Lambert J, Onderwater J, Vander Haeghen Y, Vancoillie G, Koerten H K, Mommaas A M, Naeyaert J M
Department of Dermatology, University Hospital, Gent, Belgium.
J Invest Dermatol. 1998 Nov;111(5):835-40. doi: 10.1046/j.1523-1747.1998.00395.x.
Mutations of the gene encoding myosin V can produce a dilute or silvery hair color and various neurologic defects in mice and patients with Griscelli syndrome, leading to speculations that the myosin V motor protein plays a critical role in transporting melanosomes within melanocytes and neurosecretory vesicles within neurons. Therefore, we investigated the in vitro expression of myosin V in cultured normal human melanocytes, keratinocytes, and dermal fibroblasts using reverse transcriptase-polymerase chain reaction and northern blot analysis. Subcellular distribution of myosin V and proximity to actin bundles and melanosomes were determined by double indirect immunofluorescence labeling and immunogold electron microscopy. In all studied cells myosin V is expressed and treatment of melanocytes with the cyclic AMP-inducer 3-isobutyl-1-methylxanthine causes an induction of the myosin V message. In all cells myosin V colocalizes with actin bundles, concentrating in the subcortical cell zone. In the melanocyte it is closely associated with melanosomes. Quantitative analysis of myosin V labeling in melanocytes reveals a significantly higher (p < 0.005) presence of myosin V in the periphery of dendrites. These results suggest that myosin V is important in melanosome transport in human melanocytes. Possible roles in the other skin cells remain to be elucidated.
编码肌球蛋白V的基因突变可导致小鼠和患有格里塞利综合征的患者出现毛发颜色变浅或呈银色以及各种神经学缺陷,这引发了人们的猜测,即肌球蛋白V运动蛋白在黑素细胞内运输黑素小体以及神经元内运输神经分泌小泡的过程中发挥着关键作用。因此,我们利用逆转录聚合酶链反应和Northern印迹分析,研究了肌球蛋白V在培养的正常人黑素细胞、角质形成细胞和真皮成纤维细胞中的体外表达情况。通过双重间接免疫荧光标记和免疫金电子显微镜确定了肌球蛋白V的亚细胞分布以及与肌动蛋白束和黑素小体的接近程度。在所有研究的细胞中均表达肌球蛋白V,用环磷酸腺苷诱导剂3-异丁基-1-甲基黄嘌呤处理黑素细胞会导致肌球蛋白V信息的诱导。在所有细胞中,肌球蛋白V都与肌动蛋白束共定位,集中在皮质下细胞区。在黑素细胞中,它与黑素小体密切相关。对黑素细胞中肌球蛋白V标记的定量分析显示,树突外周的肌球蛋白V含量显著更高(p < 0.005)。这些结果表明,肌球蛋白V在人类黑素细胞的黑素小体运输中很重要。它在其他皮肤细胞中的可能作用仍有待阐明。