt phosphorylation or immunoprecipitating adiponectin (found in high quantities in CR serum) completely prevented the increment in nitrite release and eNOS activation. Overall, we demonstrate that adiponectin in the serum from CR animals increases NO• signaling by activating the insulin pathway. These results suggest this hormone may be a determinant regulator of the beneficial effects of CR.

Serum from calorie-restricted rats activates vascular cell eNOS through enhanced insulin signaling mediated by adiponectin.

Caloric restriction is the most effective non-genetic intervention to enhance lifespan known to date. A major research interest has been the development of therapeutic strategies capable of promoting the beneficial results of this dietary regimen. In this sense, we propose that compounds that decrease the efficiency of energy conversion, such as mitochondrial uncouplers, can be caloric restriction mimetics. Treatment of mice with low doses of the protonophore 2,4-dinitrophenol promotes enhanced tissue respiratory rates, improved serological glucose, triglyceride and insulin levels, decrease of reactive oxygen species levels and tissue DNA and protein oxidation, as well as reduced body weight. Importantly, 2,4-dinitrophenol-treated animals also presented enhanced longevity. Our results demonstrate that mild mitochondrial uncoupling is a highly effective in vivo antioxidant strategy, and describe the first therapeutic intervention capable of effectively reproducing the physiological, metabolic and lifespan effects of caloric restriction in healthy mammals.

Mild mitochondrial uncoupling in mice affects energy metabolism, redox balance and longevity.

Undernutrition without malnutrition is an intervention that enhances laboratory animal life span, and is widely studied to uncover factors limiting longevity. In a search of the literature over a course of four years, we found that most protocols currently adopted as caloric restriction do not meet micronutrient standards set by the National Research Council for laboratory rats and mice. We provide evidence that the most commonly adopted caloric restriction protocol, a 40% restriction of the AIN-93 diet without vitamin or mineral supplementation, leads to malnutrition in both mice and rats. Furthermore, others and we find that every other day feeding, another dietary intervention often referred to as caloric restriction, does not limit the total amount of calories consumed. Altogether, we propose that the term "caloric restriction" should be used specifically to describe diets that decrease calorie intake but not micronutrient availability, and that protocols adopted should be described in detail in order to allow for comparisons and better understanding of the effects of these diets.

Commonly adopted caloric restriction protocols often involve malnutrition.

The aim of the present study was to evaluate the protective effects of the 4-anilinoquinazoline derivative PD153035 on cardiac ischemia/reperfusion and mitochondrial function.

Potent cardioprotective effect of the 4-anilinoquinazoline derivative PD153035: involvement of mitochondrial K(ATP) channel activation.

Mild mitochondrial uncoupling, or the reduction of the efficiency of energy conversion without compromising intracellular high energy phosphate levels, is a protective therapeutic strategy under many laboratory conditions. Here we discuss these conditions, which include both cell and animal models of ischemia reperfusion and complications associated with the metabolic syndrome. We also discuss drugs that promote mild mitochondrial uncoupling and naturally occurring mild mitochondrial uncoupling pathways involving free fatty acid cycling and K(+) transport.

Mild mitochondrial uncoupling as a therapeutic strategy.

Enhanced mitochondrial biogenesis promoted by eNOS activation is believed to play a central role in the beneficial effects of calorie restriction (CR). Since treatment of mice with dinitrophenol (DNP) promotes health and lifespan benefits similar to those observed in CR, we hypothesized that it could also impact biogenesis. We found that DNP and CR increase citrate synthase activity, PGC-1α, cytochrome c oxidase and mitofusin-2 expression, as well as fasting plasma levels of NO• products. In addition, eNOS and Akt phosphorylation in skeletal muscle and visceral adipose tissue was activated in fasting CR and DNP animals. Overall, our results indicate that systemic mild uncoupling activates eNOS and Akt-dependent pathways leading to mitochondrial biogenesis.

Mild mitochondrial uncoupling and calorie restriction increase fasting eNOS, akt and mitochondrial biogenesis.

