ZFIN ID: ZDB-FIG-150826-4
Auer et al., 2015 - Deletion of a kinesin I motor unmasks a mechanism of homeostatic branching control by neurotrophin-3. eLIFE   4 Full text @ Elife
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Gene:
Fish:
Anatomical Terms:
Stage Range: Prim-5 to Day 4
PHENOTYPE:
Fish:
Observed In:
Stage Range: Prim-5 to Day 5

Fig. 1 Generation of loss-of-function alleles of the anterograde motor protein Kif5aa.

(A) Employing TALENs targeting exon4 of the kif5aa open reading frame, we generated two loss-of-function alleles with a 10 bp and 13 bp deletion, respectively. These result in a frameshift at amino acid 122 and a premature stop codon after 162 of 1033aas within the motor domain of Kif5aa. L = linker region, N = neck region. (B) In situ hybridization shows a substantial downregulation of kif5aa mRNA in 24 hpf and 72 hpf old embryos. Scale bars (from left to right) = 150 µm, 100 µm, 50 µm. RGCL = Retinal Ganglion Cell layer. (C) Quantitative reverse transcription PCR confirms that only 47% of wild-type kif5aa mRNA expression levels are reached in homozygote mutant embryos at 4 dpf (p < 0.01). (D) Kif5aa mutant embryos show expanded melanosomes within their melanocytes and appear dark compared to wild-type embryos. Scale bars = 200 µm. (E) They fail to inflate their swim bladder and die 10 days post fertilization. Scale bars = 400 µm. Arrow: pointing at the respective location of the swim bladder. SB = swim bladder.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
kif5aa WT standard conditions Prim-5 brain ISH
Prim-5 whole organism ISH
Protruding-mouth brain ISH
Protruding-mouth retinal ganglion cell layer ISH
Day 4 whole organism RTPCR
kif5aaic3023/ic3023 standard conditions Prim-5 brain ISH
Protruding-mouth brain ISH
Protruding-mouth retinal ganglion cell layer ISH
Day 4 whole organism RTPCR
kif5aaic3024/ic3024 standard conditions Prim-5 brain ISH
Protruding-mouth brain ISH
Protruding-mouth retinal ganglion cell layer ISH
Day 4 whole organism RTPCR
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
kif5aaic3023/ic3023 standard conditions Prim-5 brain kif5aa expression decreased amount, abnormal
Protruding-mouth brain kif5aa expression decreased amount, abnormal
Protruding-mouth retinal ganglion cell layer kif5aa expression decreased amount, abnormal
Day 4 whole organism kif5aa expression decreased amount, abnormal
Day 5 melanocyte black, abnormal
Day 5 melanocyte melanosome distended, abnormal
Day 5 swim bladder uninflated, abnormal
kif5aaic3024/ic3024 standard conditions Prim-5 brain kif5aa expression decreased amount, abnormal
Protruding-mouth brain kif5aa expression decreased amount, abnormal
Protruding-mouth retinal ganglion cell layer kif5aa expression decreased amount, abnormal
Day 4 whole organism kif5aa expression decreased amount, abnormal
Day 5 melanocyte black, abnormal
Day 5 melanocyte melanosome distended, abnormal
Day 5 swim bladder uninflated, abnormal
Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Elife