Glia perform several energy-dependent functions that may aid neuronal survival under pathological conditions. Glycogen is the major energy reserve in brain, and it is localized almost exclusively to astrocytes. Using murine cortical cell cultures containing both glia and neurons, we examined the effect of altered glial glycogen stores on neuronal survival following glucose deprivation. As previously reported, cultures exposed for several hours to media lacking glucose developed widespread neuronal degeneration without glial degeneration. If glial astrocyte glycogen content was increased to 2–3 times control levels by a 24-h pretreatment with 1 μM insulin or 0.5 mM methionine sulfoximine (MSO), glucose deprivation-induced neuronal degeneration was attenuated. These protective effects were blocked if glycogen levels were reduced back to control levels by a 30-min exposure to 1 mM dibutyryl cyclic AMP or 20 μM norepinephrine prior to glucose deprivation. Astrocyte glycogen stores may be an important factor influencing neuronal survival under conditions of energy substrate limitation.
BradfordHFWardHKThomasAJ. (1978) Glutamine—a major substrate for nerve endings. J Neurochem30:1453–1459
2.
BrucknerRBiesoldD, (1981) Histochemistry of glycogen deposition in perinatal rat brain: Importance of radial glial cells. J Neurocytol10:749–757
3.
BrunnerEAPassoneauJVMolstadC, (1971) The effect of volatile anesthetics on levels of metabolites and on metabolic rate in brain. J Neurochem18:2301–2316
4.
BungeRPWaksmanBH. (1985) Glial development and interactions. Trends Neurosci8:424–427
5.
CataldoAMBroadwellRD. (1986) Cytochemical identification of cerebral glycogen and glucose-6-phosphate activity under normal and experimental conditions. I. Neurons and glia. J Electron Microsc Tech3:413–437
6.
ChoiDW. (1990) Cerebral hypoxia: Some new approaches to unanswered questions. J Neurosci10:2493–2501
7.
CooperAJLLaiJCK. (1987) Cerebral ammonia metabolism in normal and hyperammonemic rats. Neurochem Pathol6:67–74
8.
EricinskaMTroegerMBWilsonDFSilverIA. (1986) The role of glial cells in regulation of neurotransmitter amino acids in the external environment. II. Mechanisms of aspartate transport. Brain Res369:203–214
9.
GueldrySRochetteLBraletJ, (1987) Comparison of the effects of valproate, ethosuximide, phenytoin, and pentobarbital on cerebral energy metabolism. Epilepsia28:160–168
10.
HarrisKMRulfDRosenbergPA. (1989) Ultrastructural relationships between neurons and glia modulating glutamate toxicity in cortical cultures. Soc Neurosci Abstr15:764
11.
HertzL, (1979) Functional interactions between neurons and astrocytes I. Turnover and metabolism of putative amino acid neurotransmitters. Prog Neurobiol13:277–323
12.
HertzL, (1981) Features of astrocytic function apparently involved in the response of central nervous tissue to ischemiahypoxia. J Cereb Blood Flow Metab1:143–153
13.
HevorTKDelormeP, (1991) Biochemical and ultrastructural study of glycogen in cultured astrocytes submitted to the convulsant methionine sulfoximine. Glia4:64–69
14.
HutchinsDARogersKJ. (1970) Physiological and drug-induced changes in the glycogen content of mouse brain. Br J Pharmacol39:9–25
15.
JendelovaPSykovà (1991) Role of glia in K+ and pH homeostasis in the neonatal rat spinal cord. Glia4:56–63
16.
KnullHRKhandelwahlRL. (1982) Glycogen metabolizing enzymes in brain. Neurochem Res7:1307–1317
17.
KohJ-YChoiDW. (1987) Quantitative determination of glutamate mediated neuronal injury in cell culture by lactate dehydrogenase efflux assay. J Neurosci Meth20:83–90
18.
KohJ-YChoiDW. (1988) Vulnerability of cultured cortical neurons to damage by excitotoxins: Differential susceptibility of neurons containing NADPH-diaphorase. J Neurosci8:2153–2163
19.
KoizumiJ, (1974) Glycogen in the central nervous system. Prog Histochem Cytochem6:1–35
20.
LowryOHRosebroughNJFarrALRandallRJ. (1951) Protein measurement with the Folin phenol reagent. J Biol Chem193:265–275
21.
LowryOHPassonneauJVHasselbergerFHSchulzDW. (1964) Effect of ischemia on known substrates and cofactors of the glycolytic pathway in brain. J Biol Chem239:18–30
22.
Martinez-HernandezABellKPNorenbergMD. (1977) Glutamine synthetase: Glial localization in brain. Science195:1356–1358
MonyerHChoiDW. (1990) Glucose deprivation neuronal injury in vitro is modified by withdrawal of extracellular glutamine. J Cereb Blood Flow Metab10:337–342
25.
NelsonSRSchulzDWPassonneauJVLowryOH. (1968) Control of glycogen levels in brain. J Neurochem15:1271–1279
26.
OplerLMakmanMH. (1972) Mediation by cyclic AMP of hormone-stimulated glycogenolysis in cultured rat astrocytoma cells. Biochem Biophys Res Comm46:1142–1145
27.
PassonneauJVBrunnerEAMolstadCPassonneauR, (1971) The effects of altered endocrine states and of ether anesthesia on mouse brain. J Neurochem18:2317–2328
28.
