Eps8 regulates axonal filopodia in hippocampal neurons in response to brain-derived neurotrophic factor (BDNF)

Elisabetta Menna, Andrea Disanza, Cinzia Cagnoli, Ursula Schenk, Giuliana Gelsomino, Emanuela Frittoli, Maud Hertzog, Nina Offenhauser, Corinna Sawallisch, Hans Jürgen Kreienkamp, Frank B. Gertler, Pier Paolo Di Fiore, Giorgio Scita, Michela Matteoli

Research output: Contribution to journalArticle

59 Citations (Scopus)

Abstract

The regulation of filopodia plays a crucial role during neuronal development and synaptogenesis. Axonal filopodia, which are known to originate presynaptic specializations, are regulated in response to neurotrophic factors. The structural components of filopodia are actin filaments, whose dynamics and organization are controlled by ensembles of actin-binding proteins. How neurotrophic factors regulate these latter proteins remains, however, poorly defined. Here, using a combination of mouse genetic, biochemical, and cell biological assays, we show that genetic removal of Eps8, an actin-binding and regulatory protein enriched in the growth cones and developing processes of neurons, significantly augments the number and density of vasodilator-stimulated phosphoprotein (VASP)-dependent axonal filopodia. The reintroduction of Eps8 wild type (WT), but not an Eps8 capping-defective mutant, into primary hippocampal neurons restored axonal filopodia to WT levels. We further show that the actin barbed-end capping activity of Eps8 is inhibited by brain-derived neurotrophic factor (BDNF) treatment through MAPK-dependent phosphorylation of Eps8 residues S624 and T628. Additionally, an Eps8 mutant, impaired in the MAPK target sites (S624A/T628A), displays increased association to actin-rich structures, is resistant to BDNF-mediated release from microfilaments, and inhibits BDNF-induced filopodia. The opposite is observed for a phosphomimetic Eps8 (S624E/T628E) mutant. Thus, collectively, our data identify Eps8 as a critical capping protein in the regulation of axonal filopodia and delineate a molecular pathway by which BDNF, through MAPK-dependent phosphorylation of Eps8, stimulates axonal filopodia formation, a process with crucial impacts on neuronal development and synapse formation.

Original languageEnglish
Article numbere1000138
JournalPLoS Biology
Volume7
Issue number6
DOIs
Publication statusPublished - Jun 2009

Fingerprint

pseudopodia
Pseudopodia
neurotrophins
Brain-Derived Neurotrophic Factor
Neurons
Actins
neurons
brain
Phosphorylation
Nerve Growth Factors
microfilament proteins
Microfilament Proteins
Proteins
microfilaments
Actin Cytoskeleton
mutants
actin
Cones
Assays
phosphorylation

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)

Cite this

Eps8 regulates axonal filopodia in hippocampal neurons in response to brain-derived neurotrophic factor (BDNF). / Menna, Elisabetta; Disanza, Andrea; Cagnoli, Cinzia; Schenk, Ursula; Gelsomino, Giuliana; Frittoli, Emanuela; Hertzog, Maud; Offenhauser, Nina; Sawallisch, Corinna; Kreienkamp, Hans Jürgen; Gertler, Frank B.; Di Fiore, Pier Paolo; Scita, Giorgio; Matteoli, Michela.

In: PLoS Biology, Vol. 7, No. 6, e1000138, 06.2009.

Research output: Contribution to journalArticle

Menna, E, Disanza, A, Cagnoli, C, Schenk, U, Gelsomino, G, Frittoli, E, Hertzog, M, Offenhauser, N, Sawallisch, C, Kreienkamp, HJ, Gertler, FB, Di Fiore, PP, Scita, G & Matteoli, M 2009, 'Eps8 regulates axonal filopodia in hippocampal neurons in response to brain-derived neurotrophic factor (BDNF)', PLoS Biology, vol. 7, no. 6, e1000138. https://doi.org/10.1371/journal.pbio.1000138
Menna, Elisabetta ; Disanza, Andrea ; Cagnoli, Cinzia ; Schenk, Ursula ; Gelsomino, Giuliana ; Frittoli, Emanuela ; Hertzog, Maud ; Offenhauser, Nina ; Sawallisch, Corinna ; Kreienkamp, Hans Jürgen ; Gertler, Frank B. ; Di Fiore, Pier Paolo ; Scita, Giorgio ; Matteoli, Michela. / Eps8 regulates axonal filopodia in hippocampal neurons in response to brain-derived neurotrophic factor (BDNF). In: PLoS Biology. 2009 ; Vol. 7, No. 6.
@article{73869ad498bc44808d7c1975f1fa2438,
title = "Eps8 regulates axonal filopodia in hippocampal neurons in response to brain-derived neurotrophic factor (BDNF)",
abstract = "The regulation of filopodia plays a crucial role during neuronal development and synaptogenesis. Axonal filopodia, which are known to originate presynaptic specializations, are regulated in response to neurotrophic factors. The structural components of filopodia are actin filaments, whose dynamics and organization are controlled by ensembles of actin-binding proteins. How neurotrophic factors regulate these latter proteins remains, however, poorly defined. Here, using a combination of mouse genetic, biochemical, and cell biological assays, we show that genetic removal of Eps8, an actin-binding and regulatory protein enriched in the growth cones and developing processes of neurons, significantly augments the number and density of vasodilator-stimulated phosphoprotein (VASP)-dependent axonal filopodia. The reintroduction of Eps8 wild type (WT), but not an Eps8 capping-defective mutant, into primary hippocampal neurons restored axonal filopodia to WT levels. We further show that the actin barbed-end capping activity of Eps8 is inhibited by brain-derived neurotrophic factor (BDNF) treatment through MAPK-dependent phosphorylation of Eps8 residues S624 and T628. Additionally, an Eps8 mutant, impaired in the MAPK target sites (S624A/T628A), displays increased association to actin-rich structures, is resistant to BDNF-mediated release from microfilaments, and inhibits BDNF-induced filopodia. The opposite is observed for a phosphomimetic Eps8 (S624E/T628E) mutant. Thus, collectively, our data identify Eps8 as a critical capping protein in the regulation of axonal filopodia and delineate a molecular pathway by which BDNF, through MAPK-dependent phosphorylation of Eps8, stimulates axonal filopodia formation, a process with crucial impacts on neuronal development and synapse formation.",
author = "Elisabetta Menna and Andrea Disanza and Cinzia Cagnoli and Ursula Schenk and Giuliana Gelsomino and Emanuela Frittoli and Maud Hertzog and Nina Offenhauser and Corinna Sawallisch and Kreienkamp, {Hans J{\"u}rgen} and Gertler, {Frank B.} and {Di Fiore}, {Pier Paolo} and Giorgio Scita and Michela Matteoli",
year = "2009",
month = "6",
doi = "10.1371/journal.pbio.1000138",
language = "English",
volume = "7",
journal = "PLoS Biology",
issn = "1544-9173",
publisher = "Public Library of Science",
number = "6",

