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Development of a P2X 2 Reporter Mouse Model
Author(s) -
Kim SeolHee,
TaylorClark Thomas
Publication year - 2019
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2019.33.1_supplement.546.6
Subject(s) - cre recombinase , biology , dorsal root ganglion , nodose ganglion , sensory system , neuroscience , thoracic ganglia , superior cervical ganglion , cervical ganglia , anatomy , trigeminal ganglion , microbiology and biotechnology , population , purinergic receptor , ganglion , transgene , genetically modified mouse , vagus nerve , gene , genetics , demography , extracellular , stimulation , sociology
P2X 2 is a ligand gated ion channel and purinergic receptor that is stimulated by adenosine 5′‐triphosphate (ATP). P2X 2 is expressed in subsets of peripheral nerves and mediates various cellular responses in peripheral sensory nerve system. In particular, functional experiments have shown that P2X 2 is selectively expressed in vagal afferents derived from the placodes (nodose ganglion) but not from the neural crest (jugular ganglion). In this study, the Cre‐Lox reporter system was used to generate a mouse model enable us to target specific subtypes of sensory afferents. First, we generated a knockin P2X 2 ‐Cre strain. Here a cre recombinase was incorporated into the endogenous P2rx2 gene, downstream of the last P2rx2 exon following a 2A sequence. Expression of Cre was linked to the expression of endogenous P2X 2 via a 2A sequence. The P2X 2 ‐Cre mouse was crossed with another strain carrying Floxed Ai9 ROSA tdTomato, which results in expression of tdTomato in only P2X 2 expressing cells via Cre recombination. From these animals, sensory ganglia including vagal ganglia, trigeminal ganglia, superior cervical ganglia, carotid body and dorsal root ganglia were collected to visualize tdTomato‐expressing cells in combination with immunohistochemistry. Furthermore, brainstem was collected to map their central projections. tdTomato positive cells were observed only in the nodose and not in jugular of vagal ganglia. Some population of cells showed tdTomato expression in trigeminal ganglia, dorsal root ganglia and superior cervical ganglia. In addition, strong tdTomato expression was observed in carotid body, which appeared to surround anti‐tyrosine hydroxylase positive cells and nerves. These results imply that P2X 2 positive afferent nerves exist in different ganglia and therefore, ATP‐mediated purinergic signaling occurs in various peripheral sensory nerve systems. Lastly, tdTomato expression in sequentially sectioned medulla showed distinct patterns. The main area that has tdTomato expression is commissural nucleus of solitary tract (NTS), gelatinous NTS and dorsolateral NTS, as well as area postrema. This data suggests that the nodose‐originated sensory afferents project to specific regions of the medulla. Overall, the transgenic mouse model generated using Cre‐Lox system successfully labeled P2X 2 cells. Support or Funding Information This study is funded by National Institutes of Health Common Fund SPARC OT2 (2016–2019): “Functional mapping of peripheral and central circuits for airway protection and breathing” This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .