Cdc42 and Rac Family GTPases Regulate Mode and Speed but Not Direction of Primary Fibroblast Migration during Platelet-Derived Growth Factor-Dependent Chemotaxis

Monypenny, James; Zicha, Daniel; Higashida, Chiharu; Oceguera-Yañez, Fabian; Narumiya, Shuh; Watanabe, Naoki

doi:10.1128/mcb.01285-08

Cited by 59 publications

(64 citation statements)

References 35 publications

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“…For example, N-WASP is dispensable for haptotaxis but is required for chemotaxis (King et al, 2011). Conversely, Rac1 is necessary for haptotaxis but is dispensable for chemotaxis (Monypenny et al, 2009). In addition, we have recently mapped the pathway that fibroblasts use to sense and respond to PDGF gradients, which involves PLCγ and PKCα, and the local inactivation of myosin IIA at the leading edge of cells .…”

Section: Discussion a Working Model For Haptotaxismentioning

confidence: 99%

Lamellipodia are critical for haptotactic sensing and response

King

Asokan

Haynes

et al. 2016

Journal of Cell Science

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Haptotaxis is the process by which cells respond to gradients of substrate-bound cues, such as extracellular matrix proteins (ECM); however, the cellular mechanism of this response remains poorly understood and has mainly been studied by comparing cell behavior on uniform ECMs with different concentrations of components. To study haptotaxis in response to gradients, we utilized microfluidic chambers to generate gradients of the ECM protein fibronectin, and imaged the cell migration response. Lamellipodia are fan-shaped protrusions that are common in migrating cells. Here, we define a new function for lamellipodia and the cellular mechanism required for haptotaxisdifferential actin and lamellipodial protrusion dynamics lead to biased cell migration. Modest differences in lamellipodial dynamics occurring over time periods of seconds to minutes are summed over hours to produce differential whole cell movement towards higher concentrations of fibronectin. We identify a specific subset of lamellipodia regulators as being crucial for haptotaxis. Numerous studies have linked components of this pathway to cancer metastasis and, consistent with this, we find that expression of the oncogenic Rac1 P29S mutation abrogates haptotaxis. Finally, we show that haptotaxis also operates through this pathway in 3D environments.

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Section: Discussion a Working Model For Haptotaxismentioning

confidence: 99%

Lamellipodia are critical for haptotactic sensing and response

King

Asokan

Haynes

et al. 2016

Journal of Cell Science

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“…S3). Based on our data and previous reports, Rho GTPases are not readily activated in this system (Monypenny et al, 2009) while the use of MEFs deficient in RhoA was inconclusive due to concomitant upregulation of other GTPases (supplementary material Fig. S2B).…”

Section: Phospholipase C Activity Of Plce Is Required For Normal Chemmentioning

confidence: 99%

Activity of PLCε contributes to chemotaxis of fibroblasts towards PDGF

Martins

Warren

Kimberley

et al. 2012

Journal of Cell Science

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SummaryCell chemotaxis, such as migration of fibroblasts towards growth factors during development and wound healing, requires precise spatial coordination of signalling events. Phosphoinositides and signalling enzymes involved in their generation and hydrolysis have been implicated in regulation of chemotaxis; however, the role and importance of specific components remain poorly understood. Here, we demonstrate that phospholipase C epsilon (PLCe) contributes to fibroblast chemotaxis towards platelet-derived growth factor (PDGF-BB). Using PLCe1 null fibroblasts we show that cells deficient in PLCe have greatly reduced directionality towards PDGF-BB without detrimental effect on their basal ability to migrate. Furthermore, we show that in intact fibroblasts, signalling events, such as activation of Rac, are spatially compromised by the absence of PLCe that affects the ability of cells to enlarge their protrusions in the direction of the chemoattractant. By further application of live cell imaging and the use of FRET-based biosensors, we show that generation of Ins(1,4,5)P 3 and recruitment of PLCe are most pronounced in protrusions responding to the PDGF-BB gradient. Furthermore, the phospholipase C activity of PLCe is critical for its role in chemotaxis, consistent with the importance of Ins(1,4,5)P 3 generation and sustained calcium responses in this process. As PLCe has extensive signalling connectivity, using transgenic fibroblasts we ruled out its activation by direct binding to Ras or Rap GTPases, and suggest instead new unexpected links for PLCe in the context of chemotaxis.

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“…An alternative interpretation is that loss of Rac1 somehow affects the adhesive interactions between epidermal cells so that they can no longer maintain their normal shapes. There is considerable evidence in other systems for Rac having effects on both migration (Monypenny et al 2009;Wang et al 2010) and adhesion of epithelial cells (Eaton et al 1995;Chihara et al 2003) so these possibilities are not mutually exclusive.…”

Section: Why Are Mbcmentioning

confidence: 99%

A Targeted UAS-RNAi Screen in Drosophila Larvae Identifies Wound Closure Genes Regulating Distinct Cellular Processes

Lesch¹,

Jo²,

et al. 2010

Genetics

169

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Robust mechanisms for tissue repair are critical for survival of multicellular organisms. Efficient cutaneous wound repair requires the migration of cells at the wound edge and farther back within the epidermal sheet, but the genes that control and coordinate these migrations remain obscure. This is in part because a systematic screening approach for in vivo identification and classification of postembryonic wound closure genes has yet to be developed. Here, we performed a proof-of-principle reporter-based in vivo RNAi screen in the Drosophila melanogaster larval epidermis to identify genes required for normal wound closure. Among the candidate genes tested were kinases and transcriptional mediators of the Jun N-terminal kinase ( JNK) signaling pathway shown to be required for epithelial sheet migration during development. Also targeted were genes involved in actin cytoskeletal remodeling. Importantly, RNAi knockdown of both canonical and noncanonical members of the JNK pathway caused open wounds, as did several genes involved in actin cytoskeletal remodeling. Our analysis of JNK pathway components reveals redundancy among the upstream activating kinases and distinct roles for the downstream transcription factors DJun and DFos. Quantitative and qualitative morphological classification of the open wound phenotypes and evaluation of JNK activation suggest that multiple cellular processes are required in the migrating epidermal cells, including functions specific to cells at the wound edge and others specific to cells farther back within the epidermal sheet. Together, our results identify a new set of conserved wound closure genes, determine putative functional roles for these genes within the migrating epidermal sheet, and provide a template for a broader in vivo RNAi screen to discover the full complement of genes required for wound closure during larval epidermal wound healing.

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Cdc42 and Rac Family GTPases Regulate Mode and Speed but Not Direction of Primary Fibroblast Migration during Platelet-Derived Growth Factor-Dependent Chemotaxis

Cited by 59 publications

References 35 publications

Lamellipodia are critical for haptotactic sensing and response

Lamellipodia are critical for haptotactic sensing and response

Activity of PLCε contributes to chemotaxis of fibroblasts towards PDGF

A Targeted UAS-RNAi Screen in Drosophila Larvae Identifies Wound Closure Genes Regulating Distinct Cellular Processes

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