Preliminary Examination of Rabbit Conjunctival Mucins

Abstract

Conjunctival mucins from albino rabbits were fractionated using gel filtration and anion-exchange chromatography. Charge homogeneity of the various conjunctival mucin fractions was confirmed by polyacrylamide gel electrophoresis. The molecular weight characteristics of the isolated fractions varied with the analytical scheme employed. Two schemes yielded mucins with molecular weights of 104, 105, and 106. However, when anion-exchange chromatography was the first step in the fractionation scheme, the dominant mucin had a molecular weight of 1.7 × 105. In contrast, when gel filtration chromatography was the first step, the dominant mucin had a molecular weight of 5.7 × 104. It was postulated that during migration through the anion-exchange matrix, the low molecular weight conjunctival mucin underwent trimer formation. Comparison with the mucin fractions isolated from tear mucoid threads revealed that the scheme beginning with anion-exchange chromatography preserved the fractionation pattern seen in tear mucoid threads. This result implies that conjunctival mucins undergo an association prior to or after their entry into the tear film. The molecular event of interest in this process is self-association of the species with a molecular weight of 5.7 × 104, resulting in a trimer with a molecular weight of 1.7 × 105. This trimer appears to resist deaggregation on exposure to a medium of lesser ionic strength. Several explanations are offered for its formation as well as for its stability. The implication of multiple conjunctival (tear) mucins for tear film stability also is discussed.