---
_id: '7880'
abstract:
- lang: eng
  text: 'Following its evoked release, dopamine (DA) signaling is rapidly terminated
    by presynaptic reuptake, mediated by the cocaine-sensitive DA transporter (DAT).
    DAT surface availability is dynamically regulated by endocytic trafficking, and
    direct protein kinase C (PKC) activation acutely diminishes DAT surface expression
    by accelerating DAT internalization. Previous cell line studies demonstrated that
    PKC-stimulated DAT endocytosis requires both Ack1 inactivation, which releases
    a DAT-specific endocytic brake, and the neuronal GTPase, Rit2, which binds DAT.
    However, it is unknown whether Rit2 is required for PKC-stimulated DAT endocytosis
    in DAergic terminals or whether there are region- and/or sex-dependent differences
    in PKC-stimulated DAT trafficking. Moreover, the mechanisms by which Rit2 controls
    PKC-stimulated DAT endocytosis are unknown. Here, we directly examined these important
    questions. Ex vivo studies revealed that PKC activation acutely decreased DAT
    surface expression selectively in ventral, but not dorsal, striatum. AAV-mediated,
    conditional Rit2 knockdown in DAergic neurons impacted baseline DAT surface:intracellular
    distribution in DAergic terminals from female ventral, but not dorsal, striatum.
    Further, Rit2 was required for PKC-stimulated DAT internalization in both male
    and female ventral striatum. FRET and surface pulldown studies in cell lines revealed
    that PKC activation drives DAT-Rit2 surface dissociation and that the DAT N terminus
    is required for both PKC-mediated DAT-Rit2 dissociation and DAT internalization.
    Finally, we found that Rit2 and Ack1 independently converge on DAT to facilitate
    PKC-stimulated DAT endocytosis. Together, our data provide greater insight into
    mechanisms that mediate PKC-regulated DAT internalization and reveal unexpected
    region-specific differences in PKC-stimulated DAT trafficking in bona fide DAergic
    terminals. '
article_processing_charge: No
article_type: original
author:
- first_name: Rita R.
  full_name: Fagan, Rita R.
  last_name: Fagan
- first_name: Patrick J.
  full_name: Kearney, Patrick J.
  last_name: Kearney
- first_name: Carolyn G.
  full_name: Sweeney, Carolyn G.
  last_name: Sweeney
- first_name: Dino
  full_name: Luethi, Dino
  last_name: Luethi
- first_name: Florianne E
  full_name: Schoot Uiterkamp, Florianne E
  id: 3526230C-F248-11E8-B48F-1D18A9856A87
  last_name: Schoot Uiterkamp
- first_name: Klaus
  full_name: Schicker, Klaus
  last_name: Schicker
- first_name: Brian S.
  full_name: Alejandro, Brian S.
  last_name: Alejandro
- first_name: Lauren C.
  full_name: O'Connor, Lauren C.
  last_name: O'Connor
- first_name: Harald H.
  full_name: Sitte, Harald H.
  last_name: Sitte
- first_name: Haley E.
  full_name: Melikian, Haley E.
  last_name: Melikian
citation:
  ama: 'Fagan RR, Kearney PJ, Sweeney CG, et al. Dopamine transporter trafficking
    and Rit2 GTPase: Mechanism of action and in vivo impact. <i>Journal of Biological
    Chemistry</i>. 2020;295(16):5229-5244. doi:<a href="https://doi.org/10.1074/jbc.RA120.012628">10.1074/jbc.RA120.012628</a>'
  apa: 'Fagan, R. R., Kearney, P. J., Sweeney, C. G., Luethi, D., Schoot Uiterkamp,
    F. E., Schicker, K., … Melikian, H. E. (2020). Dopamine transporter trafficking
    and Rit2 GTPase: Mechanism of action and in vivo impact. <i>Journal of Biological
    Chemistry</i>. ASBMB Publications. <a href="https://doi.org/10.1074/jbc.RA120.012628">https://doi.org/10.1074/jbc.RA120.012628</a>'
  chicago: 'Fagan, Rita R., Patrick J. Kearney, Carolyn G. Sweeney, Dino Luethi, Florianne
    E Schoot Uiterkamp, Klaus Schicker, Brian S. Alejandro, Lauren C. O’Connor, Harald
    H. Sitte, and Haley E. Melikian. “Dopamine Transporter Trafficking and Rit2 GTPase:
    Mechanism of Action and in Vivo Impact.” <i>Journal of Biological Chemistry</i>.
