PUBLICATION
A modified acetylcholine receptor delta-subunit enables a null mutant to survive beyond sexual maturation
- Authors
- Epley, K.E., Urban, J.M., Ikenaga, T., and Ono, F.
- ID
- ZDB-PUB-081217-4
- Date
- 2008
- Source
- The Journal of neuroscience : the official journal of the Society for Neuroscience 28(49): 13223-13231 (Journal)
- Registered Authors
- Ono, Fumihito, Urban, Jason
- Keywords
- zebrafish, neuromuscular junction, acetylcholine receptor, synapse, fetal akinesia deformation sequence, fluorescent protein
- MeSH Terms
-
- Acetylcholine/metabolism
- Animals
- Animals, Genetically Modified
- Feeding Behavior/physiology
- Female
- Gene Expression Regulation, Developmental/genetics
- Longevity/genetics*
- Luminescent Proteins/genetics
- Male
- Mutation/genetics*
- Neuromuscular Junction/genetics
- Neuromuscular Junction/metabolism
- Neuromuscular Junction/physiopathology
- Neuromuscular Junction Diseases/genetics*
- Neuromuscular Junction Diseases/metabolism
- Neuromuscular Junction Diseases/physiopathology
- Protein Subunits/chemistry
- Protein Subunits/genetics
- Receptors, Cholinergic/genetics*
- Recombinant Fusion Proteins/chemistry
- Recombinant Fusion Proteins/genetics
- Sexual Behavior, Animal/physiology
- Sexual Maturation/genetics
- Synaptic Membranes/genetics
- Synaptic Membranes/metabolism
- Synaptic Membranes/ultrastructure
- Synaptic Transmission/genetics
- Transgenes/genetics
- Zebrafish/genetics*
- Zebrafish/growth & development*
- Zebrafish/metabolism
- PubMed
- 19052214 Full text @ J. Neurosci.
Citation
Epley, K.E., Urban, J.M., Ikenaga, T., and Ono, F. (2008) A modified acetylcholine receptor delta-subunit enables a null mutant to survive beyond sexual maturation. The Journal of neuroscience : the official journal of the Society for Neuroscience. 28(49):13223-13231.
Abstract
The contraction of skeletal muscle is dependent on synaptic transmission through acetylcholine receptors (AChRs) at the neuromuscular junction (NMJ). The lack of an AChR subunit causes a fetal akinesia in humans, leading to death in the first trimester and characteristic features of Fetal Akinesia Deformation Sequences (FADS). A corresponding null mutation of the delta-subunit in zebrafish (sofa potato; sop) leads to the death of embryos around 5 d postfertilization (dpf). In sop(-/-) mutants, we expressed modified delta-subunits, with one (delta1YFP) or two yellow fluorescent protein (delta2YFP) molecules fused at the intracellular loop, under the control of an alpha-actin promoter. AChRs containing these fusion proteins are fluorescent, assemble on the plasma membrane, make clusters under motor neuron endings, and generate synaptic current. We screened for germ-line transmission of the transgene and established a line of sop(-/-) fish stably expressing the delta2YFP. These delta2YFP/sop(-/-) embryos can mount escape behavior close to that of their wild-type siblings. Synaptic currents in these embryos had a smaller amplitude, slower rise time, and slower decay when compared with wild-type fish. Remarkably, these embryos grow to adulthood and display complex behaviors such as feeding and breeding. To the best of our knowledge, this is the first case of a mutant animal corresponding to first trimester lethality in human that has been rescued by a transgene and survived to adulthood. In the rescued fish, a foreign promoter drove the transgene expression and the NMJ had altered synaptic strength. The survival of the transgenic animal delineates requirements for gene therapies of NMJ.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping