The mitochondrial uncoupler DNP triggers brain cell mTOR signaling network reprogramming and CREB pathway up‐regulation

“autophagosome formation, and is thus an autophagosome marker (Kabeya et al. 2000).”

“mTOR interacts with several proteins and forms two complexes, mTORC1 and mTORC2, initially characterized in yeast (Loewith et al. 2002; Wullachleger et al. 2006).”

“Evidence that autophagy-related processes are responsive to mild mitochondrial uncoupling mTOR is a negative regulator of autophagy, a process that removes damaged proteins and organelles and recycles intracellular nutrient resources (Ravikumar et al. 2004; Yu et al. 2010).”

“…mTORC1 also regulates autophagy (Laplante and Sabatini 2012; Dibble and Manning 2013). mTORC2 responds to activation of Akt, SGK, and PKC, and controls cell survival, size, and glycolytic metabolism through FoxO transcription factors (Jacinto et al. 2004; Inoki and Guan 2006; Masui et al. 2013).”

“The mechanism whereby mTOR inhibits autophagy involves dephosphorylation of ULKs (autophagy-initiating kinase ULK1, mammalian Atg1 homologs UNC-51-like kinase 1), Atg13 (autophagy-related gene 13), and FIP200 (focal adhesion kinase family-interacting protein of 200 kDa) (Jung et al. 2010).”

“FoxO3a also plays a role in stimulating autophagy (Mammucari et al. 2007).”

“…family GTPase,), and Pdia3 (protein disulfide isomerase family A member 3, also known as ERp57), an activator of the mTORC1 complex (Ram ırez et al. 2011), and Ras homolog enriched in brain (Rheb), a GTPase and key activator of mTORC1 that is inhibited by the Tsc1-Tsc2 complex (Inoki et al. 2003).”

“mTOR interacts with several proteins and forms two complexes, mTORC1 and mTORC2, initially characterized in yeast (Loewith et al. 2002; Wullachleger et al. 2006).”

“…mTOR-related signaling networks mTOR signaling plays key roles in regulating energy metabolism, cell growth, adaptive stress responses and autophagy, and abnormalities in mTOR signaling are implicated in cancers, obesity, and age-related diseases (Laplante and Sabatini 2012; Johnson et al. 2013).”

“Evidence that autophagy-related processes are responsive to mild mitochondrial uncoupling mTOR is a negative regulator of autophagy, a process that removes damaged proteins and organelles and recycles intracellular nutrient resources (Ravikumar et al. 2004; Yu et al. 2010).”

“(2015) 134, 677--692 factor FOXO3a (or FKHRL1; Brunet et al. 2001; Sahin et al. 2013). mTORC2 can activate AKT to repress FOXO1 and FOXO3 in mammalian cells (Guertin et al. 2006).”

“…processes including thermogenesis, mitochondrial redox balance and free radical production, cellular calcium homeostasis, and autophagy/mitophagy (Enerb€ack et al. 1997; Starkov and Fiskum 2003; Maragos and Korde 2004; Andrews et al. 2005; Liu et al. 2006; Caldeira da Silva et al. 2008; Mattson…”

“…were suppressed, and Foxo3a was up-regulated transiently in the cerebral cortex in response to DNP. Down-regulation of insulin receptor signaling may contribute to the mechanism whereby caloric restriction extends longevity in animal models of aging (Bl€uher et al. 2003; Mercken et al. 2013).”

“LC3b (microtubule-associated protein 1, light chain 3), an ubiquitin-like protein and mammalian ortholog of yeast Atg8 (Nakatogawa et al. 2007), exists in a cytosolic form (LC3b-I) and an autophagosome membrane-associated form (LC3b-II).”

“Sample quality was assessed by scatter plots, principal component analysis, and gene sample z-ratio score-based hierarchy clustering to exclude possible outliers (Figure S4, and see Cheadle et al. 2003).”

