Sulfur mustard exposure disrupts transmembrane water channel proteins in rabbit corneas in vivo
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Transmembrane water channel proteins, Aquaporins (AQPs), regulate corneal homeostasis, hydration, and transparency influencing keratocyte function, inflammatory and wound healing processes in vivo. The functional role of AQPs in mustard gas keratopathy (MGK) remain elusive. Recently, for the first time, we identified involvement of AQP in MGK in vivo (Bhend et. al., Exp Eye Res 2023). This study investigated (a) the effects of sulfur mustard (SM) gas exposure on AQP1, AQP3, and AQP5 transcript and protein levels in rabbit cornea in vivo and (b) tested if SM toxicity to corneal aquaporins could be mitigated by topical eye drop (TED). New Zealand White Rabbits exposed to SM vapor (200 mg-min/m3 for 8 mins) were divided into 3 groups: Naïve, SM, and SM+TED (2 drops/day for 4-week). At 2- and 4-month post SM-exposure, corneas were collected after humane euthanasia, snap-frozen, and used to generate serial tissue sections and cDNAs. Histopathological, immunofluorescence, and qRT-PCR anal yses measured changes in AQP1, AQP3, and AQP5 transcript and protein levels. The clinical eye tests and imaging in live rabbits were performed with slit-lamp microscope, in vivo confocal microscope, specular microscope, optical coherence tomography, fluorescein staining, Schirmer’s tests, pachymetry, applanation tonometry. The SM-exposed rabbit corneas showed significant decrease AQP1, AQP3, and AQP5 mRNA and protein levels compared to naïve corneas at 2- and 4-month post-SM (p[less than]0.001). Further, the TED treatment mitigated SM toxicity to AQPs at both tested times (p[less than]0.001, p[less than]0.01, p=0.004, p=0.03). These results demand in-depth investigation of aquaporin’s role in MGK. Additional studies are underway.
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