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Calorie restriction is a dietary intervention known to improve redox state, glucose tolerance, and animal life span. Other interventions have been adopted as study models for caloric restriction, including nonsupplemented food restriction and intermittent, every-other-day feedings. We compared the short- and long-term effects of these interventions to ad libitum protocols and found that, although all restricted diets decrease body weight, intermittent feeding did not decrease intra-abdominal adiposity. Short-term calorie restriction and intermittent feeding presented similar results relative to glucose tolerance. Surprisingly, long-term intermittent feeding promoted glucose intolerance, without a loss in insulin receptor phosphorylation. Intermittent feeding substantially increased insulin receptor nitration in both intra-abdominal adipose tissue and muscle, a modification associated with receptor inactivation. All restricted diets enhanced nitric oxide synthase levels in the insulin-responsive adipose tissue and skeletal muscle. However, whereas calorie restriction improved tissue redox state, food restriction and intermittent feedings did not. In fact, long-term intermittent feeding resulted in largely enhanced tissue release of oxidants. Overall, our results show that restricted diets are significantly different in their effects on glucose tolerance and redox state when adopted long-term. Furthermore, we show that intermittent feeding can lead to oxidative insulin receptor inactivation and glucose intolerance.

Long-term intermittent feeding, but not caloric restriction, leads to redox imbalance, insulin receptor nitration, and glucose intolerance.

Restriction calorique est une intervention alimentaire connue pour am&eacute;liorer l&#39;&eacute;tat d&#39;oxydor&eacute;duction, la tol&eacute;rance au glucose et dur&eacute;e de vie animale. Autres interventions ont &eacute;t&eacute; adopt&eacute;es comme mod&egrave;les d&#39;&eacute;tude pour la restriction calorique, y compris la restriction alimentaire am&eacute;liora et intermittentes, tous les autres-jours des t&eacute;t&eacute;es. Nous avons compar&eacute; les effets &agrave; court et long terme de ces interventions aux protocoles ad libitum et constat&eacute; que, bien que tous les r&eacute;gimes restrictifs diminuent du poids corporel, alimentation intermittente ne diminue pas l&#39;adiposit&eacute; intra-abdominale. &Agrave; court terme restriction calorique et alimentation intermittente ont pr&eacute;sent&eacute; des r&eacute;sultats similaires par rapport &agrave; la tol&eacute;rance au glucose. &Eacute;tonnamment, &agrave; long terme l&#39;alimentation intermittente promu l&#39;intol&eacute;rance au glucose, sans perte de la phosphorylation du r&eacute;cepteur de l&#39;insuline. Alimentation intermittente consid&eacute;rablement augment&eacute; la nitration de r&eacute;cepteur de l&#39;insuline dans les deux le tissu adipeux intra-abdominal et musculaire, une modification associ&eacute;e &agrave; l&#39;inactivation du r&eacute;cepteur. Tous les r&eacute;gimes restrictifs am&eacute;lior&eacute; l&#39;oxyde nitrique synthase niveaux en les favorisant l&#39;insuline des tissus adipeux et les muscles squelettiques. Toutefois, consid&eacute;rant que la restriction calorique am&eacute;lior&eacute;e &eacute;tat redox des tissus, restriction alimentaire et t&eacute;t&eacute;es intermittentes n&#39;ont pas. En fait, une alimentation intermittente &agrave; long terme entra&icirc;ne largement renforc&eacute;e tissu lib&eacute;ration d&#39;oxydants. Dans l&#39;ensemble, nos r&eacute;sultats montrent que les r&eacute;gimes restrictifs sont significativement diff&eacute;rentes dans leurs effets sur l&#39;&eacute;tat de tol&eacute;rance et d&#39;oxydor&eacute;duction glucose lors de l&#39;adoption &agrave; long terme. De plus, nous montrons que l&#39;alimentation intermittente peut conduire &agrave; insuline oxydatif r&eacute;cepteur inactivation et glucose intol&eacute;rance.

Alimentation intermittente &agrave; long terme, mais la restriction calorique pas, conduit au d&eacute;s&eacute;quilibre de l&#39;oxydo-r&eacute;duction, la nitration du r&eacute;cepteur de l&#39;insuline et une intol&eacute;rance au glucose.

