Use of NAD(P)H and flavoprotein autofluorescence transients to probe neuron and astrocyte responses to synaptic activation
Open accessAbstract
Synaptic stimulation in brain slices is accompanied by changes in tissue autofluorescence, which are a consequence of changes in tissue metabolism. Autofluorescence excited by ultraviolet light has been most extensively studied, and is due to reduced pyridine nucleotides (NADH and NADPH, collectively termed NAD(P)H). Stimulation generates a characteristic compound NAD(P)H response, comprising an initial fluorescence decrease and then an overshooting increase that slowly recovers to baseline levels. Evoked NAD(P)H transients are relatively easy to record, do not require the addition of exogenous indicators and have good signal-noise ratios. These characteristics make NAD(P)H imaging methods very useful for tracking the spread of neuronal activity in complex brain tissues, however the cellular basis of synaptically-evoked autofluorescence transients has been the subject of recent debate. Of particular importance is the question of whether signals are due primarily to changes in neuronal mitochondrial function, and/or whether astrocyte metabolism triggered by glutamate uptake may be a significant contributor to the overshooting NAD(P)H fluorescence increases. This mini-review addresses the subcellular origins of NAD(P)H autofluorescence and the evidence for mitochondrial and glycolytic contributions to compound transients. It is concluded that there is no direct evidence for a contribution to NAD(P)H signals from glycolysis in astrocytes following synaptic glutamate uptake. In contrast, multiple lines of evidence, including from complimentary flavoprotein autofluorescence signals, imply that mitochondrial NADH dynamics in neurons dominate compound evoked NAD(P)H transients. These signals are thus appropriate for studies of mitochondrial function and dysfunction in brain slices, in addition to providing robust maps of postsynaptic neuronal activation following physiological activation.
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NAD(P)H autofluorescence is almost certainly dominated by mitochondrial NADH.
“The NAD(P)H autofluorescence is almost certainly dominated by mitochondrial NADH (Brennan et al., 2006; Shuttleworth, 2010; Berndt et al., 2015).”
From Neuronal Stimulation Triggers Neuronal Glycolysis and Not Lactate Uptake (2017) · cites this paper as Shuttleworth, 2010
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“, 2004), it is almost certainly due instead to replenishment of mitochondrial NADH in neurons bymultiplemeans (Brennan et al., 2006; Shuttleworth, 2010; Berndt et al., 2015): stimulation of tricarboxylic acid cycle dehydrogenases by calcium as well as increased shuttling of NADH from the cytosol (which may itself be s…”
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- Authors
- Shuttleworth, C. W.
- Affiliations
- University of New Mexico (US)
- Corresponding author
- Shuttleworth, C. William
- Venue
- Neurochemistry International
- Publisher
- Elsevier BV
- Published
- 2010-02
- Volume
- 56
- Issue
- 3
- Pages
- 379-386
- ISSN
- 0197-0186 (ISSN-L)
- PMC
- PMC3085275
- OpenAlex
- W2149009994
- Cited by
- 104
- References
- 67
- Citations by year
- 2026: 5 · 2025: 3 · 2024: 6 · 2023: 3 · 2022: 6 · 2021: 5 · 2020: 6 · 2019: 3 · 2018: 7 · 2017: 13 · 2016: 6 · 2015: 5 · 2014: 12 · 2013: 6 · 2012: 4
- FWCI
- 0.52
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Neurochemistry International (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institute of General Medical Sciences (P20GM109089)
- National Institute of Neurological Disorders and Stroke (NS051288, R01NS051288)
- Topics
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Optical Imaging and Spectroscopy Techniques
- Keywords
- NAD(P)H autofluorescence, synaptic activation, astrocyte metabolism, mitochondrial function, glycolysis, glutamate uptake, neuronal activity imaging, brain slices
- MeSH terms
- Animals, Astrocytes, Brain, Flavoproteins, Fluorescent Dyes, Humans, Mitochondria, NADP, Synaptic Transmission, Neurochemistry, Neurons, Metabolic Networks and Pathways
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