Axonal origin and purity of growth cones isolated from fetal rat brain.
The investigation of the molecular properties of nerve growth cones depends to a significant degree on their isolation from fetal brain in the form of 'growth cone particles' (GCPs). The availability of markers for developing axons and dendrites, as well as glial cells, has made it possible to characterize the GCP fraction in much greater detail than before and to optimize its yield. Marker analyses show that a member of the N-CAM family (5B4-CAM), synaptophysin, and especially GAP-43 and non-phosphorylated tau, are enriched in the GCP fraction. In contrast, MAP2 and, particularly, glial fibrillary acidic protein and vimentin are fractionated away from GCPs. Furthermore, GCP yield can be doubled relative to the original procedure, without compromising purity, by raising the sucrose concentration of the fractionation gradient's uppermost layer. The results indicate that GCPs are highly purified growth cone fragments with very little glial contamination, and that they are primarily of axonal origin.
Lohse
K
K
Department of Cellular and Structural Biology, University of Colorado School of Medicine, Denver 80262, USA.
Helmke
S M
SM
Wood
M R
MR
Quiroga
S
S
de la Houssaye
B A
BA
Miller
V E
VE
Negre-Aminou
P
P
Pfenninger
K H
KH
D000818:Animals; D001253:Astrocytes / Q000166:cytology; D001369:Axons / Q000648:ultrastructure*; D015415:Biomarkers; D001921:Brain / Q000196:embryology* / Q000648:ultrastructure; D002454:Cell Differentiation / Q000502:physiology; D002458:Cell Fractionation; D005314:Embryonic and Fetal Development / Q000502:physiology; D019922:GAP-43 Protein; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D008562:Membrane Glycoproteins / Q000032:analysis; D008869:Microtubule-Associated Proteins / Q000032:analysis; D009419:Nerve Tissue Proteins / Q000032:analysis*; D009457:Neuroglia / Q000648:ultrastructure; D009836:Oligodendroglia / Q000166:cytology; D051381:Rats; D016708:Synaptophysin / Q000032:analysis
D015415:Biomarkers; D019922:GAP-43 Protein; D005904:Glial Fibrillary Acidic Protein; D008562:Membrane Glycoproteins; D008869:Microtubule-Associated Proteins; D009419:Nerve Tissue Proteins; D016708:Synaptophysin