Generation of a glialcama knockout line in zebrafish. a An allele with a deletion of 7 nucleotides in glialcama (Δ7, from now on −/−) was generated using a TALEN nuclease. The deletion generates an early stop codon, resulting in a protein of only 28 amino acids in lenght. b Western blot of brain extracts from adult wild-type (+/+), heterozygous (+/−) or homozygous mutant (−/−) fish for the glialcama knockout allele demonstrates the absence of the glialcama protein in the homozygous zebrafish. c glialcama expression in the optic tract (Ot) (arrowheads) and diffusely in cell bodies of the preoptic region (asterisk) of wild type fish (dotted line: optic tract/preoptic region boundary). d No glialcama immunoreactivity is observed in the optic tract or brain parenchyma of glialcama knockout fish. Dotted line: optic tract/preoptic region boundary. Asterisk: preoptic region. e glialcama immunoreactivity is observed in the inner limiting membrane of the wild type retina (arrowheads). f No glialcama immunoreactivity is observed in knockout retina. Expression observed in glialcama−/− represents autofluorescence in photoreceptors that was consistently observed, even in the case of control immunostainings with secondary antibody only. Arrows point to the inner limiting membrane. Scale bars: 60 μm (e, f); 75 μm (c, d)
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