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戊四氮对未成熟大鼠海马神经元的影响

Pentylentetrazol associated changes of hippocampal neurons in immature rats.

作者信息

Jandová K, Riljak V, Pokorný J, Langmeier M

机构信息

Institute of Physiology of the First Faculty of Medicine, Charles University, Prague, Czech Republic.

出版信息

Prague Med Rep. 2007;108(1):67-74.

Abstract

Using histochemical analysis, the NADPH-diaphorase, Fluoro-Jade B and bis-benzimide (Hoechst 33342) the effect of intraperitoneal administration of pentylentetrazol (PTZ) on hippocampal neurons was studied. 18-day-old male rats of the Wistar strain received PTZ (60 mg/kg) in one dose. The next day, the 19-day-old animals were transcardially perfused with 4% paraformaldehyde under deep thiopental anaesthesia. Cryostat sections were stained to identify NADPH-diaphorase positive neurons that were then quantified in the CA1 and CA3 areas of the hippocampus, in the dorsal and ventral blades of the dentate gyrus and in the hilus of the dentate gyrus. Combination of the Fluoro-Jade B and bis-benzimide (Hoechst 33342) staining was used in the same areas, to identify possible neurodegeneration. Number of NADPH-d positive neurons was higher after pentylentetrazol administration in CA1 and CA3 areas of the hippocampus and in the hilus of the dentate gyrus, compared to the control group which we consider as baseline. Morphological alterations (cell loss) in CA3 area of the hippocampus and in the hilus of the dentate gyrus only (evaluated by Hoechst 33342) were found in animals receiving PTZ; no FJ-B positive cells were found and we can conclude that neurons were destroyed by the PTZ insult.

摘要

利用组织化学分析,通过烟酰胺腺嘌呤二核苷酸磷酸黄递酶、氟玉红B和双苯甲酰亚胺(Hoechst 33342)研究了腹腔注射戊四氮(PTZ)对海马神经元的影响。18日龄的雄性Wistar品系大鼠一次性接受60mg/kg的PTZ。第二天,在硫喷妥钠深度麻醉下,对19日龄的动物进行心脏灌注4%多聚甲醛。对冰冻切片进行染色以识别烟酰胺腺嘌呤二核苷酸磷酸黄递酶阳性神经元,然后在海马体的CA1和CA3区域、齿状回的背侧和腹侧叶片以及齿状回的hilus中对其进行定量。在相同区域使用氟玉红B和双苯甲酰亚胺(Hoechst 33342)染色组合,以识别可能的神经变性。与我们视为基线的对照组相比,在给予戊四氮后,海马体的CA1和CA3区域以及齿状回的hilus中烟酰胺腺嘌呤二核苷酸磷酸黄递酶阳性神经元的数量更高。在接受PTZ的动物中,仅在海马体的CA3区域和齿状回的hilus中发现形态学改变(细胞丢失)(通过Hoechst 33342评估);未发现氟玉红B阳性细胞,我们可以得出结论,神经元被PTZ损伤破坏。

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