Regulation of corticotropin-releasing factor-binding protein expression in amygdalar neuronal cultures.
Regulation of corticotropin-releasing factor-binding protein expression in amygdalar neuronal cultures.
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杏仁核神经元培养物中促肾上腺皮质激素释放因子结合蛋白表达的调节。
DOI：
10.1046/j.1365-2826.1999.00413.x
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发表时间：
1999
影响因子：
3.2
通讯作者：
Mulchahey,JJ
中科院分区：
文献类型：
--
作者：
Kasckow,JW;Regmi,A;Seasholtz,AF;Mulchahey,JJ
促肾上腺皮质激素释放因子结合蛋白（CRF - BP）调节CRF的生物利用度，也可能在应激行为中发挥作用。CRF‐BP已经定位于垂体以及中枢神经系统（CNS）边缘和皮质区域，包括杏仁核。调控杏仁核CRF - BP的信号转导通路尚不清楚。在本报告中，我们研究了蛋白激酶A和C激活剂、CRF、地塞米松和白细胞介素- 6 （IL6）对分离胎儿杏仁核培养物中CRF - BP mRNA和蛋白表达的影响。通过Northern分析，在以下药物作用12小时后，测定CRF - BP mRNA水平：福斯可林（1-30 μM）、CRF （1-1000 nM）、phorbol‐12‐肉豆酸酯‐13‐醋酸酯（TPA; 1-50 nM）、地塞米松（1-100 nM）和IL6 （10-500 pM）。在福斯柯林（30 mM）、CRF（100、1000 nM）、IL6（100、500 pM）、TPA （50 nM）和地塞米松（100 nM）的作用下，CRF‐BP mRNA显著升高；所有组P<0.05, n= 3-6)。我们用相同的药物治疗后，通过对细胞总蛋白进行半定量Western分析，将CRF‐BP表达的观察扩展到蛋白质水平。30 μM福斯克林、1000 nM CRF、50 nM TPA、100 pM IL6或100 nM地塞米松治疗24小时后，CRF - BP表达显著增加（P<0.05，每种治疗n=3）。然后用含有3500对CRF - BP 5 '侧链DNA的大鼠CRF - BP报告基因构建物转染原代培养物。与对照组相比，5种药物治疗均有统计学意义的增加（P<0.05，各n=3）。结果表明，杏仁核中的CRF‐BP受到多种途径的刺激，这些途径可能在促进行为改变中发挥重要作用。
Corticotropin‐releasing factor‐binding protein (CRF‐BP) is known to regulate the bioavailability of CRF and may also play a role in stress behaviours. CRF‐BP has been localized in the pituitary as well as central nervous system (CNS) limbic and cortical areas, including the amygdala. The signal transduction pathways which regulate amygdalar CRF‐BP are not well understood. In this report, we have examined the effect of protein kinase A and C activators, CRF, dexamethasone and interleukin‐6 (IL6) on CRF‐BP mRNA and protein expression in dissociated fetal amygdalar cultures. CRF‐BP mRNA levels were determined by Northern analysis following 12 h treatment with the following agents: forskolin (1–30 μM), CRF (1–1000 nM), phorbol‐12‐myristate‐13‐acetate (TPA; 1–50 nM), dexamethasone (1–100 nM) and IL6 (10–500 pM). Significant increases in CRF‐BP mRNA were observed in response to forskolin (30 mM), CRF (100, 1000 nM), IL6 (100, 500 pM), TPA (50 nM) and dexamethasone (100 nM; P<0.05 for all; n=3–6 for all). We extended our observations of CRF‐BP expression to the protein level by performing semiquantitative Western analysis of total cellular protein after treatment with the same agents. Twenty‐four hour treatment with 30 μM forskolin, 1000 nM CRF, 50 nM TPA, 100 pM IL6 or 100 nM dexamethasone significantly increased CRF‐BP expression (P<0.05, n=3 for each treatment). The primary cultures were then transfected with a rat CRF‐BP‐reporter construct containing 3500 base pairs of CRF‐BP 5′ flanking DNA. Treatment with all five agents produced statistically significant increases above control (P<0.05; n=3 for each). The results suggest that CRF‐BP in the amygdala is stimulated by numerous pathways which may play a significant role in promoting behavioural changes.