A study of gene expression profiles of cultured embryonic rat neurons induced by phenylalanine.
To have more insight into the mechanism of neuronal injury in phenylketonuria (PKU) patients, gene expression profiles were studied in cell culture of embryonic rat cortical neurons induced by phenylalanine. Randomly chose cortical cultured for 3 days were treated by 0.9-mM phenylalanine for 12 h. Control group of the same batch was treated with the same volume of medium. Total RNA was extracted and hybridized with the Affymetrix gene chip U34 according to the protocol provided by the Affymetrix Company. Real-time PCR was used to further confirm the result. We found that the hybridization signals of 167 genes were increased among the total 1323 probes plotted on the chip. The 167 increased genes could be functionally categorized into signal transduction, neuron related, cytoskeleton, metabolism, ion channels, transcription factors, cytokines, and apoptosis related. Signals of seven probes were decreased, which accounted to 0.5% of the total number. A series of genes that were not reported previously were upregulated by phenylalanine, including Ca2+/calmodulin-dependent protein kinase, Brain type II (CaMK II), ras, P38, L-voltage dependent calcium channel, some genes related to vesicle formation and transmitter release, some glutamate receptor subunits and glutamate transporters. According to the gene expression profiles, it is likely that multiprocesses are involved in the neuronal injury induced by phenylalanine, such as the activation on of the NMDR-Ca2+-CaMK II-Ras-P38 axis, the abnormality in neurotransmitter release. Our study also suggests that the excitatory neurotransmitter glutamate may play a role in the neural pathology of PKU.
Zhang
Huiwen
H
Department of Endocrinology and Genetic Metabolism, Xin Hua Hospital, Shanghai Institute for Pediatric Research, Shanghai Second Medical University, Shanghai 200092, People's Republic of China.
Gu
Xue Fan
XF
D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000187:drug effects / Q000378:metabolism*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects / Q000502:physiology*; D018698:Glutamic Acid / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D009498:Neurotoxins / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010649:Phenylalanine / Q000378:metabolism / Q000494:pharmacology*; D010661:Phenylketonurias / Q000378:metabolism*; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology
D009498:Neurotoxins; D012333:RNA, Messenger; D018698:Glutamic Acid; D010649:Phenylalanine