Anti-KRT12 (full length) polyclonal antibody

Purpose: To determine if a mutation within the coding region of the keratin 12 gene (KRT12) is responsible for a severe form of Meesmann's corneal dystrophy. Methods: A family with clinically identified Meesmann's corneal dystrophy was recruited and studied. Electron microscopy was performed on scrapings of corneal epithelial cells from the proband. Mutations in the KRT12 gene were sought using direct genomic sequencing of leukocyte DNA from two affected and two unaffected family members. Subsequently, the observed mutation was screened in all available family members using polymerase chain reaction and direct sequencing. Results: A heterozygous missense mutation (Arg430Pro) was found in exon 6 of KRT12 in all 14 affected individuals studied. Unaffected family members and 100 normal controls were negative for this mutation. Conclusions: We have identified a novel mutation in the KRT12 gene that is associated with a symptomatic phenotype of Meesmann's corneal dystrophy. This mutation results in a substitution of proline for arginine in the helix termination motif that may disrupt the normal helix, leading to a dramatic structural change of the keratin 12 protein.

The IKK beta is known to regulate transcription factor NF-kappa B activation leading to inflammatory responses. Recent gene knockout studies have shown that IKK beta can orchestrate local inflammatory responses and regulate homeostasis of epithelial tissues. To investigate whether IKK beta has an intrinsic role in epithelial cells, we established an in vivo system in the immune privileged corneal epithelium. We generated triple transgenic Krt12(rtTA/rtTAt)/tet-O-Cre/Ikk beta(F/F) (Ikk beta(Delta CE/Delta CE)) mice by crossing the Krt12-rtTA knock-in mice, which express the reverse tetracycline transcription activator in corneal epithelial cells, with the tet-O-Cre and Ikk beta(F/F) mice. Doxcycline-induced IKK beta ablation occurred in corneal epithelial cells of triple transgenic Ikk beta(Delta CE/Delta CE) mice, but loss of IKK beta did not cause ocular abnormalities in fetal development and postnatal maintenance. Instead, loss of IKK beta significantly delayed healing of corneal epithelial debridement without affecting cell proliferation, apoptosis or macrophage infiltration. In vitro studies with human corneal epithelial cells (HCEpi) also showed that IKK beta was required for cytokine-induced cell migration and wound closure but was dispensable for cell proliferation. In both in vivo and in vitro settings, IKK beta was required for optimal activation of NF-kappa B and p38 signaling in corneal epithelial cells, and p38 activation is likely mediated through formation of an IKK beta-p38 protein complex. Thus, our studies in corneal epithelium reveal a previously un-recognized role for IKK beta in the control of epithelial cell motility and wound healing.