ABSTRACT
Neurological and psychiatric side effects accompany the high-dose interferon-alpha (IFNA) therapy. The primary genes responsible for these complications are mostly unknown. Our genome-wide search in mouse and rat genomes for the conservative genes containing IFN-stimulated response elements (ISRE) in their promoters revealed a new potential target gene of IFNA, Grin3α, which encodes the 3A subunit of NMDA receptor. This study aimed to explore the impact of IFNA on the expression of Grin3α and Ifnα genes and neurotransmitters endo/exocytosis in the mouse brain. We administered recombinant human IFN-alpha 2b (rhIFN-α2b) intracranially, and 24 h later, we isolated six brain regions and used the samples for RT-qPCR and western blot analysis. Synaptosomes were isolated from the cortex to analyze endo/exocytosis with acridine orange and L-[14C]glutamate. IFNA induced an increase in Grin3α mRNA and GRIN3A protein, but a decrease in Ifnα mRNA and protein. IFNA did not affect the accumulation and distribution of L-[14C]glutamate and acridine orange between synaptosomes and the extra-synaptosomal space. It caused the more significant acridine orange release activated by NMDA or glutamate than from control mice's synaptosomes. In response to IFNA, the newly discovered association between elevated Grin3α expression and NMDA- and glutamate-evoked neurotransmitters release from synaptosomes implies a new molecular mechanism of IFNA neurotoxicity.
Subject(s)
Exocytosis/drug effects , Interferon alpha-2/toxicity , Membrane Glycoproteins/biosynthesis , N-Methylaspartate/pharmacology , Animals , Exocytosis/physiology , Female , Gene Expression , Humans , Membrane Glycoproteins/agonists , Membrane Glycoproteins/genetics , Mice , Mice, Inbred BALB C , Receptors, N-Methyl-D-Aspartate/agonists , Receptors, N-Methyl-D-Aspartate/biosynthesis , Receptors, N-Methyl-D-Aspartate/genetics , Recombinant Proteins/toxicityABSTRACT
The cellular cysteine is highly regulated in a narrow range of concentrations due to its cyto- and neurtoxicity when it is overwhelmed or its deficiency for protein synthesis and other vital metabolic reactions when its amount is restricted. The regulation of cysteine content and its metabolic products, glutathione, taurine and inorganic sulfur compounds, is scarcely explored in human placenta though cysteine metabolism is closely related to the maintenance of redox status and protection from free radical oxidation, elimination of homocysteine and detoxification. These processes are particularly important for placenta which meets substantial changes of oxygen supply during its development, and is the last metabolically active organ between mother and fetus. The abundance of CDO , CSAD , ADO , SUOX, GCLC and GCLM mRNAs was estimated by RT -qPCR and compared with the computationally analyzed microarray gene expression data from GEO , while the level of individual protein by western-blot analysis, both in placental samples from first and third trimesters of uncomplicated pregnancies. The abundance of CDO mRNA is significantly up-regulated at term compared to the first trimester, the level of GCLM and GCLC mRNAs remains almost unchanged while the abundance of other mRNAs reduces to varying degrees. Overall, the changes of gene expression in third trimester in comparison to the first one estimated by RT-qPCR and microarray coincide while the former data are more informative for the limited group of genes. The data provide the basis for further research of these genes expression and phenotype of human placenta in health and disease
