用于大鼠脑内糖皮质激素受体mRNA定位的cRNA探针原位杂交方法的优化：详细方案
Optimization of cRNA probe in situ hybridization methodology for localization of glucocorticoid receptor mRNA in rat brain: a detailed protocol.
作者信息
Whitfield H J, Brady L S, Smith M A, Mamalaki E, Fox R J, Herkenham M
机构信息
Clinical Neuroendocrinology Branch, NIMH, Bethesda, Maryland 20892.
出版信息
Cell Mol Neurobiol. 1990 Mar;10(1):145-57. doi: 10.1007/BF00733641.
Abstract
- We have described a general ribonucleotide probe in situ hybridization methodology for localization of mRNA in frozen, unfixed tissue sections of brain. 2. The most important steps in obtaining consistent and reproducible autoradiographs with ribonucleotide probes were tissue acetylation and application of the radiolabeled probe to tissue sections under unsealed, glass coverslips. 3. Variability of the hybridization signal in tissue sections has been minimized to achieve a high degree of reproducibility within a given experiment as determined by densitometric analysis of rat glucocorticoid and mineralocorticoid receptor mRNA hybridization autoradiographs. 4. Tissue quality has been optimized for high-resolution anatomical localization of mRNA species by nuclear track emulsion. 5. The protocol is amenable to rapid, batchwise processing of tissue samples.
摘要
- 我们描述了一种通用的核糖核苷酸探针原位杂交方法，用于在大脑的冷冻、未固定组织切片中定位mRNA。2. 使用核糖核苷酸探针获得一致且可重复的放射自显影照片的最重要步骤是组织乙酰化以及在未密封的盖玻片下将放射性标记探针应用于组织切片。3. 通过对大鼠糖皮质激素和盐皮质激素受体mRNA杂交放射自显影照片进行光密度分析确定，组织切片中杂交信号的变异性已降至最低，以在给定实验中实现高度的可重复性。4. 已优化组织质量，以便通过核径迹乳剂对mRNA种类进行高分辨率的解剖定位。5. 该方案适用于组织样本的快速批量处理。
相似文献
1
Optimization of cRNA probe in situ hybridization methodology for localization of glucocorticoid receptor mRNA in rat brain: a detailed protocol.用于大鼠脑内糖皮质激素受体mRNA定位的cRNA探针原位杂交方法的优化：详细方案
Cell Mol Neurobiol. 1990 Mar;10(1):145-57. doi: 10.1007/BF00733641.
2
Localization of glucocorticoid receptor mRNA in the male rat brain by in situ hybridization.通过原位杂交对雄性大鼠脑中糖皮质激素受体mRNA进行定位。
Proc Natl Acad Sci U S A. 1988 Dec;85(23):9331-5. doi: 10.1073/pnas.85.23.9331.
3
Tissue-specific regulation of glucocorticoid receptor mRNA by dexamethasone.地塞米松对糖皮质激素受体mRNA的组织特异性调控
J Biol Chem. 1987 Aug 5;262(22):10441-4.
4
The neuronal mineralocorticoid receptor as a mediator of glucocorticoid response.作为糖皮质激素反应介质的神经元盐皮质激素受体。
Neuron. 1988 Nov;1(9):887-900. doi: 10.1016/0896-6273(88)90136-5.
5
Localization of glucocorticoid receptor messenger ribonucleic acid in hippocampus of rat brain using in situ hybridization.利用原位杂交技术对大鼠脑海马体中糖皮质激素受体信使核糖核酸进行定位。
Mol Endocrinol. 1988 Aug;2(8):682-5. doi: 10.1210/mend-2-8-682.
6
Distribution of the mineralocorticoid and the glucocorticoid receptor mRNAs in the rat hippocampus.盐皮质激素和糖皮质激素受体mRNA在大鼠海马中的分布。
J Neurosci Res. 1988 Sep;21(1):88-94. doi: 10.1002/jnr.490210113.
