Analysis of Proliferation and Migration in Phenylalanine, Retinoic Acid, and 4-diethylaminobenzaldehyde Treated Cells

Abstract

Maternal phenylketonuria [MPKU] is a syndrome of multiple congenital anomalies including cardiovascular malformations [CVMs], and brain and growth restriction when a mother with Phenylketonuria [PKU] does not control her dietary intake of Phenylalanine [Phe]. However, the mechanisms responsible for Phe-induced CVMs are poorly understood. Our lab has preliminary evidence that high levels of Phe could inhibit Retinoic Acid [RA] signaling, which typically promotes the expression of genes such as proliferation, migration, and differentiation. Proliferation and migration of the neural crest cells are important in formation of the outflow tract (OFT) and aortic arch arteries (AAA). We hypothesize that Phe inhibits migration and proliferation, which may contribute to the defects seen in MPKU. We also looked at the effects of exposure to RA and 4-diethylaminobenzaldehyde [DEAB], a known RA inhibitor. We conducted in-vitro proliferation and migration assays on several cell types to determine if proliferation and migration was affected by Phe, RA, and DEAB exposure. Images were analyzed with ImageJ and GraphPad Prism. Present research suggests that Phe exposure causes a significant decrease in proliferation of cells. It is also shown that RA increases or does not affect proliferation, and that DEAB decreases cell proliferation. In this way, Phe is similar to DEAB, which suggests that it also acts as an RA inhibitor. This could contribute to the CVMs observed in MPKU. The results of mi