A remarkable permeability of canalicular tight junctions might facilitate retrograde, non-viral gene delivery to the liver via the bile duct.
OBJECTIVE
To establish the extent of retrograde bile duct infusion at an ultrastructural level, as a preliminary step before evaluating the efficacy of gene delivery to the rat liver via a branch of the bile duct.


METHODS
The extent of retrograde infusion into the biliary tree was established by light and electron microscopy, following infusion of 10 nm gold particles into the right lateral lobe. Canalicular permeability was further assessed by the infusion of a 67 kDa protein. For gene delivery, both naked DNA and a synthetic peptide vector system were evaluated. Because canalicular tight junction permeability can be compromised in damaged livers, both normal rats and rats recovering from the hepatotoxin D-galactosamine were studied.


RESULTS
The gold particles penetrated the peripheral one third of the hepatic lobules and, surprisingly, reached the space of Disse in normal rats. Equally surprisingly, blood levels of a 67 kDa protein were identical after bile duct infusion and portal vein injection. Gene delivery with peptide/DNA complexes was much more effective in rats treated with D-galactosamine. However, gene delivery with naked DNA was equally effective in normal and damaged livers. Localisation of gene expression showed a scattering of positive hepatocytes restricted to the right lateral lobe.


CONCLUSIONS
Retrograde infusion into the bile duct advances well into the hepatic lobule and reveals a remarkable permeability of the canalicular or cholangiole tight junctions in normal rats. It is an effective approach for delivering genes to a small population (approximately 1%) of hepatocytes.
Department of Hepatology and Transplantation, Guys, Kings and St Thomas School of Medicine, The Rayne Institute, London, UK.
J
J
Hu
X
X
Zhang
X
X
Dong
L
L
Collins
G J
GJ
Sawyer
J W
JW
Fabre
D000818:Animals; D001648:Bile Canaliculi; D002460:Cell Line; D002614:Chelating Agents / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006046:Gold / Q000008:administration & dosage; D007263:Infusions, Parenteral / Q000379:methods; D008099:Liver / Q000187:drug effects / Q000648:ultrastructure; D008239:Lysine / Q000235:genetics; D049329:Nanostructures; D009842:Oligopeptides / Q000235:genetics; D010316:Particle Size; D010539:Permeability; D051381:Rats; D011917:Rats, Inbred Lew; D019108:Tight Junctions / Q000502:physiology*
D002614:Chelating Agents; D009842:Oligopeptides; C418682:molossin peptide; D004533:Egtazic Acid; D006046:Gold; D008239:Lysine