Zebrafish Embryo Microinjection Protocols

Zebrafish embryos were microinjected with an Olympus SZX9 and a Picospritzer III microinjector (Parker Instrumentation). Injection mixes were composed of Danieau solution 1X (NaCl 58 mM, KCl 0.7 mM, MgSO₄ 0.4 mM, Ca(NO₃)₂ 0.6 mM, HEPES 5.0 mM, pH 7.6), Phenol red 0.1% and Morpholino (MO) antisense oligos (Gene Tools). Each embryo was injected respectively with 1.7 ng of splice blocking snap29 MO^{14 (link)}, 1.7 ng of splice-blocking snap29 together with 200 pg GFP-snap29 mRNA for the rescue, 3 ng of 5′-UTR snap25 a,b MO, or 3 ng of ATG bdnf MO^{51 (link)}.

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Mastrodonato V., Beznoussenko G., Mironov A., Ferrari L., Deflorian G, & Vaccari T. (2019). A genetic model of CEDNIK syndrome in zebrafish highlights the role of the SNARE protein Snap29 in neuromotor and epidermal development. Scientific Reports, 9, 1211.

Publication 2019

Morpholino (MO) antisense oligos

3 utr Embryo Gene Hepes Mgso4 Morpholino Mrna Nacl Snap25 Snap29 Splice Zebrafish

Corresponding Organization : University of Milan

Other organizations : European Institute of Oncology

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Variable analysis

independent variables

Injection of 1.7 ng of splice blocking snap29 MO
Injection of 1.7 ng of splice-blocking snap29 together with 200 pg GFP-snap29 mRNA for the rescue
Injection of 3 ng of 5′-UTR snap25 a,b MO
Injection of 3 ng of ATG bdnf MO

dependent variables

Unspecified

control variables

Danieau solution 1X (NaCl 58 mM, KCl 0.7 mM, MgSO4 0.4 mM, Ca(NO3)2 0.6 mM, HEPES 5.0 mM, pH 7.6)
Phenol red 0.1%

controls

Injection of 1.7 ng of splice-blocking snap29 together with 200 pg GFP-snap29 mRNA for the rescue
Unspecified

Annotations

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