Spatial quantification of single cell mRNA and ligand binding of the kappa opioid receptor in the rat hypothalamus

Maiken Krogsbaek; Nick Yin Larsen; Anne M Landau; Connie Sanchez; Jens Randel Nyengaard

doi:10.1016/j.jchemneu.2022.102205

Spatial quantification of single cell mRNA and ligand binding of the kappa opioid receptor in the rat hypothalamus

J Chem Neuroanat. 2023 Jan:127:102205. doi: 10.1016/j.jchemneu.2022.102205. Epub 2022 Dec 2.

Authors

Maiken Krogsbaek¹, Nick Yin Larsen², Anne M Landau³, Connie Sanchez⁴, Jens Randel Nyengaard⁵

Affiliations

¹ Core Centre for Molecular Morphology, Department of Clinical Medicine, Aarhus University, Aarhus, Denmark. Electronic address: mkm@clin.au.dk.
² Core Centre for Molecular Morphology, Department of Clinical Medicine, Aarhus University, Aarhus, Denmark; Center of Functionally Integrative Neuroscience, Department of Clinical Medicine, Aarhus University, Aarhus, Denmark.
³ Department of Nuclear Medicine and PET Centre, Aarhus University Hospital, Aarhus, Denmark; Translational Neuropsychiatry Unit, Department of Clinical Medicine, Aarhus University, Aarhus, Denmark.
⁴ Translational Neuropsychiatry Unit, Department of Clinical Medicine, Aarhus University, Aarhus, Denmark; Alkermes Inc., Waltham, MA, USA.
⁵ Core Centre for Molecular Morphology, Department of Clinical Medicine, Aarhus University, Aarhus, Denmark; Department of Pathology, Aarhus University Hospital, Aarhus, Denmark.

PMID: 36464066
DOI: 10.1016/j.jchemneu.2022.102205

Abstract

Detailed quantification of brain tissue provides a deeper understanding of changes in expression and function. We have created a pipeline to study the detailed expression patterns of the kappa opioid receptor in the rat hypothalamus using high resolution fluorescence microscopy and receptor autoradiography. The workflow involved structured serial sampling of rat hypothalamic nuclei, in situ detection of mRNA and receptor expression, and advanced image analysis. Our results demonstrate how maintaining spatial information can lead to increased understanding of RNA and protein expression. In addition, we show the detailed expression patterns of the kappa opioid receptor in the rat hypothalamus.

Keywords: Autoradiography; Hypothalamus; Kappa Opioid Receptor; RNAscope; Spatial image quantification.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Autoradiography
Hypothalamus* / metabolism
In Situ Hybridization
Ligands
RNA, Messenger
Rats
Receptors, Opioid, kappa* / genetics
Receptors, Opioid, kappa* / metabolism

Substances

Receptors, Opioid, kappa
RNA, Messenger
Ligands