Exposure of cultured primary rat astrocytes to hypoxia results in intracellular glucose depletion and induction of glycolytic enzymes.
Based on the neurotrophic properties of astrocytes in response to ischemia, the current work focuses on the mechanism for cultured astrocytes to adapt to a hypoxic environment. Intracellular glucose levels in primary cultured rat astrocytes exposed to hypoxia fell by 30% within 24 h, in parallel with a decrease in glycogen stores. Glycolytic metabolism was crucial for cell survival during hypoxia, as 2-deoxyglucose resulted in rapid ATP depletion and cell death. The mechanism for maintaining glucose levels under these conditions appeared to be mobilization of glycogen stores, rather than increased extracellular uptake of glucose, as gluconolactone (an inhibitor of beta1-4 amyloglucosidase) induced a rapid fall in cellular ATP in cultures subjected to hypoxia, whereas cytochalasin B was without affect. Addition of cycloheximide diminished the viability of astrocytes in hypoxia, suggesting an obligatory role of de-novo gene expression to respond to hypoxia. Consistently, the results of differential display suggested the induction of glycolytic enzymes, including aldolase A (EC 4.1.2.13), hexokinase II (ATP: D-hexose 6-phosphotransferase, EC 2.7.1.1), and triosephosphate isomerase (EC 5.3.1.1) in the hypoxic culture. Marked induction of these glycolytic enzymes in hypoxic astrocytes was confirmed by Northern blot analysis. These data provide a theoretical basis to understand the ability of astrocytes to tolerate ischemic condition.
Niitsu
Y
Y
Department of Anatomy and Neuroscience, Osaka University Medical School, 2-2 Yamada-oka, Suita, Japan.
Hori
O
O
Yamaguchi
A
A
Bando
Y
Y
Ozawa
K
K
Tamatani
M
M
Ogawa
S
S
Tohyama
M
M
D000818:Animals; D000831:Animals, Newborn; D000963:Antimetabolites / Q000494:pharmacology; D001253:Astrocytes / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D015152:Blotting, Northern; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003513:Cycloheximide / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D003847:Deoxyglucose / Q000494:pharmacology; D004790:Enzyme Induction; D004798:Enzymes / Q000235:genetics / Q000378:metabolism*; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D005947:Glucose / Q000378:metabolism* / Q000493:pharmacokinetics; D006003:Glycogen / Q000378:metabolism; D006019:Glycolysis; D006593:Hexokinase / Q000235:genetics / Q000378:metabolism; D000860:Hypoxia / Q000503:physiopathology*; D010100:Oxygen / Q000494:pharmacology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D012313:RNA / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014305:Triose-Phosphate Isomerase / Q000235:genetics / Q000378:metabolism
D000963:Antimetabolites; D018076:DNA, Complementary; D004798:Enzymes; D011500:Protein Synthesis Inhibitors; D012313:RNA; D006003:Glycogen; D003513:Cycloheximide; D003847:Deoxyglucose; D006593:Hexokinase; D005634:Fructose-Bisphosphate Aldolase; D014305:Triose-Phosphate Isomerase; D005947:Glucose; D010100:Oxygen