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An Essential and Synergistic Role of Purinergic Signaling in Guided Migration of Corneal Epithelial Cells in Physiological Electric Fields.
Cell Physiol Biochem 2019; 52(2):198-211CP

Abstract

BACKGROUND/AIMS

Directional migration of corneal epithelial cells is essential for healing of corneal wounds, which is a robust response mediated by biochemical and bioelectrical cues. Naturally occurring electric fields at corneal wounds provide a powerful guidance cue for directional cell migration, as does extracellular ATP. Our recent large-scale siRNA library screening identified a role for purinergic signaling in the electric field-guided migration (galvanotaxis/electrotaxis) of human corneal epithelial (hTCEpi) cells.

METHODS

We examined the effect of extracellular ATP on galvanotaxis of hTCEpi cells. Galvanotactic cell migration was recorded by video microscopy, and directedness and migration speed was calculated. The role of purinergic receptors in galvanotaxis regulation was evaluated by pharmacological inhibition or knocking down of P2X and P2Y receptors.

RESULTS

Addition of ATP enhanced galvanotaxis, and most remarkably sensitized galvanotaxis response to very low level of electric fields in the physiological range (10-30 mV/mm). The stimulatory effect of extracellular ATP was diminished by apyrase treatment. Importantly, cells stimulated with extracellular ATP migrated with significantly increased directedness and speed, which were diminished by knocking down or pharmacological inhibition of P2X and P2Y receptors. Inhibition of pannexin-1 (ATP permeable channel) significantly impaired galvanotaxis. Moreover, pharmacological inhibition of ectoATPase enhanced galvanotaxis.

CONCLUSION

Extracellular ATP and physiological electric fields synergistically enhanced the galvanotaxis response of hTCEpi cells. hTCEpi cells are likely to secrete ATP actively, and purinergic signaling is down-regulated by ecto-ATPases. Both P2X and P2Y receptors coordinately play a role for galvanotaxis of hTCEpi cells.

Authors+Show Affiliations

Department of Dermatology, University of California at Davis, Sacramento, CA, USA, kennakajima@ucdavis.edu.Department of Dermatology, University of California at Davis, Sacramento, CA, USA. Wakayama Medical University School of Medicine, Wakayama, Japan.Department of Dermatology, University of California at Davis, Sacramento, CA, USA. Department of Ophthalmology, University of California at Davis, Sacramento, CA, USA, minzhao@ucdavis.edu.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

30816668

Citation

Nakajima, Ken-Ichi, et al. "An Essential and Synergistic Role of Purinergic Signaling in Guided Migration of Corneal Epithelial Cells in Physiological Electric Fields." Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology, vol. 52, no. 2, 2019, pp. 198-211.
Nakajima KI, Tatsumi M, Zhao M. An Essential and Synergistic Role of Purinergic Signaling in Guided Migration of Corneal Epithelial Cells in Physiological Electric Fields. Cell Physiol Biochem. 2019;52(2):198-211.
Nakajima, K. I., Tatsumi, M., & Zhao, M. (2019). An Essential and Synergistic Role of Purinergic Signaling in Guided Migration of Corneal Epithelial Cells in Physiological Electric Fields. Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology, 52(2), pp. 198-211. doi:10.33594/000000014.
Nakajima KI, Tatsumi M, Zhao M. An Essential and Synergistic Role of Purinergic Signaling in Guided Migration of Corneal Epithelial Cells in Physiological Electric Fields. Cell Physiol Biochem. 2019;52(2):198-211. PubMed PMID: 30816668.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - An Essential and Synergistic Role of Purinergic Signaling in Guided Migration of Corneal Epithelial Cells in Physiological Electric Fields. AU - Nakajima,Ken-Ichi, AU - Tatsumi,Makiko, AU - Zhao,Min, Y1 - 2019/02/28/ PY - 2018/05/30/received PY - 2019/02/22/accepted PY - 2019/3/1/entrez PY - 2019/3/1/pubmed PY - 2019/3/9/medline KW - ATP KW - Corneal epithelial cell KW - Electric field KW - Galvanotaxis KW - Purinergic signaling SP - 198 EP - 211 JF - Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology JO - Cell. Physiol. Biochem. VL - 52 IS - 2 N2 - BACKGROUND/AIMS: Directional migration of corneal epithelial cells is essential for healing of corneal wounds, which is a robust response mediated by biochemical and bioelectrical cues. Naturally occurring electric fields at corneal wounds provide a powerful guidance cue for directional cell migration, as does extracellular ATP. Our recent large-scale siRNA library screening identified a role for purinergic signaling in the electric field-guided migration (galvanotaxis/electrotaxis) of human corneal epithelial (hTCEpi) cells. METHODS: We examined the effect of extracellular ATP on galvanotaxis of hTCEpi cells. Galvanotactic cell migration was recorded by video microscopy, and directedness and migration speed was calculated. The role of purinergic receptors in galvanotaxis regulation was evaluated by pharmacological inhibition or knocking down of P2X and P2Y receptors. RESULTS: Addition of ATP enhanced galvanotaxis, and most remarkably sensitized galvanotaxis response to very low level of electric fields in the physiological range (10-30 mV/mm). The stimulatory effect of extracellular ATP was diminished by apyrase treatment. Importantly, cells stimulated with extracellular ATP migrated with significantly increased directedness and speed, which were diminished by knocking down or pharmacological inhibition of P2X and P2Y receptors. Inhibition of pannexin-1 (ATP permeable channel) significantly impaired galvanotaxis. Moreover, pharmacological inhibition of ectoATPase enhanced galvanotaxis. CONCLUSION: Extracellular ATP and physiological electric fields synergistically enhanced the galvanotaxis response of hTCEpi cells. hTCEpi cells are likely to secrete ATP actively, and purinergic signaling is down-regulated by ecto-ATPases. Both P2X and P2Y receptors coordinately play a role for galvanotaxis of hTCEpi cells. SN - 1421-9778 UR - https://www.unboundmedicine.com/medline/citation/30816668/An_Essential_and_Synergistic_Role_of_Purinergic_Signaling_in_Guided_Migration_of_Corneal_Epithelial_Cells_in_Physiological_Electric_Fields L2 - https://www.cellphysiolbiochem.com/Articles/?DOI=10.33594/000000014 DB - PRIME DP - Unbound Medicine ER -