Dynamin triple knockout cells reveal off target effects of commonly used dynamin inhibitors

J Cell Sci. 2013 Nov 15;126(Pt 22):5305-12. doi: 10.1242/jcs.138578. Epub 2013 Sep 17.

Abstract

Dynamin, which is encoded by three genes in mammals, is a GTPase implicated in endocytic membrane fission. Dynamin 1 and 3 are predominantly expressed in brain, whereas dynamin 2 is ubiquitously expressed. With the goal of assessing the impact of the lack of dynamin on cell physiology, we previously generated and characterized dynamin 1 and 2 double knockout (DKO) fibroblasts. These DKO cells were unexpectedly viable in spite of a severe impairment of clathrin-mediated endocytosis. As low-level expression of the dynamin 3 gene in these cells could not be excluded, we have now engineered dynamin 1, 2 and 3 triple KO (TKO) fibroblasts. These cells did not reveal any additional defects beyond what was previously observed in DKO fibroblasts. Surprisingly, although fluid-phase endocytosis and peripheral membrane ruffling were not impaired by the lack of all three dynamins, two structurally similar, widely used dynamin inhibitors, dynasore and Dyngo-4a, robustly inhibited these two processes both in wild-type and TKO cells. Dynamin TKO cells will be useful tools for the further exploration of dynamin-dependent processes and the development of more specific dynamin inhibitors.

Keywords: Actin; Dynamin; Dynasore; Dyngo; Synapse; Synaptic vesicle.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • Dynamin I / genetics
  • Dynamin I / metabolism*
  • Dynamin II / genetics
  • Dynamin II / metabolism*
  • Dynamin III / genetics
  • Dynamin III / metabolism*
  • Endocytosis / genetics
  • Fibroblasts / metabolism
  • Gene Expression Regulation / drug effects
  • Gene Knockout Techniques
  • Hydrazones / pharmacology
  • Mice
  • Naphthols / pharmacology
  • Synapses / metabolism*

Substances

  • Hydrazones
  • N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide
  • Naphthols
  • dyngo-4a
  • Dynamin I
  • Dynamin II
  • Dynamin III