}

TY - JOUR

T1 - Eps8 regulates axonal filopodia in hippocampal neurons in response to brain-derived neurotrophic factor (BDNF)

AU - Menna, Elisabetta

AU - Disanza, Andrea

AU - Cagnoli, Cinzia

AU - Schenk, Ursula

AU - Gelsomino, Giuliana

AU - Frittoli, Emanuela

AU - Hertzog, Maud

AU - Offenhauser, Nina

AU - Sawallisch, Corinna

AU - Kreienkamp, Hans Jürgen

AU - Gertler, Frank B.

AU - Di Fiore, Pier Paolo

AU - Scita, Giorgio

AU - Matteoli, Michela

PY - 2009/6

Y1 - 2009/6

N2 - The regulation of filopodia plays a crucial role during neuronal development and synaptogenesis. Axonal filopodia, which are known to originate presynaptic specializations, are regulated in response to neurotrophic factors. The structural components of filopodia are actin filaments, whose dynamics and organization are controlled by ensembles of actin-binding proteins. How neurotrophic factors regulate these latter proteins remains, however, poorly defined. Here, using a combination of mouse genetic, biochemical, and cell biological assays, we show that genetic removal of Eps8, an actin-binding and regulatory protein enriched in the growth cones and developing processes of neurons, significantly augments the number and density of vasodilator-stimulated phosphoprotein (VASP)-dependent axonal filopodia. The reintroduction of Eps8 wild type (WT), but not an Eps8 capping-defective mutant, into primary hippocampal neurons restored axonal filopodia to WT levels. We further show that the actin barbed-end capping activity of Eps8 is inhibited by brain-derived neurotrophic factor (BDNF) treatment through MAPK-dependent phosphorylation of Eps8 residues S624 and T628. Additionally, an Eps8 mutant, impaired in the MAPK target sites (S624A/T628A), displays increased association to actin-rich structures, is resistant to BDNF-mediated release from microfilaments, and inhibits BDNF-induced filopodia. The opposite is observed for a phosphomimetic Eps8 (S624E/T628E) mutant. Thus, collectively, our data identify Eps8 as a critical capping protein in the regulation of axonal filopodia and delineate a molecular pathway by which BDNF, through MAPK-dependent phosphorylation of Eps8, stimulates axonal filopodia formation, a process with crucial impacts on neuronal development and synapse formation.

AB - The regulation of filopodia plays a crucial role during neuronal development and synaptogenesis. Axonal filopodia, which are known to originate presynaptic specializations, are regulated in response to neurotrophic factors. The structural components of filopodia are actin filaments, whose dynamics and organization are controlled by ensembles of actin-binding proteins. How neurotrophic factors regulate these latter proteins remains, however, poorly defined. Here, using a combination of mouse genetic, biochemical, and cell biological assays, we show that genetic removal of Eps8, an actin-binding and regulatory protein enriched in the growth cones and developing processes of neurons, significantly augments the number and density of vasodilator-stimulated phosphoprotein (VASP)-dependent axonal filopodia. The reintroduction of Eps8 wild type (WT), but not an Eps8 capping-defective mutant, into primary hippocampal neurons restored axonal filopodia to WT levels. We further show that the actin barbed-end capping activity of Eps8 is inhibited by brain-derived neurotrophic factor (BDNF) treatment through MAPK-dependent phosphorylation of Eps8 residues S624 and T628. Additionally, an Eps8 mutant, impaired in the MAPK target sites (S624A/T628A), displays increased association to actin-rich structures, is resistant to BDNF-mediated release from microfilaments, and inhibits BDNF-induced filopodia. The opposite is observed for a phosphomimetic Eps8 (S624E/T628E) mutant. Thus, collectively, our data identify Eps8 as a critical capping protein in the regulation of axonal filopodia and delineate a molecular pathway by which BDNF, through MAPK-dependent phosphorylation of Eps8, stimulates axonal filopodia formation, a process with crucial impacts on neuronal development and synapse formation.

UR - http://www.scopus.com/inward/record.url?scp=67649935286&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67649935286&partnerID=8YFLogxK

U2 - 10.1371/journal.pbio.1000138

DO - 10.1371/journal.pbio.1000138

M3 - Article

C2 - 19564905

AN - SCOPUS:67649935286

VL - 7

JO - PLoS Biology

JF - PLoS Biology

SN - 1544-9173

IS - 6

M1 - e1000138

ER -