Kalorienrestriktion ist eine di&auml;tetische Intervention zur Verbesserung der Redox-Zustand, Glukosetoleranz und tierischen Lebensdauer bekannt. Sonstige Eingriffe wurden als Studie Modelle f&uuml;r kalorische Einschr&auml;nkung, einschlie&szlig;lich nonsupplemented Essen Einschr&auml;nkung und selten auftretende, Sonstiges-t&auml;glichen F&uuml;tterungen angenommen. Wir verglichen die kurz- und langfristigen Auswirkungen dieser Interventionen auf ad libitum Protokolle und gefunden, dass, obwohl alle eingeschr&auml;nkte Di&auml;ten K&ouml;rpergewicht verringern, intermittierende F&uuml;tterung nicht intraabdominellen Adipositas abnehmen. Kurzfristige Kalorienreduktion und intermittierende F&uuml;tterung pr&auml;sentiert &auml;hnliche Ergebnisse relativ Glukosetoleranz. Langfristige intermittierende F&uuml;tterung gef&ouml;rdert &uuml;berraschend, Glukoseintoleranz, ohne dass die Insulin-Rezeptor-Phosphorylierung. Intermittierende F&uuml;tterung betr&auml;chtlich Insulin-Rezeptor-Nitrierung in intra-abdominalen Fettgewebe und Muskeln, eine &Auml;nderung, die zugeordneten Rezeptor-Inaktivierung. Alle eingeschr&auml;nkte Di&auml;ten verbessert Stickoxidspiegel Synthase in den Insulin-responsive Fettgewebe und Skelettmuskulatur. Jedoch w&auml;hrend die Kalorienrestriktion Gewebe Redox-Zustand verbessert, nicht essen-Beschr&auml;nkung und intermittierende F&uuml;tterungen. In der Tat bewirkte langfristige intermittierende F&uuml;tterung weitgehend verbesserte Gewebe Freisetzung von Oxidantien. Insgesamt zeigen unsere Ergebnisse, dass eingeschr&auml;nkte Di&auml;ten in ihrer Wirkung auf Glukose Toleranz und Redox-Zustand als langfristige verabschiedet signifikant unterscheiden. Dar&uuml;ber hinaus zeigen wir, dass die intermittierende F&uuml;tterung zu oxidativen Insulin-Rezeptor-Inaktivierung und Glukose-Intoleranz f&uuml;hren kann.

Langfristige intermittierende F&uuml;tterung, aber nicht kalorische Einschr&auml;nkung f&uuml;hrt zu Redox-Ungleichgewicht, Insulin-Rezeptor-Nitrierung und Glukoseintoleranz.

Restrizione calorica &egrave; un intervento dietetico noto al miglioramento dello stato redox, tolleranza al glucosio e durata di vita degli animali. Altri interventi sono stati adottati come modelli di studio per la restrizione calorica, tra cui la restrizione alimentare nonsupplemented e poppate intermittente, ogni altro giorno. Abbiamo confrontato gli effetti di breve e lungo termine di questi interventi ai protocolli ad libitum e ha trovato che, nonostante tutte le diete con restrizioni diminuiscono il peso corporeo, alimentazione intermittente ha fatto non diminuire adiposit&agrave; intra-addominale. A breve termine restrizione calorica e alimentazione intermittente presentato risultati analoghi rispetto alla tolleranza al glucosio. Sorprendentemente, a lungo termine di alimentazione intermittente promosso intolleranza al glucosio, senza perdita di fosforilazione del recettore insulina. Alimentazione intermittente aumentato sostanzialmente nitrazione del recettore insulina sia intra-addominale del tessuto adiposo e muscolare, una modifica associata con l&#39;inattivazione del recettore. Tutte le diete con restrizioni enhanced livelli di ossido nitrico sintasi in insulino-sensibli a reagire del tessuto adiposo e muscolo scheletrico. Tuttavia, considerando che la restrizione calorica migliorato stato redox di tessuto, restrizione alimentare e poppate intermittente non ha fatto. In realt&agrave;, alimentazione intermittente a lungo termine ha portato nella versione tessuto largamente migliorata di ossidanti. Nel complesso, i nostri risultati mostrano che diete con restrizioni sono significativamente differenti nei loro effetti sullo stato di tolleranza e redox glucosio quando adottata a lungo termine. Inoltre, mostriamo che alimentazione intermittente pu&ograve; portare a ossidativo insulina recettore inattivazione e glucosio intolleranza.

Alimentazione intermittente a lungo termine, ma la restrizione calorica non, porta ad uno squilibrio redox, nitrazione del recettore insulina e intolleranza al glucosio.