“…energetic challenges including exercise and intermittent fasting, which were previously reported to enhance autophagy and stimulate CREB signaling (Altarejos and Montminy 2011; Mattson 2012; Jamart et al. 2013; Yang et al. 2014), even though DNP has the specific effect to enhance energy (a)…”

“…(GABA-A receptor associated protein, LC3 family), p62 (an E3 ubiquitin ligase), Ulk1 (a protein critical for the formation of the isolation membrane in autophagy), and lysosomal heat-shock protein of 70 kDa (hsp70) which plays a key role in chaperone-mediated autophagy (Chiang et al. 1989) (Fig.”

“Up-regulated genes included JUNB, EGE1, Sgk1, and Egr1. mTORC2 has been reported to activate SGK1(serum and glucocorticoid regulated serine/threonine protein kinase 1) and control cellular ion homeostasis (Garcia-Martinez and Alessi 2008).”

“(2015) 134, 677--692 factor FOXO3a (or FKHRL1; Brunet et al. 2001; Sahin et al. 2013). mTORC2 can activate AKT to repress FOXO1 and FOXO3 in mammalian cells (Guertin et al. 2006).”

“Similar to other cell types, mitochondria generate the majority of energy substrates (ATP and NAD) in neurons, and energy availability influences cellular metabolism via core regulatory networks including those involving mammalian target of rapamycin (mTOR), AMPactivated protein kinase (AMPK) and insulin signaling pat…”

“Beclin/Atg6, a Bcl-2- interacting protein involved in the early stages of the autophagic complex formation (Liang et al. 1999; Kihara et al. 2001), was increased at both the mRNA and protein levels in cortical tissue samples from DNP-treated mice (Figure 5a and c).”

“Previous studies have demonstrated a complex interplay between PI3K/Akt, TSC and mTOR complexes (Shah et al. 2004; Huang and Manning 2009).”

“…in neurons, and energy availability influences cellular metabolism via core regulatory networks including those involving mammalian target of rapamycin (mTOR), AMP-activated protein kinase (AMPK), and insulin signaling pathways (Cheng et al. 2010; Inoki et al. 2012; Salminen andKaarniranta 2012).”

“While mitochondrial uncoupling can generate heat in brown fat cells (Brand 2000), the roles of mild mitochondrial uncoupling in neurons are unclear and likely complex.”

“…response to increased amino acid and energy substrate availability and growth factors. mTORC1 also regulates autophagy (Laplante and Sabatini 2012; Dibble and Manning 2013). mTORC2 responds to activation of Akt, SGK, and PKC, and controls cell survival, size, and glycolytic metabolism through…”

“Previous studies have provided evidence that impaired autophagy occurs in neurons in Alzheimer’s and Parkinson’s diseases (Nixon and Yang 2011; Manzoni and Lewis 2013), and that the mTOR inhibitor rapamycin can up-regulate autophagy and protect neurons against dysfunction and degeneration in…”

“Abnormalities in mitochondrial uncoupling are implicated in pathological conditions including obesity, insulin resistance/diabetes, cardiovascular disease, and neurodegenerative disorders (Vidal-Puig 2000; Chan and Harper 2006; Colman 2007; Tseng et al. 2010).”

“CREB is activated in response to excitatory synapse activation, BDNF, and exercise, and plays important roles in synaptic plasticity and learning and memory (for review see Benito and Barco 2010; Sakamoto et al. 2011).”

“The Tsc1/Tsc2 complex interacts with PI3K/Akt signaling such that inhibition of TSC1/TSC2 activates mTOR and disrupts PI3K-Akt signaling (Zhang et al. 2003, 2011).”

“…thermogenesis, mitochondrial redox balance and free radical production, cellular calcium homeostasis, and autophagy/mitophagy (Enerb€ack et al. 1997; Starkov and Fiskum 2003; Maragos and Korde 2004; Andrews et al. 2005; Liu et al. 2006; Caldeira da Silva et al. 2008; Mattson 2010; Mookerjee et al.…”

“ATF4 is a transcriptional regulator that increases in cells exposed to various stressful conditions (Baird and Wek 2012).”

