Journal: Naunyn-Schmiedeberg's Archives of Pharmacology
Article Title: Initial characterization of a transgenic mouse with overexpression of the human D 1 -dopamine receptor in the heart
doi: 10.1007/s00210-023-02901-y
Figure Lengend Snippet: A Autoradiography. Representative images of autoradiography of D 1 -TG and WT mouse heart sections labeled with 5 nM [ 3 H]SCH23390 (total binding, TB) or 5 nM [ 3 H]SCH23390 + 10 M butaclamol (unspecific binding, UB). B In situ hybridization. Expression of D 1 -dopamine receptor mRNA detected by DIG-labelled human D 1 -dopamine receptor antisense probes (A-X) and DIG-labelled mouse D 1 -dopamine receptor antisense probes (a-l). DIG-labeled sense probes were used as controls. (A-H): Low magnification microscopic images, scale bar = 500 µm. mRNA expression of D 1 -dopamine receptor was not detected in WT mouse atria (A) and ventricles (C); whereas expression of human D 1 -dopamine receptor mRNA was detected in D 1 -Tg mouse atria (E) and ventricles including interventricular septum (G). (B, D, F, H) are corresponding control sections hybridized with DIG-labeled sense probes. (I-X): High magnification microscopic images, scale bar = 50 µm. No or very scarce and weak D 1 -dopamine mRNA expression was detected in WT mouse atria (I) and interventricular septum (K), while abundant human D 1 -dopamine mRNA expression was detected in atriums (M), interventricular septum (O), and ventricle walls (Q, S). (J, L, N, P, R, T) are corresponding control sections hybridized with DIG-labeled sense probes. D 1 -dopamine mRNA expression was also detected in human cardiomyocytes (U, W); while no signal was seen in corresponding control sections hybridized with DIG-labeled sense probes (V, X). (a-d): Low magnification microscopic images, scale bar = 500 µm. (e-h): High magnification microscopic images, scale bar = 50 µm. Expression of mouse D 1 -dopamine receptor mRNA was not detected in WT mouse atria (a, e) and ventricles (c, g); (b, d, f, h) are corresponding control sections hybridized with DIG-labeled sense probes. Low magnification images (i and j, scale bar = 500 µm) and high magnification images (k and l, scale bar = 50 µm) showed that the mouse antisense probe also detected human D 1 -dopamine mRNA expression in D 1 -TG mouse interventricular septum (i, k), while the mouse sense probe (control) did not detect any specific signal (j, l). C : Histology. Histological staining of myocardial sections with hematoxylin-eosin (HE) or Masson-Goldner trichome (MG) staining revealed no apparent abnormalities or signs of fibrosis, neither in WT nor in D 1 -TG. Scale bar = 100 µm. D : PCR. Scatter plots with means for the expression of exogenous transgenic D 1 -dopamine receptors (human-DRD1, left hand side) or endogenous mouse D 1 -dopamine receptors. Ordinates indicate the amount of the mRNA as inferred from the amplification cycles (Cq). *indicates significant differences between the mRNA from the whole heart of D 1 -TG versus WT
Article Snippet: First antibodies were rabbit polyclonal anti-calsequestrin (CSQ) antibody, #ab3516, Abcam, Cambridge, UK (diluted 1:20000), rabbit polyclonal anti-DRD1, #bs-1007R, Bioss, Woburn, MA, USA (dilution 1:500), and rabbit polyclonal anti-phospho-troponin I (P-TnI) antibody, #4004, Cell Signaling Technology Europe, Leiden, The Netherlands (diluted 1:5000).
Techniques: Autoradiography, Labeling, Binding Assay, In Situ Hybridization, Expressing, Control, Staining, Transgenic Assay, Amplification
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![A Autoradiography. Representative images of autoradiography of D 1 -TG and WT mouse heart sections labeled with 5 nM [ 3 H]SCH23390 (total binding, TB) or 5 nM [ 3 H]SCH23390 + 10 M butaclamol (unspecific binding, UB). B In situ hybridization. Expression of D 1 -dopamine receptor mRNA detected by DIG-labelled human D 1 -dopamine receptor antisense probes (A-X) and DIG-labelled mouse D 1 -dopamine receptor antisense probes (a-l). DIG-labeled sense probes were used as controls. (A-H): Low magnification microscopic images, scale bar = 500 µm. mRNA expression of D 1 -dopamine receptor was not detected in WT mouse atria (A) and ventricles (C); whereas expression of human D 1 -dopamine receptor mRNA was detected in D 1 -Tg mouse atria (E) and ventricles including interventricular septum (G). (B, D, F, H) are corresponding control sections hybridized with DIG-labeled sense probes. (I-X): High magnification microscopic images, scale bar = 50 µm. No or very scarce and weak D 1 -dopamine mRNA expression was detected in WT mouse atria (I) and interventricular septum (K), while abundant human D 1 -dopamine mRNA expression was detected in atriums (M), interventricular septum (O), and ventricle walls (Q, S). (J, L, N, P, R, T) are corresponding control sections hybridized with DIG-labeled sense probes. D 1 -dopamine mRNA expression was also detected in human cardiomyocytes (U, W); while no signal was seen in corresponding control sections hybridized with DIG-labeled sense probes (V, X). (a-d): Low magnification microscopic images, scale bar = 500 µm. (e-h): High magnification microscopic images, scale bar = 50 µm. Expression of mouse D 1 -dopamine receptor mRNA was not detected in WT mouse atria (a, e) and ventricles (c, g); (b, d, f, h) are corresponding control sections hybridized with DIG-labeled sense probes. Low magnification images (i and j, scale bar = 500 µm) and high magnification images (k and l, scale bar = 50 µm) showed that the mouse antisense probe also detected human D 1 -dopamine mRNA expression in D 1 -TG mouse interventricular septum (i, k), while the mouse sense probe (control) did not detect any specific signal (j, l). C : Histology. Histological staining of myocardial sections with hematoxylin-eosin (HE) or Masson-Goldner trichome (MG) staining revealed no apparent abnormalities or signs of fibrosis, neither in WT nor in D 1 -TG. Scale bar = 100 µm. D : PCR. Scatter plots with means for the expression of exogenous transgenic D 1 -dopamine receptors (human-DRD1, left hand side) or endogenous mouse D 1 -dopamine receptors. Ordinates indicate the amount of the mRNA as inferred from the amplification cycles (Cq). *indicates significant differences between the mRNA from the whole heart of D 1 -TG versus WT](https://pub-med-central-images-cdn.bioz.com/pub_med_central_ids_ending_with_6794/pmc11166794/pmc11166794__210_2023_2901_Fig2a_HTML.jpg)