The interaction of DNR and glutaraldehyde with cell membrane proteins leads to morphological changes in erythrocytes

In this study, the effects of DNR and glutaraldehyde on isolated erythrocyte membrane proteins were examined. For this purpose, SDS-gel electrophoresis was carried out. Additionally, analyses of the disturbances in erythrocyte shape and size, accompanied by the application of flow cytometry and microscopy examination, were undertaken. The amount of DNR linked to erythrocyte cell membranes was measured by a fluorimetric technique. It was observed that glutaraldehyde caused in concentration dependent manner an increase of percent of DNR linked to cell membrane proteins. After this incorporation, perturbations in the protein content of cell membranes were observed. The protein aggregates and changes in the level of spectrin, actin and band 3 protein were noted. Due to the changes in spectrin, which is mainly responsible for maintenance of the discocyte shape of erythrocytes, flow cytometry and microscopy techniques were used to control the size and shape of erythrocytes after treatment with DNR and glutaraldehyde. The disturbances in the shape and size of erythrocytes were observed for all tested concentrations of glutaraldehyde. For all tested concentrations of glutaraldehyde, the changes were statistically significant.