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. Author manuscript; available in PMC: 2013 Feb 1.
Published in final edited form as: Alcohol. 2011 Sep 17;46(1):3–16. doi: 10.1016/j.alcohol.2011.08.003

Table 1.

Chronic Ethanol Exposure (CEE) experimental paradigm and ethanol concentrations

Days of
Study 		Day 1 Day 2 Day 3 Day 4 Day 5 Day 6
(24+W)		 Day 7
(48+W)		 Day 8
(72+W)
EtOH aliquot (Time of day) NA 1530 hrs 1530 hrs 1530 hrs 0900 hrs NA NA 0900 hrs
CEE treatment (Time of day) 100 mM @ 1600 hrs 100 mM @ 1600 hrs 100 mM @ 1600 hrs 100 mM @ 1600 hrs NA NA NA NA
EtOH (mM) NA 45.4±1.63 60.7±3.9a 74.3±2.4a 77.8±2.4a NA NA 14.2±4.1a,b
Mean EtOH (mM) 57.3 ± 2.1
Post-CEE experiments conducted between 1000 – 1700 hrs

  • ICC

  • Trypan-blue staining

  • Patch clamp recording

  • ICC

  • Trypan-blue staining

  • Patch clamp recording

  • ICC

  • Trypan-blue staining

  • Patch clamp recording

  • ICC

  • Trypan-blue staining

  • Patch clamp recording

Table represents the CEE experimental paradigm with the days of study represented across the columns and various experimental parameters along the rows. Within the CEE paradigm, the CGC astrocytic co-cultures were treated with ethanol every day for four days (Day 1 to Day 4) at 1600 hrs. Before replacing the media, an aliquot of the media was collected for ethanol concentration determination (Day 2 to Day 4), with the values represented as mean ± sem above. Similarly, aliquots for the determination of ethanol concentration were collected on Day 5 (at a time during which ethanol levels were high) and Day 8 (72+W) prior to the start of the post-CEE experiments: immunocytochemistry (ICC), trypan-blue staining and patch clamp recordings.

a

Ethanol concentrations were significantly greater at Days 3 (q = 4.9; P ≤ 0.01); Day 4 (q = 8.6; P ≤ 0.001) and Day 5 (q = 4.9; P ≤ 0.01); compared to end of Day 2 values. There were no significant differences in ethanol concentrations among Days 3 – 5.

(b)

At the 72W time point, ethanol values were significantly lower compared to Day 2 (q = 6.4; P ≤ 0.001) as well as all other time points, data not shown.