“Previous studies have provided evidence that impaired autophagy occurs in neurons in Alzheimer’s and Parkinson’s diseases (Nixon and Yang 2011; Manzoni and Lewis 2013), and that the mTOR inhibitor rapamycin can up-regulate autophagy and protect neurons against dysfunction and degeneration in experimental models of the…”

“Similar Ca responses are known to trigger long-lasting (hours to days) changes in CREB-mediated gene expression and associated synaptic plasticity (Benito and Barco 2010; Suzuki et al. 2011).”

“…in neurons, and energy availability influences cellular metabolism via core regulatory networks including those involving mammalian target of rapamycin (mTOR), AMP-activated protein kinase (AMPK), and insulin signaling pathways (Cheng et al. 2010; Inoki et al. 2012; Salminen andKaarniranta 2012).”

“…including exercise and intermittent fasting, which were previously reported to enhance autophagy and stimulate CREB signaling (Altarejos and Montminy 2011; Mattson 2012; Jamart et al. 2013; Yang et al. 2014), even though DNP has the specific effect to enhance energy (a) (b) (c) (d) (e) Fig.”

“mTORC2 responds to activation of Akt, SGK, and PKC, and controls cell survival, size, and glycolytic metabolism through FoxO transcription factors (Jacinto et al. 2004; Inoki and Guan 2006; Masui et al. 2013).”

“BDNF can enhance glucose uptake and mitochondrial biogenesis in neurons, which may contribute to its important roles in synaptic plasticity and neuroprotection (Burkhalter et al. 2003; Cheng et al. 2012).”

“5b and e), suggesting the activation of chaperone-mediated autophagy and mitochondrial dynamics (Lee et al. 2011; Liesa and Shirihai 2013) in response to mild mitochondrial uncoupling.”

“…radical production, cellular calcium homeostasis, and autophagy/mitophagy (Enerb€ack et al. 1997; Starkov and Fiskum 2003; Maragos and Korde 2004; Andrews et al. 2005; Liu et al. 2006; Caldeira da Silva et al. 2008; Mattson 2010; Mookerjee et al. 2010; Youle and Narendra 2011; Ramsden et al.…”

“DNP has been reported to improve brain bioenergetics when administered systemically (Caldeira da Silva et al. 2008).”

“In addition, pharmacological agents that inhibit mTOR or activate AMPK can protect neurons against dysfunction and degeneration in animal models of acute brain injury and neurodegenerative disorders (Culmsee et al. 2001; Tain et al. 2009).”

“Mitochondrial uncoupling imposes an energetic stress on cells by causing a proton leak across the inner membrane which reduces the membrane potential and uncouples substrate oxidation from ADP phosphorylation, thereby increasing energy expenditure (Starkov 2006; Echtay 2007).”

“Cortical neuronal cell culture and cell viability assay Dissociated embryonic rat cerebral cortical cell cultures were established and maintained as described previously (Mattson et al. 1995).”

“In this view, the effects of DNP on brain cell gene expression documented in the present study may represent prolonged adaptive responses to a metabolic challenge similar to the adaptive responses of brain cells to vigorous exercise (Tong et al., 2001; Stranahan et al., 2010) and preconditioning ischemia (Stenzel-Poor…”

“Levels of 4-hydroxynonenal (HNE) lysine and histidine adducts (lipid peroxidation products) were measured using tandem mass spectrometry as described previously (Cutler et al. 2002).”

“In addition, pharmacological agents that inhibit mTOR or activate AMPK can protect neurons against dysfunction and degeneration in animal models of acute brain injury and neurodegenerative disorders (Culmsee et al. 2001; Tain et al. 2009).”

