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Show SCHOOL OF MEDICINE AND HEALTH SCIENCES UNDERGRADUATE RESEARCH ABSTRACTS DETERMINING EXPRESSION OF CD64 SURFACE RECEPTOR GENE IN NEURONS Haley Carlson (John Greenlee) Departments of Bioengineering, Neurology University of Utah Paraneoplastic cerebllar degeneration (PCD) is a neurological syndrome characterized by loss of cerebellar function. PCD is most often seen in w o m e n with a gynecological or breast cancer, but can occur in patients with other types of cancer. The symptoms of PCD are motor and speech impairment, as well as vertigo, and PCD eventually can lead to death. Currently, little is known about the mechanism of this disease. It has been observed that a certain antibody, called "anti-Yo", which attacks tumor cells, presents in the Purkinje cells in the cerebellum. The anti-Yo antibody was found to be a type of antibody called immunoglobulin G (IgG). Through previous research, it has been observed that these antibodies are taken up into Purkinje cells and cause subsequent death, which implies that these observed antibodies are the cause of PCD. To better understand the mechanism of PCD, we want to know what antibody receptors could be responsible for the uptake of the anti-Yo antibodies. There are three types of receptors that bind to IgG antibodies: CD64, CD32, and CD16. The CD64 receptor has the highest affinity for IgG, and is thought to be a likely candidate for uptake in neurons. However, expression of these proteins in neurons has not been characterized. Our hypothesis was CD64 was present in neurons, implying CD64 could be responsible for antibody uptake. Here, we cultured neurons and assayed for the presence of CD64 receptor expression using RT-PCR and compared this expression to that seen in cultures of microglia that are known to express the CD64 receptor, and served as a positive control for our assay. W e found by analyzing the PCR product by gel electrophoresis, that the CD64 receptor is present in neurons. Negative Microglia Neurons CD64 GAPDH 25 cycles Figure 1 -Gei of P C R products for the C D 6 4 gene. Equivalent amounts of cDSiA for microglia and neurons were used as templates for a P C R to amplify the C D 6 4 gene. The P C R products were fractionated on an agar/JST gel and visualized. Note the lack of a band in the negative control, and the presence of a band for the neurons. John Greenlee |
