Experimental Analysis of Spherocytosis

BIOC 211
Experiment 2
Introduction
Spherocytosis is an auto-hemolytic anemia marked by the production of spherical erythrocytes instead of normal biconcave cells. Even though these spherical cells are capable of efficacious oxygen transport, they are vulnerable to subtle changes in tonicity and can degrade much more easily than normal erythrocytes. This genesis of this condition can traced to four abnormalities that occur in the proteins of the erythrocyte membrane: defects in only spectrin, in both spectrin and ankyrin, flaws with the anion exchanger, and defects of palladin. In some strains of spherocytosis, erythrocytes are so fragile that hemolysis occurs within the bloodstream. This creates microemboli that are capable of blocking capillaries, leading to cumulative brain, heart, retinal, and kidney damage.  
Spherocytosis is currently a terminal condition. Therapies being developed to extend the patients’ lifespan or eliminate the disease altogether rely on removing affected bone marrow stem cells from individuals with spherocytosis, culturing the cells, and replacing defective genes with normal genes. It is fundamental that the bone marrow removed actually produced abnormal erythrocytes and the treated marrow produces normal erythrocytes. Therefore, it is crucial that we establish what differences, if any, exist between in vitro and in vivo cells, both abnormal and normal.
The experimental objective was to determine which proteins were present in the erythrocyte skeleton. The results of the experiment could then be used to provide a basis for the comparison discussed previously.
The model blood sample used for this study was taken from the domestic horse (Equus caballus). Horse blood was an ideal experimental model for this study due to numerous factors: it is readily available, its components may be isolated easily, and resolves clearly in gel electrophoresis. Furthermore, horses and humans are mammals, allowing potential extrapolations of data to...