Antigen retrieval pre-treatment causes a different expression pattern of Cav3.2 in rat and mouse spinal dorsal horn
Cav3 channels consist of three isoforms, Cav3.1 (α1G), Cav3.2 (α1H), and Cav3.3 (α1I), which produce low-threshold spikes that trigger burst firings in nociceptive neurons of the spinal dorsal horn (SDH) and dorsal root ganglion (DRG). Although Cav3.2 plays a crucial role in pathological pain, its...
Main Authors: | , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
PAGEPress Publications
2019-01-01
|
Series: | European Journal of Histochemistry |
Subjects: | |
Online Access: | https://www.ejh.it/index.php/ejh/article/view/2988 |
id |
doaj-c0fb7ec4653545c5a19bb7aab1103106 |
---|---|
record_format |
Article |
spelling |
doaj-c0fb7ec4653545c5a19bb7aab11031062020-11-25T03:29:24ZengPAGEPress PublicationsEuropean Journal of Histochemistry 1121-760X2038-83062019-01-0163110.4081/ejh.2019.2988Antigen retrieval pre-treatment causes a different expression pattern of Cav3.2 in rat and mouse spinal dorsal hornXiao E Cheng0Long Xian Ma1Xiao Jin Feng2Meng Ye Zhu3Da Ying Zhang4Lin Lin Xu5Tao Liu6The First Affiliated Hospital of Nanchang University, Center for Experimental MedicineThe First Affiliated Hospital of Nanchang University, Department of AnesthesiologyThe First Affiliated Hospital of Nanchang University, Center for Experimental MedicineThe First Affiliated Hospital of Nanchang University, Department of Pain ClinicThe First Affiliated Hospital of Nanchang University, Department of Pain ClinicThe First Affiliated Hospital of Nanchang University, Center for Experimental MedicineThe First Affiliated Hospital of Nanchang University, Center for Experimental Medicine Cav3 channels consist of three isoforms, Cav3.1 (α1G), Cav3.2 (α1H), and Cav3.3 (α1I), which produce low-threshold spikes that trigger burst firings in nociceptive neurons of the spinal dorsal horn (SDH) and dorsal root ganglion (DRG). Although Cav3.2 plays a crucial role in pathological pain, its distribution in SDH still remains controversial. One study showed that Cav3.2 is ubiquitously expressed in neurons, but another study implied that Cav3.2 is expressed restricted to astrocytes. To unravel these discrepancies, we used methods of immunohistochemistry either with or without antigen retrieval (AR) pre-treatment to detect Cav3 in SDH and DRG from both rats and mice. Moreover, Cav3.2 mRNA was detected in mice SDH using in situ hybridization. We found that the expression pattern of Cav3.2 but not Cav3.1 and Cav3.3 in SDH were largely different with or without AR pre-treatment, which showed a neuron-like and an astrocyte-like appearance, respectively. Double staining further demonstrated that Cav3.2 was mainly co-stained with the neuronal marker NeuN in the presence of AR but was with glial fibrillary acidic protein (GFAP, marker for astrocytes) in the absence of AR pre-treatment. Importantly, Cav3.2 mRNA was mainly co-localized with Cav3.2 but not GFAP. Together, our findings indicate that AR pre-treatment or not impacts the expression pattern of Cav3.2, which may make a significant contribution to the future study of Cav3.2 in SDH. https://www.ejh.it/index.php/ejh/article/view/2988Cav3.2immunohistochemistryantigen retrievalspinal dorsal horn |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Xiao E Cheng Long Xian Ma Xiao Jin Feng Meng Ye Zhu Da Ying Zhang Lin Lin Xu Tao Liu |
spellingShingle |
Xiao E Cheng Long Xian Ma Xiao Jin Feng Meng Ye Zhu Da Ying Zhang Lin Lin Xu Tao Liu Antigen retrieval pre-treatment causes a different expression pattern of Cav3.2 in rat and mouse spinal dorsal horn European Journal of Histochemistry Cav3.2 immunohistochemistry antigen retrieval spinal dorsal horn |
