Phospholipase D is essential for keratocyte-like migration of NBT-II cells.
NBT-II cells on collagen-coated substrates move rapidly and persistently, maintaining a semi-circular shape with a large lamellipodium, in a manner similar to fish keratocytes. The inhibitor of phospholipase D (PLD), n-butanol, completely blocked the migration and disturbed the characteristic localization of actin along the edge of lamellipodia. To investigate the functional difference between the two isozymes of PLD (PLD1 and PLD2), we transfected NBT-II cells with vectors expressing shRNA to deplete PLD1 or PLD2. Depletion of both PLD1 and 2 by RNA interference reduced the velocity of the migration, but depletion of PLD2 inhibited motility more severely than that of PLD1. Furthermore, GFP-PLD2 was localized to the protruding regions of lamellipodia in migrating cells. Thus, PLD is essential for the maintenance of keratocyte-like locomotion of NBT-II cells, presumably by regulating the actin cytoskeleton.
Nagasaki
Akira
A
Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 4, Tsukuba, Ibaraki 305-8562, Japan. a-nagasaki@aist.go.jp
Inotsume
Kimiko
K
Kanada
Masamitsu
M
Uyeda
Taro Q P
TQ
D020001:1-Butanol / Q000494:pharmacology; D008841:Actin Cytoskeleton / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology*; D018076:DNA, Complementary / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D010739:Phospholipase D / Q000235:genetics / Q000378:metabolism*; D011554:Pseudopodia / Q000378:metabolism; D034622:RNA Interference; D051381:Rats; D014162:Transfection
D018076:DNA, Complementary; D049452:Green Fluorescent Proteins; D020001:1-Butanol; C105606:phospholipase D2; D010739:Phospholipase D; C106801:phospholipase D1