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利用原位杂交技术对大鼠中枢神经系统中生长抑素mRNA分布进行半定量分析。

Semi-quantitative analysis of somatostatin mRNA distribution in the rat central nervous system using in situ hybridization.

作者信息

Priestley J V, Réthelyi M, Lund P K

机构信息

Department of Physiology, UMDS St Thomas's Campus, London, UK.

出版信息

J Chem Neuroanat. 1991 Mar-Apr;4(2):131-53. doi: 10.1016/0891-0618(91)90037-d.

Abstract

The distribution of somatostatin mRNA in the rat brain has been examined by in situ hybridization using 32P-labelled oligonucleotide probes. Numerous telencephalic and diencephalic areas contained labelled cells with the largest numbers of cells occurring in the anterior olfactory nucleus, olfactory and entorhinal cortices, hippocampus, neocortex, caudate nucleus, accumbens, septum, amygdala and periventricular nucleus. Fewer labelled cells occurred in the mesencephalon and rhombencephalon but groups were seen in the region of the central grey, lateral lemniscus, parabrachial and tegmental nuclei, medial longitudinal fasciculus and nucleus of the solitary tract. This distribution closely matches published maps of the distribution of somatostatin-immunoreactive cell bodies. The intensity of individual cell labelling has also been quantified using image analysis and compared with the intensity of somatostatin immunocytochemical cell staining. In situ hybridization cell labelling varied both within different regions and from region to region. Highest labelling was seen in the periventricular nucleus of the hypothalamus followed by telencephalic regions such as cortex, hippocampus and the medial nucleus of the amygdala. In contrast all brainstem areas had low levels of labelling with the lowest levels of the brain occurring in the dorsolateral tegmental nucleus. Somatostatin immunocytochemistry showed similar variations such that the intensity of cell immunostaining broadly paralleled the intensity of cell in situ hybridization labelling. Thus both peptide and mRNA levels were much lower in brainstem cells than in forebrain, although a close correlation between immunocytochemistry and in situ hybridization was not seen in all brain regions.

摘要

利用32P标记的寡核苷酸探针,通过原位杂交技术检测了生长抑素mRNA在大鼠脑内的分布情况。许多端脑和间脑区域含有标记细胞,其中标记细胞数量最多的区域为前嗅核、嗅皮质和内嗅皮质、海马体、新皮质、尾状核、伏隔核、隔区、杏仁核和室周核。中脑和后脑的标记细胞较少,但在中央灰质、外侧丘系、臂旁核和被盖核、内侧纵束和孤束核区域可见细胞群。这种分布与已发表的生长抑素免疫反应性细胞体分布图非常吻合。还利用图像分析对单个细胞标记的强度进行了量化,并与生长抑素免疫细胞化学细胞染色的强度进行了比较。原位杂交细胞标记在不同区域内以及不同区域之间均存在差异。下丘脑室周核的标记最强,其次是端脑区域,如皮质、海马体和杏仁核内侧核。相比之下,所有脑干区域的标记水平都很低,其中背外侧被盖核的标记水平最低。生长抑素免疫细胞化学显示出类似的差异,即细胞免疫染色的强度与细胞原位杂交标记的强度大致平行。因此,尽管并非在所有脑区都能看到免疫细胞化学和原位杂交之间的密切相关性,但脑干细胞中的肽和mRNA水平均远低于前脑。

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