Abstract
Low-density cultures of embryonic rat hippocampal neurons have been widely used to investigate localization and function of neuronal proteins using immunocytochemistry and electrophysiology. These cultures provide a relatively homogeneous population of hippocampal pyramidal neurons and interneurons compared to post-natal mixed neuron/glial cultures from hippocampus, cerebral cortex, and cerebellum. However, the limited quantity of neurons and the difficulty in harvesting adequate amounts makes biochemical analyses of endogenous neuronal proteins in these low-density cultured neurons difficult. Here, we provide detailed methods to prepare cultures of embryonic rat hippocampal neurons suitable for biochemical analyses of both endogenously and exogenously expressed proteins. The procedures described here are also suitable for comprehensive studies of expression, localization, post-translational modification, and function of neuronal proteins in the same neuronal culture system.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 165-173 |
| Number of pages | 9 |
| Journal | Journal of Neuroscience Methods |
| Volume | 144 |
| Issue number | 2 |
| DOIs | |
| State | Published - Jun 15 2005 |
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Keywords
- Ion channels
- Mammalian cell culture
- Membrane proteins
- Neurons
ASJC Scopus subject areas
- Neuroscience(all)
Cite this
A primary culture system for biochemical analyses of neuronal proteins. / Misonou, Hiroaki; Trimmer, James.
In: Journal of Neuroscience Methods, Vol. 144, No. 2, 15.06.2005, p. 165-173.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - A primary culture system for biochemical analyses of neuronal proteins
AU - Misonou, Hiroaki
AU - Trimmer, James
PY - 2005/6/15
Y1 - 2005/6/15
N2 - Low-density cultures of embryonic rat hippocampal neurons have been widely used to investigate localization and function of neuronal proteins using immunocytochemistry and electrophysiology. These cultures provide a relatively homogeneous population of hippocampal pyramidal neurons and interneurons compared to post-natal mixed neuron/glial cultures from hippocampus, cerebral cortex, and cerebellum. However, the limited quantity of neurons and the difficulty in harvesting adequate amounts makes biochemical analyses of endogenous neuronal proteins in these low-density cultured neurons difficult. Here, we provide detailed methods to prepare cultures of embryonic rat hippocampal neurons suitable for biochemical analyses of both endogenously and exogenously expressed proteins. The procedures described here are also suitable for comprehensive studies of expression, localization, post-translational modification, and function of neuronal proteins in the same neuronal culture system.
AB - Low-density cultures of embryonic rat hippocampal neurons have been widely used to investigate localization and function of neuronal proteins using immunocytochemistry and electrophysiology. These cultures provide a relatively homogeneous population of hippocampal pyramidal neurons and interneurons compared to post-natal mixed neuron/glial cultures from hippocampus, cerebral cortex, and cerebellum. However, the limited quantity of neurons and the difficulty in harvesting adequate amounts makes biochemical analyses of endogenous neuronal proteins in these low-density cultured neurons difficult. Here, we provide detailed methods to prepare cultures of embryonic rat hippocampal neurons suitable for biochemical analyses of both endogenously and exogenously expressed proteins. The procedures described here are also suitable for comprehensive studies of expression, localization, post-translational modification, and function of neuronal proteins in the same neuronal culture system.
KW - Ion channels
KW - Mammalian cell culture
KW - Membrane proteins
KW - Neurons
UR - http://www.scopus.com/inward/record.url?scp=19344373938&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=19344373938&partnerID=8YFLogxK
U2 - 10.1016/j.jneumeth.2004.11.007
DO - 10.1016/j.jneumeth.2004.11.007
M3 - Article
C2 - 15910974
AN - SCOPUS:19344373938
VL - 144
SP - 165
EP - 173
JO - Journal of Neuroscience Methods
JF - Journal of Neuroscience Methods
SN - 0165-0270
IS - 2
ER -