Diao W-F, Afjehi-Sadat L, Chen W-Q, Höger J, Höger H, Pollak A, Lubec G
Department of Pediatrics, Medical University of Vienna, Waehringer Guertel 18, A-1090, Vienna, Austria.
Neuroscience. 2008 Jun 26;154(3):1009-20. doi: 10.1016/j.neuroscience.2008.04.008. Epub 2008 Apr 11.
Hippocampal functions vary across the estrous cycle but metabolic changes at the protein level have not been systematically studied so far. It was therefore the aim of the study to screen expression of metabolic proteins mainly represented by metabolic enzymes in the hippocampus over the estrous cycle and in males. Female and male OFA Sprague-Dawley rats were used and female estrous phases were determined by vaginal smears, according to which females were separated into groups of proestrous, estrous, early and late metestrous and diestrous. Proteins were extracted from hippocampal tissue and separated on two-dimensional gel electrophoresis followed by identification with mass spectrometry methods (MALDI-TOF-TOF and nano-LC-ESI-MS/MS). Comparative analysis of protein levels was carried out by quantifying protein spot volumes by means of specific software. Levels of one expression form of gamma-enolase were different between diestrous and early metestrous; C-1-tetrahydrofolate synthase levels were elevated in proestrous as compared with estrous and serotransferrin levels were increased in diestrous as compared with proestrous, estrous, metestrous and in males. The outcome of estrous cycle- and gender-dependent protein fluctuations is relevant for the interpretation of previous and future work as well as for the design of further studies at the protein level in the hippocampus.
海马体的功能在发情周期中会有所不同,但迄今为止,蛋白质水平的代谢变化尚未得到系统研究。因此,本研究的目的是筛选在发情周期和雄性大鼠中海马体中主要由代谢酶代表的代谢蛋白的表达情况。使用雌性和雄性OFA斯普拉格-道利大鼠,通过阴道涂片确定雌性的发情阶段,并据此将雌性分为动情前期、发情期、动情后期早期和晚期以及动情间期组。从海马体组织中提取蛋白质,在二维凝胶电泳上进行分离,然后用质谱方法(基质辅助激光解吸电离飞行时间串联质谱和纳升级液相色谱-电喷雾电离串联质谱)进行鉴定。通过特定软件对蛋白质斑点体积进行定量,对蛋白质水平进行比较分析。γ-烯醇化酶的一种表达形式在动情间期和动情后期早期之间存在差异;与发情期相比,C-1-四氢叶酸合酶水平在动情前期升高,与动情前期、发情期、动情后期以及雄性相比,血清转铁蛋白水平在动情间期升高。发情周期和性别依赖性蛋白质波动的结果对于解释之前和未来的研究工作以及设计海马体蛋白质水平的进一步研究具有重要意义。