Mechanisms and Consequences of Dopamine Depletion-Induced Attenuation of the Spinophilin/Neurofilament Medium Interaction

Figure 1

STRING map of proteins with altered expression in spinophilin immunoprecipitates isolated from DA lesioned compared to intact striatal lysates. The substantia nigra of adult C57Bl6 mice was unilaterally lesioned with 6-OHDA. Striata from the intact and lesioned hemispheres were collected, fractionated into a membrane and synaptic fraction, and immunoprecipitated for spinophilin using a goat spinophilin antibody. (a) The regions of the gel containing spinophilin and NF-M (red boxes) and spinophilin interacting proteins (black boxes) were excised and analyzed by mass spectrometry. (b) Striatal lysates from the intact (I) and lesioned (L) hemispheres were stained for a total protein stain (Ponceau S) and subsequently immunoblotted for tyrosine hydroxylase. The Uniprot ID for proteins detected in spinophilin immunoprecipitates that had a >18% decrease or increase in spectral counts isolated from either the extrasynaptic (c and d, respectively) or synaptic (e and f, respectively) fractions was inputted into the string database (http://www.string-db.org) to demonstrate connectivity between proteins. Proteins were assigned to categories based on protein function. The gene names for the proteins are used as identifiers. Mass spectrometry was performed on an Orbitrap Velos mass spectrometer.

(a)

(b)

(c) S2 decreased

(d) S2 increased

(e) S3 decreased

(f) S3 increased