PassonneauJVSchwartzJPLustWD. (1978) Some aspects of intermediary metabolism in glioma cells in culture. In: Dynamic Properties of Glial Cells (SchoffenielsEFranckGHertzLTowerDB, eds), Oxford, Pergamon Press, pp 133–142
29.
PentreathVWSealLHMorrisonJHMagistrettiPJ. (1986) Transmitter mediated regulation of energy metabolism in nervous tissue at the cellular level. Neurochem Int9:1–10
30.
PhelpsCH. (1975) An ultrastructural study of methionine sulphoximine-induced glycogen accumulation in astrocytes of the mouse cerebral cortex. J Neurocytol4:479–490
31.
PrasannanKGSubrahmanyamK, (1966) Effect of insulin on the synthesis of glycogen in cerebral cortical slices of alloxan diabetic rats. Endocrinology82:1–6
32.
RaoSHNMeisterA, (1972) In vivo formation of methionine sulfoximine phosphate, a protein bound metabolite of methionine sulfoximine. Biochemistry11:1123–1127
33.
ReinhartPHPfeifferBSpenglerSHamprechtB, (1990) Purification of glycogen phosphorylase from bovine brain and immunocytochemical examination of rat glial primary cultures using monoclonal antibodies raised against this enzyme. J Neurochem54:1474–1483
34.
RoseKGoldbergMPChoiDW. (1993) Cytotoxicity in murine cell culture. In: Methods in Enzymology (TysonCAFrazierJM, eds), San Diego, Academic Press, pp. 46–60 (TysonCAWitschH, eds. In vitro biologic systems; vol. 1)
35.
RosenbergPADichterMA. (1985) Glycogen accumulation in rat cerebral cortex in dissociated cell culture. J Neurosci Meth15:101–112
36.
SchurrAWestCARigorBM. (1988) Lactate-supported synaptic function in the rat hippocampal slice preparation. Science240:1326–1328
37.
SelakISkaperSDVaronS, (1985) Pyruvate participation in the low molecular weight trophic activity for central nervous system neurons in glia-conditioned media. J Neurosci5:23–28
38.
SiesjöBK. (1978) Brain Energy Metabolism, New York, John Wiley and Sons, pp 196–209
39.
SiesjöBKBengtssonF, (1989) Calcium fluxes, calcium antagonists, and calcium-related pathology in brain ischemia, hypoglycemia, and spreading depression. A unifying hypothesis. J Cereb Blood Flow Metab9:127–140
40.
SomersDLBecksteadRM. (1990) Chronic methionine sulfoximine administration reduces synaptosomal aspartate and glutamate in rat striatum. Neurosci Lett11:335–340
41.
SubbaraoKVHertzL, (1990) Effect of adrenergic agonists or glycogenolysis in primary cultures of astrocytes. Brain Res36:220–226
42.
SugiyamaKBrunoriAMayerML. (1989) Glial uptake of excitatory amino acids influences neuronal survival in culture of mouse hippocampus. Neuroscience32:779–791
43.
SwansonRAYuACHSharpFRChanPH. (1989) Regulation or glycogen content in primary astrocyte culture: Effects of glucose analogues, phenobarbital, and methionine sulfoximineJ Neurochem52:1359–1365
44.
SwansonRAYuACHChanPHSharpFR. (1990a) Glutamat increases glycogen content and reduces glucose utilization i primary astrocyte culture. J Neurochem54:490–496
45.
SwansonRAShiraishiKMortonMTSharpFR. (1990b) Methionine sulfoximine reduces cortical infarct size in rats after middle cerebral artery occlusion. Stroke21:322–327
46.
TewsJKStoneWE. (1964) Effects of methionine sulfoximine c levels of free amino acids and related substances in brainBiochem Biophys Res Comm13:543–545
47.
TombaughGCYangSSwansonRASapolskyRM. (1992) Glucocorticoids exacerbate hypoxic and hypoglycemic hippocampal injury in vitro: Biochemical correlates and a role for astrocytes. J Neurochem, 59:137–146
48.
TsacopoulosMEvequoz-MercierVPerrottetPBuchnerE, (1988) Honeybee retinal glial cells transform glucose and supply the neurons with metabolic substrate. Proc Natl Acad Sci USA85:8727–8731
49.
VaronSSomjenGG. (1979) Neuron-glia interactions. Neurosci Res Prog Bull17:1–239
50.
VibulsrethSHeftiFGinsbergMDDietrichWDBustoR, (1987) Astrocytes protect cultured neurons from degeneration induced by anoxia. Brain Res422:303–311
51.
WalzW, (1989) Role of glial cells in the regulation of the brain ion microenvironment. Prog Neurobiol33:309–333
52.
WalzWMukerjiS, (1988) Lactate production and release in cultured astrocytes. Neurosci Lett86:296–300
53.
WaniewskiRAMartinDL. (1986) Exogenous glutamate is metabolized to glutamine and exported by rat primary astrocyte cultures. J Neurochem47:304–313
54.
WenzelJLammertGMeyerUKrugM, (1991) The influence of long-term potentiation on the spatial relationship between astrocyte processes and potentiated synapses in the dentate gyrus of rat brain. Brain Res560:122–131
55.
YudkoffMNissimIPleasureD, (1988) Astrocyte metabolism of [15N]glutamine: Implications for the glutamine-glutamate cycle. J Neurochem51:843–850
56.
ZielkeHRTildonJTBaabPJHopkinsIB. (1989) Synthesis of glutamate and glutamine in dibutyryl cyclic AMP-treated astrocytes. Neurosci Lett97:209–214
