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Expression of mRNAs of multiple growth factors and receptors by neuronal cell lines: Detection with RT-PCR

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Abstract

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.

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Abbreviations

BDNF:

brain-derived neurotrophic factor

CNTF:

ciliary neurotrophic factor

EGF:

epidermal growth factor

bFGF:

basic fibroblast growth factor

GMF-β:

glia maturation factor β

IGF-1:

insulin-like growth factor 1

IGF-2:

insulin-like growth factor 2

IL-1:

interleukin 1

IL-6:

interleukin 6

INS:

insulin

LIF:

leukemia inhibitory factor

NGF:

nerve growth factor

NT-3:

neurotrophin 3

NT-4:

neurotrophin 4

PDGF:

platelet-derived growth factor

RT-PCR:

reverse transcription—polymerase chain reaction

SCF:

stem cell factor

TGF-β:

transforming growth factor β

“R”:

following a growth factor desiguates its receptor

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  1. Department of Neurology (Division of Neurochemistry and Neurobiology), The University of Iowa College of Medicine and Veterans Affairs Medical Center, 52242, Iowa City, IA

    Asgar Zaheer, Weixiong Zhong & Ramon Lim

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  1. Asgar Zaheer
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Zaheer, A., Zhong, W. & Lim, R. Expression of mRNAs of multiple growth factors and receptors by neuronal cell lines: Detection with RT-PCR. Neurochem Res 20, 1457–1463 (1995). https://doi.org/10.1007/BF00970594

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