New blood for hemoglobin in the lens: roles in stem cell differentiation and fibre cell organelle loss?

Abstract

Purpose Evidence is emerging for haemoglobin (Hb) expression outside the vascular system. We previously demonstrated Hb expression in the mouse lens during post-natal development and cataract progression. Here, we extended this work by carrying out a comprehensive spatio-temporal analysis of Hb subunit expression during mouse lens development and maturation. Methods We used RT-PCR, Western blotting and immunofluorescence to analyze Hb expression in mouse eyes (E16.5 to 9 wks). We also used a sensitive heme assay to test for the presence of heme in the lens by colourimetric assay and histological staining of paraffin-embedded sections. Results Hb subunits were expressed in lens epithelial cells and cortical lens fibre cells. However, the heme assay revealed negligible levels of this prosthetic group in the lens. Hb immunofluorescence was also observed in other regions of the developing eye including the cornea, the retinal ganglion cell layer and the retinal pigment epithelium. Finally, we also observed Hb expression in early embryos by microarray and during differentiation of embryonic stem (ES) cells into embryoid bodies (EBs) in vitro. Conclusion These results suggest a paradigm shift: Hb subunits are expressed in the eye during development and in the adult and, therefore, may have novel roles in ocular development, physiology and pathophysiology. The absence of heme from the lens indicates that at least some of these functions may be independent of oxygen metabolism. The pattern of expression of Hb in lens epithelial cells and cortical lens fibre cells may indicate an involvement for Hb subunits in lens epithelial cell differentiation into lens fibre cells and/or lens fibre cell organelle loss.