“…as fasting and exercise can be beneficial for neurons and brain health (Mattson 2012), and are neuroprotective in animal models of stroke, Parkinson’s disease, and Alzheimer’s disease (Halagappa et al. 2007; Arumugam et al. 2010; Zhang et al. 2011; Griffioen et al. 2013; Meller and Simon 2013).”

“Moreover, crosstalk between cAMP-mediated CREB signaling and the mTOR pathway have been reported previously, although the underlying molecular mechanisms are unclear (Carlon et al. 2010).”

“…were suppressed, and Foxo3a was up-regulated transiently in the cerebral cortex in response to DNP. Down-regulation of insulin receptor signaling may contribute to the mechanism whereby caloric restriction extends longevity in animal models of aging (Bl€uher et al. 2003; Mercken et al. 2013).”

“The Bdnf gene is a target of CREB and, conversely, BDNF signaling can activate CREB (Suzuki et al. 2011).”

“Cell viability was quantified using the dye Alamar blue (resazurin) as described previously (Liu et al. 2009).”

“In a recent study (Perry et al., 2013), when DNP was administered to rats at the same dose (5 mg/kg) and by the same route (i.”

“Ca2+ and CREB signaling play critical roles in synaptic plasticity and learning and memory (Sakamoto et al. 2011; Chen et al. 2012).”

“It was reported that rapamycin, an inhibitor of mTORC1, improves cognitive function in a mouse model of Alzheimer’s disease (Spilman et al. 2010; Majumder et al. 2012).”

“Mitochondrial uncoupling proteins regulate multiple physiological processes including thermogenesis, mitochondrial redox balance and free radical production, cellular calcium homeostasis, and autophagy/mitophagy (Enerb€ack et al. 1997; Starkov and Fiskum 2003; Maragos and Korde 2004; Andrews et al. 2005; Liu et al. 20…”

“…radical production, cellular calcium homeostasis, and autophagy/mitophagy (Enerb€ack et al. 1997; Starkov and Fiskum 2003; Maragos and Korde 2004; Andrews et al. 2005; Liu et al. 2006; Caldeira da Silva et al. 2008; Mattson 2010; Mookerjee et al. 2010; Youle and Narendra 2011; Ramsden et al. 2012).”

“CREB also mediates adaptive responses to metabolic and oxidative stress, including stimulation of DNA repair (Lee et al. 2009; Sebollela et al. 2010; Yang et al. 2010).”

“BDNF regulates neuronal energy metabolism and stress responses (Nakagawa et al. 2002; Arancibia et al. 2003; Larsen et al. 2010; Marosi and Mattson 2014).”

“Abnormalities in mitochondrial uncoupling are implicated in pathological conditions including obesity, insulin resistance/diabetes, cardiovascular disease, and neurodegenerative disorders (Vidal-Puig 2000; Chan and Harper 2006; Colman 2007; Tseng et al. 2010).”

“…including exercise and intermittent fasting, which were previously reported to enhance autophagy and stimulate CREB signaling (Altarejos and Montminy 2011; Mattson 2012; Jamart et al. 2013; Yang et al. 2014), even though DNP has the specific effect to enhance energy (a) (b) (c) (d) (e) Fig.”

“The gene encoding the tumor suppressor protein TSC2 (tuberous sclerosis complex 2), an important upstream regulator of mTOR (Russell et al. 2011; Yoshida et al. 2011), was upregulated at 24 h (red, z-ratio = 1.”

“Because such improvements in peripheral energy metabolism can enhance neuronal plasticity and can protect neurons against dysfunction and degeneration in animal models of Alzheimer’s disease and stroke (Mattson, 2012; Searcy et al., 2012), it is possible that some of the molecular responses of brain cells to DNP docum…”

“…in the present study may represent prolonged adaptive responses to a metabolic challenge similar to the adaptive responses of brain cells to vigorous exercise (Tong et al. 2001; Stranahan et al. 2010) and preconditioning ischemia (Stenzel-Poore et al. 2003) reported previously.”