&#x30AB;&#x30ED;&#x30EA;&#x30FC;&#x5236;&#x9650;&#x306E;&#x9178;&#x5316;&#x9084;&#x5143;&#x72B6;&#x614B;&#x3001;&#x8010;&#x7CD6;&#x80FD;&#x304A;&#x3088;&#x3073;&#x52D5;&#x7269;&#x306E;&#x5BFF;&#x547D;&#x3092;&#x6539;&#x5584;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x77E5;&#x3089;&#x308C;&#x3066;&#x98DF;&#x990C;&#x7642;&#x6CD5;&#x306E;&#x4ECB;&#x5165;&#x3067;&#x3059;&#x3002;&#x4ED6;&#x306E;&#x4ECB;&#x5165;&#x306F;&#x30E2;&#x30C7;&#x30EB; nonsupplemented &#x306E;&#x98DF;&#x990C;&#x5236;&#x9650;&#x3068;&#x6388;&#x4E73;&#x306E;&#x65AD;&#x7D9A;&#x7684;&#x306A;&#x3059;&#x3079;&#x3066;&#x4ED6;&#x306E;&#x65E5;&#x3092;&#x542B;&#x3080;&#x3001;&#x30AB;&#x30ED;&#x30EA;&#x30FC;&#x5236;&#x9650;&#x306B;&#x95A2;&#x3059;&#x308B;&#x4E00;&#x8003;&#x5BDF;&#x3068;&#x3057;&#x3066;&#x63A1;&#x7528;&#x3055;&#x308C;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x4ECB;&#x5165;&#x306E;&#x30A2;&#x30C9;&#x30EA;&#x30D6; &#x30D7;&#x30ED;&#x30C8;&#x30B3;&#x30EB;&#x3078;&#x306E;&#x77ED;&#x671F;&#x304A;&#x3088;&#x3073;&#x9577;&#x671F;&#x52B9;&#x679C;&#x3092;&#x6BD4;&#x8F03;&#x3057;&#x3001;&#x3059;&#x3079;&#x3066;&#x5236;&#x9650;&#x30C0;&#x30A4;&#x30A8;&#x30C3;&#x30C8;&#x306F;&#x4F53;&#x91CD;&#x6E1B;&#x5C11;&#x304C;&#x3001;&#x9593;&#x6B20;&#x9001;&#x308A;&#x8179;&#x8154;&#x5185;&#x8102;&#x80AA;&#x6E1B;&#x5C11;&#x3057;&#x306A;&#x3044;&#x3053;&#x3068;&#x304C;&#x898B;&#x3064;&#x304B;&#x308A;&#x307E;&#x3057;&#x305F;&#x3002;&#x77ED;&#x671F;&#x7684;&#x306A;&#x30AB;&#x30ED;&#x30EA;&#x30FC;&#x5236;&#x9650;&#x3068;&#x9593;&#x6B20;&#x9001;&#x308A;&#x30D6;&#x30C9;&#x30A6;&#x7CD6;&#x306E;&#x8A31;&#x5BB9;&#x57FA;&#x6E96;&#x306B;&#x3057;&#x3066;&#x3001;&#x540C;&#x69D8;&#x306E;&#x7D50;&#x679C;&#x3092;&#x63D0;&#x793A;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x9A5A;&#x3044;&#x305F;&#x3053;&#x3068;&#x306B;&#x3001;&#x9577;&#x671F;&#x9593;&#x6B20;&#x9001;&#x308A;&#x306E;&#x30A4;&#x30F3;&#x30B9;&#x30EA;&#x30F3;&#x53D7;&#x5BB9;&#x4F53;&#x306E;&#x30EA;&#x30F3;&#x9178;&#x5316;&#x306B;&#x640D;&#x5931;&#x306A;&#x3057;&#x306E;&#x30B0;&#x30EB;&#x30B3;&#x30FC;&#x30B9;&#x4E0D;&#x5BDB;&#x5BB9;&#x3092;&#x63A8;&#x9032;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x9593;&#x6B20;&#x9001;&#x308A;&#x30A4;&#x30F3;&#x30B9;&#x30EA;&#x30F3;&#x53D7;&#x5BB9;&#x4F53;&#x306E;&#x30CB;&#x30