Abstract
Phenylketonuria is a genetic defect that, without strict dietary control, results in the accumulation of phenylalanine (Phe) in body fluids. If a low-Phe diet is not maintained during pregnancy, the offspring of phenylketonuric women are born with mental retardation and microcephaly. Primary cultures of rat cerebellar granule cells, rat cortical astrocytes, human fetal astrocytes, and human neuroblastoma (SY5Y) cells and human astrocytoma (1321N1) cells were used to test the hypothesis that the microencephaly may be a result of neuronal cell death and reduced astrocyte proliferation. Exposure to Phe or to six Phe metabolites [phenylacetic acid (PAA), phenyllactic acid, hydroxyphenylacetic acid, phenylpyruvic acid, phenylethylamine (PEA), and mandelic acid] did not result in astroglial or neuronal cell cytotoxicity. Treatment of 1321N1 cells, human fetal astrocytes, or rat astrocytes with 5 mM Phe for 24 h decreased DNA synthesis 19 ± 4, 30 ± 4, and 60 ± 6%, respectively. This effect was concentration dependent, and flow cytometry revealed that Phe treatment resulted in the accumulation of cells in the G0/G1 phase of the cell cycle. In addition, in 1321N1 cells, exposure to 5 mM PAA, and in rat astrocytes, exposure to 0.5 mM PEA inhibited cell proliferation 42 ± 4 and 55 ± 4%, respectively. These metabolites also resulted in the accumulation of cells in the G0/G1 phase of the cell cycle. In human fetal astrocytes, 0.5 mM PEA and 0.5 mM PAA resulted in a 41 ± 12 and 52 ± 11% reduction proliferation, respectively.
Footnotes
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Send reprint requests to: Dr. Lucio G. Costa, Department of Environmental Health, University of Washington, 4225 Roosevelt Way NE, #100, Seattle, WA 98105-6099. E-mail:lgcosta{at}u.washington.edu
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↵1 This study was supported in part by Grants T32 ES07032 and P30 ES07033 from the National Institute of Environmental Health Sciences.
- Abbreviations:
- PKU
- phenylketonuria
- Phe
- phenylalanine
- cPKU
- classic PKU
- mPKU
- maternal PKU
- CNS
- central nervous system
- PEA
- phenylethylamine
- PAA
- phenylacetic acid
- MA
- mandelic acid
- PPA
- phenylpyruvic acid
- PLA
- phenyllactic acid
- HPA
- hydroxyphenylacetic acid
- FBS
- fetal bovine serum
- MTT
- 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
- LDH
- lactate dehydrogenase
- BrdU
- 5-bromodeoxyuridine
- DMEM
- Dulbecco's modified Eagle's medium
- MPPD
- mevalonate pyrophosphate decarboxylase
- Received May 2, 2000.
- Accepted June 15, 2000.
- The American Society for Pharmacology and Experimental Therapeutics
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