“…including exercise and intermittent fasting, which were previously reported to enhance autophagy and stimulate CREB signaling (Altarejos and Montminy 2011; Mattson 2012; Jamart et al. 2013; Yang et al. 2014), even though DNP has the specific effect to enhance energy (a) (b) (c) (d) (e) Fig.”

“It is, therefore, important to determine the relative contributions of mTOR cascade signaling reprogramming to the neuroprotective actions of DNP in models of neurodegenerative disorders (Korde et al. 2005; Pandya et al. 2007; da Costa et al. 2010).”

“The gene encoding the tumor suppressor protein TSC2 (tuberous sclerosis complex 2), an important upstream regulator of mTOR (Russell et al. 2011; Yoshida et al. 2011), was upregulated at 24 h (red, z-ratio = 1.59, p < 0.05) and became more prominent at 72 h (z-ratio = 3.47, p < 0.01) in DNPtreated…”

“However, DNP can also reverse diet-induced obesity, hypertriglyceridemia, and insulin resistance in diet-induced obesity (Perry et al. 2013; Goldgof et al. 2014).”

“BDNF can enhance glucose uptake and mitochondrial biogenesis in neurons, which may contribute to its important roles in synaptic plasticity and neuroprotection (Burkhalter et al. 2003; Cheng et al. 2012).”

“BDNF regulates neuronal energy metabolism and stress responses (Nakagawa et al. 2002; Arancibia et al. 2003; Larsen et al. 2010; Marosi and Mattson 2014).”

“…radical production, cellular calcium homeostasis, and autophagy/mitophagy (Enerb€ack et al. 1997; Starkov and Fiskum 2003; Maragos and Korde 2004; Andrews et al. 2005; Liu et al. 2006; Caldeira da Silva et al. 2008; Mattson 2010; Mookerjee et al. 2010; Youle and Narendra 2011; Ramsden et al. 2012).”

“It is therefore likely that up-regulation of autophagy contributes to the neuroprotective effects of DNP in animal models of stroke (Korde et al. 2005) and traumatic nerve injury (da Costa et al. 2010).”

“Mild intermittent metabolic challenges such as fasting and exercise can be beneficial for neurons and brain health (Mattson 2012), and are neuroprotective in animal models of stroke, Parkinson’s disease, and Alzheimer’s disease (Halagappa et al. 2007; Arumugam et al. 2010; Zhang et al. 2011; Griffioen et al. 2013; Mel…”

“…of mitochondrial uncoupling proteins (UCP2 and UCP4) were genetically manipulated in neurons provided evidence that mitochondrial uncoupling affects Ca2+ signaling (Chan et al. 2006; Mehta and Li 2009) suggesting a mechanism whereby DNP treatment could activate the Ca2+– Arc–CaMKII–CREB pathway.”

“(2015) 134, 677--692 that BDNF contributes to bioenergetic responses of neurons to DNP as we found that cortical neurons treated with DNP exhibit enhanced glucose uptake (Liu et al. 2008).”

“Levels of cyoplasmic free calcium [Ca2+]i and mitochondrial calcium [Ca 2+]m were evaluated using the fluorescent probes Fluo-4 and Rhod-2, respectively, as described previously (Liu et al. 2010).”

“…radical production, cellular calcium homeostasis, and autophagy/mitophagy (Enerb€ack et al. 1997; Starkov and Fiskum 2003; Maragos and Korde 2004; Andrews et al. 2005; Liu et al. 2006; Caldeira da Silva et al. 2008; Mattson 2010; Mookerjee et al. 2010; Youle and Narendra 2011; Ramsden et al. 2012).”

“CREB also mediates adaptive responses to metabolic and oxidative stress, including stimulation of DNA repair (Lee et al. 2009; Sebollela et al. 2010; Yang et al. 2010).”

