Tissue-specific expression of FoxD reporter constructs in amphioxus embryos

Cephalochordates (amphioxus), the closest living invertebrate relatives of the vertebrates, are key to understanding the evolution of developmental mechanisms during the invertebrate-to-vertebrate transition. However, a major impediment to amphioxus as a model organism for developmental biology has...

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Bibliographic Details
Published in:Developmental biology 2004-10, Vol.274 (2), p.452-461
Main Authors: Yu, Jr-Kai, Holland, Nicholas D., Holland, Linda Z.
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Branchiostoma floridae
Chordata, Nonvertebrate - embryology
Cis-regulatory sequence
DNA-Binding Proteins - genetics
DNA-Binding Proteins - metabolism
Embryo, Nonmammalian - physiology
Evolution
Evolution, Molecular
Female
Forebrain
Fox domain transcription factor
Gene Expression Regulation, Developmental
Gene Transfer Techniques
Genes, Reporter
In Situ Hybridization
Male
Marine
Microinjections
Molecular Sequence Data
Neural crest
Notochord
Oocytes - cytology
Oocytes - physiology
Repetitive DNA
Reporter gene construct
Somite
Space life sciences
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Summary:Cephalochordates (amphioxus), the closest living invertebrate relatives of the vertebrates, are key to understanding the evolution of developmental mechanisms during the invertebrate-to-vertebrate transition. However, a major impediment to amphioxus as a model organism for developmental biology has been the inability to introduce transgenes or other macromolecules into the embryos. Here, we report the development of a reproducible method for microinjection of amphioxus eggs. Specifically, we show that expression of a LacZ reporter construct including 6.3 kb of AmphiFoxD upstream regulatory DNA recapitulates expression of the endogenous gene in the nerve cord, somites, and notochord. We have also identified the 1.6 kb at the 5′ end of this region as essential for expression in the first two of these domains and the 4.7 kb at the 3′ end as sufficient for expression in the notochord. This study, which is the first report of a method for introduction of large molecules such as DNA into amphioxus embryos, opens the way for studies of gene regulation and function in amphioxus and for comparative studies with vertebrates to understand the relationship between the extensive gene duplications that occurred within the vertebrate lineage and the evolution of vertebrate innovations such as neural crest.
ISSN:0012-1606
1095-564X
DOI:10.1016/j.ydbio.2004.07.010