Expression of mRNAs of multiple growth factors and receptors by neuronal cell lines: detection with RT-PCR.
Neurons and glia are capable of both secreting and responding to a large variety of growth factors. However, information on multiple expression of growth factors and their receptors was usually obtained from uncorrelated observations, using cells from various animals of origin, developmental stages, growth phases, culture ages and culture conditions. Because of its specificity and extreme sensitivity, reverse transcription-polymerase chain reaction (RT-PCR) is uniquely suitable to study a large panel of growth factors and their receptors from a limited cell sample, free of these intervening variables. In this paper we evaluate the expression of mRNA of a total of 35 growth factor-related proteins by conducting RT-PCR on three neuronal cell lines: the PC12 rat pheochromocytoma line, the MAH rat sympathoadrenal progenitor line, and the N18 mouse neuroblastoma line. Three types of results are presented. The first confirms the existing knowledge such as the presence of Trk-A (NFG receptor) in PC12. The second consists of new information that expands and extends earlier observations, such as the presence of CNTF receptor complex in PC12, which explains our previous report that CNTF enhances the biological effects of NGF on these cells. The third consists of novel information that leads the way to further experimentation by the more conventional methods. These include the strong expression of Trk-B by MAH, predicting the biological responsiveness of MAH to BDNF and NT-4, and the expression of CNTF receptor in N18. Our results also suggest that CNTF is an autocrine factor for PC12 and MAH, since both lines express the growth factor as well as the receptor. Thus, RT-PCR is a valuable tool in growth factor research that can be used in complement to, and interactively with, other approaches such as bioassay, receptor binding, and immunochemical determination. It will be particularly useful for screening a large number of growth factors in minute areas of the brain in patients suffering from neurodegenerative diseases such as Parkinson's and Alzheimer's.
Zaheer
A
A
Department of Neurology, University of Iowa College of Medicine and Veterans Affairs Medical Center, Iowa City 52242, USA.
Zhong
W
W
Lim
R
R
D000818:Animals; D001483:Base Sequence; D019208:Brain-Derived Neurotrophic Factor; D020934:Ciliary Neurotrophic Factor; D015870:Gene Expression; D006133:Growth Substances / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000494:pharmacology; D009447:Neuroblastoma; D009474:Neurons / Q000378:metabolism*; D016716:PC12 Cells; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism*; D012194:RNA-Directed DNA Polymerase; D051381:Rats; D020801:Receptor, Ciliary Neurotrophic Factor; D017978:Receptors, Growth Factor / Q000235:genetics*; D017475:Receptors, Nerve Growth Factor / Q000235:genetics / Q000378:metabolism; D014407:Tumor Cells, Cultured
D019208:Brain-Derived Neurotrophic Factor; D020934:Ciliary Neurotrophic Factor; D006133:Growth Substances; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D020801:Receptor, Ciliary Neurotrophic Factor; D017978:Receptors, Growth Factor; D017475:Receptors, Nerve Growth Factor; D012194:RNA-Directed DNA Polymerase