Immunofluorescence signal intensity measurements as a semi-quantitative tool to assess sarcoglycan complex expression in muscle biopsy
Sarcoglycanopathies are highly heterogeneous in terms of disease progression, muscular weakness, loss of ambulation and cardiac/respiratory involvement. Their clinical severity usually correlates with the residual protein amount, which makes protein quantification extremely relevant. Sarcoglycanopa...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
PAGEPress Publications
2022-09-01
|
| Series: | European Journal of Histochemistry |
| Subjects: | |
| Online Access: | https://www.ejh.it/index.php/ejh/article/view/3418 |
| _version_ | 1811338154286776320 |
|---|---|
| author | Simona Zanotti Francesca Magri Francesca Poggetti Michela Ripolone Daniele Velardo Francesco Fortunato Patrizia Ciscato Maurizio Moggio Stefania Corti Giacomo Pietro Comi Monica Sciacco |
| author_facet | Simona Zanotti Francesca Magri Francesca Poggetti Michela Ripolone Daniele Velardo Francesco Fortunato Patrizia Ciscato Maurizio Moggio Stefania Corti Giacomo Pietro Comi Monica Sciacco |
| author_sort | Simona Zanotti |
| collection | DOAJ |
| description |
Sarcoglycanopathies are highly heterogeneous in terms of disease progression, muscular weakness, loss of ambulation and cardiac/respiratory involvement. Their clinical severity usually correlates with the residual protein amount, which makes protein quantification extremely relevant. Sarcoglycanopathy diagnosis is genetic, but skeletal muscle analysis - by both immunohistochemistry and Western blot (WB) - is still mandatory to establish the correct diagnostic process. Unfortunately, however, WB analysis cannot be performed if the bioptic specimen is scarce. This study provides a sensitive tool for semi-quantification of residual amount of sarcoglycans in patients affected by sarcoglycanopathies, based on immunofluorescence staining on skeletal muscle sections, image acquisition and software elaboration. We applied this method to eleven sarcoglycanopathies, seven Becker muscular dystrophies and four age-matched controls. Fluorescence data analysed in patients and compared to age-matched controls showed a significant reduction of the mutated sarcoglycan expression and a variable reduction of the other sarcoglycans. Fluorescence normalized data analysed in relation to the age of onset of the disease, showed a negative correlation of α-sarcoglycan fluorescent signal versus fibrosis in patients with an early age of onset and a negative correlation between δ-sarcoglycan signal and fibrosis in both intermediate and late age of onset groups. The availability of a method that allows objective quantification of the sarcolemmal proteins, faster and less consuming than WB analysis and able to detect low residual sarcoglycan expression with great sensitivity, proves useful to better define both patient prognosis and expected disease evolution. The proposed method could be employed also to monitor the efficacy of therapeutic interventions and during clinical trials.
|
| first_indexed | 2024-04-13T18:05:45Z |
| format | Article |
| id | doaj.art-c0ecdf06f8ce4cba9e60afd0f3eca236 |
| institution | Directory Open Access Journal |
| issn | 1121-760X 2038-8306 |
| language | English |
| last_indexed | 2024-04-13T18:05:45Z |
| publishDate | 2022-09-01 |
| publisher | PAGEPress Publications |
| record_format | Article |
| series | European Journal of Histochemistry |
| spelling | doaj.art-c0ecdf06f8ce4cba9e60afd0f3eca2362022-12-22T02:36:04ZengPAGEPress PublicationsEuropean Journal of Histochemistry1121-760X2038-83062022-09-0166310.4081/ejh.2022.3418Immunofluorescence signal intensity measurements as a semi-quantitative tool to assess sarcoglycan complex expression in muscle biopsySimona Zanotti0Francesca Magri1Francesca Poggetti2Michela Ripolone3Daniele Velardo4Francesco Fortunato5Patrizia Ciscato6Maurizio Moggio7Stefania Corti8Giacomo Pietro Comi9Monica Sciacco10Neuromuscular and Rare Disease Unit, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, MilanNeurology Unit, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, MilanNeuromuscular and Rare Disease Unit, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, MilanNeuromuscular and Rare Disease Unit, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, MilanNeuromuscular and Rare Disease Unit, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, MilanNeurology Unit, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, MilanNeuromuscular and Rare Disease Unit, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, MilanNeuromuscular and Rare Disease Unit, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, MilanNeurology Unit, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, MilanNeuromuscular and Rare Disease Unit, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, MilanNeuromuscular and Rare Disease Unit, Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milan Sarcoglycanopathies are highly heterogeneous in