Phospholipase D is essential for keratocyte-like migration of NBT-II cells

Akira Nagasaki; Kimiko Inotsume; Masamitsu Kanada; Taro Q.P. Uyeda

doi:10.1247/csf.07042

Phospholipase D is essential for keratocyte-like migration of NBT-II cells

Akira Nagasaki^*, Kimiko Inotsume, Masamitsu Kanada, Taro Q.P. Uyeda

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

11 Citations (Scopus)

Abstract

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.

Original language	English
Pages (from-to)	27-33
Number of pages	7
Journal	Cell Structure and Function
Volume	33
Issue number	1
DOIs	https://doi.org/10.1247/csf.07042
Publication status	Published - 2008
Externally published	Yes

Keywords

Actin
Cell migration
Knock down
NBT-II cell
PLD

ASJC Scopus subject areas

Physiology
Molecular Biology
Cell Biology

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10.1247/csf.07042

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title = "Phospholipase D is essential for keratocyte-like migration of NBT-II cells",

abstract = "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.",

keywords = "Actin, Cell migration, Knock down, NBT-II cell, PLD",

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year = "2008",

doi = "10.1247/csf.07042",

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T1 - Phospholipase D is essential for keratocyte-like migration of NBT-II cells

AU - Nagasaki, Akira

AU - Inotsume, Kimiko

AU - Kanada, Masamitsu

AU - Uyeda, Taro Q.P.

PY - 2008

Y1 - 2008

N2 - 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.

AB - 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.

KW - Actin

KW - Cell migration

KW - Knock down

KW - NBT-II cell

KW - PLD

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JO - Cell Structure and Function

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Phospholipase D is essential for keratocyte-like migration of NBT-II cells

Abstract

Keywords

ASJC Scopus subject areas

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