“…of mitochondrial uncoupling proteins (UCP2 and UCP4) were genetically manipulated in neurons provided evidence that mitochondrial uncoupling affects Ca2+ signaling (Chan et al. 2006; Mehta and Li 2009) suggesting a mechanism whereby DNP treatment could activate the Ca2+– Arc–CaMKII–CREB pathway.”

“Abnormalities in mitochondrial uncoupling are implicated in pathological conditions including obesity, insulin resistance/diabetes, cardiovascular disease, and neurodegenerative disorders (Vidal-Puig 2000; Chan and Harper 2006; Colman 2007; Tseng et al. 2010).”

“…as fasting and exercise can be beneficial for neurons and brain health (Mattson 2012), and are neuroprotective in animal models of stroke, Parkinson’s disease, and Alzheimer’s disease (Halagappa et al. 2007; Arumugam et al. 2010; Zhang et al. 2011; Griffioen et al. 2013; Meller and Simon 2013).”

“…redox balance and free radical production, cellular calcium homeostasis, and autophagy/mitophagy (Enerb€ack et al. 1997; Starkov and Fiskum 2003; Maragos and Korde 2004; Andrews et al. 2005; Liu et al. 2006; Caldeira da Silva et al. 2008; Mattson 2010; Mookerjee et al. 2010; Youle and Narendra…”

“The energy demand of neurons is acutely responsive to synaptic activity, and the resulting Ca2+ influx and ion-motive ATPase activities (Gellerich et al. 2013).”

“(2015) 134, 677--692 factor FOXO3a (or FKHRL1; Brunet et al. 2001; Sahin et al. 2013). mTORC2 can activate AKT to repress FOXO1 and FOXO3 in mammalian cells (Guertin et al. 2006).”

“It was previously reported that low concentrations of DNP increase cyclic AMP levels in cultured neurons (Wasilewska-Sampaio et al. 2005) which, in addition to Ca2+, would also be expected to activate CREB and up-regulate BDNF expression.”

“Mitochondrial uncoupling imposes an energetic stress on cells by causing a proton leak across the inner membrane which reduces the membrane potential and uncouples substrate oxidation from ADP phosphorylation, thereby increasing energy expenditure (Starkov 2006; Echtay 2007).”

“It is therefore likely that up-regulation of autophagy contributes to the neuroprotective effects of DNP in animal models of stroke (Korde et al. 2005) and traumatic nerve injury (da Costa et al. 2010).”

“CREB also mediates adaptive responses to metabolic and oxidative stress, including stimulation of DNA repair (Lee et al. 2009; Sebollela et al. 2010; Yang et al. 2010).”

“The DNP dose was chosen based on neuronal cell survival experiments (Figure S1a) and previous reports showing that it is non-toxic or neuroprotective in rodents (Kaiser 1964; Korde et al. 2005; Pandya et al. 2007; da Costa et al. 2010).”

“GLP-1 receptor agonists increase insulin sensitivity and are neuroprotective in animal models of AD, PD, and stroke (Li et al., 2009, 2010; Liu et al., 2015b).”

“…mitochondrial uncoupling agents), they become resistant to subsequent exposure to a high level of stress (e.g., metabolic, excitotoxic, or oxidative stressor) that would have killed them had they not been previously exposed to the mild stress (Marini & Paul, 1992; Lee et al, 1999; Liu et al, 2015).”

“Injection of DNP in C57BL/6 mice leads to the suppression of this pathway [94,251] and to the upregulation of genes involved in autophagy, such as LC3B, p62, or Ulk1 [94].”

“Both nuclear and mitochondrial CREB pathways promote neuronal survival in the brain [132, 133, 135], which is consistent with their positive roles in mitochondrial biogenesis.”

“This causes a lower influx of Ca2+ into the matrix via closing the uniporter, but simultaneously a rise in the cytosolic Ca2+ (see Figure 1) [30].”

“When mice were treated with DNP (5 mg/kg) once each day for 14 days, their performance on a learning and memory test was significantly enhanced compared with vehicle-treated control mice [56].”

“Moreover, DNP treatment improves the performance of mice in a learning and memory task (Liu et al., 2015).”

“The expression of neuronal uncoupling proteins (UCP) is induced by metabolic and oxidative challenges such as physical exercise and caloric restriction (Liu et al., 2015).”

“Importantly, the mild uncoupling induced by MP201 does not translate into fully uncoupled isolated mitochondria as evidenced in our assays, but is consistent in DNP chronic administration improving cognition and neuroprotection (Geisler et al., 2017; Liu et al., 2015; Wu et al., 2017).”

“A survey of the literature at this juncture indicates that uncouplers of mitochondrial respiration not only mitigate mitochondrial free radicals but, during this process, also activate cellular autophagy (39, 40).”

“This is consistent with previous studies that found low sustained levels of DNP administration ameliorate neurological disease processes and improve functional outcomes, without reducing body weight as seen at much higher doses [25, 58, 61, 70].”

“40,64,65 Collectively, this pharmacology appears pan-neuroprotective at low doses and may be neuroprotective with aging.”

“CR, suppresses mTOR leading to increased autophagy in mouse brain homogenates (Liu et al., 2015).”

“45,46 However, since amycretin did not increase energy expenditure above the vehicle group, this points towards a counter-regulatory effect on metabolic adaptation rather than an energy expenditure-increasing effect per se , which is consistent with that observed with administration of semaglutide…”

“Mild mitochondrial uncoupling in neurons has been shown to reprogram mTOR and insulin signaling, up-regulate signaling pathways associated with adaptive stress responses and synaptic plasticity, and improve learning and memory [50].”

“The inhibition of mTOR could have neuroprotective effects in AD by promoting the removal of damaged cells or mitochondria [54].”

“2,4-dinitrophenol (DNP) C6H4N2O5 Dissipation of the ΔΨm No [88] BAM15 C16H10F2N6O Dissipation of the ΔΨm No [89] Valinomycin C54H90N6O18 ΔΨm collapse due to K+ influx No [90] Salinomycin C42H70O11 Induce mitophagy, mitoptosis and increased ∆Ψ and reduced ATP level No [91]”

“Next, we administered DNP (5mg/kg, i.p.) in vivo, a low dose reported to be nontoxic in vivo (Liu et al., 2015).”

“…DNP suppresses the mammalian target of rapamycin (mTOR) pathway and stimulates the cAMP-response element-binding (CREB) protein–BDNF cascade, which is essential for synaptic plasticity, adaptive stress responses and cognitive function (Fernanda et al., 2007; Kandel et al., 2014; Liu et al., 2015).”

“The volume, count and size of individual mitochondria detected using Imaris were exported to an Excel file for further analysis. can stimulate autophagic responses in the cerebral cortex (Liu et al., 2015), which may be mediated via mTOR signaling.”

“Of interest, these effects are similar to those seen in caloric restriction [12, 15, 19] and are consistent with the “uncoupling to survive” hypothesis [20].”

“In mice, DNP treatment has been shown to improve learning and memory, consistent with molecular evidence of enhanced synaptic plasticity (Liu et al. 2015).”

“Microarray studies of DNP exposure in rat hippocampal neuronal cultures [35] and intact mouse cortex [36] revealed an up-regulation of genes involved in both cyclic adenosine 50monophosphate and phosphoinositide 3-kinase signaling, suggesting a neuroprotective role for DNP at low doses.”

“2014), as well as within cap-dependent translation (Liu et al. 2015); the possible influence of time of day or night on the magnitude of learning-induced enhanced polymerase I-driven nucleolar rRNA transcription is an important question, particularly, as recent reports show a daytime dependency of spatial learning (Ji…”