author_facet |
Xiao E Cheng Long Xian Ma Xiao Jin Feng Meng Ye Zhu Da Ying Zhang Lin Lin Xu Tao Liu |
author_sort |
Xiao E Cheng |
title |
Antigen retrieval pre-treatment causes a different expression pattern of Cav3.2 in rat and mouse spinal dorsal horn |
title_short |
Antigen retrieval pre-treatment causes a different expression pattern of Cav3.2 in rat and mouse spinal dorsal horn |
title_full |
Antigen retrieval pre-treatment causes a different expression pattern of Cav3.2 in rat and mouse spinal dorsal horn |
title_fullStr |
Antigen retrieval pre-treatment causes a different expression pattern of Cav3.2 in rat and mouse spinal dorsal horn |
title_full_unstemmed |
Antigen retrieval pre-treatment causes a different expression pattern of Cav3.2 in rat and mouse spinal dorsal horn |
title_sort |
antigen retrieval pre-treatment causes a different expression pattern of cav3.2 in rat and mouse spinal dorsal horn |
publisher |
PAGEPress Publications |
series |
European Journal of Histochemistry |
issn |
1121-760X 2038-8306 |
publishDate |
2019-01-01 |
description |
Cav3 channels consist of three isoforms, Cav3.1 (α1G), Cav3.2 (α1H), and Cav3.3 (α1I), which produce low-threshold spikes that trigger burst firings in nociceptive neurons of the spinal dorsal horn (SDH) and dorsal root ganglion (DRG). Although Cav3.2 plays a crucial role in pathological pain, its distribution in SDH still remains controversial. One study showed that Cav3.2 is ubiquitously expressed in neurons, but another study implied that Cav3.2 is expressed restricted to astrocytes. To unravel these discrepancies, we used methods of immunohistochemistry either with or without antigen retrieval (AR) pre-treatment to detect Cav3 in SDH and DRG from both rats and mice. Moreover, Cav3.2 mRNA was detected in mice SDH using in situ hybridization. We found that the expression pattern of Cav3.2 but not Cav3.1 and Cav3.3 in SDH were largely different with or without AR pre-treatment, which showed a neuron-like and an astrocyte-like appearance, respectively. Double staining further demonstrated that Cav3.2 was mainly co-stained with the neuronal marker NeuN in the presence of AR but was with glial fibrillary acidic protein (GFAP, marker for astrocytes) in the absence of AR pre-treatment. Importantly, Cav3.2 mRNA was mainly co-localized with Cav3.2 but not GFAP. Together, our findings indicate that AR pre-treatment or not impacts the expression pattern of Cav3.2, which may make a significant contribution to the future study of Cav3.2 in SDH.
|
topic |
Cav3.2 immunohistochemistry antigen retrieval spinal dorsal horn |
url |
https://www.ejh.it/index.php/ejh/article/view/2988 |
work_keys_str_mv |
AT xiaoecheng antigenretrievalpretreatmentcausesadifferentexpressionpatternofcav32inratandmousespinaldorsalhorn AT longxianma antigenretrievalpretreatmentcausesadifferentexpressionpatternofcav32inratandmousespinaldorsalhorn AT xiaojinfeng antigenretrievalpretreatmentcausesadifferentexpressionpatternofcav32inratandmousespinaldorsalhorn AT mengyezhu antigenretrievalpretreatmentcausesadifferentexpressionpatternofcav32inratandmousespinaldorsalhorn AT dayingzhang antigenretrievalpretreatmentcausesadifferentexpressionpatternofcav32inratandmousespinaldorsalhorn AT linlinxu antigenretrievalpretreatmentcausesadifferentexpressionpatternofcav32inratandmousespinaldorsalhorn AT taoliu antigenretrievalpretreatmentcausesadifferentexpressionpatternofcav32inratandmousespinaldorsalhorn |
_version_ |
1724579480607916032 |