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Acoustic detection of melanosome transport in Xenopus laevis melanophores

Journal article
Authors Rickard Frost
Elisabeth Norström
Lovisa Bodin
Christoph Langhammer
Joachim Sturve
Margareta Wallin
Sofia Svedhem
Published in Analytical Biochemistry
Volume 435
Issue 1
Pages 10-18
ISSN 0003-2697
Publication year 2013
Published at Department of Biological and Environmental Sciences
Pages 10-18
Language en
Links dx.doi.org/10.1016/j.ab.2012.12.004
Keywords Xenopus laevis; Melanophores; Melanosomes; QCM-D; Latrunculin; Nocodazole
Subject categories Chemical Sciences

Abstract

Organelle transport studies are often performed using melanophores from lower vertebrates due to the ease of inducing movements of pigment granules (melanosomes) and visualizing them by optical microscopy. Here, we present a novel methodology to monitor melanosome translocation (which is a light-sensitive process) in the dark using the quartz crystal microbalance with dissipation monitoring (QCM-D) technique. This acoustic sensing method was used to study dispersion and aggregation of melanosomes in Xenopus laevis melanophores. Reversible sensor responses, correlated to optical reflectance measurements, were obtained by alternating addition and removal of melatonin (leading to melanosome aggregation) and melanocyte-stimulating hormone (MSH) (leading to melanosome dispersion). By confocal microscopy, it was shown that a vertical redistribution of melanosomes occurred during the dispersion/aggregation processes. Furthermore, the transport process was studied in the presence of cytoskeleton-perturbing agents disrupting either actin filaments (latrunculin) or microtubules (nocodazole). Taken together, these experiments suggest that the acoustic responses mainly originate from melanosome transport along actin filaments (located close to the cell membrane), as expected based on the penetration depth of the QCM-D technique. The results clearly indicate the potential of QCM-D for studies of intracellular transport processes in melanophores.

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