TY - JOUR
T1 - Effective synaptome analysis of itch-mediating neurons in the spinal cord
T2 - A novel immunohistochemical methodology using high-voltage electron microscopy
AU - Satoh, Keita
AU - Takanami, Keiko
AU - Murata, Kazuyoshi
AU - Kawata, Mitsuhiro
AU - Sakamoto, Tatsuya
AU - Sakamoto, Hirotaka
N1 - Publisher Copyright:
© 2015 Elsevier Ireland Ltd.
PY - 2015/7/1
Y1 - 2015/7/1
N2 - Transmission electron microscopy (TEM) is used for three-dimensional (3-D) analysis of synaptic connections in neuroscience research. However, 3-D reconstruction of the synapses using serial ultrathin sections is a powerful but tedious approach requiring advanced technical skills. High-voltage electron microscopy (HVEM) allows examination of thicker sections of biological specimens due to the increased penetration of the more accelerated electrons, which is useful to analyze the 3-D structure of biological specimens. However, it is still difficult to visualize the neural networks and synaptic connections in 3-D using HVEM because of insufficient and non uniform heavy metal staining in the membranous structures in semi-thin sections. Here, we present the successful chemical 3-D neuroanatomy of the rat spinal dorsal horn at the ultrastructural level as a first step for effective synaptome analysis by applying a high-contrast en bloc staining method to immune-HVEM tomography. Our new approach made it possible to examine many itch-mediating synaptic connections and neural networks in the spinal cord simultaneously using HVEM tomography. This novel 3-D electron microscopy is very useful for the analysis of synaptic structure and the chemical neuroanatomy at the 3-D ultrastructural level.
AB - Transmission electron microscopy (TEM) is used for three-dimensional (3-D) analysis of synaptic connections in neuroscience research. However, 3-D reconstruction of the synapses using serial ultrathin sections is a powerful but tedious approach requiring advanced technical skills. High-voltage electron microscopy (HVEM) allows examination of thicker sections of biological specimens due to the increased penetration of the more accelerated electrons, which is useful to analyze the 3-D structure of biological specimens. However, it is still difficult to visualize the neural networks and synaptic connections in 3-D using HVEM because of insufficient and non uniform heavy metal staining in the membranous structures in semi-thin sections. Here, we present the successful chemical 3-D neuroanatomy of the rat spinal dorsal horn at the ultrastructural level as a first step for effective synaptome analysis by applying a high-contrast en bloc staining method to immune-HVEM tomography. Our new approach made it possible to examine many itch-mediating synaptic connections and neural networks in the spinal cord simultaneously using HVEM tomography. This novel 3-D electron microscopy is very useful for the analysis of synaptic structure and the chemical neuroanatomy at the 3-D ultrastructural level.
KW - High-voltage electron microscopy
KW - Itch sensation
KW - Spinal cord
KW - Synapse
KW - Three-dimensional analysis
KW - Tomography
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U2 - 10.1016/j.neulet.2015.05.031
DO - 10.1016/j.neulet.2015.05.031
M3 - Article
C2 - 26007703
AN - SCOPUS:84930629124
VL - 599
SP - 86
EP - 91
JO - Neuroscience Letters
JF - Neuroscience Letters
SN - 0304-3940
ER -