The effect of active serum albumin on PC12 cells: I. Neurite retraction and activation of the phosphoinositide second messenger system.
Vertebrate blood sera contain a factor that triggers oscillatory chloride currents in Xenopus oocytes through activation of the phosphoinositide/Ca2+ second system. The active serum component consists of lipids bound to an isoform of serum albumin that we have named active serum albumin (ASA). In undifferentiated PC12 cells, micromolar concentrations of ASA inhibit the early morphological changes induced by NGF, whereas in differentiated PC12 cells ASA caused a rapid withdrawal of neurites, which was reversible and dependent upon culture age. In contrast to normal serum, plasma and thrombin did not cause neurite retraction. Preincubation of ASA with monospecific antibodies to serum albumin suppressed its ability to induce neurite retraction in a dose dependent fashion. As in the oocyte, ASA activated the phosphatidylinositol second messenger system of PC12 cells, causing a several fold increase in Ins1,4,5P3 levels within minutes of application. The Ins1,4,5P3 increase was also blocked, in a titratable fashion, when ASA was preincubated with monospecific antibodies to serum albumin. This suggests that ASA-induced neurite retraction in PC12 cells may depend, at least in part, on activation of the phosphatidylinositol second messenger system. Results involving albumin-depleted sera show that ASA is the main factor responsible for serum vulnerability of neurites in PC12 cells. These findings point to some limitations in the use of serum in culture media, and raise the possibility that the serum factor may impair neuronal plasticity in disorders that are accompanied by the activation of blood coagulation together with a breakdown of the blood-brain barrier.
Dyer
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D
Department of Psychobiology, University of California, Irvine 92717.
Tigyi
G
G
Miledi
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R
D000818:Animals; D000906:Antibodies / Q000276:immunology; D002454:Cell Differentiation / Q000187:drug effects; D006801:Humans; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D009414:Nerve Growth Factors / Q000037:antagonists & inhibitors*; D016501:Neurites / Q000187:drug effects*; D016716:PC12 Cells; D010716:Phosphatidylinositols / Q000378:metabolism*; D015290:Second Messenger Systems / Q000502:physiology*; D012709:Serum Albumin / Q000276:immunology / Q000494:pharmacology*
D000906:Antibodies; D009414:Nerve Growth Factors; D010716:Phosphatidylinositols; D012709:Serum Albumin; D015544:Inositol 1,4,5-Trisphosphate