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首页> 外文期刊>Journal of Neuroscience Methods >In situ hybridization for vasopressin mRNA in the human supraoptic and paraventricular nucleus; quantitative aspects of formalin-fixed paraffin-embedded tissue sections as compared to cryostat sections.
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In situ hybridization for vasopressin mRNA in the human supraoptic and paraventricular nucleus; quantitative aspects of formalin-fixed paraffin-embedded tissue sections as compared to cryostat sections.

机译：原位杂交人视上核和心室旁核中血管加压素的mRNA;与冷冻切片机相比，福尔马林固定石蜡包埋的组织切片的定量方面。

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In order to study the suitability of formalin-fixed paraffin-embedded brain tissue for vasopressin (AVP)-mRNA detection, we used symmetric halves of 5 human hypothalami. In every case, one half was formalin fixed for 10-35 days and paraffin embedded while the other half was frozen rapidly. Following in situ hybridization (ISH) histochemistry on systematically obtained sections of the supraoptic (SON) and paraventricular nucleus (PVN) of both halves, total amounts of AVP-mRNA in these nuclei were estimated using densitometry of film autoradiographs. Total amounts of radioactivity were found to vary considerably between patients and amounted to 1297 +/- 302 arbitrary units (AU) (PVN) (mean +/- SEM) and 2539 +/- 346 (SON) for the cryostat sections and 868 +/- 94 (PVN) and 1259 +/- 126 (SON) for the paraffin tissue. Variations introduced by the method itself yielded a coefficient of variation of only 0.19. Furthermore, a non-significant negative trend with postmortem delay was found in cryostat tissue, butnot in paraffin sections. No effect of fixation time was observed in the paraffin tissue. Both ways of tissue treatment have specific advantages and disadvantages that may be different for other probes or other brain areas. For ISH of a highly abundant mRNA like AVP in a very heterogeneous brain area such as the human hypothalamus, formalin-fixed paraffin-embedded tissue sections can be used for quantitative analysis of entire brain nuclei because of the small variation in this tissue, the remarkably good signal recovery (some 75% as compared to cryostat sections) and its practical advantages with regards to anatomical orientation, storage and sampling of the tissue.
机译：为了研究福尔马林固定石蜡包埋的脑组织对血管加压素（AVP）-mRNA检测的适用性，我们使用了对称的两半人类下丘脑。在每种情况下，一半用福尔马林固定10-35天，石蜡包埋，另一半快速冷冻。在系统获取的两半的视上（SON）和室旁核（PVN）切片上进行原位杂交（ISH）组织化学后，使用胶片放射自显影仪的光密度法估算了这些核中AVP-mRNA的总量。发现患者之间的总放射性差异很大，低温恒温器切片的总放射性为1297 +/- 302任意单位（AU）（PVN）（平均+/- SEM）和2539 +/- 346（SON），868 + /-石蜡组织为94（PVN）和1259 +/- 126（SON）。该方法本身引入的变化产生的变异系数仅为0.19。此外，在低温恒温器组织中发现了不显着的负趋势，具有事后延迟，但在石蜡切片中未发现。在石蜡组织中未观察到固定时间的影响。两种组织治疗方式都具有特定的优点和缺点，这些优点和缺点可能与其他探针或其他大脑区域不同。对于在高度异质的大脑区域（例如人下丘脑）中高度丰富的mRNA（如AVP）的ISH，由于该组织的细微变化，福尔马林固定石蜡包埋的组织切片可用于定量分析整个脑核。信号恢复良好（与低温恒温器切片相比约为75％），并且在解剖学定位，组织存储和采样方面具有实际优势。

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