Cx31.1 is associated with apoptosis of scrape-wounded HaCaTs in diabetic conditions

Catherine Wright, Iain Robertson, Narges Elgaseai , Patricia Martin

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Modulating connexin43 (Cx43) mediated communication via gap junctions and/or hemichannels increases the wound-healing capacity of skin models in simulated normal and diabetic conditions. Cx31.1 expression is associated with apoptosis in eye and ovary. This study investigated whether Cx31.1 expression was altered in HaCaT scrape-wounded monolayers cultured in normal (NGI) and high glucose and insulin (HGI) and if this was associated with apoptosis.
HaCaT cells were treated for 5 days with normal or high glucose (5.5 or 25 mM) and insulin (1 or 10 nM). 600 µm scrape-wounds were introduced and cell movement into the denuded area monitored over 48 h. Cx31.1 expression was determined by immunocytochemistry. Conditioned media was assessed for caspase 3/7 activity, with cells exposure to UV 18 h before caspase measurement (positive control). MTT assay measured cell viability.
Cx31.1 expression was increased in HGI compared to NGI, and at higher levels on the membranes of apoptotic rather than healthy cells. Cx31.1 appeared to increase in wound edges at 24 and 48 h after scrape-wounding. Caspase activity in conditioned media from the scrape-wounds showed that apoptosis in NGI reduced at 24 h post-wounding and was similar to control levels at 48 h, although caspase in HGI did not alter. UV exposure significantly increased caspase in both NGI and HGI (P < 0.05) although HGI cells showed less caspase than NGI cells pre-UV (P < 0.01). In parallel cell viability was reduced post-UV exposure in NGI cells (P < 0.001), although not in HGI. Cell viability was not significantly different in unexposed cells under NGI or HGI.
Cx31.1 expression altered during wound closure and was associated with ‘diabetic’ levels of glucose, insulin and apoptosis in HaCaT cells. Despite Cx31.1 being higher in HGI, caspase activity did not alter in HGI conditioned media. Cells in HGI retained viability after UV exposure to a greater extent than those in NGI. Cx31.1 is modulated in wound healing, and may be associated with tissue turn-over in diabetic wounds.
Original languageEnglish
Title of host publicationJournal of Investigative Dermatology
PagesS256
Number of pages1
Volume133
DOIs
Publication statusPublished - 2013

Keywords

  • connexin
  • wound healing
  • diabetes

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    Wright, C., Robertson, I., Elgaseai , N., & Martin, P. (2013). Cx31.1 is associated with apoptosis of scrape-wounded HaCaTs in diabetic conditions. In Journal of Investigative Dermatology (Vol. 133, pp. S256). [1508] https://doi.org/10.1038/jid.2013.107