In situ hybridization histochemistry for messenger ribonucleic acid (mRNA) encoding gonadotropin-releasing hormone (GnRH): effect of estrogen on cellular levels of GnRH mRNA in female rat brain.
In situ hybridization histochemistry for messenger ribonucleic acid (mRNA) encoding gonadotropin-releasing hormone (GnRH): effect of estrogen on cellular levels of GnRH mRNA in female rat brain.
复制标题
编码促性腺激素释放激素 (GnRH) 的信使核糖核酸 (mRNA) 的原位杂交组织化学：雌激素对雌性大鼠脑中 GnRH mRNA 细胞水平的影响。
作者：
R. Zoeller;P. Seeburg;W. Young
利用原位杂交组织化学技术，我们在大鼠脑核周围检测到含有编码GnRH和GnRH相关肽（GAP）的mRNA。合成的DNA低聚物与编码GnRH结构区和GAP结构区的大鼠cDNA序列互补，杂交到甲醛固定的冠状切片上。含有GnRH/GAP mRNA的细胞分布和数量与免疫细胞化学研究结果相似。核周围显示GnRH mRNA的区域包括内侧间隔区、Broca斜带、视前区和下丘脑前部。在一个包含对角线带和终层血管器官（OVLT）的单个12微米切片中检测到多达55个细胞，而在更多的尾端区域细胞数量减少。此外，两种探针标记相邻切片中包含的相同细胞。我们使用该技术检测了雌激素对正常和雄激素不育雌性大鼠OVLT区域GnRH mRNA水平的影响，使用了雌激素治疗模式，该模式先前在研究下丘脑对雌激素负反馈和正反馈的调节的研究中具有特征。我们发现卵巢切除术后7天，雌激素治疗2天导致正常和雄激素不育雌性GnRH mRNA的平均细胞水平显著降低。对标记强度与细胞丰度相关的直方图的分析表明，一小部分GnRH细胞对雌激素治疗有反应。然而，我们没有发现证据表明在OVLT区域内存在这样一个gnrh应答细胞亚群的离散神经解剖学分离。
Using in situ hybridization histochemistry, we have detected perikarya containing mRNA encoding GnRH and GnRH-associated peptide (GAP) in rat brains. Synthetic DNA oligomers with sequences complementary to the rat cDNA encoding the GnRH structural region and the GAP structural region were hybridized to formaldehyde-fixed coronal sections. The distribution and number of cells containing GnRH/GAP mRNA were similar to those shown by immunocytochemical studies. The areas in which GnRH mRNA perikarya were shown included the medial septal area, the diagonal band of Broca, the preoptic area, and the anterior hypothalamus. Up to 55 cells were detected in a single 12-micron section containing the diagonal band and organum vasculosum lamina terminalis (OVLT) whereas cell numbers diminished in more caudal regions. In addition, both probes labeled the same cells contained within adjacent sections. We used this technique to examine the effect of estrogen on GnRH mRNA levels in the area of the OVLT of normal and androgen-sterilized female rats, using an estrogen treatment paradigm previously characterized in studies investigating the hypothalamic regulation of negative and positive estrogen feedback. We found that 7 days after ovariectomy, 2 days of estrogen treatment resulted in a significant reduction in the average cellular level of GnRH mRNA in both normal and androgen-sterilized females. Analysis of histograms relating the intensity of labeling to the abundance of cells suggested that a small population of GnRH cells responded to the estrogen treatment. However, we found no evidence for a discrete neuroanatomical segregation of such a subpopulation of GnRH-responsive cells within the area of the OVLT.