Journal: Pflugers Archiv
Article Title: Tachykinergic signaling mediated by NK1 receptors in the respiratory parafacial region drives respiratory output and contributes to the chemoreflex response to CO 2
doi: 10.1007/s00424-025-03147-8
Figure Lengend Snippet: Anatomical distribution of the ChR2-expressing respiratory parafacial neurons. ( A ) Photomicrography showing the injection site of the AAV-EF1a-double-floxed-hChR2(H134R)-mCherry-WPRE-HGHpA into the pFRG/RTN region. ( B ) Cell counts were performed on 1:4 sections at the level of the pFRG/RTM region to quantify the average number of mCherry-positive (red, circle), the double-labeled mCherry and Tac1 (mCherry + /Tac1 + , green, square), the double-labeled mCherry and Phox2b (mCherry + /Phox2b + , triangle), and the labeled mCherry, Phox2b and Tac1 that is not TH (mCherry + /Tac1+/Phox2b + /TH − , inverted triangle). C-F) Photomicrography showing the immunohistochemistry for ( C ) Tac-1-expressing cells (Tac1 cre/+ ::Ai6 mice), ( D ) mCherry (Alexa 647, red), ( E ) Phox2b (Alexa 594, magenta, F) TH (DyLight, blue). Ci-Fi) Represents a higher magnification of the yellow square depicted in C-F. Abbreviations: py, pyramid tract; Sp5, spinal trigeminal nucleus; VII, facial motor nucleus. N = 4/group of mice). Scale bar = 1 mm in A; 20 μm in Fi applied to Ci-Fi
Article Snippet: For the immunofluorescence technique, NK1-R was detected using a polyclonal rabbit anti-NK1 antibody (1:2000; Sigma-Aldrich, Saint Louis, MO, USA); tyrosine hydroxylase (TH) was detected using a polyclonal mouse anti-TH antibody (MAB 318; Millipore; 1:1000); Phox2b was detected using a polyclonal rabbit anti-Phox2b antibody (AB 2813765, Santa Cruz Biotechnology; 1:100); mCherry protein was detected using a polyclonal rabbit anti-DsRed antibody (Takakara 632496; 1:1000); and GFP was detected using a polyclonal chicken-anti-GFP (AB 13970, Abcam; 1:10,000).
Techniques: Expressing, Injection, Labeling, Immunohistochemistry
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