We developed the Alb-DsRed2 transgenic (Tg) rat designed with liver-specific expression of the red fluorescent protein, DsRed2. Herein, we report high expression of DsRed2 in neonate liver of both sexes, although they were sexually dimorphic and exhibited a male-specific pattern in adult rats. In an effort to examine the expression in each animal under development, we employed an <i>in vivo</i> Bio-imaging system to quantitatively estimate hepatic DsRed2 expression levels. The temporal profiles pertaining to DsRed expression were similar in male and female Tg rats until 28 days old. The levels in both sexes decreased gradually following birth, and were not detectable at 21 days. Subsequently, expression in males increased again at 35 days and was maintained at a persistently high level thereafter. On the other hand, expression in females disappeared steadily. Although hepatic DsRed expression levels in gonadectomized Tg rats was not significantly different, DsRed expression in hypophysectomized female Tg rats appeared dramatically 72 hr following operation. Hepatocytes were collected from adult Tg rats and cultured in conditioning medium. DsRed expression in female hepatocytes could be detected 72 hr following culturing. These results suggest that hepatic DsRed expression in female rats is regulated <i>in vivo</i> by the pituitary. This report is shows use of Alb-DsRed2 Tg rats in conjunction with a novel bio-imaging system represents a powerful experimental system.
The genetic modification for organ transplantation is one of the most promising strategies to regulate allogeneic immune response. Organ-selective gene transfer has especially benefit to control local immune responses. Based on the catheter technique, we tested to deliver naked plasmid DNA to target graft organs of rats (liver and limbs) by a rapid injection (hydrodynamics-based transfection). Recent advances in transplantation have been achieved by visualization of cellular process and delivered gene expression during the inflammatory process by using non-invasive in vivo imaging. Herein, we examined the fate of genetically modified grafts using a firefly luciferase expression plasmid. For liver modification before transplantation, 6.25% of body weight PBS containing plasmid DNA was injected into the liver through the inferior vena cava using a catheter, and the liver was subsequently transplanted to the recipient rat. For limb modification, the femoral caudal epigastric vein was used. In the rat liver transplantation model, substantial luciferase expression was visualized and sustained for only a few days in the grafted liver. We also addressed stress responses by this hydrodynamics procedure using reporter plasmids containing cis-acting enhancer binding site such as NF-kappa B, cAMP, or heat shock response element. In contrast to hepatic transduction, this genetic limb targeting achieved long lasting luciferase expression in the muscle for 2 months or more. Thus, our results suggest that this catheter-based in vivo transfection technique provides an effective strategy for organ-selective gene modification in transplantation, and the bioluminescent imaging is broadening its potential for evaluation to various preclinical studies.