7
Localization and regulation of glucocorticoid and mineralocorticoid receptor messenger RNAs in the hippocampal formation of the rat.大鼠海马结构中糖皮质激素和盐皮质激素受体信使核糖核酸的定位与调节
Mol Endocrinol. 1989 Nov;3(11):1886-94. doi: 10.1210/mend-3-11-1886.
8
Localization of insulin-like growth factor-I mRNA in rat brain by in situ hybridization--relationship to IGF-I receptors.大鼠脑中胰岛素样生长因子-I mRNA的原位杂交定位——与IGF-I受体的关系
Mol Endocrinol. 1990 May;4(5):773-8. doi: 10.1210/mend-4-5-773.
9
Tissue localization of 11 beta-hydroxysteroid dehydrogenase and its relationship to the glucocorticoid receptor.11β-羟类固醇脱氢酶的组织定位及其与糖皮质激素受体的关系。
J Steroid Biochem Mol Biol. 1992 Jan;41(1):21-8. doi: 10.1016/0960-0760(92)90220-d.
10
Localization of prodynorphin messenger rna in rat brain by in situ hybridization using a synthetic oligonucleotide probe.
Neurosci Lett. 1986 Aug 15;69(1):104-8. doi: 10.1016/0304-3940(86)90423-4.
引用本文的文献
1
Neural activation following offensive aggression in Japanese quail.日本鹌鹑攻击性攻击后的神经激活
Biol Open. 2018 Dec 10;7(12):bio038026. doi: 10.1242/bio.038026.
2
Effects of Immune Activation during Early or Late Gestation on -Methyl-d-Aspartate Receptor Measures in Adult Rat Offspring.孕期早期或晚期免疫激活对成年大鼠后代N-甲基-D-天冬氨酸受体指标的影响。
Front Psychiatry. 2017 Sep 4;8:77. doi: 10.3389/fpsyt.2017.00077. eCollection 2017.
3
Neuroendocrine correlates of sex-role reversal in barred buttonquails.斑纹纽扣鹌鹑性别角色反转的神经内分泌关联
Proc Biol Sci. 2016 Nov 30;283(1843). doi: 10.1098/rspb.2016.1969.
4
The effect of adolescent testosterone on hippocampal BDNF and TrkB mRNA expression: relationship with cell proliferation.青少年睾酮对海马脑源性神经营养因子（BDNF）和酪氨酸激酶受体B（TrkB）mRNA表达的影响：与细胞增殖的关系。
BMC Neurosci. 2015 Feb 18;16:4. doi: 10.1186/s12868-015-0142-x.
5
Breeding status and social environment differentially affect the expression of sex steroid receptor and aromatase mRNA in the brain of female Damaraland mole-rats.繁殖状况和社会环境对雌性沙漠棉鼠大脑中性类固醇受体和芳香化酶 mRNA 的表达有不同影响。
Front Zool. 2014 May 8;11:38. doi: 10.1186/1742-9994-11-38. eCollection 2014.
6
Decreased BDNF and TrkB mRNA expression in multiple cortical areas of patients with schizophrenia and mood disorders.精神分裂症和心境障碍患者多个皮质区域中脑源性神经营养因子（BDNF）和酪氨酸激酶受体B（TrkB）mRNA表达降低。
Transl Psychiatry. 2014 May 6;4(5):e389. doi: 10.1038/tp.2014.26.
7
Developmental trajectory of the endocannabinoid system in human dorsolateral prefrontal cortex.人类背外侧前额叶皮层中环腺苷酸系统的发育轨迹。
BMC Neurosci. 2012 Jul 24;13:87. doi: 10.1186/1471-2202-13-87.
8
Social status affects the degree of sex difference in the songbird brain.社会地位影响鸣禽大脑的性别差异程度。
PLoS One. 2011;6(6):e20723. doi: 10.1371/journal.pone.0020723. Epub 2011 Jun 8.
