In this work we compared “specialized” muscles as masticatory (masseter), extraocular (rectus lateralis) and laryngeal (vocalis portion of tyroaritenoideus, interarytenoideus, cricothyroideus and the posterior portion of cricoarytenoideus) with vastus lateralis, atrial myocardium and foetal psoas muscle (21 weeks) which are used as internal controls since their specific MyHC expression profile (1, 2A, 2X, -cardiac and developmental isoforms, respectively). Combining isoform-specific MyHC TaqMan probes with SDS-PAGE electrophoresis we provided a reliable MyHC classification in all muscles studied, both in term of mRNA and protein expression. However, we encountered a high MyHC expression diversity depending on the sampling site within the muscle and individual variability. Our results indicate that specialized muscles express skeletal isoforms (1, 2A and 2X) together with MyHC peculiar isoforms: -masseter expresses -cardiac and perinatal/neonatal; -laryngeal muscles express -cardiac, laryngeal, perinatal and embryonic; -extraocular muscles express all MyHC isoforms except for the perinatal. Overall, this study shows that the flux of MyHC mRNA examined by Real Time PCR is very irregular compared to the corresponding protein confirming that it is essential to investigate both molecules to gain consistent results. Interestingly our data show for the first time the expression, at protein level, of the 2B isoform in rectus lateralis muscle and confirm the presence of a novel laryngeal isoform.
Identification of 2B, 2EO and laryngeal myosin heavy chain isoforms in human pre-somitic skeletal muscles
MACCATROZZO, LISA;TONIOLO, LUANA;PATRUNO, MARCO VINCENZO;REGGIANI, CARLO;MASCARELLO, FRANCESCO
2008
Abstract
In this work we compared “specialized” muscles as masticatory (masseter), extraocular (rectus lateralis) and laryngeal (vocalis portion of tyroaritenoideus, interarytenoideus, cricothyroideus and the posterior portion of cricoarytenoideus) with vastus lateralis, atrial myocardium and foetal psoas muscle (21 weeks) which are used as internal controls since their specific MyHC expression profile (1, 2A, 2X, -cardiac and developmental isoforms, respectively). Combining isoform-specific MyHC TaqMan probes with SDS-PAGE electrophoresis we provided a reliable MyHC classification in all muscles studied, both in term of mRNA and protein expression. However, we encountered a high MyHC expression diversity depending on the sampling site within the muscle and individual variability. Our results indicate that specialized muscles express skeletal isoforms (1, 2A and 2X) together with MyHC peculiar isoforms: -masseter expresses -cardiac and perinatal/neonatal; -laryngeal muscles express -cardiac, laryngeal, perinatal and embryonic; -extraocular muscles express all MyHC isoforms except for the perinatal. Overall, this study shows that the flux of MyHC mRNA examined by Real Time PCR is very irregular compared to the corresponding protein confirming that it is essential to investigate both molecules to gain consistent results. Interestingly our data show for the first time the expression, at protein level, of the 2B isoform in rectus lateralis muscle and confirm the presence of a novel laryngeal isoform.Pubblicazioni consigliate
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