Department of Molecular & Cellular Biology, Baylor College of Medicine, Houston, TX 77030-3411, USA.
J Neuroendocrinol. 2009 Nov;21(11):954-60. doi: 10.1111/j.1365-2826.2009.01917.x. Epub 2009 Sep 1.
We have previously demonstrated that dopamine agonist, SKF38396 (SKF), can substitute for progesterone in the facilitation of female reproductive behaviour in oestradiol benzoate-primed female rats and mice. We also reported that both progesterone- and SKF-initiated signalling were mediated by the cAMP-dependent protein kinase A signal transduction cascade. As the rapid effects of progesterone are also mediated by calcium-dependent kinases, calcium- and calmodulin-dependent kinase (CaMKII) and protein kinase (PKC), we sought to determine whether SKF-initiated signalling also recruited calcium as a second messenger. We measured the changes in the activation of CaMKII and PKC in the ventromedial nucleus (VMN) of the hypothalamus and preoptic area (POA) of the rat brain, which are the two regions implicated in the regulation of female reproductive behaviour in rodents. We measured the basal activities representing the activation of the kinases by in vivo treatments, as well as the total kinase activities assayed in the presence of exogenous cofactors in vitro. We report that, in contrast to progesterone-initiated signalling, there was no recruitment of calcium by SKF in the hypothalamus, as shown by the absence of changes in CaMKII activities in the VMN and POA. Furthermore, SKF-treatment resulted in a rapid increase in calcium-independent basal PKC activity in the VMN but not the POA. These rapid changes were not the result of changes in PKC protein levels or phosphorylation status. These data indicate that progesterone- and SKF-recruit distinct signalling molecules within the same regions of the brain to activate region-specific signal transduction pathways.
我们之前已经证明,多巴胺激动剂 SKF38396(SKF)可以在苯甲酸雌二醇预处理的雌性大鼠和小鼠中替代孕激素促进雌性生殖行为。我们还报告说,孕激素和 SKF 引发的信号都由 cAMP 依赖性蛋白激酶 A 信号转导级联介导。由于孕激素的快速作用也由钙依赖性激酶介导,包括钙和钙调蛋白依赖性激酶(CaMKII)和蛋白激酶(PKC),我们试图确定 SKF 引发的信号是否也招募钙作为第二信使。我们测量了下丘脑腹内侧核(VMN)和视前区(POA)中 CaMKII 和 PKC 的激活变化,这两个区域与啮齿动物雌性生殖行为的调节有关。我们通过体内处理测量了代表激酶激活的基础活性,以及在体外存在外源性辅助因子时测定的总激酶活性。我们报告说,与孕激素引发的信号不同,SKF 在下丘脑没有招募钙,因为 VMN 和 POA 中的 CaMKII 活性没有变化。此外,SKF 处理导致 VMN 中钙非依赖性基础 PKC 活性的快速增加,但 POA 中没有。这些快速变化不是 PKC 蛋白水平或磷酸化状态变化的结果。这些数据表明,孕激素和 SKF 在大脑的相同区域招募不同的信号分子,以激活特定区域的信号转导途径。