Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY 40536-0596, USA.
Neuropharmacology. 2018 May 1;133:276-288. doi: 10.1016/j.neuropharm.2018.01.032. Epub 2018 Jan 31.
Neural stem cell-driven adult neurogenesis contributes to the integrity of the hippocampus. Excessive alcohol consumption in alcoholism results in hippocampal degeneration that may recover with abstinence. Reactive, increased adult neurogenesis during abstinence following alcohol dependence may contribute to recovery, but the mechanism driving reactive neurogenesis is not known. Therefore, adult, male rats were exposed to alcohol for four days and various markers were used to examine cell cycle dynamics, the percentage and number of neural progenitor cell subtypes, and the percentage of quiescent versus activated progenitors. Using a screen for cell cycle perturbation, we showed that the cell cycle is not likely altered at 7 days in abstinence. As the vast majority of Bromodeoxyuridine-positive (+) cells were co-labeled with progenitor cell marker, Sox2, we then developed a quadruple fluorescent labeling scheme to examine Type-1, -2a, -2b and -3 progenitor cells simultaneously. Prior alcohol dependence indiscriminately increased all subtypes at 7 days, the peak of the reactive proliferation. An evaluation of the time course of reactive cell proliferation revealed that cells begin proliferating at 5 days post alcohol, where only actively dividing Type 2 progenitors were increased by alcohol. Furthermore, prior alcohol increased the percentage of actively dividing Sox2+ progenitors, which supported that reactive neurogenesis is likely due to the activation of progenitors out of quiescence. These observations were associated with granule cell number returning to normal at 28 days. Therefore, activating stem and progenitor cells out of quiescence may be the mechanism underlying hippocampal recovery in abstinence following alcohol dependence.
神经干细胞驱动的成年神经发生有助于海马体的完整性。酗酒导致的酒精中毒会导致海马体退化,而戒酒后可能会恢复。酒精依赖戒断后,反应性、增加的成年神经发生可能有助于恢复,但驱动反应性神经发生的机制尚不清楚。因此,雄性成年大鼠暴露于酒精中四天,并使用各种标志物来检查细胞周期动力学、神经祖细胞亚型的百分比和数量,以及静止与激活祖细胞的百分比。通过细胞周期扰动筛选,我们表明在戒断 7 天时细胞周期不太可能改变。由于绝大多数 BrdU 阳性(+)细胞与祖细胞标志物 Sox2 共标记,因此我们开发了一种四重荧光标记方案来同时检查 Type-1、-2a、-2b 和-3 祖细胞。先前的酒精依赖在 7 天时不加区分地增加了所有亚型,这是反应性增殖的高峰期。对反应性细胞增殖的时间过程的评估表明,细胞在酒精后 5 天开始增殖,只有活跃分裂的 Type 2 祖细胞被酒精增加。此外,先前的酒精增加了活跃分裂的 Sox2+祖细胞的百分比,这表明反应性神经发生可能是由于祖细胞从静止中激活所致。这些观察结果与 28 天时颗粒细胞数量恢复正常有关。因此,使静止的干细胞和祖细胞激活可能是酒精依赖戒断后海马体恢复的机制。