Expression of transfected genes by differentiated, postmitotic neurons and photoreceptors in primary cell cultures

Expression of transfected genes by differentiated, postmitotic neurons and photoreceptors in... We report here that differentiated, primary, postmitotic neurons and photoreceptors in cultures obtained from embryonic chicks can express foreign genes after transfection by the calcium phosphate method. A variety of viral promoters were tested by using either β‐galactosidase or chloramphenicol acetyltransferase as reporter genes. Histochemical and immunocytochemical analysis showed β‐galactosidase expression by both neurons and photoreceptors. As commonly observed with dividing cells, transfection efficiencies showed inter‐experimental variability, with efficiencies ranging from 2% to 20% using the same plasmid. On the other hand, intra‐experimental variability between replicate dishes was much smaller. Analysis using the highly sensitive enzymatic assay for chloramphenicol acetyl transferase (CAT) showed that all of the promoter/enhancers‐CAT constructs tested, with the exception of a construct containing the Maloney sarcoma virus promoter, led to the expression of detectable activity when transfected into cultured retinal cells. The calcium phosphate treatment used for cell transactions did not show detectable effects on overall cell survival, although it caused selective decreases in some metabolic activities of the cells. The studies demonstrate that it is possible to obtain expression of genes transfected into primary, postmitotic neuronal cells. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Neuroscience Research Wiley

Expression of transfected genes by differentiated, postmitotic neurons and photoreceptors in primary cell cultures

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Publisher
Wiley
Copyright
Copyright © 1990 Wiley‐Liss, Inc.
ISSN
0360-4012
eISSN
1097-4547
DOI
10.1002/jnr.490250107
Publisher site
See Article on Publisher Site

Abstract

We report here that differentiated, primary, postmitotic neurons and photoreceptors in cultures obtained from embryonic chicks can express foreign genes after transfection by the calcium phosphate method. A variety of viral promoters were tested by using either β‐galactosidase or chloramphenicol acetyltransferase as reporter genes. Histochemical and immunocytochemical analysis showed β‐galactosidase expression by both neurons and photoreceptors. As commonly observed with dividing cells, transfection efficiencies showed inter‐experimental variability, with efficiencies ranging from 2% to 20% using the same plasmid. On the other hand, intra‐experimental variability between replicate dishes was much smaller. Analysis using the highly sensitive enzymatic assay for chloramphenicol acetyl transferase (CAT) showed that all of the promoter/enhancers‐CAT constructs tested, with the exception of a construct containing the Maloney sarcoma virus promoter, led to the expression of detectable activity when transfected into cultured retinal cells. The calcium phosphate treatment used for cell transactions did not show detectable effects on overall cell survival, although it caused selective decreases in some metabolic activities of the cells. The studies demonstrate that it is possible to obtain expression of genes transfected into primary, postmitotic neuronal cells.

Journal

Journal of Neuroscience ResearchWiley

Published: Jan 1, 1990

References

  • Expression of cone‐like properties by chick embryo neural retina cells in glial‐free monolayer cultures
    Adler, Adler; Lindsey, Lindsey; Elsner, Elsner
  • A rapid radiochemical method for the determination of choline acetyltransferase
    Fonnum, Fonnum

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