Alcohol-drinking procedure
Alcohol preferring P rats, HAD rats, and AA rats were given ad libitum access to tap water and to 5%, and 20% ethanol solution (v/v). All rats underwent a two-week deprivation cycle after 8 weeks of continuous alcohol availability. After the deprivation period, rats were given access to alcohol again and 3 more deprivation periods were introduced similar to a previous report from our group (Vengeliene et al., 2003 link). The long-term voluntary alcohol drinking procedure including all deprivation phases lasted in total 52 weeks. For comparison, 3 age- and weight-matched control P, HAD and AA rat groups underwent the exact same handling procedures for the entire time of the experiment but did not receive alcohol.
Establishment of a brain / organ bank
Animals were deprived from food and liquids for 6 hours to ensure that blood ethanol concentrations were zero (in order to avoid direct acute alcohol effects on gene expression); then animals were sacrificed by exposing them to CO2 and decapitated. The brains were quickly removed and put into a glass with iso-pentane (Sigma-Aldrich Co., St. Louis, MO,USA) kept on dry ice for 5 minutes. Afterwards, the brains were wrapped with parafilm and aluminum folia and stored at -80°C. Other organs, which are also known to be affected by chronic alcohol intake were also quickly removed and stored in liquid nitrogen. These include esophagus, salivary glands, stomach, heart, liver, blood, gut and pancreas.
Vengeliene V, Siegmund S, Singer MV, Sinclair JD, Li TK, Spanagel R (2003) A comparative study on alcohol-preferring rat lines: effects of deprivation and stress phases on voluntary alcohol intake. Alcohol Clin Exp Res 27:1048-1054 hier als pdf-Datei ... |