C8;&#x30ED;&#x5316;&#x8179;&#x8154;&#x5185;&#x8102;&#x80AA;&#x7D44;&#x7E54;&#x3068;&#x7B4B;&#x8089;&#x3001;&#x53D7;&#x5BB9;&#x4F53;&#x306E;&#x4E0D;&#x6D3B;&#x6027;&#x5316;&#x3068;&#x95A2;&#x9023;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B;&#x5909;&#x66F4;&#x3067;&#x5927;&#x5E45;&#x306B;&#x5897;&#x52A0;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x3059;&#x3079;&#x3066;&#x306E;&#x5236;&#x9650;&#x3055;&#x308C;&#x305F;&#x98DF;&#x4E8B;&#x30A4;&#x30F3;&#x30B9;&#x30EA;&#x30F3;&#x5FDC;&#x7B54;&#x6027;&#x8102;&#x80AA;&#x7D44;&#x7E54;&#x3068;&#x9AA8;&#x683C;&#x7B4B;&#x306B;&#x304A;&#x3051;&#x308B;&#x4E00;&#x9178;&#x5316;&#x7A92;&#x7D20;&#x5408;&#x6210;&#x9175;&#x7D20;&#x30EC;&#x30D9;&#x30EB;&#x5F37;&#x5316;&#x3002;&#x30AB;&#x30ED;&#x30EA;&#x30FC;&#x5236;&#x9650;&#x306F;&#x30C6;&#x30A3;&#x30C3;&#x30B7;&#x30E5;&#x306E;&#x9178;&#x5316;&#x9084;&#x5143;&#x72B6;&#x614B;&#x6539;&#x5584;&#x306B;&#x5BFE;&#x3057;&#x3001;&#x305F;&#x3060;&#x3057;&#x3001;&#x98DF;&#x4E8B;&#x5236;&#x9650;&#x306F;&#x3001;&#x65AD;&#x7D9A;&#x7684;&#x306A;&#x6388;&#x4E73;&#x3057;&#x307E;&#x305B;&#x3093;&#x3067;&#x3057;&#x305F;&#x3002;&#x5B9F;&#x969B;&#x306B;&#x306F;&#x3001;&#x9577;&#x671F;&#x9593;&#x6B20;&#x9001;&#x308A;&#x30AA;&#x30AD;&#x30B7;&#x30C0;&#x30F3;&#x30C8;&#x306E;&#x5927;&#x304D;&#x304F;&#x5F37;&#x5316;&#x3055;&#x308C;&#x305F;&#x7D44;&#x7E54;&#x306E;&#x30EA;&#x30EA;&#x30FC;&#x30B9;&#x3067;&#x8D77;&#x56E0;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x5168;&#x4F53;&#x7684;&#x306B;&#x3001;&#x6211;&#x3005; &#x306E;&#x7D50;&#x679C;&#x306F;&#x3001;&#x7279;&#x5225;&#x306A;&#x98DF;&#x4E8B;&#x5236;&#x9650;&#x304C;&#x9577;&#x671F;&#x63A1;&#x7528;&#x6642;&#x306E;&#x30B0;&#x30EB;&#x30B3;&#x30FC;&#x30B9;&#x5BDB;&#x5BB9;&#x3068;&#x9178;&#x5316;&#x9084;&#x5143;&#x72B6;&#x614B;&#x3078;&#x306E;&#x5F71;&#x97FF;&#x3067;&#x5927;&#x5E45;&#x306B;&#x7570;&#x306A;&#x308B;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;&#x3055;&#x3089;&#x306B;, &#x9593;&#x6B20;&#x9001;&#x308A;&#x9178;&#x5316;&#x30A4;&#x30F3;&#x30B9;&#x30EA;&#x30F3;&#x53D7;&#x5BB9;&#x4F53;&#x306E;&#x4E0D;&#x6D3B;&#x6027;&#x5316;&#x304A;&#x3088;&#x3073;&#x30B0;&#x30EB;&#x30B3;&#x30FC;&#x30B9;&#x4E0D;&#x5BDB;&#x5BB9;&#x306B;&#x3064;&#x306A;&#x304C;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;

