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PAC1 receptor mRNA and protein distribution in rat and human trigeminal and sphenopalatine ganglia, spinal trigeminal nucleus and in dura mater

Hensley, Kelly; Pretorius, Jim; Chan, Brian; Page, Keith; Liu, Hantao; Choi, Chang; Shi, Di; Xu, Cen; Edvinsson, Lars LU and Miller, Silke (2019) In Cephalalgia 39(7). p.827-840
Abstract

Background: To further understand the role of pituitary adenylate cyclase-activating polypeptide 1 (PAC1) receptors in headache disorders, we mapped their expression in tissues of the trigemino-autonomic system by immunohistochemistry and in situ hybridization. Methods: To optimize screening for monoclonal antibodies suitable for immunohistochemistry on formalin-fixed, paraffin-embedded tissues, we developed a new enzyme-linked immunosorbent assay using formalin-fixed, paraffin-embedded cells overexpressing human PAC1 receptors. 169G4.1 was selected from these studies for analysis of rat and human tissues and chimerized onto a mouse backbone to avoid human-on-human cross-reactivity. Immunoreactivity was compared to PAC1 receptor mRNA by... (More)

Background: To further understand the role of pituitary adenylate cyclase-activating polypeptide 1 (PAC1) receptors in headache disorders, we mapped their expression in tissues of the trigemino-autonomic system by immunohistochemistry and in situ hybridization. Methods: To optimize screening for monoclonal antibodies suitable for immunohistochemistry on formalin-fixed, paraffin-embedded tissues, we developed a new enzyme-linked immunosorbent assay using formalin-fixed, paraffin-embedded cells overexpressing human PAC1 receptors. 169G4.1 was selected from these studies for analysis of rat and human tissues and chimerized onto a mouse backbone to avoid human-on-human cross-reactivity. Immunoreactivity was compared to PAC1 receptor mRNA by in situ hybridization in both species. Results: 169G4.1 immunoreactivity delineated neuronal cell bodies in the sphenopalatine ganglion in both rat and human, whereas no staining was detected in the trigeminal ganglion. The spinal trigeminal nucleus in both species showed immunoreactivity as especially strong in the upper laminae with both cell bodies and neuropil being labelled. No immunoreactivity was seen in either rat or human dura mater vessels. In situ hybridization in both species revealed mRNA in sphenopalatine ganglion neurons and the spinal trigeminal nucleus, a weak signal in the trigeminal nucleus and no signal in dural vessels. Conclusion: Taken together, these data support a role for PAC1 receptors in the trigemino-autonomic system as it relates to headache pathophysiology.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
headache, in situ hybridization, monoclonal antibody, PAC1, Pituitary adenylate cyclase-activating polypeptide
in
Cephalalgia
volume
39
issue
7
pages
827 - 840
publisher
Wiley-Blackwell
external identifiers
  • scopus:85060342794
ISSN
0333-1024
DOI
10.1177/0333102418821621
language
English
LU publication?
yes
id
42d246f4-4b34-4c33-9081-e4d2fc59f5e9
date added to LUP
2019-02-04 13:19:29
date last changed
2019-11-08 11:48:16
@article{42d246f4-4b34-4c33-9081-e4d2fc59f5e9,
  abstract     = {<p>Background: To further understand the role of pituitary adenylate cyclase-activating polypeptide 1 (PAC1) receptors in headache disorders, we mapped their expression in tissues of the trigemino-autonomic system by immunohistochemistry and in situ hybridization. Methods: To optimize screening for monoclonal antibodies suitable for immunohistochemistry on formalin-fixed, paraffin-embedded tissues, we developed a new enzyme-linked immunosorbent assay using formalin-fixed, paraffin-embedded cells overexpressing human PAC1 receptors. 169G4.1 was selected from these studies for analysis of rat and human tissues and chimerized onto a mouse backbone to avoid human-on-human cross-reactivity. Immunoreactivity was compared to PAC1 receptor mRNA by in situ hybridization in both species. Results: 169G4.1 immunoreactivity delineated neuronal cell bodies in the sphenopalatine ganglion in both rat and human, whereas no staining was detected in the trigeminal ganglion. The spinal trigeminal nucleus in both species showed immunoreactivity as especially strong in the upper laminae with both cell bodies and neuropil being labelled. No immunoreactivity was seen in either rat or human dura mater vessels. In situ hybridization in both species revealed mRNA in sphenopalatine ganglion neurons and the spinal trigeminal nucleus, a weak signal in the trigeminal nucleus and no signal in dural vessels. Conclusion: Taken together, these data support a role for PAC1 receptors in the trigemino-autonomic system as it relates to headache pathophysiology.</p>},
  author       = {Hensley, Kelly and Pretorius, Jim and Chan, Brian and Page, Keith and Liu, Hantao and Choi, Chang and Shi, Di and Xu, Cen and Edvinsson, Lars and Miller, Silke},
  issn         = {0333-1024},
  keyword      = {headache,in situ hybridization,monoclonal antibody,PAC1,Pituitary adenylate cyclase-activating polypeptide},
  language     = {eng},
  number       = {7},
  pages        = {827--840},
  publisher    = {Wiley-Blackwell},
  series       = {Cephalalgia},
  title        = {PAC1 receptor mRNA and protein distribution in rat and human trigeminal and sphenopalatine ganglia, spinal trigeminal nucleus and in dura mater},
  url          = {http://dx.doi.org/10.1177/0333102418821621},
  volume       = {39},
  year         = {2019},
}