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2014 ; 15
(7
): 762-71
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A biophysical analysis of mitochondrial movement: differences between transport
in neuronal cell bodies versus processes
#MMPMID24673933
Narayanareddy BR
; Vartiainen S
; Hariri N
; O'Dowd DK
; Gross SP
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2014[Jul]; 15
(7
): 762-71
PMID24673933
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There is an increasing interest in factors that can impede cargo transport by
molecular motors inside the cell. Although potentially relevant (Yi JY,
Ori-McKenney KM, McKenney RJ, Vershinin M, Gross SP, Vallee RB. High-resolution
imaging reveals indirect coordination of opposite motors and a role for LIS1 in
high-load axonal transport. J Cell Biol 2011;195:193-201), the importance of
cargo size and subcellular location has received relatively little attention.
Here we address these questions taking advantage of the fact that
mitochondria - a common cargo - in Drosophila neurons exhibit a wide distribution
of sizes. In addition, the mitochondria can be genetically marked with green
fluorescent protein (GFP) making it possible to visualize and compare their
movement in the cell bodies and in the processes of living cells. Using total
internal reflection microscopy coupled with particle tracking and analysis, we
quantified the transport properties of GFP-positive mitochondria as a function of
their size and location. In neuronal cell bodies, we find little evidence for
significant opposition to motion, consistent with a previous study on lipid
droplets (Shubeita GT, Tran SL, Xu J, Vershinin M, Cermelli S, Cotton SL, Welte
MA, Gross SP. Consequences of motor copy number on the intracellular transport of
kinesin-1-driven lipid droplets. Cell 2008;135:1098-1107). However, in the
processes, we observe an inverse relationship between the mitochondrial size and
velocity and the run distances. This can be ameliorated via hypotonic treatment
to increase process size, suggesting that motor-mediated movement is impeded in
this more-confined environment. Interestingly, we also observe local
mitochondrial accumulations in processes but not in cell bodies. Such
accumulations do not completely block the transport but do increase the
probability of mitochondria-mitochondria interactions. They are thus particularly
interesting in relation to mitochondrial exchange of elements.