ZFIN ID: ZDB-FIG-190711-31
Cox et al., 2019 - The Metalloproteinase adam19b Is Required for Sensory Axon Guidance in the Hindbrain. Frontiers in neural circuits   13:14 Full text @ Front. Neural Circuits
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Anatomical Terms:
Stage Range: Prim-5 to Day 4
PHENOTYPE:
Fish:
Observed In:
Stage: Day 4

Fig. 1

Characterization of the sl19 mutant line. (A) Wild type larva at 4 days post fertilization (4 dpf) showing normal pattern of cranial sensory ganglia (CSG) projections. sV (also llf-lateral longitudinal fascicle), sVII, sIX and sX are the projections of the trigeminal (gV), facial (VII), glossopharyngeal (IX) and vagus (X) ganglia, respectively. White arrowhead shows normal hindbrain plexus, where sVII, sIX and sX sensory axons terminate. (B) CSG projections in a 4dpf sl19 mutant. White arrow indicates malformed hindbrain plexus and dotted arrow shows the misrouted and defasciculated sVII failing to terminate in the plexus. sIX and sX fail to form normal terminal fields. Projections from gV appear normal. (C) Positional cloning of sl19. The lesion was mapped to a critical region on chromosome 14 between 6 and 7 Mb. Transcripts of all genes in this region (20 in total) were sequenced (for clarity, only some of these genes are shown in the Figure). Only the adam19b gene showed an altered coding sequence—a single base mutation in exon 17. This C > T conversion causes an arginine to cysteine switch at amino acid 673 in the cysteine-rich domain (CRD) of A Disintegrin and Metalloproteinase (ADAM19b (CRD). (D) Wholemount in situ hybridization (ISH) of adam19b in the zebrafish head. At 24 hpf, adam19b is expressed in the trigeminal ganglion (gV), the anterior lateral line ganglion (allg) and the posterior lateral line ganglion (pllg). At 48 hpf, expression has expanded to include regions of the hindbrain. By 72 hpf, adam19b is also expressed in the epibranchial ganglia (gVII, gIX and gX) and in the hindbrain (hb); by 96 hpf, expression has increased in all these cranial structures. Sense probe did not produce staining at any time point (data not shown). (E) adam19b full length RNA injected into sl19 mutant. Over-expression of adam19b did not rescue the mutant phenotype, but resulted in a higher incidence of the mutant phenotype (see text). White arrow indicates malformed plexus; dotted arrow shows misrouted sVII and sIX. (F) adam19b full length RNA injected into wild type. As can be seen, over-expression of adam19b induced aberrant epibranchial projections in 4 dpf wild type fish, identical to that found in the sl19mutant. White arrow: malformed plexus, showing how sX forms a dense clump of axons, instead of the organized branching of the axons seen in un-injected controls (A). Dotted arrow: misrouted sVII and sIX. Panels (A,B) are confocal z-stacks, panels (E,F) are epifluorescent micrographs. All images are oriented anterior to the left and dorsal at top. Scale bars = 100 μm.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
adam19b WT standard conditions Prim-5 anterior lateral line ganglion ISH
Prim-5 posterior lateral line ganglion ISH
Prim-5 trigeminal ganglion ISH
Long-pec anterior lateral line ganglion ISH
Long-pec cerebellar plate ISH
Long-pec hindbrain ISH
Long-pec optic tectum ISH
Long-pec posterior lateral line ganglion ISH
Long-pec trigeminal ganglion ISH
Protruding-mouth anterior lateral line ganglion ISH
Protruding-mouth cerebellar plate ISH
Protruding-mouth facial ganglion ISH
Protruding-mouth glossopharyngeal ganglion ISH
Protruding-mouth hindbrain ISH
Protruding-mouth olfactory epithelium ISH
Protruding-mouth posterior lateral line ganglion ISH
Protruding-mouth retina ISH
Protruding-mouth trigeminal ganglion ISH
Protruding-mouth vagal ganglion ISH
Day 4 anterior lateral line ganglion ISH
Day 4 cerebellar plate ISH
Day 4 facial ganglion ISH
Day 4 glossopharyngeal ganglion ISH
Day 4 hindbrain ISH
Day 4 olfactory epithelium ISH
Day 4 posterior lateral line ganglion ISH
Day 4 retina ISH
Day 4 trigeminal ganglion ISH
Day 4 vagal ganglion ISH
EGFP sl1Tg standard conditions Day 4 cranial nerve IX ISH
Day 4 cranial nerve V ISH
Day 4 cranial nerve VII ISH
Day 4 cranial nerve X ISH
Day 4 facial ganglion ISH
Day 4 glossopharyngeal ganglion ISH
Day 4 medulla oblongata ISH
Day 4 trigeminal ganglion ISH
Day 4 vagal ganglion ISH
sl1Tg ; adam19bsl19/sl19 standard conditions Day 4 cranial nerve IX ISH
Day 4 cranial nerve V ISH
Day 4 cranial nerve VII ISH
Day 4 cranial nerve X ISH
Day 4 facial ganglion ISH
Day 4 glossopharyngeal ganglion ISH
Day 4 medulla oblongata ISH
Day 4 trigeminal ganglion ISH
Day 4 vagal ganglion ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
sl1Tg ; adam19bsl19/sl19 standard conditions Day 4 cranial nerve IX axon arborization decreased occurrence, abnormal
Day 4 cranial nerve IX axon guidance disrupted, abnormal
Day 4 cranial nerve IX EGFP expression decreased amount, abnormal
Day 4 cranial nerve V axon extension process quality, normal
Day 4 cranial nerve VII defasciculated, abnormal
Day 4 cranial nerve VII EGFP expression spatial pattern, abnormal
Day 4 cranial nerve VII misrouted, abnormal
Day 4 cranial nerve X axon arborization decreased occurrence, abnormal
Day 4 cranial nerve X EGFP expression decreased amount, abnormal
Day 4 hindbrain axonogenesis decreased magnitude, abnormal
Day 4 medulla oblongata EGFP expression decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Frontiers in neural circuits for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Front. Neural Circuits