Generation and Genotyping of SOD1H46R Transgenic Rats

All Sprague-Dawley rats were obtained from SLC Japan (Shizuoka, Japan). Transgenic rats carrying the human mutant SOD1 gene for H46R (SOD1^H46R) were housed and genotyped as previously.²¹ (link) The DNA of newborn rats was extracted from their tails, and PCR amplification (forward primer: 5′-TTGGGAGGAGGTAGTGATTA; reverse primer: 5′-AGCTAGCAGGATAACAGATGA; 94°C for 30 s, 55°C for 30 s, 72°C for 30 s, 30 cycles) was performed to identify the exogenous human SOD1 transgene DNA. Founder rats were mated with Sprague-Dawley rats. For western blotting or cDNA microarray experiments, rats were anesthetized intraperitoneally using the combination of medetomidine, midazolam, and butorphanol tartrate. Subsequently, they were intracardially perfused with PBS. For immunohistochemistry, 4% PFA (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) was additionally used for tissue fixation.