A Morpholino Strategy to Assess TSC Gene Function in Zebrafish
Abstract
Tuberous sclerosis complex (TSC) is a genetic disorder associated with severe neurological symptoms including mental retardation, autism, infantile spasms, and epilepsy. Seizures occur in the majority of TSC patients and are often refractory to antiepileptic drugs. Cortical tubers, the primary neuropathology in these patients, were identified as epileptogenic in several clinical studies. However, the contribution of TSC gene inactivation to altered excitability (in the absence of tuber formation) is not fully understood. Here we used antisense morpholino oligonucleotides to knockdown TSC genes in developing zebrafish (Danio rerio). Using intact agar-immobilized zebrafish larvae and extracellular recording techniques, we present evidence for neuronal hyperexcitability following loss of Tsc1. Importantly, Tsc1 morphants show no evidence of tuber formation in histological studies. In addition, rapamycin effectively inhibited target of rapamycin (TOR) kinase activity in zebrafish with no amelioration of the hyperexcitable phenotype. Our study demonstrates that Tsc1 knockdown in a relatively simple brain structure can result in hyperexcitability.
Document Details
- Document Type
- Technical Report
- Publication Date
- Nov 01, 2006
- Accession Number
- ADA462962
Entities
People
- Scott C. Baraban
Organizations
- University of California, San Francisco