Universität zu Köln

Jakobsen, Jens R., Schjerling, Peter ORCID: 0000-0001-7138-3211, Svensson, Rene B., Buhl, Rikke ORCID: 0000-0002-8201-0186, Carstensen, Helena ORCID: 0000-0001-8952-9637, Koch, Manuel ORCID: 0000-0002-2962-7814, Krogsgaard, Michael R., Kjaer, Michael and Mackey, Abigail L. (2021). RNA sequencing and immunofluorescence of the myotendinous junction of mature horses and humans. Am. J. Physiol.-Cell Physiol., 321 (3). S. C453 - 18. BETHESDA: AMER PHYSIOLOGICAL SOC. ISSN 1522-1563

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Abstract

The myotendinous junction (MTJ) is a specialized interface for transmitting high forces between the muscle and tendon and yet the MTJ is a common site of strain injury with a high recurrence rate. The aim of this study was to identify previously unknown MTJ components in mature animals and humans. Samples were obtained from the superficial digital flexor (SDF) muscle-tendon interface of 20 horses, and the tissue was separated through a sequential cryosectioning approach into muscle, MTJ (muscle tissue enriched in myofiber tips attached to the tendon), and tendon fractions. RT-PCR was performed for genes known to be expressed in the three tissue fractions and t-distributed stochastic neighbor embedding (t-SNE) plots were used to select the muscle, MTJ, and tendon samples from five horses for RNA sequencing. The expression of previously known and unknown genes identified through RNA sequencing was studied by immunofluorescence on human hamstring MTJ tissue. The main finding was that RNA sequencing identified the expression of a panel of 61 genes enriched at the MTJ. Of these, 48 genes were novel for the MTJ and 13 genes had been reported to be associated with the MTJ in earlier studies. The expression of known [COL22A1 (collagen XXII), NCAM (neural cell adhesion molecule), POSTN (periostin), NES (nestin), OSTN (musclin/osteocrin)] and previously undescribed [MNS1 (meiosis-specific nuclear structural protein 1), and LCT (lactase)] MTJ genes was confirmed at the protein level by immunofluorescence on tissue sections of human MTJ. In conclusion, in muscle-tendon interface tissue enriched with myofiber tips, we identified the expression of previously unknown MTJ genes representing diverse biological processes, which may be important in the maintenance of the specialized MTJ.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Jakobsen, Jens R. UNSPECIFIED UNSPECIFIED UNSPECIFIED Schjerling, Peter UNSPECIFIED orcid.org/0000-0001-7138-3211 UNSPECIFIED Svensson, Rene B. UNSPECIFIED UNSPECIFIED UNSPECIFIED Buhl, Rikke UNSPECIFIED orcid.org/0000-0002-8201-0186 UNSPECIFIED Carstensen, Helena UNSPECIFIED orcid.org/0000-0001-8952-9637 UNSPECIFIED Koch, Manuel UNSPECIFIED orcid.org/0000-0002-2962-7814 UNSPECIFIED Krogsgaard, Michael R. UNSPECIFIED UNSPECIFIED UNSPECIFIED Kjaer, Michael UNSPECIFIED UNSPECIFIED UNSPECIFIED Mackey, Abigail L. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-580768
DOI:	10.1152/ajpcell.00218.2021
Journal or Publication Title:	Am. J. Physiol.-Cell Physiol.
Volume:	321
Number:	3
Page Range:	S. C453 - 18
Date:	2021
Publisher:	AMER PHYSIOLOGICAL SOC
Place of Publication:	BETHESDA
ISSN:	1522-1563
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language SKELETAL-MUSCLE REGENERATION; MUSCULAR-DYSTROPHY; ACHILLES-TENDON; TENOMODULIN; EXPRESSION; MARKER; ACETYLCHOLINESTERASE; ANGIOGENESIS; ADAPTATION; MYOGENESIS Multiple languages Cell Biology; Physiology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/58076