RT-PCR ELISA method for the analysis of neurotrophin mRNA expression in brain and peripheral tissues

The levels of nerve growth factor (NGF) and brain derived neurotrophic fact or (BDNF) in brain and periphery are susceptible to changes during developm ent and as result of different physiopathological conditions, such as stres s and aging and during the onset and progression of neurological and autoim mune diseases. Despite the sensitive methods for measurement of neurotrophi n protein levels in different tissues, no easily applicable methods to eval uate changes in the level of NGF and BDNF mRNA expression within physiologi cal range have been described. This study reports the development of a repr oducible and simple procedure for measurement of neurotrophin mRNA expressi on in brain and peripheral tissues based upon an enzyme linked immunosorben t assay (ELISA) detection system of reverse transcriptase-polymerase chain reaction (RT-PCR) products. The major advantages of this RT-PCR ELISA proce dure is to allow the co-amplification of diverse mRNAs starting from small amounts of tissues; to contemporaneously test a large number of samples to be rapid and to use only commercial reagents and widely available equipment . The procedure could also be useful in studies addressed to measure the pa ttern of expression of molecules involved in the pathogenesis of neurodegen erative and inflammatory diseases, such as neuropeptides and cytokines. (C) 2000 Elsevier Science B.V. All rights reserved.