Glutamine synthetase deficiency in murine astrocytes results in neonatal death.
Glutamine synthetase (GS) is a key enzyme in the "glutamine-glutamate cycle" between astrocytes and neurons, but its function in vivo was thus far tested only pharmacologically. Crossing GS(fl/lacZ) or GS(fl/fl) mice with hGFAP-Cre mice resulted in prenatal excision of the GS(fl) allele in astrocytes. "GS-KO/A" mice were born without malformations, did not suffer from seizures, had a suckling reflex, and did drink immediately after birth, but then gradually failed to feed and died on postnatal day 3. Artificial feeding relieved hypoglycemia and prolonged life, identifying starvation as the immediate cause of death. Neuronal morphology and brain energy levels did not differ from controls. Within control brains, amino acid concentrations varied in a coordinate way by postnatal day 2, implying an integrated metabolic network had developed. GS deficiency caused a 14-fold decline in cortical glutamine and a sevenfold decline in cortical alanine concentration, but the rising glutamate levels were unaffected and glycine was twofold increased. Only these amino acids were uncoupled from the metabolic network. Cortical ammonia levels increased only 1.6-fold, probably reflecting reduced glutaminolysis in neurons and detoxification of ammonia to glycine. These findings identify the dramatic decrease in (cortical) glutamine concentration as the primary cause of brain dysfunction in GS-KO/A mice. The temporal dissociation between GS(fl) elimination and death, and the reciprocal changes in the cortical concentration of glutamine and alanine in GS-deficient and control neonates indicate that the phenotype of GS deficiency in the brain emerges coincidentally with the neonatal activation of the glutamine-glutamate and the associated alanine-lactate cycles.
He
Youji
Y
AMC Liver Center and Department of Anatomy and Embryology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.
Hakvoort
Theodorus B M
TB
Vermeulen
Jacqueline L M
JL
Labruyère
Wilhelmina T
WT
De Waart
D Rudi
DR
Van Der Hel
W Saskia
WS
Ruijter
Jan M
JM
Uylings
Harry B M
HB
Lamers
Wouter H
WH
D000263:Adenylate Kinase / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism; D000641:Ammonia / Q000378:metabolism; D000704:Analysis of Variance; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000502:physiology*; D001786:Blood Glucose / Q000378:metabolism; D001921:Brain / Q000378:metabolism / Q000473:pathology; D053148:Caspase 3 / Q000378:metabolism; D016923:Cell Death / Q000235:genetics; D004195:Disease Models, Animal; D004622:Embryo, Mammalian; D004734:Energy Metabolism / Q000235:genetics; D005452:Fluoresceins; D015870:Gene Expression / Q000235:genetics; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics; D005974:Glutamate-Ammonia Ligase / Q000172:deficiency* / Q000235:genetics; D006801:Humans; D008659:Metabolic Diseases / Q000235:genetics* / Q000401:mortality*; D051379:Mice; D018345:Mice, Knockout; D009930:Organic Chemicals; D012333:RNA, Messenger / Q000378:metabolism
D000596:Amino Acids; D001786:Blood Glucose; D005452:Fluoresceins; D005904:Glial Fibrillary Acidic Protein; D009930:Organic Chemicals; D012333:RNA, Messenger; C435731:fluoro jade; D000641:Ammonia; D000263:Adenylate Kinase; D053148:Caspase 3; D005974:Glutamate-Ammonia Ligase