Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 158-167 |
| Number of pages | 10 |
| Journal | Experimental Neurobiology |
| Volume | 26 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - 2017 Jun 1 |
| Externally published | Yes |
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Keywords
- 3D culture
- Adenovirus
- Functional characterization
- Passive conductance
- Reactive astrocyte
- Tonic GABA current
ASJC Scopus subject areas
- Clinical Neurology
- Cellular and Molecular Neuroscience
Cite this
Functional characterization of resting and adenovirus-induced reactive astrocytes in three-dimensional culture. / Woo, Junsung; Im, Sun Kyoung; Chun, Heejung; Jung, Soon Young; Oh, Soo Jin; Choi, Nakwon; Lee, Changjoon; Hur, Eun Mi.
In: Experimental Neurobiology, Vol. 26, No. 3, 01.06.2017, p. 158-167.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Functional characterization of resting and adenovirus-induced reactive astrocytes in three-dimensional culture
AU - Woo, Junsung
AU - Im, Sun Kyoung
AU - Chun, Heejung
AU - Jung, Soon Young
AU - Oh, Soo Jin
AU - Choi, Nakwon
AU - Lee, Changjoon
AU - Hur, Eun Mi
PY - 2017/6/1
Y1 - 2017/6/1
N2 - 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.
AB - 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.
KW - 3D culture
KW - Adenovirus
KW - Functional characterization
KW - Passive conductance
KW - Reactive astrocyte
KW - Tonic GABA current
UR - http://www.scopus.com/inward/record.url?scp=85021362313&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85021362313&partnerID=8YFLogxK
U2 - 10.5607/en.2017.26.3.158
DO - 10.5607/en.2017.26.3.158
M3 - Article
AN - SCOPUS:85021362313
VL - 26
SP - 158
EP - 167
JO - Experimental Neurobiology
JF - Experimental Neurobiology
SN - 1226-2560
IS - 3
ER -