Introduction of the human growth hormone gene into the guinea pig mammary gland by in vivo transfection promotes sustained expression of human growth hormone in the milk throughout lactation.
We tested the feasibility of transfecting mammary tissue in vivo with an expression plasmid encoding the human growth hormone (hGH) gene, under the control of the cytomegalovirus promoter. Guinea pig mammary glands were transfected with plasmid DNA infused through the nipple canal and expression was monitored in control and transfected glands by radioimmunoassay of milk samples for hGH. Sustained expression of hGH throughout lactation was attained with a polyion transfection complex shown to be optimal for the transfection of bovine mammary cells, in vitro. However, contrary to expectations, hGH expression was consistently 5- to 10-fold higher when DEAE-dextran was used alone for transfection. Thus polyion complexes which are optimal for the transfection of cells in vitro may not be optimal in vivo. The highest concentrations of hGH in milk were obtained when glands were transfected within 3 days before parturition. This method may have application for studying the biological role or physical properties of recombinant proteins expressed in low quantities, or for investigating the regulation of gene promoters without the need to construct viral vectors or produce transgenic animals.
Hens
J R
JR
Department of Animal and Avian Sciences, University of Maryland, College Park 20742, USA.
Amstutz
M D
MD
Schanbacher
F L
FL
Mather
I H
IH
D000818:Animals; D005260:Female; D006168:Guinea Pigs; D019382:Human Growth Hormone / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D007774:Lactation / Q000502:physiology*; D008321:Mammary Glands, Animal / Q000502:physiology*; D008892:Milk / Q000737:chemistry*; D010957:Plasmids; D011247:Pregnancy; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D014162:Transfection / Q000379:methods
D011994:Recombinant Proteins; D019382:Human Growth Hormone