Effect of graded hypoxia on the high-affinity CPP binding site of the NMDA receptor in the cerebral cortex of newborn piglets.
Previous studies have shown that the N-methyl-D-aspartate (NMDA) receptor is modified during hypoxia in the cerebral cortex of newborn piglets. The present study tests the hypothesis that the NMDA receptor 3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid (CPP) high-affinity binding site is modified during hypoxia and that the degree of modification correlates with the progressive decrease in cerebral cellular energy metabolism and increase in lipid peroxidation induced by hypoxia. Studies were conducted in twelve anesthetized, ventilated newborn piglets, five normoxic and seven hypoxic which were exposed to decreased fraction of inspired oxygen (FiO2) to achieve varying phosphocreatine (PCr) levels. 3[H]-CPP binding was performed with CPP concentrations ranging from 0.5 to 1500 nM at 23 degrees C for 40 min in P2 membrane fractions. Brain tissue PCr levels were determined biochemically. Conjugated dienes (CDs) were measured as an index of lipid peroxidation. In the normoxic group, B(max) (receptor number) for the CPP binding site was 329+/-93 fmol/mg protein and Kd (dissociation constant) 137+/-44 nM, the mean PCr value was 2.5+/-0.4 micromol/g brain and the CD level was 0.0 nmol/g brain. As tissue hypoxia worsened, there was a gradual decline in tissue PCr as well as receptor B(max) and K(d) values, and there was an increase in conjugated dienes. Both the receptor B(max) (r=0.90) and Kd (r=0.72) decreased in a linear relationship as PCr decreased. As the levels of CDs increased both the receptor B(max) (r=0.88) and Kd (r=0.68) decreased in a linear fashion. The data show that there is not a critical hypoxic threshold for modification of the CPP binding site of the NMDA receptor, but that modification is coupled to a gradual decrease in brain cell energy metabolism and increase in lipid peroxidation. We speculate that hypoxia-induced modification of the NMDA receptor is mediated not only by changes in the receptor recognition site but also by an alteration of brain cell membrane structure secondary to conjugated diene formation.
Fritz
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KI
Department of Pediatrics, St. Christopher's Hospital for Children, MCP Hahnemann University, Medical College of Pennsylvania, Neonatology Research Laboratory, 7th Floor Heritage Building, 3300 Henry Ave., Philadelphia, PA 19129, USA. fritzki@hotmail.com
Zanelli
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S
Mishra
O P
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Delivoria-Papadopoulos
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M
D000818:Animals; D000831:Animals, Newborn / Q000378:metabolism; D001238:Asphyxia Neonatorum / Q000378:metabolism / Q000503:physiopathology; D001665:Binding Sites / Q000187:drug effects / Q000502:physiology; D002462:Cell Membrane / Q000378:metabolism; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism* / Q000503:physiopathology; D004195:Disease Models, Animal; D004734:Energy Metabolism / Q000502:physiology; D018691:Excitatory Amino Acid Antagonists / Q000378:metabolism* / Q000493:pharmacokinetics; D006801:Humans; D002534:Hypoxia, Brain / Q000378:metabolism* / Q000503:physiopathology; D007231:Infant, Newborn; D015227:Lipid Peroxidation / Q000502:physiology; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D010879:Piperazines / Q000378:metabolism* / Q000493:pharmacokinetics; D011869:Radioligand Assay; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects / Q000378:metabolism*; D013552:Swine; D014316:Tritium
D018691:Excitatory Amino Acid Antagonists; D010879:Piperazines; D016194:Receptors, N-Methyl-D-Aspartate; D014316:Tritium; C050592:3-(2-carboxypiperazin-4-yl)propyl-1-phosphonic acid