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ZFIN ID: ZDB-FIG-100506-43
Nakano et al., 2010 - Biogenesis of GPI-anchored proteins is essential for surface expression of sodium channels in zebrafish Rohon-Beard neurons to respond to mechanosensory stimulation. Development (Cambridge, England)   137(10):1689-1698 Full text @ Development
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Antibodies:
Fish:
Anatomical Terms:
Stage: Long-pec
PHENOTYPE:
Fish:
Observed In:
Stage: Long-pec

Fig. 4 Surface expression of sodium channels is impaired in mutant RB neurons. (A-F) Wholemount immunostaining showed Nav expression in wild-type and mutant RB neurons at 2 dpf. (A) Anti-HuC labeled cell bodies of wild-type RB neurons at the dorsal spinal cord. Dotted lines represent somite boundaries. (B) Immunostaining with anti-pan Nav displayed Nav expression in wild-type spinal cord. (C) A merged image showed that all HuC-positive RB cells expressed Nav in wild-type. (D) Labeling with anti-HuC showed RB neurons in mutant spinal cord. (E) Anti-pan Nav labeled dorsally located spinal neurons in mutants. (F) All of the HuC-positive mutant RB neurons expressed Nav. (G-R) Dissociated RB neurons (GFP-positive) from 2 dpf embryos were labeled with anti-Scn1bb, with or without TX-100, which permeabilizes the plasma membrane. Note that anti-Scn1bb reacts with the extracellular domain of Scn1bb. (G-I) Scn1bb staining in the permeable condition (+TX-100) demonstrated Scn1bb expression in a wild-type RB neuron. (J-L) Labeling of Scn1bb in the impermeable condition (-TX-100) displayed surface distribution of Scn1bb in a wild-type RB cell. (M-O) Scn1bb staining with TX-100 showed Scn1bb expression in a wild-type RB neuron. (P-R) In the absence of TX-100, the surface labeling of Scn1bb was missing in a mutant RB cell, suggesting that Scn1bb exists inside the mutant cell without expression at the surface.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
elavl3 Ab1-elavl WT standard conditions Long-pec Rohon-Beard neuron IHC
Ab1-elavl pigumi310/mi310 standard conditions Long-pec Rohon-Beard neuron IHC
GFP pigumi310/mi310; rw011aTg standard conditions Long-pec Rohon-Beard neuron IHC
rw011aTg standard conditions Long-pec Rohon-Beard neuron IHC
scn1bb Ab1-scn1bb pigumi310/mi310; rw011aTg standard conditions Long-pec Rohon-Beard neuron cell body IHC
Ab1-scn1bb Long-pec Not Detected Rohon-Beard neuron plasma membrane IHC
Ab1-scn1bb rw011aTg standard conditions Long-pec Rohon-Beard neuron cell body IHC
Ab1-scn1bb Long-pec Rohon-Beard neuron plasma membrane IHC
Antibody Labeling Details
Antibody Assay Fish Conditions Stage Anatomy
Ab2-scn IHC WT standard conditions Long-pec Rohon-Beard neuron
IHC pigumi310/mi310 standard conditions Long-pec Rohon-Beard neuron
Phenotype Details
Fish Conditions Stage Phenotype
pigumi310/mi310; rw011aTg standard conditions Long-pec protein localization to cell surface arrested, abnormal
Long-pec Rohon-Beard neuron voltage-gated sodium channel complex mislocalised, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Development (Cambridge, England) for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Development