Miller M A, Zoeller R T, Dorsa D M
Geriatric Research, Education and Clinical Center, Veterans Administration Medical Center, Seattle, WA 98108.
Neurosci Lett. 1988 Dec 5;94(3):264-8. doi: 10.1016/0304-3940(88)90028-6.
In situ hybridization histochemistry and quantitative autoradiography were used to confirm the presence of cells within the bed nucleus of the stria terminalis (BNST) which express the vasopressin (VP) gene and to assess the biosynthetic capacity of these cells throughout the rostrocaudal extent of the nucleus. Brain sections from adult male Wistar rats were hybridized with a 35S-labeled 48-base oligonucleotide probe. Clusters of grains were present over cells in the BNST. Cells were parvocellular in appearance and signal over cells was determined to be specific since it was abolished by RNase pretreatment or incubation with 100-fold excess unlabeled probe. The distribution of VP-mRNA containing cells in the BNST corresponds closely to that previously reported by immunocytochemistry. No clear-cut rostral to caudal gradient was found for gene expression as measured by grains/cell. In situ hybridization techniques can provide a powerful tool to study the regulation of central VP pathways in the BNST.
采用原位杂交组织化学和定量放射自显影技术，以证实终纹床核（BNST）内存在表达血管加压素（VP）基因的细胞，并评估这些细胞在整个核的前后范围内的生物合成能力。取自成年雄性Wistar大鼠的脑切片与35S标记的48碱基寡核苷酸探针进行杂交。BNST中的细胞上出现了颗粒簇。细胞外观为小细胞，细胞上的信号被确定为特异性的，因为它被核糖核酸酶预处理或与100倍过量的未标记探针孵育所消除。BNST中含VP-mRNA细胞的分布与先前免疫细胞化学报道的分布密切对应。通过颗粒/细胞测量，未发现基因表达有明显的前后梯度。原位杂交技术可为研究BNST中中枢VP通路的调节提供有力工具。
