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

✍ Scribed by M. Werner; S. Madreperla; P. Lieberman; Dr. R. Adler

• Publisher: John Wiley and Sons
• Year: 1990
• Tongue: English
• Weight: 731 KB
• Volume: 25
• Category: Article
• ISSN: 0360-4012
• DOI: 10.1002/jnr.490250107

✦ Synopsis

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.