terms of disease progression, muscular weakness, loss of ambulation and cardiac/respiratory involvement. Their clinical severity usually correlates with the residual protein amount, which makes protein quantification extremely relevant. Sarcoglycanopathy diagnosis is genetic, but skeletal muscle analysis - by both immunohistochemistry and Western blot (WB) - is still mandatory to establish the correct diagnostic process. Unfortunately, however, WB analysis cannot be performed if the bioptic specimen is scarce. This study provides a sensitive tool for semi-quantification of residual amount of sarcoglycans in patients affected by sarcoglycanopathies, based on immunofluorescence staining on skeletal muscle sections, image acquisition and software elaboration. We applied this method to eleven sarcoglycanopathies, seven Becker muscular dystrophies and four age-matched controls. Fluorescence data analysed in patients and compared to age-matched controls showed a significant reduction of the mutated sarcoglycan expression and a variable reduction of the other sarcoglycans. Fluorescence normalized data analysed in relation to the age of onset of the disease, showed a negative correlation of α-sarcoglycan fluorescent signal versus fibrosis in patients with an early age of onset and a negative correlation between δ-sarcoglycan signal and fibrosis in both intermediate and late age of onset groups. The availability of a method that allows objective quantification of the sarcolemmal proteins, faster and less consuming than WB analysis and able to detect low residual sarcoglycan expression with great sensitivity, proves useful to better define both patient prognosis and expected disease evolution. The proposed method could be employed also to monitor the efficacy of therapeutic interventions and during clinical trials. https://www.ejh.it/index.php/ejh/article/view/3418sarcoglycansimmunofluorescenceprotein quantificationhistologyfibrosis |
| spellingShingle | Simona Zanotti Francesca Magri Francesca Poggetti Michela Ripolone Daniele Velardo Francesco Fortunato Patrizia Ciscato Maurizio Moggio Stefania Corti Giacomo Pietro Comi Monica Sciacco Immunofluorescence signal intensity measurements as a semi-quantitative tool to assess sarcoglycan complex expression in muscle biopsy European Journal of Histochemistry sarcoglycans immunofluorescence protein quantification histology fibrosis |
| title | Immunofluorescence signal intensity measurements as a semi-quantitative tool to assess sarcoglycan complex expression in muscle biopsy |
| title_full | Immunofluorescence signal intensity measurements as a semi-quantitative tool to assess sarcoglycan complex expression in muscle biopsy |
| title_fullStr | Immunofluorescence signal intensity measurements as a semi-quantitative tool to assess sarcoglycan complex expression in muscle biopsy |
| title_full_unstemmed | Immunofluorescence signal intensity measurements as a semi-quantitative tool to assess sarcoglycan complex expression in muscle biopsy |
| title_short | Immunofluorescence signal intensity measurements as a semi-quantitative tool to assess sarcoglycan complex expression in muscle biopsy |
| title_sort | immunofluorescence signal intensity measurements as a semi quantitative tool to assess sarcoglycan complex expression in muscle biopsy |
| topic | sarcoglycans immunofluorescence protein quantification histology fibrosis |
| url | https://www.ejh.it/index.php/ejh/article/view/3418 |
| work_keys_str_mv | AT simonazanotti immunofluorescencesignalintensitymeasurementsasasemiquantitativetooltoassesssarcoglycancomplexexpressioninmusclebiopsy AT francescamagri immunofluorescencesignalintensitymeasurementsasasemiquantitativetooltoassesssarcoglycancomplexexpressioninmusclebiopsy AT francescapoggetti immunofluorescencesignalintensitymeasurementsasasemiquantitativetooltoassesssarcoglycancomplexexpressioninmusclebiopsy AT michelaripolone immunofluorescencesignalintensitymeasurementsasasemiquantitativetooltoassesssarcoglycancomplexexpressioninmusclebiopsy AT danielevelardo immunofluorescencesignalintensitymeasurementsasasemiquantitativetooltoassesssarcoglycancomplexexpressioninmusclebiopsy AT francescofortunato immunofluorescencesignalintensitymeasurementsasasemiquantitativetooltoassesssarcoglycancomplexexpressioninmusclebiopsy AT patriziaciscato immunofluorescencesignalintensitymeasurementsasasemiquantitativetooltoassesssarcoglycancomplexexpressioninmusclebiopsy AT mauriziomoggio immunofluorescencesignalintensitymeasurementsasasemiquantitativetooltoassesssarcoglycancomplexexpressioninmusclebiopsy AT stefaniacorti immunofluorescencesignalintensitymeasurementsasasemiquantitativetooltoassesssarcoglycancomplexexpressioninmusclebiopsy AT giacomopietrocomi immunofluorescencesignalintensitymeasurementsasasemiquantitativetooltoassesssarcoglycancomplexexpressioninmusclebiopsy AT monicasciacco immunofluorescencesignalintensitymeasurementsasasemiquantitativetooltoassesssarcoglycancomplexexpressioninmusclebiopsy |