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High-resolution imaging of protein phosphorylation in squid axons and synapse by electron energy loss spectroscopy

R Martin, R Door and D Breitig

Electron Microscopy Section, University of Ulm, Germany.

We demonstrate that clusters of phosphorus atoms can be detected in energy loss spectroscopic images (ESI) of cytoskeletal proteins of squid axons. In series of images taken at four energy windows below and three windows above the phosphorus P-L2,3 ionization edge, signal-to- background intensity differences were analyzed by videodensitometry. A distinct increase of relative intensities was recorded above the phosphorus edge in neurofilaments of the peripheral giant axon and in those of the presynaptic terminal. A high level of neurofilament phosphorylation in the peripheral axon supports previous biochemical and immunochemical results, but our finding of phosphorylated neurofilaments in the presynaptic axon conflicts with these studies. Our method may be advantageous for analysis with high elemental and spatial resolution of the phosphorylation state of cytoskeletal protein molecules in situ.

Volume 41, Issue 8, pp. 1133-1139, 08/01/1993
Copyright © 1993 by The Histochemical Society


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