    ASBMB Publications, 2020. <a href="https://doi.org/10.1074/jbc.RA120.012628">https://doi.org/10.1074/jbc.RA120.012628</a>.'
  ieee: 'R. R. Fagan <i>et al.</i>, “Dopamine transporter trafficking and Rit2 GTPase:
    Mechanism of action and in vivo impact,” <i>Journal of Biological Chemistry</i>,
    vol. 295, no. 16. ASBMB Publications, pp. 5229–5244, 2020.'
  ista: 'Fagan RR, Kearney PJ, Sweeney CG, Luethi D, Schoot Uiterkamp FE, Schicker
    K, Alejandro BS, O’Connor LC, Sitte HH, Melikian HE. 2020. Dopamine transporter
    trafficking and Rit2 GTPase: Mechanism of action and in vivo impact. Journal of
    Biological Chemistry. 295(16), 5229–5244.'
  mla: 'Fagan, Rita R., et al. “Dopamine Transporter Trafficking and Rit2 GTPase:
    Mechanism of Action and in Vivo Impact.” <i>Journal of Biological Chemistry</i>,
    vol. 295, no. 16, ASBMB Publications, 2020, pp. 5229–44, doi:<a href="https://doi.org/10.1074/jbc.RA120.012628">10.1074/jbc.RA120.012628</a>.'
  short: R.R. Fagan, P.J. Kearney, C.G. Sweeney, D. Luethi, F.E. Schoot Uiterkamp,
    K. Schicker, B.S. Alejandro, L.C. O’Connor, H.H. Sitte, H.E. Melikian, Journal
    of Biological Chemistry 295 (2020) 5229–5244.
date_created: 2020-05-24T22:00:59Z
date_published: 2020-04-17T00:00:00Z
date_updated: 2023-08-21T06:26:22Z
day: '17'
department:
- _id: SaSi
doi: 10.1074/jbc.RA120.012628
external_id:
  isi:
  - '000530288000006'
  pmid:
  - '32132171'
intvolume: '       295'
isi: 1
issue: '16'
language:
- iso: eng
main_file_link:
- open_access: '1'
  url: https://escholarship.umassmed.edu/oapubs/4187
month: '04'
oa: 1
oa_version: Submitted Version
page: 5229-5244
pmid: 1
publication: Journal of Biological Chemistry
publication_identifier:
  eissn:
  - 1083351X
  issn:
  - '00219258'
publication_status: published
publisher: ASBMB Publications
quality_controlled: '1'
scopus_import: '1'
status: public
title: 'Dopamine transporter trafficking and Rit2 GTPase: Mechanism of action and
  in vivo impact'
type: journal_article
user_id: 4359f0d1-fa6c-11eb-b949-802e58b17ae8
volume: 295
year: '2020'
...
---
_id: '668'
abstract:
- lang: eng
  text: Macrophage filopodia, finger-like membrane protrusions, were first implicated
    in phagocytosis more than 100 years ago, but little is still known about the involvement
    of these actin-dependent structures in particle clearance. Using spinning disk
    confocal microscopy to image filopodial dynamics in mouse resident Lifeact-EGFP
    macrophages, we show that filopodia, or filopodia-like structures, support pathogen
    clearance by multiple means. Filopodia supported the phagocytic uptake of bacterial
    (Escherichia coli) particles by (i) capturing along the filopodial shaft and surfing
    toward the cell body, the most common mode of capture; (ii) capturing via the
    tip followed by retraction; (iii) combinations of surfing and retraction; or (iv)
    sweeping actions. In addition, filopodia supported the uptake of zymosan (Saccharomyces
    cerevisiae) particles by (i) providing fixation, (ii) capturing at the tip and
    filopodia-guided actin anterograde flow with phagocytic cup formation, and (iii)
    the rapid growth of new protrusions. To explore the role of filopodia-inducing
    Cdc42, we generated myeloid-restricted Cdc42 knock-out mice. Cdc42-deficient macrophages
    exhibited rapid phagocytic cup kinetics, but reduced particle clearance, which
    could be explained by the marked rounded-up morphology of these cells. Macrophages
    lacking Myo10, thought to act downstream of Cdc42, had normal morphology, motility,
    and phagocytic cup formation, but displayed markedly reduced filopodia formation.
    In conclusion, live-cell imaging revealed multiple mechanisms involving macrophage
    filopodia in particle capture and engulfment. Cdc42 is not critical for filopodia
    or phagocytic cup formation, but plays a key role in driving macrophage lamellipodial
    spreading.
article_type: original
author:
- first_name: Markus
  full_name: Horsthemke, Markus
  last_name: Horsthemke
- first_name: Anne
  full_name: Bachg, Anne
  last_name: Bachg
- first_name: Katharina
  full_name: Groll, Katharina
  last_name: Groll
- first_name: Sven
  full_name: Moyzio, Sven
  last_name: Moyzio
- first_name: Barbara
  full_name: Müther, Barbara
  last_name: Müther
- first_name: Sandra
  full_name: Hemkemeyer, Sandra
  last_name: Hemkemeyer
- first_name: Roland
  full_name: Wedlich Söldner, Roland
  last_name: Wedlich Söldner
- first_name: Michael K
  full_name: Sixt, Michael K
  id: 41E9FBEA-F248-11E8-B48F-1D18A9856A87
  last_name: Sixt
  orcid: 0000-0002-6620-9179
- first_name: Sebastian
  full_name: Tacke, Sebastian
  last_name: Tacke
- first_name: Martin
  full_name: Bähler, Martin
  last_name: Bähler
- first_name: Peter
  full_name: Hanley, Peter
  last_name: Hanley
citation:
  ama: Horsthemke M, Bachg A, Groll K, et al. Multiple roles of filopodial dynamics
    in particle capture and phagocytosis and phenotypes of Cdc42 and Myo10 deletion.
