JG Selenate and LiCl-induced pMAPK and c-Fos

Expt Code: JG

TODO: get dates and number of rats, DOBs

IACAUC Protocol: 1113

1. Animals

• Adult male Sprague Dawley rats (n=12)
• weight 300 - 500g
• Source: Charles River
• DOB:

1. Procedure (rev 2013-7-24)

Weight rats

1. 1. Schedule:

(Stagger pairs of rats by 20 min to allow time for perfusion)

at 0h, inject with either 1 ml/kg vehicle (0.15 M NaCl) or 0.5 mg/kg Na2SeO4

at 1 h, inject with either 12 ml/kg NaCl or 12 ml/kg LiCl (0.15 M)

at 2 h, overdose with Sleepaway and perfuse

Dissect brains, postfix for 2 h in 4%PFA, rinse with 0.1 M PB, cyroprotect in 30% sucrose

1. Immunohistochemistry

Process brains for immunohistochemistry with anti-cFos and anti-pMAPK:

From Kwon and Houpt:

anti-phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) rabbit mAb (#4370, 1:500, Cell Signaling Technology)

From Kimbrough TORC-1 paper:

anti-c-Fos goat polyclonal antibodies (#SC-52-G, 1:10,000 Santa Cruz Biotechnology)