March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
Plasma Membrane Calcium-ATPase Isoform-1 Plays a Role in Corneal Epithelial Wound Healing
Author Affiliations & Notes
  • Ernest F. Talarico, Jr.
    Anatomy & Cell Biology, Indiana Univ Sch of Med-Northwest, Gary, Indiana
  • Stephen M. Koveck
    Chemistry, Indiana University Northwest, Gary, Indiana
  • Footnotes
    Commercial Relationships  Ernest F. Talarico, Jr., Affinity BioReagents, Inc. (F); Stephen M. Koveck, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 3539. doi:
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      Ernest F. Talarico, Jr., Stephen M. Koveck; Plasma Membrane Calcium-ATPase Isoform-1 Plays a Role in Corneal Epithelial Wound Healing. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3539.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : Plasma Membrane Calcium-ATPase (PMCA) is essential in the control of intracellular Ca2+ concentration. In humans, four genes encode PMCA proteins termed PMCA1 - PMCA4. PMCA4 is the major PMCA isoform expressed in human corneal epithelium (CE); with lesser predominance of PMCA1, 2 and 3, respectively, however, little is known about their role(s) in corneal biology. Prior work demonstrated a role for PMCA4 in CE wound healing. The present work examined the functional role of PMCA1 during CE wound healing.

Methods: : The distribution of PMCA1 in postmortem and surgical sections of CE was determined by immunohistochemistry using isoform specific antibodies (Abs) and a panPMCA Ab that recognizes all PMCAs. The role of PMCA1 in wound healing was studied in human telomerase-immortalized corneal epithelial cells (hTCEpi) after transfection with siRNAPMCA1. Cell cultures were wounded 48 h after transfection, and the wound area was measured at 3-h intervals for 48 h. The wound area was normalized to 0 h, and plotted as wound area vs. time post-wounding.

Results: : PMCA1 immunoreactivity (IR) was found mainly on basal and wing cells. In contrast to PMCA4 (located mainly on the apical portions of basal cell plasma membranes), PMCA1 IR was located on portions of basal cell plasma membranes adjacent to the stroma. siRNAPMCA1-transfected hTCEpi cells failed to completely seal the wound area, whereas wounds in cultures transfected with a scrambled construct were completely closed.

Conclusions: : PMCA1 is expressed in human CE and exhibits immunolocalization mainly on basal and wing cells, with a predominance along the basal cell plasma membrane adjacent to the corneal stroma. Knockdown of PMCA1 expression in hTCEpi cells decreases wound healing. The present findings suggest that PMCA1 is one factor in mediating calcium-regulated events necessary for cell migration in regenerating CE.

Keywords: cornea: epithelium • cornea: basic science • wound healing 
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