ZFIN ID: ZDB-FIG-181005-16
Jardin et al., 2018 - BMP- and Neuropilin-1-mediated motor axon navigation relies on spastin alternative translation. Development (Cambridge, England)   145(17) Full text @ Development
ADDITIONAL FIGURES
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage Range: Prim-25 to Protruding-mouth

Fig. 7

DrM61 acts as a downstream effector of neuropilin 1a signalling in motor neurons. (A-C) Neuropilin 1a (nrp1a) morphants show misrouted motor neuron somata similar to ATG2 morphants. (A) Analysis of secondary motor neurons (sMN) in 72 hpf Tg(Hb9:GFP) transgenic larvae injected with MOnrp1a, MOspATG2 or MOscr morpholinos using zn-5 and GFP antibodies. (B) Representative stills of time-lapse recordings showing SMN behaviour in 40-72 hpf Tg(Hb9:GFP) transgenic larvae injected with Nrp1a (n=5 larvae; n=32 spinal hemisegment monitored). Arrows track the aberrant migration of SMN somata along motor axon tracts of nrp1a morphants. Scale bar: 10 µm. (D-F) Rescue experiments of nrp1a morphant phenotypes with mouse Nrp1 mRNA (Ms_nrp1 mRNA). (D) Overall morphology (upper panels) and SMN analysis (bottom panels) of 72 hpf larvae injected with MOscr, MONrp1a or MONrp1a and Ms_nrp1 mRNA. (E) Distribution of wild-type and nrp1a morphant phenotypes in each injection group. ***P≤0.001; χ2 test. (F) Mean number of misplaced SMN somata per larvae. (G) SMN analysis in 72 hpf Tg(Hb9:GFP) transgenic larvae injected with sub-efficient doses of MOspATG1, MOspATG2 or MONrp1a or co-injected with the same sub-efficient doses of MONrp1a and MOspATG1 or MOspATG2 morpholinos. (I) SMN analysis in 72 hpf Tg(Hb9:GFP) transgenic larvae injected with control (MOscr) or MONrp1a morpholinos or co-injected with MONrp1a and DrM1- or DrM61-spastin mRNA. (J) SMN analysis in 72 hpf Tg(mnGFF7;UAS:GFP), Tg(mnGFF7;UAS:GFP;UAS:DrM61-HA) and Tg(mnGFF7; UAS:GFP;UAS:DrM1-HA) transgenic larvae using HA and GFP antibodies. (A,D,G,I,J) Lateral views of the trunk, anterior towards the left. Arrows indicate mispositioned SMN somata. Scale bars: 50 µm. (J) Empty arrowheads indicate misplaced HA-positive SMN. (C,F,H,K,L) Mean number of mispositioned SMN per larva (quantified on 24 spinal hemisegments/larva). (M) Percentage of misplaced HA-positive or -negative SMN per larva (quantified on 24 spinal hemisegments/larva). (C,F,H,K-M) The number of larvae or SMN analysed per group (n) is mentioned under each corresponding histogram bar. *P≤0.05, **P≤0.01, ***P≤0.001 versus internal controls; $$$P≤0.001 versus single injection of each morpholino; ###P≤0.001 versus morphant larvae; ††P≤0.01, †††P≤0.001 HA-positive versus -negative SMN; Kruskal–Wallis ANOVA test with Dunn's post test. Error bars are s.e.m.

Gene Expression Details No data available
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
ml2Tg + MO1-nipa1 control Prim-25 - Protruding-mouth secondary motor neuron cell body mislocalised, abnormal
Protruding-mouth secondary motor neuron axon guidance process quality, abnormal
Protruding-mouth secondary motor neuron cell body mislocalised, abnormal
ml2Tg + MO1-nipa1 + MO4-spast control Protruding-mouth secondary motor neuron cell body mislocalised, exacerbated
ml2Tg + MO4-spast control Protruding-mouth secondary motor neuron axon guidance process quality, abnormal
Protruding-mouth secondary motor neuron cell body mislocalised, abnormal
nkmnGFF7Tg; nkuasgfp1aTg + MO1-nipa1 control Protruding-mouth secondary motor neuron cell body mislocalised, abnormal
nkmnGFF7Tg; nkuasgfp1aTg; zf2200Tg + MO1-nipa1 control Protruding-mouth secondary motor neuron cell body mislocalised, ameliorated
nkmnGFF7Tg; nkuasgfp1aTg; zf2201Tg + MO1-nipa1 control Protruding-mouth secondary motor neuron cell body mislocalised, 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 @ Development