Targeted Mutation of the Gene Encoding Prion Protein in Zebrafish reveals a Conserved Role in Neuron Excitability
- Authors
- Fleisch, V.C., Leighton, P.L., Wang, H., Pillay, L.M., Ritzel, R.G., Bhinder, G., Roy, B., Tierney, K.B., Ali, D.W., Waskiewicz, A.J., and Allison, W.T.
- ID
- ZDB-PUB-130408-21
- Date
- 2013
- Source
- Neurobiology of disease 55: 11-25 (Journal)
- Registered Authors
- Allison, Ted, Fleisch, Valerie, Leighton, Patricia, Wang, Hao, Waskiewicz, Andrew
- Keywords
- NMDA receptor, prion protein, seizure susceptibility, targeted mutagenesis
- MeSH Terms
-
- Age Factors
- Animals
- Animals, Genetically Modified
- Disease Models, Animal
- Epilepsy/chemically induced
- Epilepsy/physiopathology
- Gene Expression Regulation, Developmental/genetics*
- Gene Library
- Green Fluorescent Proteins/genetics
- Green Fluorescent Proteins/metabolism
- Larva
- Mice
- Mutagenesis, Site-Directed
- Mutation/genetics*
- Neurons/metabolism*
- Pentylenetetrazole/toxicity
- Phenotype
- Prions/genetics*
- Receptors, N-Methyl-D-Aspartate/metabolism
- Zebrafish/genetics
- Zebrafish/growth & development
- Zinc Fingers/genetics
- PubMed
- 23523635 Full text @ Neurobiol. Dis.
The function of the cellular prion protein (PrPC) in healthy brains remains poorly understood, in part because Prnp knockout mice are viable. On the other hand, transient knockdown of Prnp homologs in zebrafish (including two paralogs, prp1 and prp2) has suggested that PrPC is required for CNS development, cell adhesion, and neuroprotection. It has been argued that zebrafish Prp2 is most similar to mammalian PrPC, yet it has remained intransigent to the most thorough confirmations of reagent specificity during knockdown. Thus we investigated the role of prp2 using targeted gene disruption via zinc finger nucleases. Prp2/ zebrafish were viable and did not display overt developmental phenotypes. Back-crossing female prp2/ fish ruled out a role for maternal mRNA contributions. Prp2/ larvae were found to have increased seizure-like behavior following exposure to the convulsant pentylenetetrazol (PTZ), as compared to wild type fish. In situ recordings from intact hindbrains demonstrated that prp2 regulates closing of N-Methyl-d-aspartate (NMDA) receptors, concomitant with neuroprotection during glutamate excitotoxicity. Overall, the knockout of Prp2 function in zebrafish independently confirmed hypothesized roles for PrP, identifying deeply conserved functions in post-developmental regulation of neuron excitability that are consequential to the etiology of prion and Alzheimer diseases.