Determination of the metabolic index using the fluorescence lifetime of free and bound nicotinamide adenine dinucleotide using the phasor approach

Ranjit, Suman; Malacrida, Leonel; Stakic, Milka; Gratton, Enrico

doi:10.1002/jbio.201900156

Cited by 48 publications

(58 citation statements)

References 50 publications

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“…In order to directly assess whether PD-1 + B cells are more reliant on either glycolysis or oxidative phosphorylation (OXPHOS), FLIM was utilized. FLIM is based on the principle of endogenous fluorescence molecules such as NAD and as a result requires no staining, fixation, or other type of processing of the target cells (26). FLIM's capacity to distinguish between bound and unbound NAD is based on the self-quenching ability of NAD.…”

Section: Cxcr3 Is An Important Mediator For the Accumulation Of Memormentioning

confidence: 99%

Pathogenic, glycolytic PD-1+ B cells accumulate in the hypoxic RA joint

Floudas¹,

Neto²,

Marzaioli³

et al. 2020

JCI Insight

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Section: Cxcr3 Is An Important Mediator For the Accumulation Of Memormentioning

confidence: 99%

Pathogenic, glycolytic PD-1+ B cells accumulate in the hypoxic RA joint

Floudas¹,

Neto²,

Marzaioli³

et al. 2020

JCI Insight

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“…Here, we clearly show that free NADH increased in LPL deficient PVN neurons ( Figure 4 D). Typically, this would suggest an increased flux through glycolysis [ 35 , 36 , 37 ]. However, since neurons have a limited capacity to upregulate glycolysis [ 38 ], increased glucose utilization through the PPP in neurons is more likely [ 21 ].…”

Section: Discussionmentioning

confidence: 99%

Neuronal Lipoprotein Lipase Deficiency Alters Neuronal Function and Hepatic Metabolism

et al. 2020

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The autonomic regulation of hepatic metabolism offers a novel target for the treatment of non-alcoholic fatty liver disease (NAFLD). However, the molecular characteristics of neurons that regulate the brain-liver axis remain unclear. Since mice lacking neuronal lipoprotein lipase (LPL) develop perturbations in neuronal lipid-sensing and systemic energy balance, we reasoned that LPL might be a component of pre-autonomic neurons involved in the regulation of hepatic metabolism. Here, we show that, despite obesity, mice with reduced neuronal LPL (NEXCreLPLflox (LPL KD)) show improved glucose tolerance and reduced hepatic lipid accumulation with aging compared to wilt type (WT) controls (LPLflox). To determine the effect of LPL deficiency on neuronal physiology, liver-related neurons were identified in the paraventricular nucleus (PVN) of the hypothalamus using the transsynaptic retrograde tracer PRV-152. Patch-clamp studies revealed reduced inhibitory post-synaptic currents in liver-related neurons of LPL KD mice. Fluorescence lifetime imaging microscopy (FLIM) was used to visualize metabolic changes in LPL-depleted neurons. Quantification of free vs. bound nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) revealed increased glucose utilization and TCA cycle flux in LPL-depleted neurons compared to controls. Global metabolomics from hypothalamic cell lines either deficient in or over-expressing LPL recapitulated these findings. Our data suggest that LPL is a novel feature of liver-related preautonomic neurons in the PVN. Moreover, LPL loss is sufficient to cause changes in neuronal substrate utilization and function, which may precede changes in hepatic metabolism.

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“…4D). Typically, this would suggest an increased flux through glycolysis (49)(50)(51). However, since neurons have a limited capacity to up-regulate glycolysis (52), increased glucose utilization through the PPP in neurons is more likely (35).…”

Section: Dicussionmentioning

confidence: 99%

Neuronal Lipoprotein Lipase Deficiency alters Neuronal Function and Hepatic Metabolism

Bruce

Wang

Dobrinskikh

et al. 2020

Preprint

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Background: The autonomic regulation of hepatic metabolism offers a novel target for the treatment of non-alcoholic fatty liver disease (NAFLD). However, the molecular characteristics of neurons that regulate the brain-liver axis remain unclear. Since mice lacking neuronal lipoprotein lipase (LPL) develop perturbations in neuronal lipid-sensing and systemic energy balance, we reasoned that LPL may be a component of pre-autonomic neurons involved in the regulation of hepatic metabolism. Methods: Measures of glucose homeostasis in mice homozygous (NEXLPL-/-) and heterozygous (NEXLPL+/-) for neuronal LPL deficiency were compared to that of WT mice. A detailed analysis of hepatic glucose and lipid metabolism was also determined in NEXLPL+/- at 6-18 mo. To determine the effect of neuronal LPL deficiency on neuronal physiology, liver-related neurons were identified in the paraventricular nucleus (PVN) of the hypothalamus using the transsynaptic retrograde tracer PRV-152. In addition, we used Fluorescence Lifetime Imaging Microscopy (FLIM) as a novel method to visualize changes in neuronal metabolism following LPL-depletion directly in the PVN. Results: Here we show that despite obesity, mice with reduced neuronal LPL also show improved glucose tolerance and reduced hepatic lipid accumulation with aging, concomitant with reduced hepatic lipogenic gene expression (e.g. SCD1 and FADS2). Retroviral tracing and patch clamp studies revealed reduced inhibitory post-synaptic currents in liver-related neurons lacking LPL. Quantification of the free versus bound Nicotinamide Adenine Dinucleotide (NADH) and Flavin Adenine Dinucleotide (FAD), revealed that LPL loss resulted in altered substrate utilization characterized by increased glucose utilization and TCA cycle flux. These findings were recapitulated by analysis of global metabolites from hypothalamic cell lines either deficient in, or over-expressing, LPL. Conclusions: Our data suggest that LPL is a novel feature of liver–related preautonomic neurons in the PVN. Moreover, LPL loss is sufficient to alter neuronal metabolism and function, leading to changes in systemic glucose metabolism including improved hepatic function with age.

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Determination of the metabolic index using the fluorescence lifetime of free and bound nicotinamide adenine dinucleotide using the phasor approach

Cited by 48 publications

References 50 publications

Pathogenic, glycolytic PD-1+ B cells accumulate in the hypoxic RA joint

Pathogenic, glycolytic PD-1+ B cells accumulate in the hypoxic RA joint

Neuronal Lipoprotein Lipase Deficiency Alters Neuronal Function and Hepatic Metabolism

Neuronal Lipoprotein Lipase Deficiency alters Neuronal Function and Hepatic Metabolism

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