Expression and distribution of the serum and glucocorticoid regulated kinase and the epithelial sodium channel subunits in the human cornea.
The sodium transporting capacity of the corneal endothelium is vital for preserving corneal transparency, and has traditionally been attributed to the endothelial pump transporting sodium and bicarbonate across the corneal endothelium, maintaining the cornea in a dehydrated state. Recent studies have shown that the enzyme, serum and glucocorticoid regulated kinase isoform 1 (SGK1), plays a pivotal role in the corticosteroid induction of epithelial sodium transport in tissues such as the distal nephron, through activation of the epithelial sodium channels (ENaC). This study was designed to identify whether these elements were present within the human cornea. In situ hybridisation studies were conducted on paraffin embedded sections from six human eyes, using in-house generated cRNA antisense probes for human SGK1 and ENaC subunits (alpha, beta, gamma), and confirmed expression of SGK1 and all ENaC subunits in the corneal endothelial cytoplasm. Although ENaC subunits were not demonstrated in the corneal epithelium, SGK1 mRNA was identified in the nuclear region of central basal cells of the corneal epithelium, and limbal epithelial cells. Minimal chromagen precipitation was seen in the Bowman's membrane, corneal stroma, or Descemet's membrane. Control experiments consisted of no antisense probe, competition of the labelled antisense cRNA probe by a 60-fold excess unlabelled antisense cRNA, and use of labelled sense cRNA probes, revealing minimal or no hybridisation signal throughout the corneal layers. These data define components of the mineralocorticoid regulatory pathways of sodium transport in human corneal endothelium, and provide evidence for an additional mechanism contributing to corneal transparency and the 'metabolic' sodium pump.
Rauz
Saaeha
S
Academic Unit of Ophthalmology, Division of Immunity and Infection, University of Birmingham, Birmingham and Midland Eye Centre, Dudley Road, Birmingham B18 7QU, UK.
Walker
Elizabeth A
EA
Murray
Philip I
PI
Stewart
Paul M
PM
D003315:Cornea / Q000737:chemistry*; D003593:Cytoplasm / Q000737:chemistry; D004728:Endothelium, Corneal / Q000737:chemistry / Q000648:ultrastructure; D004847:Epithelial Cells / Q000737:chemistry; D053503:Epithelial Sodium Channels; D019573:Epithelium, Corneal / Q000737:chemistry; D006801:Humans; D017874:Immediate-Early Proteins; D017403:In Situ Hybridization / Q000379:methods; D009687:Nuclear Proteins; D020033:Protein Isoforms / Q000032:analysis; D017346:Protein Serine-Threonine Kinases / Q000032:analysis*; D015222:Sodium Channels / Q000032:analysis*; D000099184:Serum-Glucocorticoid Regulated Kinases
D053503:Epithelial Sodium Channels; D017874:Immediate-Early Proteins; D009687:Nuclear Proteins; D020033:Protein Isoforms; D017346:Protein Serine-Threonine Kinases; D015222:Sodium Channels; D000099184:Serum-Glucocorticoid Regulated Kinases