Exaggerated Response of a Vasopressin-Enhanced Green Fluorescent Protein Transgene to Nociceptive Stimulation in the Rat
Exaggerated Response of a Vasopressin-Enhanced Green Fluorescent Protein Transgene to Nociceptive Stimulation in the Rat
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DOI：
10.1523/jneurosci.2624-09.2009
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发表时间：
2009-10-21
影响因子：
5.3
通讯作者：
Ueta, Yoichi
中科院分区：
文献类型：
--
作者：
Suzuki, Hitoshi;Kawasaki, Makoto;Ueta, Yoichi
痛觉刺激引起神经内分泌反应，如精氨酸抗利尿激素（AVP）的释放以及下丘脑-垂体-肾上腺轴的激活。我们培育了表达avp增强绿色荧光蛋白（eGFP）融合基因的新型转基因大鼠，并在这些大鼠的双侧后肢皮下注射生理盐水或福尔马林后，检测了伤害性刺激对下丘脑转基因表达的影响。我们利用原位杂交组织化学技术评估了(1)血浆中AVP水平以及视上核（SON）和室旁核（PVN）中eGFP mRNA和AVP异核RNA （hnRNA）的变化；(2)PVN不同的大细胞和旁细胞分裂中基因表达的变化；(3)SON、PVN、中位突起（ME）和垂体后腺（PP）中eGFP荧光的变化。注射福尔马林15 min后血浆AVP水平显著升高。在同一时间段内，注射福尔马林大鼠PVN中AVP hnRNA水平升高，尤其是PVN的细细胞分裂。在同一区域，注射福尔马林后eGFP mRNA水平也显著高于AVP hnRNA水平。注射福尔马林后，大鼠SON、PVN、ME和PP的eGFP荧光明显增强，尤其是PVN的细细胞分裂时eGFP荧光增强。总之，我们的研究结果表明，在急性伤害性刺激后，大鼠下丘脑中AVP-eGFP转基因的表达发生了强劲而快速的变化。
Nociceptive stimulation elicits neuroendocrine responses such as arginine vasopressin (AVP) release as well as activation of the hypothalamo-pituitary-adrenal axis. We have generated novel transgenic rats expressing an AVP-enhanced green fluorescent protein (eGFP) fusion gene, and we examined the effects of nociceptive stimulation on transgene expression in the hypothalamus after subcutaneous injection of saline or formalin into the bilateral hindpaws in these rats. We have assessed (1) AVP levels in plasma and the changes of eGFP mRNA and AVP heteronuclear RNA (hnRNA) in the supraoptic nucleus (SON) and the paraventricular nucleus (PVN) using in situ hybridization histochemistry, (2) gene expression changes in distinct magnocellular and parvocellular divisions of the PVN, (3) eGFP fluorescence in the SON, the PVN, the median eminence (ME), and the posterior pituitary gland (PP). Plasma AVP levels were significantly increased 15 min after formalin injection. In the same time period, the AVP hnRNA levels in the PVN were increased, especially in the parvocellular division of the PVN in formalin-injected rats. In the same region, eGFP mRNA levels after formalin injection were also significantly increased to a much greater extent than those of AVP hnRNA. The eGFP fluorescence in the SON, the PVN, the ME, and the PP was markedly increased in formalin-injected rats and especially increased in the parvocellular divisions of the PVN. Together, our results demonstrate robust and rapid changes in the expression of the AVP-eGFP transgene in the rat hypothalamus after acute nociceptive stimulation.