Functional Characterization of Resting and Adenovirus-Induced Reactive Astrocytes in Three-Dimensional Culture
Functional Characterization of Resting and Adenovirus-Induced Reactive Astrocytes in Three-Dimensional Culture
复制标题
三维培养中静息和腺病毒诱导的反应性星形胶质细胞的功能表征
作者：
Junsung Woo;Sun Kyoung Im;Heejung Chun;Soon Young Jung;Soo Jin Oh;Nakwon Choi;Nakwon Choi;C. J. Lee;Eun Mi Hur;Eun Mi Hur
脑是一个丰富的环境，神经元和胶质细胞与邻近细胞以及细胞外基质在三维空间中相互作用。星形胶质细胞是哺乳动物大脑中最丰富的细胞，存在于3D空间中，并延伸形成微区和接触突触的高度分支的过程。有人建议，星形胶质细胞培养在3D可能会保持在一个较低的反应状态相比，在传统的二维（2D）单层培养生长。然而，在三维培养的星形胶质细胞的功能表征一直缺乏。在这里，我们共同培养的神经元和星形胶质细胞在3D和检查的形态，分子生物学和电生理特性的3D培养的海马星形胶质细胞。在我们的3D神经元-星形胶质细胞共培养中，星形胶质细胞表现出典型的具有许多分支的小索马形态，并表现出被动传导的独特膜特性，更接近于它们在体内的天然对应物。此外，我们还通过感染高滴度腺病毒来模拟体内病理生理条件，在培养物中诱导反应性星形胶质细胞增多。腺病毒感染诱导星形胶质细胞的形态学变化，增加被动传导，增加GABA含量以及紧张性GABA释放，这是反应性胶质增生的特征。总之，我们的研究提出了一个强大的体外模型，类似于体内的生理和病理生理条件，从而为研究伴随反应性星形胶质细胞的各种神经系统疾病提供了一个通用的实验工具。
Brain is a rich environment where neurons and glia interact with neighboring cells as well as extracellular matrix in three-dimensional (3D) space. Astrocytes, which are the most abundant cells in the mammalian brain, reside in 3D space and extend highly branched processes that form microdomains and contact synapses. It has been suggested that astrocytes cultured in 3D might be maintained in a less reactive state as compared to those growing in a traditional, two-dimensional (2D) monolayer culture. However, the functional characterization of the astrocytes in 3D culture has been lacking. Here we cocultured neurons and astrocytes in 3D and examined the morphological, molecular biological, and electrophysiological properties of the 3D-cultured hippocampal astrocytes. In our 3D neuron-astrocyte coculture, astrocytes showed a typical morphology of a small soma with many branches and exhibited a unique membrane property of passive conductance, more closely resembling their native in vivo counterparts. Moreover, we also induced reactive astrocytosis in culture by infecting with high-titer adenovirus to mimic pathophysiological conditions in vivo. Adenoviral infection induced morphological changes in astrocytes, increased passive conductance, and increased GABA content as well as tonic GABA release, which are characteristics of reactive gliosis. Together, our study presents a powerful in vitro model resembling both physiological and pathophysiological conditions in vivo, and thereby provides a versatile experimental tool for studying various neurological diseases that accompany reactive astrocytes.
影响因子：
2.5
作者：
Zhou, M;Kimelberg, HK
通讯作者：
Kimelberg, HK