Microinjection of zebrafish embryos to analyze gene function.
One of the advantages of studying zebrafish is the ease and speed of manipulating protein levels in the embryo. Morpholinos, which are synthetic oligonucleotides with antisense complementarity to target RNAs, can be added to the embryo to reduce the expression of a particular gene product. Conversely, processed mRNA can be added to the embryo to increase levels of a gene product. The vehicle for adding either mRNA or morpholino to an embryo is microinjection. Microinjection is efficient and rapid, allowing for the injection of hundreds of embryos per hour. This video shows all the steps involved in microinjection. Briefly, eggs are collected immediately after being laid and lined up against a microscope slide in a Petri dish. Next, a fine-tipped needle loaded with injection material is connected to a microinjector and an air source, and the microinjector controls are adjusted to produce a desirable injection volume. Finally, the needle is plunged into the embryo's yolk and the morpholino or mRNA is expelled.
Rosen
Jonathan N
JN
Department of Genetics, Harvard Medical School, USA. jrosen@fas.harvard.edu
Sweeney
Michael F
MF
Mably
John D
JD
D000818:Animals; D005260:Female; D018507:Gene Expression Regulation, Developmental; D008297:Male; D008845:Microinjections / Q000379:methods*; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage; D012333:RNA, Messenger / Q000008:administration & dosage; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics*
D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger