title: Insertion of MP20 into lens fibre cell plasma membranes correlates with the formation of an extracellular diffusion barrier.
abstract: It is known that during lens differentiation a number of fibre cell specific membrane proteins change their expression profiles. In this study we have investigated how the profiles of the two most abundant fibre cell membrane proteins AQP0 (formerly known as Major Intrinsic Protein, MIP) and MP20 change as a function of fibre cell differentiation. While AQP0 was always found associated with fibre cell membranes, MP20 was initially found in the cytoplasm of peripheral fibre cells before becoming inserted into the membranes of deeper fibre cells. To determine at what stage in fibre cell differentiation MP20 becomes inserted into the membrane, sections were double-labelled with an antibody against MP20, and propidium iodide, a marker of cell nuclei. This showed that membrane insertion of MP20 occurs in a discrete transition zone that coincided with the degradation of cell nuclei. To test the significance of the membrane insertion of MP20 to overall lens function, whole lenses were incubated for varying times in a solution containing either Texas Red-dextran or Lucifer yellow as markers of extracellular space. Lenses were fixed and then processed for immunocytochemistry. Analysis of these sections showed that both tracer dyes were excluded from the extracellular space in an area that coincided with insertion of MP20 into the plasma membrane. Our results suggest that the insertion of MP20 into fibre cell membranes coincides with the creation of a barrier that restricts the diffusion of molecules into the lens core via the extracellular space.
authors.affiliation: Department of Physiology, School of Medical Sciences, University of Auckland, Private Bag 92019, Auckland, New Zealand.
authors.forename: Angus C
authors.initials: AC
authors.lastname: Grey
authors.forename: Marc D
authors.initials: MD
authors.lastname: Jacobs
authors.forename: Tamir
authors.initials: T
authors.lastname: Gonen
authors.forename: Joerg
authors.initials: J
authors.lastname: Kistler
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Donaldson
mesh_terms: D000818:Animals; D020346:Aquaporins; D002454:Cell Differentiation / Q000502:physiology; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D005110:Extracellular Space / Q000378:metabolism; D005136:Eye Proteins / Q000378:metabolism / Q000502:physiology*; D007908:Lens, Crystalline / Q000166:cytology / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000378:metabolism / Q000502:physiology; D008565:Membrane Proteins; D051381:Rats; D012756:Sheep
chemical_list: D020346:Aquaporins; D005136:Eye Proteins; C489026:Lim2 protein, rat; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; C030328:aquaporin 0