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. 2021 Sep 4;8(9):183.
doi: 10.3390/vetsci8090183.

TRPV1 Receptor Identification in Bovine and Canine Mitral Valvular Interstitial Cells

Cristina Vercelli  1 , Graziana Gambino  1 , Michela Amadori  1 , Giovanni Re  1 , Eugenio Martignani  1 , Rossella V Barberis  2 , Izabela Janus  3 , Massimiliano Tursi  1
Affiliations

TRPV1 Receptor Identification in Bovine and Canine Mitral Valvular Interstitial Cells

Cristina Vercelli et al. Vet Sci. .

Abstract

Myxomatous mitral valve degeneration (MMVD) is the most common acquired cardiac disease in canine species, and valvular interstitial cells (VICs) are considered the main responsible for the development of this pathology. The scientific interest is focused on isolating and characterizing these cells. The aims of the present study were to verify a novel VICs mechanical isolation method and to characterize isolated cells using immunocytochemistry and immunofluorescence, with parallel histological and immunohistochemistry assays on bovine and canine healthy and MMVD mitral valves. Antibodies against vimentin (VIM), smooth muscle actin (SMA), von Willebrand (vW) factor, Transforming Growth Factor (TGF) β1, and Transient Receptor Potential Vanilloid 1 (TRPV1) were used. The isolation method was considered reliable and able to isolate only VICs. The different assays demonstrated a different expression of SMA in healthy and MMVD mitral valves, and TRPV1 was isolated for the first time from bovine and canine VICs and the correspondent mitral valve leaflets. The novelties of the present study are the new isolation method, that may allow correlations between laboratory and clinical conditions, and the identification of TRPV1, which will lead to further investigations to understand its function and possible role in the etiology of MMVD and to the design of new therapeutic strategies.

Keywords: Transient Receptor Vanilloid 1; bovine; canine; immunocytochemistry; immunohistochemistry; mitral valve degeneration; valvular interstitial cells.

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Conflict of interest statement

The authors declare no conflict of interest. The funder had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Immunocytochemistry analysis of bovine valvular interstitial cells (VICs) to visualize vimentin (A) (magnification: 10×), smooth muscle actin (B) (magnification: 40×), and von Willebrand factor (C) (magnification: 10×).
Figure 2
Figure 2
Results of immunocytochemistry (ICC) (A,C,E,G,I) and immunofluorescence (B,D,F,H,J) assays using vimentin (VIM), actin smooth muscle (SMA), von Willembrand (vW) factor, Trasforming growth Factor (TGF) β1, and Transient Receptor Potential Vanilloid 1 (TRPV1) antibodies in cells derived from a healthy mitral valve.
Figure 3
Figure 3
Results of immunocytochemistry (ICC) (A,C,E,G,I) and immunofluorescence (B,D,F,H,J) assays using VIM, SMA, vW factor, TGF β1, and TRPV1 antibodies in cells derived from an MMVD mitral valve.
Figure 4
Figure 4
The picture shows positivity to vimentin (VIM) in bovine (A) (magnification: 5×) and healthy (B) (magnification: 5×) and pathological (C) (magnification: 5×) canine mitral valves. Black arrows indicate the atrial aspect, while yellow arrows indicate the origin of the cordae tendinae (ventricular aspect). *: area of myxomatous degeneration.
Figure 5
Figure 5
The picture shows positivity to smooth muscle actin (SMA) in bovine (A) (magnification: 5×) and healthy (B) (magnification: 10×) and pathological (C) (magnification: 5×) canine mitral valves. Black arrows indicate the atrial aspect, while yellow arrows indicate small arteries of the spongiosa layer. *: area of myxomatous degeneration.
Figure 6
Figure 6
The picture shows positivity to von Willebrand (vW) factor in the endocardial layer of bovine (A) (magnification: 5×) and healthy (B) (magnification: 10×) and pathological (C) (magnification: 10×) canine mitral valves. Black arrows indicate the atrial aspect. In (A), black circles delimitate small arteries in the spongiosa layer.
Figure 7
Figure 7
The picture shows positivity to Transforming Growth Factor (TGF) β1 in bovine (A) (magnification: 5×) and healthy (B) (magnification: 5×) and pathological (C) (magnification: 5×) canine mitral valves. Black arrows indicate the atrial aspect. In (A), the black circle delimitates a small artery in the spongiosa layer. *: area of myxomatous degeneration.
Figure 8
Figure 8
The picture shows positivity to Transient Receptor Potential Vanilloid 1 (TRPV1) in bovine (A) (magnification: 5×) and healthy (B) (magnification: 10×) and pathological (C) (magnification: 20×) canine mitral leaflets. Black arrows indicate the atrial aspect, while the yellow arrow indicates a strong positivity in myocardial fibers that normally penetrate into the proximal portion of the caudal leaflet. In (A), the black circle delimitates small arteries in the spongiosa layer.
See this image and copyright information in PMC

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