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&#xCE7C;&#xB85C;&#xB9AC; &#xC81C;&#xD55C; &#xC2DD;&#xC774;&#xC694;&#xBC95; &#xC911;&#xC7AC; redox &#xC0C1;&#xD0DC;, &#xD3EC;&#xB3C4; &#xB2F9; &#xD5C8;&#xC6A9; &#xBC0F; &#xB3D9;&#xBB3C; &#xC218;&#xBA85;&#xC744; &#xAC1C;&#xC120; &#xD558;&#xAE30; &#xC704;&#xD574; &#xC54C;&#xB824;&#xC9C4; &#xC774;&#xB2E4;. &#xB2E4;&#xB978; &#xAC1C;&#xC785; nonsupplemented &#xC74C;&#xC2DD; &#xC81C;&#xD55C; &#xD558; &#xACE0; &#xAC04;&#xD5D0;&#xC801;&#xC73C;&#xB85C;, &#xB9E4;&#xC77C; &#xB2E4;&#xB978; &#xD558;&#xB8E8; &#xC218; &#xC720;&#xB97C; &#xD3EC;&#xD568; &#xD55C; &#xC5F4; &#xB7C9; &#xC81C;&#xD55C;&#xC5D0; &#xB300; &#xD55C; &#xC5F0;&#xAD6C; &#xBAA8;&#xB378;&#xB85C; &#xCC44;&#xD0DD; &#xB418;&#xC5C8;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xB294; &#xB2E8;&#xAE30; &#xBC0F; &#xC7A5;&#xAE30; &#xD6A8;&#xACFC; ad libitum &#xD504;&#xB85C;&#xD1A0;&#xCF5C;&#xC744; &#xC774;&#xB7EC;&#xD55C; &#xAC1C;&#xC785;&#xC758; &#xBE44;&#xAD50; &#xD558; &#xACE0;, &#xBAA8;&#xB4E0; &#xC81C;&#xD55C; &#xB2E4;&#xC774;&#xC5B4;&#xD2B8;&#xB294; &#xCCB4;&#xC911; &#xAC10;&#xC18C;, &#xBE44;&#xB85D; &#xAC04;&#xD5D0;&#xC801;&#xC73C;&#xB85C; &#xBA39;&#xC774; &#xC54A;&#xC558;&#xB2E4; &#xC904;&#xC5B4;&#xB4E4;&#xC9C0; &#xB0B4;&#xBD80;&#xC758; &#xAC83; &#xBC1C;&#xACAC;. &#xB2E8;&#xAE30; &#xCE7C;&#xB85C;&#xB9AC; &#xC81C;&#xD55C; &#xD558; &#xACE0; &#xAC04;&#xD5D0;&#xC801;&#xC73C;&#xB85C; &#xBA39;&#xC774; &#xD3EC;&#xB3C4; &#xB2F9; &#xB0B4;&#xC131; &#xAE30;&#xC900;&#xC73C;&#xB85C; &#xBE44;&#xC2B7;&#xD55C; &#xACB0;&#xACFC; &#xBC1C;&#xD45C; &#xD569;&#xB2C8;&#xB2E4;. &#xB180;&#xB78D;&#xAC8C;&#xB3C4;, &#xD3EC;&#xB3C4; &#xB2F9; &#xC639; &#xC878;, &#xC778;&#xC290;&#xB9B0; &#xC218;&#xC6A9; &#xCCB4; &#xC778; &#xC0B0;&#xD654; &#xC190;&#xC2E4; &#xC5C6;&#xC774; &#xC2B9;&#xC9C4; &#xC7A5;&#xAE30; &#xAC04;&#xD5D0;&#xC801;&#xC73C;&#xB85C; &#xBA39;&#xC774;. &#xB0B4;&#xBD80;-&#xBCF5; &#xBD80; &#xC9C0;&#xBC29; &#xC870;&#xC9C1; &#xBC0F; &#xADFC;&#xC721;, &#xC218;&#xC6A9; &#xCCB4; &#xBE44;&#xD65C;&#xC131;&#xD654;&#xC640; &#xAD00;&#xB828; &#xB41C; &#xC218;&#xC815;&#xC5D0;&#xC11C; &#xC778;&#xC290;&#xB9B0; &#xC218;&#xC6A9; &#xCCB4; nitration &#xC99D;&#xAC00; &#xC2E4;&#xC9C8;&#xC801;&#xC73C;&#xB85C; &#xAC04;&#xD5D0;&#xC801;&#xC73C;&#xB85C; &#xBA39;&#xC774;. &#xBAA8;&#xB4E0; &#xC81C;&#xD55C; &#xB41C; &#xB2E4;&#xC774;&#xC5B4;&#xD2B8; &#xC778;&#xC290;&#xB9B0; &#xBC18;&#xC751; &#xC9C0;&#xBC29; &#xC870;&#xC9C1; &#xBC0F; &#xACE8;&#xACA9; &#xADFC;&#xC721;&#xC5D0; &#xC0B0;&#xD654; &#xC9C8;&#xC18C; synthase &#xC218;&#xC900; &#xD5A5;&#xC0C1;. &#xADF8;&#xB7EC;&#xB098;, &#xBC18;&#xBA74;&#xC5D0; &#xCE7C;&#xB85C;&#xB9AC; &#xC81C;&#xD55C; &#xC870;&#xC9C1;&#xC758; &#xC0B0;&#xD654; &#xC0C1;&#xD0DC; &#xAC1C;&#xC120;, &#xC74C;&#xC2DD; &#xC81C;&#xD55C; &#xD558; &#xACE0; &#xAC04;&#xD5D0;&#xC801;&#xC73C;&#xB85C; &#xC218; &#xC720; &#xD558;&#xC9C0; &#xC54A;&#xC558;&#xB2E4;. &#xC0AC;&#xC2E4;, &#xC7A5;&#xAE30; &#xAC04;&#xD5D0;&#xC801;&#xC778; &#xBA39;&#xC774; oxidants&#xC758; &#xD06C;&#xAC8C; &#xD5A5;&#xC0C1; &#xB41C; &#xC870;&#xC9C1; &#xB9B4;&#xB9AC;&#xC2A4; &#xACB0;&#xACFC;. &#xC804;&#xBC18;&#xC801;&#xC73C;&#xB85C;, &#xC6B0;&#xB9AC;&#xC758; &#xACB0;&#xACFC; &#xD45C;&#xC2DC; &#xC81C;&#xD55C; &#xB2E4;&#xC774;&#xC5B4;&#xD2B8;&#xB294; &#xD3EC;&#xB3C4; &#xB2F9; &#xB0B4;&#xC131; &#xBC0F; redox &#xC0C1;&#xD0DC; &#xC7A5;&#xAE30; &#xCC44;&#xD0DD; &#xD558;&#xB294; &#xACBD;&#xC6B0;&#xC5D0; &#xADF8;&#xB4E4;&#xC758; &#xD6A8;&#xACFC;&#xC5D0; &#xD06C;&#xAC8C; &#xB2E4;&#xB985;&#xB2C8;&#xB2E4;. &#xB610;&#xD55C;, &#xC6B0;&#xB9AC;&#xB294; &#xAC04;&#xD5D0;&#xC801;&#xC73C;&#xB85C; &#xC218; &#xC720; &#xC0B0;&#xD654; &#xC778;&#xC290;&#xB9B0; &#xC218;&#xC6A9; &#xCCB4; &#xBE44;&#xD65C;&#xC131;&#xD654; &#xBC0F; &#xD3EC;&#xB3C4; &#xB2F9; &#xC639; &#xC878; &#xBC1C;&#xC0DD;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4; &#xBCF4;&#xC5EC;&#xC90D;&#xB2C8;&#xB2E4;.

