ZFIN ID: ZDB-FIG-150826-9
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: Day 5 to Days 7-13
PHENOTYPE:
Fish:
Observed In:
Stage Range: Day 5 to Days 7-13

Fig. 4 Kif5aa mutant larvae show no activity in RGCs and no synaptic transmission to tectal cells.

(A) 5–7 dpf larvae were visually stimulated by bars on an LED screen running in caudal-to-rostral direction across the larva′s visual field. Wild-type and kif5aa mutant larvae expressing genetically encoded calcium indicators (GCaMPs) in different subsets of neurons of the visual system were confocally imaged in the tectum contralaterally to the stimulated eye (dashed box inset). RGCs = Retinal Ganglion Cells, PVNs = periventricular neurons, NP = neuropil. (B) The activity of visual system neurons in response to visual stimuli is shown as normalized GCaMP fluorescence intensity changes (deltaF/F0) over time. GCaMP intensity was averaged over manually determined regions of interest (ROIs) that corresponded to well-distinguishable anatomical regions in the larval tectum, the neuropil (NP) and the periventricular cell bodies area (PVNs). In Tg(HuC:GCaMP5) fish (left), GCaMP5 is expressed pan-neuronaly, that is, in both neuropil and PVNs, whereas in Tg(Isl2b:Gal4) × Tg(UAS:GCaMP3) fish (right), it is expressed in all RGCs and their processes. Scale bars = 20 µm. Asterisk = pigment cell in the skin. (C) Averaged deltaF/F0 ratio over time in response to a moving bar visual stimulation (black vertical line denotes the time point of the stimulus onset) in fish with pan-neuronal GCaMP5G expression. Four sequential rounds of stimulus presentation and the time-courses of Ca2+-transients in the neuropil ROIs (left panel) and periventricular cell bodies ROIs (right panel) of wild-type/heterozygous (red curve) and kif5aa mutant larvae (blue curve) (n = 7 each) are shown. Activity in the visual system of kif5aa mutants was almost absent between 5 and 7 dpf compared to wild-type larvae. Light red and blue zones indicate the 95% confidence intervals around the averaged deltaF/F0 curves for each region of interest (NP and PVN), respectively. (D) Averaged deltaF/F0 ratio over time in response to a moving bar visual stimulation (black vertical line denotes stimulus onset) in fish with GCaMP3 expression in RGC axons. Five sequential rounds of stimulus presentation and time-courses of Ca2+-transients in the neuropil of wild-type/heterozygous (red curve) and kif5aa mutant larvae (blue) (n = 9 each). RGC arbor activity in kif5aa mutants was strongly diminished. Light red and blue zones indicate the 95% confidence intervals around the averaged deltaF/F0 curves for the neuropil region of interest (NP).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
GCaMP a4598Tg standard conditions Day 5 to Days 7-13 optic tectum dendrite IFL
Day 5 to Days 7-13 optic tectum neuron IFL
ic3025Tg; zf350Tg standard conditions Day 5 to Days 7-13 retinal ganglion cell axon IFL
kif5aaic3023/ic3023; a4598Tg standard conditions Day 5 to Days 7-13 optic tectum dendrite IFL
Day 5 to Days 7-13 optic tectum neuron IFL
kif5aaic3023/ic3023; ic3025Tg; zf350Tg standard conditions Day 5 to Days 7-13 retinal ganglion cell axon IFL
kif5aaic3024/ic3024; a4598Tg standard conditions Day 5 to Days 7-13 optic tectum dendrite IFL
Day 5 to Days 7-13 optic tectum neuron IFL
kif5aaic3024/ic3024; ic3025Tg; zf350Tg standard conditions Day 5 to Days 7-13 retinal ganglion cell axon IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
kif5aaic3023/ic3023; a4598Tg standard conditions Day 5 - Days 7-13 optic tectum anterograde trans-synaptic signaling decreased occurrence, abnormal
Day 5 - Days 7-13 optic tectum dendrite GCaMP expression decreased amount, abnormal
Day 5 - Days 7-13 optic tectum neuron GCaMP expression decreased amount, abnormal
Day 5 - Days 7-13 optomotor response arrested, abnormal
kif5aaic3023/ic3023; ic3025Tg; zf350Tg standard conditions Day 5 - Days 7-13 cranial nerve II anterograde trans-synaptic signaling decreased occurrence, abnormal
Day 5 - Days 7-13 optomotor response arrested, abnormal
Day 5 - Days 7-13 retinal ganglion cell axon GCaMP expression decreased amount, abnormal
kif5aaic3024/ic3024; a4598Tg standard conditions Day 5 - Days 7-13 optic tectum anterograde trans-synaptic signaling decreased occurrence, abnormal
Day 5 - Days 7-13 optic tectum dendrite GCaMP expression decreased amount, abnormal
Day 5 - Days 7-13 optic tectum neuron GCaMP expression decreased amount, abnormal
Day 5 - Days 7-13 optomotor response arrested, abnormal
kif5aaic3024/ic3024; ic3025Tg; zf350Tg standard conditions Day 5 - Days 7-13 cranial nerve II anterograde trans-synaptic signaling decreased occurrence, abnormal
Day 5 - Days 7-13 optomotor response arrested, abnormal
Day 5 - Days 7-13 retinal ganglion cell axon GCaMP expression decreased amount, 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