TY - JOUR
T1 - Analysis of the growth cone turning assay for studying axon guidance
AU - Pujic, Zac
AU - Giacomantonio, Clare E.
AU - Unni, Divya
AU - Rosoff, William J.
AU - Goodhill, Geoffrey J.
N1 - Funding Information:
This work was supported by grants to GJG from the Australian Research Council (DP0666126) and the Australian Mental Health and Medical Research Council (project grant 456003), and by the Queensland Brain Institute, School of Physical Sciences and Institute for Molecular Bioscience, The University of Queensland.
PY - 2008/5/30
Y1 - 2008/5/30
N2 - The "pipette" or "growth cone turning" assay is widely used for studying how axons respond to diffusible guidance cues in their environment. However, little quantitative analysis has been presented of the gradient shapes produced by this assay, or how they depend on parameters of the assay. Here we used confocal microscopy of fluorescent gradients to characterize these shapes in 3 dimensions. We found that the shape, and more specifically the concentration at the position usually occupied by the growth cone in this assay, varied in sometimes unexpected ways with the molecular weight of the diffusible factor, charge, pulse duration and pulse frequency. These results suggest that direct observation of the gradient of the particular guidance factor under consideration may be necessary to quantitatively determine the signal to which the growth cone is responding.
AB - The "pipette" or "growth cone turning" assay is widely used for studying how axons respond to diffusible guidance cues in their environment. However, little quantitative analysis has been presented of the gradient shapes produced by this assay, or how they depend on parameters of the assay. Here we used confocal microscopy of fluorescent gradients to characterize these shapes in 3 dimensions. We found that the shape, and more specifically the concentration at the position usually occupied by the growth cone in this assay, varied in sometimes unexpected ways with the molecular weight of the diffusible factor, charge, pulse duration and pulse frequency. These results suggest that direct observation of the gradient of the particular guidance factor under consideration may be necessary to quantitatively determine the signal to which the growth cone is responding.
KW - Axon guidance
KW - Pipette assay
KW - Turning assay
UR - http://www.scopus.com/inward/record.url?scp=41949085887&partnerID=8YFLogxK
U2 - 10.1016/j.jneumeth.2008.01.014
DO - 10.1016/j.jneumeth.2008.01.014
M3 - Article
C2 - 18313760
AN - SCOPUS:41949085887
VL - 170
SP - 220
EP - 228
JO - Journal of Neuroscience Methods
JF - Journal of Neuroscience Methods
SN - 0165-0270
IS - 2
ER -