    <i>Journal of Biological Chemistry</i>. 2017;292(17):7258-7273. doi:<a href="https://doi.org/10.1074/jbc.M116.766923">10.1074/jbc.M116.766923</a>
  apa: Horsthemke, M., Bachg, A., Groll, K., Moyzio, S., Müther, B., Hemkemeyer, S.,
    … Hanley, P. (2017). Multiple roles of filopodial dynamics in particle capture
    and phagocytosis and phenotypes of Cdc42 and Myo10 deletion. <i>Journal of Biological
    Chemistry</i>. American Society for Biochemistry and Molecular Biology. <a href="https://doi.org/10.1074/jbc.M116.766923">https://doi.org/10.1074/jbc.M116.766923</a>
  chicago: Horsthemke, Markus, Anne Bachg, Katharina Groll, Sven Moyzio, Barbara Müther,
    Sandra Hemkemeyer, Roland Wedlich Söldner, et al. “Multiple Roles of Filopodial
    Dynamics in Particle Capture and Phagocytosis and Phenotypes of Cdc42 and Myo10
    Deletion.” <i>Journal of Biological Chemistry</i>. American Society for Biochemistry
    and Molecular Biology, 2017. <a href="https://doi.org/10.1074/jbc.M116.766923">https://doi.org/10.1074/jbc.M116.766923</a>.
  ieee: M. Horsthemke <i>et al.</i>, “Multiple roles of filopodial dynamics in particle
    capture and phagocytosis and phenotypes of Cdc42 and Myo10 deletion,” <i>Journal
    of Biological Chemistry</i>, vol. 292, no. 17. American Society for Biochemistry
    and Molecular Biology, pp. 7258–7273, 2017.
  ista: Horsthemke M, Bachg A, Groll K, Moyzio S, Müther B, Hemkemeyer S, Wedlich
    Söldner R, Sixt MK, Tacke S, Bähler M, Hanley P. 2017. Multiple roles of filopodial
    dynamics in particle capture and phagocytosis and phenotypes of Cdc42 and Myo10
    deletion. Journal of Biological Chemistry. 292(17), 7258–7273.
  mla: Horsthemke, Markus, et al. “Multiple Roles of Filopodial Dynamics in Particle
    Capture and Phagocytosis and Phenotypes of Cdc42 and Myo10 Deletion.” <i>Journal
    of Biological Chemistry</i>, vol. 292, no. 17, American Society for Biochemistry
    and Molecular Biology, 2017, pp. 7258–73, doi:<a href="https://doi.org/10.1074/jbc.M116.766923">10.1074/jbc.M116.766923</a>.
  short: M. Horsthemke, A. Bachg, K. Groll, S. Moyzio, B. Müther, S. Hemkemeyer, R.
    Wedlich Söldner, M.K. Sixt, S. Tacke, M. Bähler, P. Hanley, Journal of Biological
    Chemistry 292 (2017) 7258–7273.
date_created: 2018-12-11T11:47:49Z
date_published: 2017-04-28T00:00:00Z
date_updated: 2021-01-12T08:08:34Z
day: '28'
ddc:
- '570'
department:
- _id: MiSi
doi: 10.1074/jbc.M116.766923
file:
- access_level: open_access
  checksum: d488162874326a4bb056065fa549dc4a
  content_type: application/pdf
  creator: dernst
  date_created: 2019-10-24T15:25:42Z
  date_updated: 2020-07-14T12:47:37Z
  file_id: '6971'
  file_name: 2017_JBC_Horsthemke.pdf
  file_size: 5647880
  relation: main_file
file_date_updated: 2020-07-14T12:47:37Z
has_accepted_license: '1'
intvolume: '       292'
issue: '17'
language:
- iso: eng
month: '04'
oa: 1
oa_version: Published Version
page: 7258 - 7273
publication: Journal of Biological Chemistry
publication_identifier:
  issn:
  - '00219258'
publication_status: published
publisher: American Society for Biochemistry and Molecular Biology
publist_id: '7059'
quality_controlled: '1'
scopus_import: 1
status: public
title: Multiple roles of filopodial dynamics in particle capture and phagocytosis
  and phenotypes of Cdc42 and Myo10 deletion
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 292
year: '2017'
...