Dickson S L, Luckman S M
Anatomy and Human Biology Group, King's College London, United Kingdom.
Endocrinology. 1997 Feb;138(2):771-7. doi: 10.1210/endo.138.2.4907.
In this study we investigated the neurochemical identity of the arcuate cells activated following GH-releasing peptide-6 (GHRP-6) injection by comparing, on consecutive sections, the distribution c-fos messenger RNA (mRNA) with that of mRNAs for peptides synthesized in arcuate cells, including neuropeptide Y (NPY), GH-releasing factor (GRF), tyrosine hydroxylase, POMC, and somatostatin. Rats bearing chronically implanted jugular catheters were injected with either 50 micrograms GHRP-6 or vehicle. Thirty minutes later they were terminally anesthetized and perfused with fixative. Paraffin-embedded sections of 7 microns thickness were processed using in situ hybridization for either c-fos mRNA or mRNAs for the neurochemical markers. In GHRP-6-treated rats the mean (+/-SEM) number of cells expressing c-fos mRNA in the arcuate nucleus (23 +/- 2 cells/section per rat; n = 5) was significantly higher than for vehicle-treated controls (2 +/- 1 cells/section per rat; n = 5; P < 0.001, Mann-Whitney U test). Superimposed camera lucida maps indicated that, in GHRP-6-injected rats, neurochemically identifiable cells expressing c-fos mRNA also express NPY mRNA (51 +/- 4%), GRF mRNA (23 +/- 1%) tyrosine hydroxylase mRNA (11 +/- 3%), POMC mRNA (11 +/- 2%), or somatostatin mRNA (4 +/- 1%). Thus, the majority of cells expressing c-fos mRNA following GHRP-6 injection are NPY and GRF-containing cells.
在本研究中,我们通过在连续切片上比较c-fos信使核糖核酸(mRNA)与弓状核细胞中合成的肽类mRNA的分布,来研究注射生长激素释放肽-6(GHRP-6)后被激活的弓状核细胞的神经化学特性,这些肽类包括神经肽Y(NPY)、生长激素释放因子(GRF)、酪氨酸羟化酶、阿黑皮素原(POMC)和生长抑素。给长期植入颈静脉导管的大鼠注射50微克GHRP-6或赋形剂。30分钟后,将它们深度麻醉并灌注固定剂。对7微米厚的石蜡包埋切片进行原位杂交,检测c-fos mRNA或神经化学标志物的mRNA。在GHRP-6处理的大鼠中,弓状核中表达c-fos mRNA的细胞平均数量(±标准误)为每只大鼠23±2个细胞/切片(n = 5),显著高于赋形剂处理的对照组(每只大鼠2±1个细胞/切片;n = 5;P < 0.001,曼-惠特尼U检验)。叠加的明场绘图显示,在注射GHRP-6的大鼠中,表达c-fos mRNA的神经化学可识别细胞也表达NPY mRNA(51±4%)、GRF mRNA(23±1%)、酪氨酸羟化酶mRNA(11±3%)、POMC mRNA(11±2%)或生长抑素mRNA(4±1%)。因此,注射GHRP-6后表达c-fos mRNA的大多数细胞是含NPY和GRF的细胞。