9
Decreased BDNF, trkB-TK+ and GAD67 mRNA expression in the hippocampus of individuals with schizophrenia and mood disorders.精神分裂症和心境障碍患者海马脑源性神经营养因子、trkB-TK+ 和 GAD67mRNA 表达降低。
J Psychiatry Neurosci. 2011 May;36(3):195-203. doi: 10.1503/jpn.100048.
10
Increased interstitial white matter neuron density in the dorsolateral prefrontal cortex of people with schizophrenia.精神分裂症患者背外侧前额叶皮质的细胞间白质神经元密度增加。
Biol Psychiatry. 2011 Jan 1;69(1):63-70. doi: 10.1016/j.biopsych.2010.08.020. Epub 2010 Oct 25.
本文引用的文献
1
Binding of radiolabeled opiates to slide-mounted sections of molded minced rat brain: a novel method for conducting radioreceptor assays.放射性标记阿片类药物与大鼠脑模制碎块载玻片切片的结合：一种进行放射受体测定的新方法。
Neuropeptides. 1983 Oct;3(6):493-9. doi: 10.1016/0143-4179(83)90039-2.
2
Light microscopic localization of brain opiate receptors: a general autoradiographic method which preserves tissue quality.脑阿片受体的光镜定位：一种保留组织质量的通用放射自显影方法。
J Neurosci. 1982 Aug;2(8):1129-49. doi: 10.1523/JNEUROSCI.02-08-01129.1982.
3
AChE-stained horizontal sections of the rat brain in stereotaxic coordinates.以立体定位坐标表示的大鼠脑乙酰胆碱酯酶染色水平切片。
J Neurosci Methods. 1980 Dec;3(2):129-49. doi: 10.1016/0165-0270(80)90021-7.
4
Mapping of glucocorticoid receptor immunoreactive neurons in the rat tel- and diencephalon using a monoclonal antibody against rat liver glucocorticoid receptor.使用抗大鼠肝脏糖皮质激素受体的单克隆抗体对大鼠端脑和间脑中糖皮质激素受体免疫反应性神经元进行定位。
Endocrinology. 1985 Nov;117(5):1803-12. doi: 10.1210/endo-117-5-1803.
5
Genetic complementation of a glucocorticoid receptor deficiency by expression of cloned receptor cDNA.通过克隆受体cDNA的表达对糖皮质激素受体缺陷进行基因互补。
Cell. 1986 Aug 1;46(3):389-99. doi: 10.1016/0092-8674(86)90659-8.
6
Adrenal steroid receptors and actions in the nervous system.肾上腺类固醇受体及其在神经系统中的作用。
Physiol Rev. 1986 Oct;66(4):1121-88. doi: 10.1152/physrev.1986.66.4.1121.
7
Localization of glucocorticoid receptor messenger ribonucleic acid in hippocampus of rat brain using in situ hybridization.利用原位杂交技术对大鼠脑海马体中糖皮质激素受体信使核糖核酸进行定位。
Mol Endocrinol. 1988 Aug;2(8):682-5. doi: 10.1210/mend-2-8-682.
8
Localization of glucocorticoid receptor mRNA in the male rat brain by in situ hybridization.通过原位杂交对雄性大鼠脑中糖皮质激素受体mRNA进行定位。
Proc Natl Acad Sci U S A. 1988 Dec;85(23):9331-5. doi: 10.1073/pnas.85.23.9331.
9
The neuronal mineralocorticoid receptor as a mediator of glucocorticoid response.作为糖皮质激素反应介质的神经元盐皮质激素受体。
Neuron. 1988 Nov;1(9):887-900. doi: 10.1016/0896-6273(88)90136-5.
10
Distribution of the mineralocorticoid and the glucocorticoid receptor mRNAs in the rat hippocampus.盐皮质激素和糖皮质激素受体mRNA在大鼠海马中的分布。
J Neurosci Res. 1988 Sep;21(1):88-94. doi: 10.1002/jnr.490210113.
Zotero 插件