Gene expression screening for hypoxia inducible factor-1 (HIF-1) and nuclear factor-kB in Human cDNA libraries.

VICENTE BERMÚDEZ; W.J. LUKIW; N.G. BAZAN

Buenos Aires

Congreso; Joint Meeting Eighteenth Biennal Meeting of the International Society for Neurochemistry (ISN) and Thirty-Second Annual Meeting of the American Society for Neurochemistry; 2001

International Society for Neurochemistry y American Society for Neurochemistry

Hypoxia inducible factor (HIF-1) and nuclear factor-kB (NF-kB) are key oxygen sensitive transcription factors (TFs) involved in the rapid induction of oxygen sensitive genes. The aim of this study was to determine the expression patterns of HIF-1 and NF-kB in human tissue samples by screening expressed genes in cDNA libraries. Determinations of B-Actin, glyceraldehyde phosphate dehydrogenase (GAPDH) and ribosomal protein S9 expression patterns were used as internal controls. ATCC Express-Check human cDNA libraries consisting of 63 RT-PCR amplified samples from tissue specific cDNA libraries were used; RT-PCR assay was performed for each sample and amplified products were separated according to their size on 1.5% agarose 1 × Tris-Borate-EDTA gels stained with ethidium bromide. Stained gels were imaged using a Bio-Rad GS-250 Molecular Imager. High expression of NF-kB was detected mainly in brain embryonic tissue cDNA at different stages of development and in kidney proximal tubules. In nearly all the brain tissues cDNAs of the library significant expression of NF-kB was found. HIF-1 RNA message was detected in abundance in 75 day embryonic brain, in B lymphocyte and in kidney proximal tubules. HIF-1 and NF-kB RNA message levels were both abundant in 1-day-old brain stem, basal ganglia, tonsil, B lymphocyte and also in kidney proximal tubule cDNA pools. These results suggest that elevated expression of these transcripts may be important during the development of these tissues and in particular during embryonic brainstem and basal ganglia development. Supported by NS230002 and the EENTFoundation.