Comparative analysis of cytosolic and mitochondrial ATP synthesis in embryonic and postnatal hippocampal neuronal cultures

Show full item record



Permalink

http://hdl.handle.net/10138/42504

Citation

Surin , A M , Khiroug , S , Gorbacheva , L R , Khodorov , B I , Pinelis , V G & Khiroug , L 2013 , ' Comparative analysis of cytosolic and mitochondrial ATP synthesis in embryonic and postnatal hippocampal neuronal cultures ' , Frontiers in Molecular Neuroscience , vol. 5 , no. January , Article 102 . https://doi.org/10.3389/fnmol.2012.00102

Title: Comparative analysis of cytosolic and mitochondrial ATP synthesis in embryonic and postnatal hippocampal neuronal cultures
Author: Surin, Alexander M.; Khiroug, Serguei; Gorbacheva, Lubov R.; Khodorov, Boris I.; Pinelis, Vsevolod G.; Khiroug, Leonard
Other contributor: University of Helsinki, Neuroscience Center
Date: 2013
Language: eng
Number of pages: 15
Belongs to series: Frontiers in Molecular Neuroscience
ISSN: 1662-5099
DOI: https://doi.org/10.3389/fnmol.2012.00102
URI: http://hdl.handle.net/10138/42504
Abstract: ATP in neurons is commonly believed to be synthesized mostly by mitochondria via oxidative phosphorylation. Neuronal mitochondria have been studied primarily in culture, i.e., in neurons isolated either from embryos or from neonatal pups. Although it is generally assumed that both embryonic and postnatal cultured neurons derive their ATP from mitochondrial oxidative phosphorylation, this has never been tested experimentally. We expressed the FRET-based ATP sensor AT1.03 in cultured hippocampal neurons isolated either from E17 to E18 rat embryos or from P1 to P2 rat pups and monitored [ATP]c simultaneously with mitochondrial membrane potential ΔΨm; TMRM) and NAD(P)H autofluorescence. In embryonic neurons, transient glucose deprivation induced a near-complete decrease in [ATP]c, which was partially reversible and was accelerated by inhibition of glycolysis with 2-deoxyglucose. In the absence of glucose, pyruvate did not cause any significant increase in [ATP]c in 84% of embryonic neurons, and inhibition of mitochondrial ATP synthase with oligomycin failed to decrease [ATP]c. Moreover, ΔΨm was significantly reduced by oligomycin, indicating that mitochondria acted as consumers rather than producers of ATP in embryonic neurons. In sharp contrast, in postnatal neurons pyruvate added during glucose deprivation significantly increased [ATP]c (by 54 ± 8%), whereas oligomycin induced a sharp decline in [ATP]c and increased ΔΨm. These signs of oxidative phosphorylation were observed in all tested P1-P2 neurons. Measurement of ΔΨm with the potential-sensitive probe JC-1 revealed that neuronal mitochondrial membrane potential was significantly reduced in embryonic cultures compared to the postnatal ones, possibly due to increased proton permeability of inner mitochondrial membrane. We conclude that, in embryonic, but not postnatal neuronal cultures, ATP synthesis is predominantly glycolytic and the oxidative phosphorylation-mediated synthesis of ATP by mitochondrial F1Fo-ATPase is insignificant. © 2013 Surin, Khiroug, Gorbacheva, Khodorov, Pinelis and Khiroug.
Subject: Adenosine triphosphate
F1Fo-ATPase
Fluorescent protein
OxPhos
TMRM
adenosine triphosphate
deoxyglucose
glucose
oligomycin
proton transporting adenosine triphosphate synthase
pyruvic acid
reduced nicotinamide adenine dinucleotide phosphate oxidase
animal cell
animal tissue
article
autofluorescence imaging
brain mitochondrion
cell function
controlled study
developmental stage
embryo
enzyme inhibition
fluorescence resonance energy transfer
glycolysis
mitochondrial membrane potential
nerve cell culture
nonhuman
nucleotide metabolism
oxidative phosphorylation
perinatal period
protein expression
rat
3112 Neurosciences
Rights:


Files in this item

Total number of downloads: Loading...

Files Size Format View
fnmol_05_00102.pdf 2.766Mb PDF View/Open

This item appears in the following Collection(s)

Show full item record