&#xC7A5;&#xAE30; &#xAC04;&#xD5D0;&#xC801;&#xC778; &#xBA39;&#xC774;, &#xD558;&#xC9C0; &#xC5F4; &#xB7C9; &#xC81C;&#xD55C; &#xD558;&#xC9C0;&#xB9CC; redox &#xBD88;&#xADE0;&#xD615;, &#xC778;&#xC290;&#xB9B0; &#xC218;&#xC6A9; &#xCCB4; nitration &#xBC0F; &#xD3EC;&#xB3C4; &#xB2F9; &#xC639; &#xC878;&#xC744; &#xB9AC;&#xB4DC;.

Restricci&oacute;n cal&oacute;rica es una intervenci&oacute;n diet&eacute;tica conocida para mejorar estado redox, tolerancia a la glucosa y vida animal. Otras intervenciones se han adoptado como modelo de estudio para la restricci&oacute;n cal&oacute;rica, incluyendo la restricci&oacute;n de alimentos nonsupplemented y alimentaci&oacute;n intermitente, diarios-otros. Compararon los efectos a corto y largo plazo de estas intervenciones a protocolos ad libitum y encontr&oacute; que, aunque todas las dietas restringidas disminuyen peso corporal, alimentaci&oacute;n intermitente no disminuyeron adiposidad intraabdominal. La restricci&oacute;n cal&oacute;rica a corto plazo y alimentaci&oacute;n intermitente presentan resultados similares en relaci&oacute;n con la tolerancia a la glucosa. Sorprendentemente, alimentaci&oacute;n intermitente a largo plazo promueve la intolerancia a la glucosa, sin una p&eacute;rdida en la fosforilaci&oacute;n del receptor de insulina. Alimentaci&oacute;n intermitente increment&oacute; sustancialmente la nitraci&oacute;n de receptor de insulina en tanto el tejido adiposo intraabdominal y m&uacute;sculo, una modificaci&oacute;n asociados con la inactivaci&oacute;n del receptor. Todas las dietas restringidas hab&iacute;an mejorado los niveles de &oacute;xido n&iacute;trico sintasa en la insulina sensible al tejido adiposo y m&uacute;sculo esquel&eacute;tico. Sin embargo, Considerando que la restricci&oacute;n cal&oacute;rica mejora estado redox de tejido, restricci&oacute;n alimentaria y alimentaci&oacute;n intermitente no. De hecho, alimentaci&oacute;n intermitente a largo plazo result&oacute; en el lanzamiento de mejorada en gran parte del tejido de oxidantes. En general, nuestros resultados muestran que las dietas restringidas son significativamente diferentes en sus efectos sobre el estado con tolerancia y redox de glucosa cuando adopt&oacute; a largo plazo. Adem&aacute;s, demostramos que la alimentaci&oacute;n intermitente puede conducir a la intolerancia de inactivaci&oacute;n y glucosa de receptor de insulina oxidativo.

