uu.seUppsala University Publications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Characterization of preproenkephalin expressing neurons in the nervous system using a transgenic line
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Developmental Genetics. (Sensory Circuits)ORCID iD: 0000-0002-1386-1307
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Developmental Genetics. (Sensory Circuits)
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Developmental Genetics. (Sensory Circuits)
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Developmental Genetics. (Sensory Circuits)
(English)Manuscript (preprint) (Other academic)
Abstract [en]

Here we have constructed a Cre line to target preproenkephalin expressing cells (Penk-Cre) using a BAC cloning strategy. By crossing our Penk-Cre line with the tdTomato reporter, tdTomato;Penk-Cre expressing cells could be visualized. Penk-Cre was expressed throughout the dorsal spinal cord, where approximately 50% of the neurons were residing within lamina III. Furthermore, single-cell analysis of spinal Penk-Cre expressing cells showed that 41% was positive for Penk mRNA and that a majority (94%) of the population expressed Vglut2 mRNA. Immunohistochemical analysis showed that only 7% and 13% expressed the inhibitory markers Viaat-egfp and Pax2, respectively, hence identifying spinal Penk-Cre expressing neurons as mainly excitatory. The expression of Penk-Cre in the brain was extensive, including dense expression in the striatum, nucleus accumbens and several amygdaloid nuclei. Furthermore, Penk-Cre expression was also shown in insular cortex, cingulated cortex, primary and secondary somatosensory cortices and the trigeminal nuclei. The Penk-Cre line represents a useful genetic tool for future analysis of PENK expressing neurons in the nervous system.

 

Keyword [en]
characterization, Cre, preproenkephalin, Penk
National Category
Medical and Health Sciences
Research subject
Medical Science
Identifiers
URN: urn:nbn:se:uu:diva-284068OAI: oai:DiVA.org:uu-284068DiVA: diva2:919706
Available from: 2016-04-14 Created: 2016-04-14 Last updated: 2016-06-01Bibliographically approved
In thesis
1. Functional Aspects of Peripheral and Spinal Cord Neurons Involved in Itch and Pain
Open this publication in new window or tab >>Functional Aspects of Peripheral and Spinal Cord Neurons Involved in Itch and Pain
2016 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

We have investigated the role of the metabotropic glutamate receptor 7 (mGluR7) and the gastrin releasing peptide receptor (Grpr) population that are involved at different levels of itch transmission. We found that mGuR7 deficient mice displayed an anaphylaxis-like behavior when provoked with histamine. Analysis of blood revealed elevated plasma levels of histamine and mouse mast cell protease-1 (mMCP1), two indicators of anaphylaxis, in mGluR7 deficient mice compared with control mice. Inhibition of the neurokinin 1 receptor, by preventing binding of the corresponding ligand substance P (SP), prior to provocation with histamine prevented the development of anaphylaxis in mGluR7 deficient animals. However, blocking GRPR (gastrin releasing peptide receptor) only resulted in decreased itch levels in mGluR7 deficient mice but did not prevent the systemic anaphylaxis-like behavior. Our findings indicate that mGluR7 normally functions as a brake on histaminergic itch that is mediated through GRPR as well as anaphylaxis through Substance P.

Grpr has previously been shown to mediate both histaminergic and non-histaminergic itch but little is known about the GRPR neuronal population. We used a BAC cloning strategy to construct a Grpr-Cre line, which we crossed with the reporter lines tdTomato and Viaat-egfp as well as with Vglut2-lox. We could conclude that Grpr-Cre neurons are mainly excitatory interneurons located in lamina II-IV, that convey itch using VGLUT2-mediated glutamatergic transmission to the next, currently unknown, step in the labeled line of chemical itch.

To eventually deduce the function of the endogenous opioids dynorphin and enkephalin, which are hypothesized to be involved in gating pain and itch in the spinal cord, we constructed two Cre lines using BAC cloning that targeted the precursor proteins preprodynorphin and preproenkephalin, respectively. Preprodynorphin-Cre neurons were mainly located in lamina II-IV and overlapped to 47% with Vglut2 mRNA, while the co-expression with the inhibitory markers Viaat-egfp and PAX2 was 13% and 28% respectively in the spinal cord. Preproenkephalin neurons were more localized to lamina III in the dorsal horn, furthermore single cell analysis showed that they overlapped to 94% with Vglut2 mRNA while 7% and 13% expressed Viaat-egfp and PAX2 respectively.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2016. 48 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine, ISSN 1651-6206 ; 1222
Keyword
mGluR7, anaphylaxis, Grpr, Penk, Pdyn, Cre line, BAC cloning, spinal cord, transgenic line
National Category
Medical and Health Sciences
Research subject
Medical Science
Identifiers
urn:nbn:se:uu:diva-284070 (URN)978-91-554-9568-8 (ISBN)
Public defence
2016-06-04, C2:301, Husargatan 3, Uppsala, 09:00 (English)
Opponent
Supervisors
Available from: 2016-05-13 Created: 2016-04-14 Last updated: 2016-06-01

Open Access in DiVA

No full text

Authority records BETA

Aresh, Bejan

Search in DiVA

By author/editor
Aresh, Bejan
By organisation
Developmental Genetics
Medical and Health Sciences

Search outside of DiVA

GoogleGoogle Scholar

urn-nbn

Altmetric score

urn-nbn
Total: 264 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf