Department of Zoology and Physiology, University of Wyoming, Laramie, WY 82071, USA.
Brain Res. 2013 Jun 26;1518:9-25. doi: 10.1016/j.brainres.2013.04.042. Epub 2013 Apr 28.
To facilitate the study of the CaMKIIα function in vivo, a CaMKIIα-GFP transgenic mouse line was generated. Here, our goal is to provide the first neuroanatomical characterization of GFP expression in the CNS of this line of mouse. Overall, CaMKIIα-GFP expression is strong and highly heterogeneous, with the dentate gyrus of the hippocampus as the most abundantly expressed region. In the hippocampus, around 70% of granule and pyramidal neurons expressed strong GFP. In the neocortex, presumed pyramidal neurons were GFP positive: around 32% of layer II/III and 35% of layer VI neurons expressed GFP, and a lower expression rate was found in other layers. In the thalamus and hypothalamus, strong GFP signals were detected in the neuropil. GFP-positive cells were also found in many other regions such as the spinal trigeminal nucleus, cerebellum and basal ganglia. We further compared the GFP expression with specific antibody staining for CaMKIIα and GABA. We found that GFP+ neurons were mostly positive for CaMKIIα-IR throughout the brain, with some exceptions throughout the brain, especially in the deeper layers of neocortex. GFP and GABA-IR marked distinct neuronal populations in most brain regions with the exception of granule cells in the olfactory bulb, purkinje cells in the cerebellar, and some layer I cells in neocortex. In conclusion, GFP expression in the CaMKIIα-GFP mice is similar to the endogenous expression of CaMKIIα protein, thus these mice can be used in in vivo and in vitro physiological studies in which visualization of CaMKIIα- neuronal populations is required.
为了便于研究 CaMKIIα 在体内的功能，我们构建了 CaMKIIα-GFP 转基因小鼠品系。在这里，我们的目标是首次对该小鼠品系中枢神经系统中 GFP 表达的神经解剖结构进行特征描述。总的来说，CaMKIIα-GFP 的表达很强且具有高度异质性，其中以海马齿状回的表达最为丰富。在海马中，约 70%的颗粒细胞和锥体细胞表达强烈的 GFP。在大脑皮层中，假定的锥体细胞呈 GFP 阳性：约 32%的 II/III 层和 35%的 VI 层神经元表达 GFP，而在其他层中表达率较低。在丘脑和下丘脑，神经胶质中检测到强烈的 GFP 信号。GFP 阳性细胞还存在于许多其他区域，如三叉神经脊核、小脑和基底神经节。我们进一步将 GFP 表达与 CaMKIIα 和 GABA 的特异性抗体染色进行比较。我们发现 GFP+神经元在整个大脑中大多对 CaMKIIα-IR 呈阳性，除了大脑的某些区域（尤其是大脑皮层的深层）有一些例外。GFP 和 GABA-IR 在大多数脑区标记了不同的神经元群体，除了嗅球的颗粒细胞、小脑的浦肯野细胞和新皮层的一些 I 层细胞。总之，CaMKIIα-GFP 小鼠中的 GFP 表达与 CaMKIIα 蛋白的内源性表达相似，因此这些小鼠可用于需要可视化 CaMKIIα-神经元群体的体内和体外生理研究。