Alimentaci&oacute;n intermitente a largo plazo, pero la restricci&oacute;n cal&oacute;rica no conduce al desequilibrio redox, nitraci&oacute;n de receptor de insulina y la intolerancia a la glucosa.

Calorie restriction (CR) enhances animal life span and prevents age-related diseases, including neurological decline. Recent evidence suggests that a mechanism involved in CR-induced life-span extension is NO(•)-stimulated mitochondrial biogenesis. We examine here the effects of CR on brain mitochondrial content. CR increased eNOS and nNOS and the content of mitochondrial proteins (cytochrome c oxidase, citrate synthase, and mitofusin) in the brain. Furthermore, we established an in vitro system to study the neurological effects of CR using serum extracted from animals on this diet. In cultured neurons, CR serum enhanced nNOS expression and increased levels of nitrite (a NO(•) product). CR serum also enhanced the levels of cytochrome c oxidase and increased citrate synthase activity and respiratory rates in neurons. CR serum effects were inhibited by L-NAME and mimicked by the NO(•) donor SNAP. Furthermore, both CR sera and SNAP were capable of improving neuronal survival. Overall, our results indicate that CR increases mitochondrial biogenesis in a NO(•)-mediated manner, resulting in enhanced reserve respiratory capacity and improved survival in neurons.

Calorie restriction increases cerebral mitochondrial respiratory capacity in a NO•-mediated mechanism: impact on neuronal survival.

Mitochondrial energy metabolism and mitochondrially-derived oxidants have, for many years, been recognized as central toward the effects of aging. A body of recent work has focused on the relationship between mitochondrial redox state, aging and dietary interventions that affect lifespan. These studies have uncovered mechanisms through which diet alters mitochondrial metabolism, in addition to determining how these changes affect oxidant generation, which in itself has an impact on mitochondrial function in aged animals. Many of the studies conducted to date, however, are correlative, and it remains to be determined which of the energy metabolism and redox modifications induced by diet are central toward lifespan extent. Furthermore, dietary interventions used for laboratory animals are often unequal, and of difficult comparison with humans (for whom, by nature, no long-term sound scientific information on the effects of diet on mitochondrial redox state and aging is available). We hope future studies will be able to mechanistically characterize which energy metabolism and redox changes promoted by dietary interventions have positive lifespan effects, and translate these findings into human prevention and treatment of age-related disease.

Fernanda M. Cerqueira

Department of Medicine

Boston University Medical Center

A Faster, High Resolution, mtPA-GFP-based Mitochondrial Fusion Assay Acquiring Kinetic Data of Multiple Cells in Parallel Using Confocal Microscopy

Fernanda M. Cerqueira

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Medicine

Boston University Medical Center

A Faster, High Resolution, mtPA-GFP-based Mitochondrial Fusion Assay Acquiring Kinetic Data of Multiple Cells in Parallel Using Confocal Microscopy

Department of Medicine