Evidence that the small GTPase Rab8 is involved in melanosome traffic and dendrite extension in B16 melanoma cells.
One of the major activities of melanocytes in skin is to produce melanin and transport it via dendrites to neighboring keratinocytes. Here, we present evidence that Rab8, a member of the small GTPase superfamily, is present in purified melanosomal fractions, and is upregulated by pigmentogenic agents like melanocyte-stimulating hormone/isobutylmethyl xanthine (MSH/IBMX) and ultraviolet radiation B (UVB). Confocal immunofluorescence microscopic studies revealed that Rab8 is colocalized with Mel5, a melanosomal protein, at the trans-Golgi area and in the cytoplasmic vesicles of B16 cells. During MSH/IBMX treatment, while a number of dendrites with numerous processes are formed, colocalization is extended towards the tips of protrusions. Since process formation is supported by cytoskeletal assembly as well as membrane transport, we tested the colocalization of Rab8 with actin filaments in B16 cells. Rab8, indeed, colocalized with phalloidin, mostly at the periphery, but when irradiated with UVB, cells were rounded instead of dendritic, and colocalization was found predominantly at the cytoplasmic area. Further, suppression of Rab8 expression by its antisense oligonucleotide revealed the reduction in staining intensity of Rab8 but not of Mel5, dendrite formation and melanosome transport towards the tips of the dendrites in B16 melanoma cells. Taken together, it is suggestive that Rab8, in B16 melanoma cells, might have a role in melanosome traffic and dendrite extension, both in constitutive and regulated fashion.
Chakraborty
Ashok K
AK
Department of Dermatology, Yale University School of Medicine, New Haven, CT 06520, USA. ashok.chakraborty@yale.edu
Funasaka
Yoko
Y
Araki
Keishi
K
Horikawa
Tatsuya
T
Ichihashi
Masamitsu
M
D008841:Actin Cytoskeleton / Q000378:metabolism; D000818:Animals; D045744:Cell Line, Tumor; D022081:Cell Surface Extensions / Q000201:enzymology* / Q000528:radiation effects / Q000648:ultrastructure; D006023:Glycoproteins; D009074:Melanocyte-Stimulating Hormones / Q000378:metabolism / Q000494:pharmacology; D008544:Melanocytes / Q000166:cytology / Q000201:enzymology* / Q000528:radiation effects; D008546:Melanoma, Experimental / Q000201:enzymology* / Q000473:pathology; D020460:Melanosomes / Q000187:drug effects / Q000201:enzymology* / Q000528:radiation effects; D051379:Mice; D009368:Neoplasm Transplantation; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010590:Phalloidine / Q000378:metabolism; D021381:Protein Transport / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D013871:Thiones / Q000494:pharmacology; D022161:Transport Vesicles / Q000201:enzymology / Q000648:ultrastructure; D014466:Ultraviolet Rays; D015854:Up-Regulation / Q000502:physiology / Q000528:radiation effects; D020691:rab GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D021601:trans-Golgi Network / Q000201:enzymology
D006023:Glycoproteins; C094218:MEL-5 protein, human; D016376:Oligonucleotides, Antisense; D013871:Thiones; D010590:Phalloidine; C027826:isobutyl xanthate; D009074:Melanocyte-Stimulating Hormones; D020691:rab GTP-Binding Proteins