Development of a constant pressure perfused ex vivo model of the equine larynx.

Distal axonopathy is seen in a broad range of species including equine patients. In horses, this degenerative disorder of the recurrent laryngeal nerve is described as recurrent laryngeal neuropathy (RLN). The dysfunctional innervation of the cricoarytenoideus dorsalis muscle (CAD) leads to a loss o...

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Main Authors: Sven Otto, Jule K Michler, Stefan Dhein, Christoph K W Mülling
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2021-01-01
Series:PLoS ONE
Online Access:https://doi.org/10.1371/journal.pone.0251530
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author Sven Otto
Jule K Michler
Stefan Dhein
Christoph K W Mülling
author_facet Sven Otto
Jule K Michler
Stefan Dhein
Christoph K W Mülling
author_sort Sven Otto
collection DOAJ
description Distal axonopathy is seen in a broad range of species including equine patients. In horses, this degenerative disorder of the recurrent laryngeal nerve is described as recurrent laryngeal neuropathy (RLN). The dysfunctional innervation of the cricoarytenoideus dorsalis muscle (CAD) leads to a loss of performance in affected horses. In general, ex vivo models of the larynx are rare and for equine patients, just one short report is available. To allow for testing new therapy approaches in an isolated organ model, we examined equine larynges in a constant pressure perfused setup. In order to check the vitality and functionality of the isolated larynx, the vessels´ reaction to norepinephrine (NE) and sodium nitroprusside (NP) as vasoactive agents was tested. Additionally, the contractility of the CAD was checked via electrical stimulation. To determine the extent of hypoxic alterations, lactate dehydrogenase (LDH) and lactate were measured and an immunofluorescent analysis of hypoxia-inducible factor (HIF-1α), a key transcription factor in hypoxia, was performed. For this, a hypoxia-induced cell culture for HIF-1α was developed. The application of NE led to an expected vasoconstriction while NP caused the expected vasodilation. During a perfusion period of 352 ±20.78 min, LDH values were in the reference range and lactate values slightly exceeded the reference range at the end of the perfusion. HIF-1α nuclear translocation could reliably be detected in the hypoxia-induced cell cultures, but not in sections of the perfused CAD. With the approach presented here, a solid basis for perfusing equine larynges was established and may serve as a tool for further investigations of equine larynx disorders as well as a transferrable model for other species.
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spelling doaj.art-f196e68bd9b14c2daba0e2af63ffdcd62022-12-21T18:43:39ZengPublic Library of Science (PLoS)PLoS ONE1932-62032021-01-01165e025153010.1371/journal.pone.0251530Development of a constant pressure perfused ex vivo model of the equine larynx.Sven OttoJule K MichlerStefan DheinChristoph K W MüllingDistal axonopathy is seen in a broad range of species including equine patients. In horses, this degenerative disorder of the recurrent laryngeal nerve is described as recurrent laryngeal neuropathy (RLN). The dysfunctional innervation of the cricoarytenoideus dorsalis muscle (CAD) leads to a loss of performance in affected horses. In general, ex vivo models of the larynx are rare and for equine patients, just one short report is available. To allow for testing new therapy approaches in an isolated organ model, we examined equine larynges in a constant pressure perfused setup. In order to check the vitality and functionality of the isolated larynx, the vessels´ reaction to norepinephrine (NE) and sodium nitroprusside (NP) as vasoactive agents was tested. Additionally, the contractility of the CAD was checked via electrical stimulation. To determine the extent of hypoxic alterations, lactate dehydrogenase (LDH) and lactate were measured and an immunofluorescent analysis of hypoxia-inducible factor (HIF-1α), a key transcription factor in hypoxia, was performed. For this, a hypoxia-induced cell culture for HIF-1α was developed. The application of NE led to an expected vasoconstriction while NP caused the expected vasodilation. During a perfusion period of 352 ±20.78 min, LDH values were in the reference range and lactate values slightly exceeded the reference range at the end of the perfusion. HIF-1α nuclear translocation could reliably be detected in the hypoxia-induced cell cultures, but not in sections of the perfused CAD. With the approach presented here, a solid basis for perfusing equine larynges was established and may serve as a tool for further investigations of equine larynx disorders as well as a transferrable model for other species.https://doi.org/10.1371/journal.pone.0251530
spellingShingle Sven Otto
Jule K Michler
Stefan Dhein
Christoph K W Mülling
Development of a constant pressure perfused ex vivo model of the equine larynx.
PLoS ONE
title Development of a constant pressure perfused ex vivo model of the equine larynx.
title_full Development of a constant pressure perfused ex vivo model of the equine larynx.
title_fullStr Development of a constant pressure perfused ex vivo model of the equine larynx.
title_full_unstemmed Development of a constant pressure perfused ex vivo model of the equine larynx.
title_short Development of a constant pressure perfused ex vivo model of the equine larynx.
title_sort development of a constant pressure perfused ex vivo model of the equine larynx
url https://doi.org/10.1371/journal.pone.0251530
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