Cyclic AMP mediates keratinocyte directional migration in an electric field

Christine E. Pullar, Roslyn Rivkah Isseroff

Research output: Contribution to journalArticle

69 Citations (Scopus)

Abstract

Re-epithelialization of wounded skin is necessary for wound closure and restoration of barrier function and requires directional keratinocyte migration towards the center of the wound. The electric field (EF) generated immediately upon wounding could be the earliest signal keratinocytes receive to initiate directional migration and healing. Keratinocytes express many β2-adrenergic receptors (β2-ARs), but their role in the epidermis is unknown. We have previously shown that β-AR agonists decrease keratinocyte migration in a cyclic AMP (cAMP) independent mechanism involving the activation of protein phosphatase 2A (PP2A). Here, we ask whether β2-ARs play a role in keratinocyte galvanotaxis. We report a bimodal response. When keratinocytes were exposed to higher concentrations of β-AR agonist (0.1 μM), their tracked migratory speed was inhibited, in both the presence (directional migration) and the absence (random migration) of a 100 mV mm-1 EF, as expected. At lower agonist concentrations (0.1 pM to 0.1 nM), there was no effect on migratory speed; however, all directionality was lost - essentially, cells were 'blinded' to the directional cue. Preincubating the cells with β-antagonist restored directional migration, demonstrating that the 'blindness' was β2-AR mediated. Incubation of keratinocytes with agents known to increase intracellular cAMP levels, such as sp-cAMP, pertussis toxin and forskolin, resulted in similar 'blinding' to the EF, whereas random migration was unaffected. The inactive cAMP analog rp-cAMP had no effect on keratinocyte migration, whether directional or random. However, rp-cAMP pretreatment before β-agonist addition fully restored galvanotaxis, demonstrating the complete cAMP dependence of the attenuation of keratinocyte directional migration. This is the first report that cAMP is capable of mediating keratinocyte galvanotaxis. β-AR agonists and antagonists could be valuable tools for modulating re-epithelialization, an essential step in the wound-healing process. Thus, β-ARs regulate the two distinct components of keratinocyte directional migration differently: migration speed via a cAMP-independent mechanism and galvanotaxis by a cAMP-dependent one.

Original languageEnglish (US)
Pages (from-to)2023-2034
Number of pages12
JournalJournal of Cell Science
Volume118
Issue number9
DOIs
StatePublished - May 1 2005

Fingerprint

Keratinocytes
Cyclic AMP
Adrenergic Receptors
Re-Epithelialization
Protein Phosphatase 2
Pertussis Toxin
Wounds and Injuries
Colforsin
Blindness
Epidermis
Wound Healing
Cues
Skin

Keywords

  • β-adrenergic receptor
  • Adenylyl cyclase
  • Electrotaxis
  • Galvanotaxis
  • Wound healing

ASJC Scopus subject areas

  • Cell Biology

Cite this

Cyclic AMP mediates keratinocyte directional migration in an electric field. / Pullar, Christine E.; Isseroff, Roslyn Rivkah.

In: Journal of Cell Science, Vol. 118, No. 9, 01.05.2005, p. 2023-2034.

Research output: Contribution to journalArticle

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AB - Re-epithelialization of wounded skin is necessary for wound closure and restoration of barrier function and requires directional keratinocyte migration towards the center of the wound. The electric field (EF) generated immediately upon wounding could be the earliest signal keratinocytes receive to initiate directional migration and healing. Keratinocytes express many β2-adrenergic receptors (β2-ARs), but their role in the epidermis is unknown. We have previously shown that β-AR agonists decrease keratinocyte migration in a cyclic AMP (cAMP) independent mechanism involving the activation of protein phosphatase 2A (PP2A). Here, we ask whether β2-ARs play a role in keratinocyte galvanotaxis. We report a bimodal response. When keratinocytes were exposed to higher concentrations of β-AR agonist (0.1 μM), their tracked migratory speed was inhibited, in both the presence (directional migration) and the absence (random migration) of a 100 mV mm-1 EF, as expected. At lower agonist concentrations (0.1 pM to 0.1 nM), there was no effect on migratory speed; however, all directionality was lost - essentially, cells were 'blinded' to the directional cue. Preincubating the cells with β-antagonist restored directional migration, demonstrating that the 'blindness' was β2-AR mediated. Incubation of keratinocytes with agents known to increase intracellular cAMP levels, such as sp-cAMP, pertussis toxin and forskolin, resulted in similar 'blinding' to the EF, whereas random migration was unaffected. The inactive cAMP analog rp-cAMP had no effect on keratinocyte migration, whether directional or random. However, rp-cAMP pretreatment before β-agonist addition fully restored galvanotaxis, demonstrating the complete cAMP dependence of the attenuation of keratinocyte directional migration. This is the first report that cAMP is capable of mediating keratinocyte galvanotaxis. β-AR agonists and antagonists could be valuable tools for modulating re-epithelialization, an essential step in the wound-healing process. Thus, β-ARs regulate the two distinct components of keratinocyte directional migration differently: migration speed via a cAMP-independent mechanism and galvanotaxis by a cAMP-dependent one.

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