Actin-dependent motility of melanosomes from fish retinal pigment epithelial (RPE) cells investigated using in vitro motility assays
Abstract
Melanosomes (pigment granules) within retinal pigment epithelial (RPE) cells of fish and amphibians undergo massive migrations in response to light conditions to control light flux to the retina. Previous research has shown that melanosome motility within apical projections of dissociated fish RPE cells requires an intact actin cytoskeleton, but the mechanisms and motors involved in melanosome transport in RPE have not been identified. Two in vitro motility assays, the Nitella assay and the sliding filament assay, were used to characterize actin-dependent motor activity of RPE melanosomes. Melanosomes applied to dissected filets of the Characean alga, Nitella, moved along actin cables at a mean rate of 2 microm/min, similar to the rate of melanosome motility in dissociated, cultured RPE cells. Path lengths of motile melanosomes ranged from 9 to 37 microm. Melanosome motility in the sliding filament assay was much more variable, ranging from 0.4-33 microm/min; 70% of velocities ranged from 1-15 microm/min. Latex beads coated with skeletal muscle myosin II and added to Nitella filets moved in the same direction as RPE melanosomes, indicating that the motility is barbed-end directed. Immunoblotting using antibodies against myosin VIIa and rab27a revealed that both proteins are enriched on melanosome membranes, suggesting that they could play a role in melanosome transport within apical projections of fish RPE.
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In fish RPE, myosin-7a appears responsible for dispersion of melanosomes along apical processes in light, whereas in darkness retrograde movement out of apical processes appears to require myosin-2.
“In fish RPE, myosin-7a appears to be responsible for the dispersion of melanosomes along the apical processes in the light (McNeil et al., 2004), whereas, in the dark, retrograde movement out of the apical processes appears to require myosin-2 (Barsoum and King-Smith, 2007).”
From The cell biology of the retinal pigment epithelium (2020) · cites this paper as McNeil et al., 2004
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- Authors
- McNeil, E. L., Tacelosky, D., Basciano, P., Biallas, B., Williams, R., Damiani, P., Deacon, S., Fox, C., Stewart, B., Petruzzi, N., Osborn, C., Klinger, K., Sellers, J. R., & Smith, C. K.
- Affiliations
- Saint Joseph's University (US)
- National Institutes of Health (US)
- Corresponding author
- Smith, C. King
- Venue
- Cell Motility
- Publisher
- Wiley
- Published
- 2004-03-10
- Volume
- 58
- Issue
- 2
- Pages
- 71-82
- ISSN
- 0886-1544 (ISSN-L)
- OpenAlex
- W2097076748
- Cited by
- 20
- References
- 37
- Citations by year
- 2026: 1 · 2021: 1 · 2020: 1 · 2016: 2 · 2015: 1 · 2014: 1 · 2012: 1
- FWCI
- 1.14
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Cell Motility and the Cytoskeleton (publishedVersion)
- PubMed (publishedVersion)
- Funders
- University of Oregon
- National Institutes of Health (1r15gm059460-01)
- National Institute of General Medical Sciences (R15 GM59460, R15GM066961)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › melanin and skin pigmentation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Keywords
- retinal pigment epithelium, actin cytoskeleton, myosin II, MYO7A, Rab27a
- MeSH terms
- Myosin VIIa, Actins, Animals, Biological Assay, Biological Transport, Dyneins, Microspheres, Myosins, Perciformes, Pigment Epithelium of Eye, Immunoblotting, Molecular Motor Proteins, Melanosomes, rab GTP-Binding Proteins, Myosin Type II, Nitella, In Vitro Techniques
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