Fig. 4. lbk larvae display vesicular phenotypes in liver and intestinal
cells. (A,B) Longitudinal sections showing the liver and
part of the intestine. There is a substantial increase in liver size
(outlined) in lbk. Sections were derived from the same region of the
body and images are reproduced at the same magnification. (C) Confocal
LSM images of larvae expressing GFP under the intestinal promoter ef1 reveal
an increase in liver size (top row, outlined) in lbk. Nuclei are
stained with DAPI (blue). Bottom row shows higher magnifications of regions
from livers shown in the top row. Individual liver cells are outlined showing
the increase in hepatocyte size in lbk. The arrangement of
hepatocytes in lbk is disorganised. (D) Quantification of the
average cell area per nucleus in histological sections of livers revealed that
the lbk liver shows on average an area 30% larger than that of
age-matched siblings [n=5 (sibs.), n=7 (lbk);
P=0.0001]. (E-H) Transverse TEM sections of livers show that
lbk hepatocytes contain numerous large vesicles (v). Asterisks
indicate erythrocytes. At 7 dpf, lbk hepatocytes display necrotic
changes: e.g. a condensation of nuclear chromatin; an increase in the space
between the nuclear membranes (arrowheads); a massively inflated ER lumen (#),
which is continuous with the nuclear membranes; and numerous vesicles often
displaying an electron-dense lumen. N, nucleus. (I,J) Oil Red O
staining of the liver (I) and RPE (J) reveals lipid-containing vesicles
(arrowheads) in hepatocytes and the RPE in lbk. (K,L)
Transverse TEM sections of the anterior intestine show an increase in number
and decrease in size of vesicles in the cells lining the intestine in
lbk. Scale bars: 50 µm in A,B; 50 µm in C; 25 µm in insets;
10 µm in E,F,I,J; 5 µm in G,H; 2 µm in K,L.