Quach et al., 2015 - A Multifunctional Mutagenesis System for Analysis of Gene Function in Zebrafish. G3 (Bethesda)   5(6):1283-99 Full text @ G3 (Bethesda)

Fig. 3

Reporter expression in various cell types. (A–I) Examples of enhancer trap reporter GFP expression patterns in the liver (A), pancreas (B), intestine (C), central nervous system (D), swim bladder (D), muscle cells (E), heart tube (F), branchial arches (G), fins (H), and cranial cartilage (I). (J–O) Examples of protein trap reporter mCherry expression patterns in notochord (J), anterior pronephric duct (K), mouth (L), inner ear (M), olfactory placode (N), and pineal gland (O). Ventral views are shown in (A) and (G). Lateral views are shown in (B–F), (H), (J), (K), and (M), with anterior to the left. Dorsal views are shown in (I), (L), (N), and (O). CNS, central nervous system; SB, swim bladder. Scale bars represent 100 µm in (B), 200 µm in (D), 20 µm in (J) and (M), and 50 µm in other images.

Fig. 4

Tissue-specific expression patterns may reveal activities of trapped loci. (A–D) Bright field (A) and fluorescent images of Tg(DsDELGT4)ws2036 embryos at 52 hr after fertilization (A–C), and 5 d after fertilization (D). Arrows show GFP expression in venous sprouts from posterior cardinal vein (C) and inter-segmental veins (ISVs) (D). (E–J) Bright field (I) and fluorescent images of Tg(DsDELGT4)ws0449 embryos at 5.25 hr after fertilization (E, G), 2 d after fertilization (F, H), and 6 d after fertilization (I, J). GFP expresses in CNS and mCherry expresses in posterior cardinal vein (PCV), caudal vein (CV) (arrowheads in H), and intersegmental veins (arrows in J), but not in dorsal aorta (DA in J). (K, L) Fluorescent images of a CU928220.1ws21321Tg embryo with mCherry expression in the inner ear of a larva 7 d after fertilization (K, arrowhead) and in neuromasts of the posterior lateral line primordium (pLLP) (L, arrows) at 6 d after fertilization. Scale bars, 50 µm in (A, C, D, E, I), 250 µm in (F) and (L), and 10 µm in (K).

Fig. 6

Representative mutant phenotypes in DsDELGT4 insertion lines. The Tg(DsDELGT4)ws0069 mutant shows cell death in the brain (B) compared to siblings (A) (bright field images). (C, D) Acridine Orange labeling to show cell death in the brain of mutant embryos (D) compared to siblings (C). White arrows in (B) and (D) show cell death in mutants. DIC images of Tg(DsDELGT4)ws01962 mutants showing smaller head and edema (F) compared to siblings (E). (G) and (H) In situ hybridization staining results with a probe to detect the endothelial marker flk1. Expression of flk1 is dramatically reduced in mutants (H) compared to siblings (G). Tg(DsDELGT4)ws21322 mutants exhibit fat metabolism defects (J) compared to siblings (I). (K) and (L) Oil red O staining in sibling (K) and mutant (L) embryos. Nonmetabolized fat is observed below the swim bladder in mutant embryos (arrows in J and L). Scale bars, 100 µm. Lateral views shown.

Fig. 7

Protein trap-induced mutant phenotype affecting the dhx37 gene. (A–D) Bright field images of dhx37ws0977Tg/+ sibling (A), homozygous dhx37ws0977Tg/ws0977Tg mutant (B), dhx37 ATG morpholino-injected (C), or full-length dhx37 RNA-injected homozygous mutant embryo (D). (E) The Ds insertion in chr8:4765207-4765215 is linked to the mutant phenotype. Black arrows indicate the position of primers used for genotyping PCRs. (F–H) RT-PCR results from homozygous dhx37ws0977Tg/ws0977Tg embryos show that the bcl2 splice acceptor sequence trapped endogenous upstream splice donor sites in the dhx37 gene, producing multiple fusion transcripts (F, G), leading to the disruption of wild-type transcripts (H). (G) The main fusion products, 1 and 2, are out of frame with respect to the reading sequence. Black arrows in the schematic in (F) indicate the positions of primers used for RT-PCR. No evidence of wild-type transcripts was found, and only mutant fusion transcripts were detected. The fusion transcripts always have a 71-bp linker sequence (SA) originated from bcl2 splice acceptor site left between upstream exons and the mCherry reporter sequence. The major fusion transcripts 1 and 2 are out of frame for mCherry coding sequences (G).

Fig. 8 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage: Day 5

Fig. 9

Remobilization of DsDELGT4 leads to imprecise excision and generates integrations with new reporter expression patterns. (A) Schematic representation of the remobilization experiment. Ac transposase mRNA was injected into one-cell stage embryos that show reporter expression. The injected embryos are raised to adulthood and mated with wild-type fish. Their progeny were examined for sequences flanking the original integration site and screened for new reporter expression patterns. (B–E) Remobilization of Ds in embryos of the single insertion line Tg(DsDELGT4)ws01961 (antisense strand, chr1:20205236-20205244) generates novel expression patterns (C–E) distinct from the original expression pattern (B). (F–H) Imprecise excision of DsDELGT4 by Ac-mediated remobilization causes mutations at the original integration site (location of the original insert is shown by black arrowhead in F), including deletions (F), indels (G), and insertions (H). Genomic DNA for sequence analysis (F–H) was obtained from the embryos of Tg(DsDELGT4)ws0310 (sense strand, chr21:17731630–17731722) and collected 24 hr after injection of Ac transposase mRNA.

EXPRESSION / LABELING:
Gene:
Fish:
Anatomical Term:
Stage: Day 5

Table S1 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Table S2 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Table S3 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Table S4 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

EXPRESSION / LABELING:
Gene:
Fish:
Anatomical Term:
Stage Range: Day 4 to Day 6

Table S5 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Table S6 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Table S7 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Table S8 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Table S9 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Table S10 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

EXPRESSION / LABELING:
Gene:
Fish:
Anatomical Term:
Stage: Day 6

Table S11 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Table S12 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

EXPRESSION / LABELING:
Gene:
Fish:
Anatomical Term:
Stage Range: Long-pec to Day 6

Table S13 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Term:
Stage Range: Protruding-mouth to Day 6

Table S14 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Table S15 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Table S16 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Term:
Stage Range: Day 5 to Days 7-13

Table S17 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Term:
Stage Range: Long-pec to Days 7-13

Table S18 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Table S19 ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

Acknowledgments:
ZFIN wishes to thank the journal G3 (Bethesda) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ G3 (Bethesda)