Fig. 1.
Progranulin expression in the intestine and neurons begins during development. Light (Left) and fluorescent (Right) images of animals expressing a fluorescent mCherry transcriptional reporter (Ppgrn-1::pgrn-1::polycistronic mCherry) (A–F) or a translational RFP reporter (Ppgrn-1::pgrn-1::RFP) (G–J). (A and B) Confocal images of representative mixed-stage embryos demonstrate that progranulin expression begins at approximately 180 min after the first cleavage event in embryogenesis (at approximately the time programmed cell death begins). Left embryo is at approximately 150 min of development and displays no fluorescent signal. Center embryo is at approximately 180 min of development, during early to mid-gastrulation when intestinal precursor cells are seen clearly in the interior of the embryo. These four intestinal precursor cells contain the mCherry fluorescent protein (dashed line). Right: Comma-stage embryo (~390 min) demonstrates strong mCherry expression (See also Fig. S3 A and B). (C–F) Nomarski and fluorescent micrographs show a representative late larval (L4) animal producing PGRN-1 in the intestine (C and D) and select neurons (E and F). (G–J) Nomarski and fluorescent micrographs of a comma-stage animal (G and H) and day 1 adult animal (I and J) expressing a PGRN-1::RFP translational reporter show the protein diffusing throughout the animal. Arrowheads mark a refractile corpse (G and H) and secreted PGRN-1::RFP taken up by coelomocytes (I and J). (Scale bars: A, B, E–J, 10 μm; C and D, 100 μm.)