PMC5248598_01
Analysis of patient material. Microscopical investigations (1A): (I) M. vastus lateralis biopsy of patient 1 showing a myopathic pattern with minor muscle fiber atrophy and hypertrophy as well as frequent nonsubsarcolemmal nuclei (arrows). Cryostat section, H&E. Scale bar: 80 mum. (II) Low-grade muscle fiber atrophy and solitary rimmed vacuole (arrow) in cryostat section of the rectus femoris muscle biopsy of patient 2. Cryostat section, H&E. Scale bar: 50 mum. (III) M. quadriceps femoris biopsy of patient 3 showing minor atrophic as well as hypertrophic muscle fibers. Cryostat section, H&E. Scale bar: 80 mum Electron microscopic studies (1B): (I) Subsarcolemmal membrane-bound vacuoles, presumably corresponding to enlarged caveolae (arrows) in the muscle biopsy of patient 1. Scale bar: 250 nm. (II-VI) Patient 2; (II) subsarcolemmal membrane-bound vacuoles (arrows) similar to the one depicted in Fig. 1A. Scale bar: 200 nm. (III) Further abnormal membrane bound subsarcolemmal vacuoles as well as tubular structures (arrows). Scale bar: 2 mum. (IV) Large vacuole (asterisk) associated with a myonucleus. Scale bar: 4 mum. (V) Large (asterisk) and small (arrowheads) vacuoles in the vicinity of a myonucleus. Scale bar: 2 mum. (VI) Large vacuole containing autophagic material (arrows) associated with a deformed, possibly degenerating myonucleus. Arrowheads: Membranous material, presumably corresponding to a focal outfolding of the sarcoplasmic reticulum. Scale bar: 4 mum. (VII) Patient 3: subsarcolemmal membrane-bound vacuoles (arrows). Scale bar: 800 nm. Immunoblots of myopathy-associated proteins (1C): comparison of beta-Spectrin, Calpain-3, Lamin A/C, Myotilin, alpha- and gamma-Sarcoglycan, beta-Dystroglycan and Emerin in G56S Caveolin-3 patient and control muscle revealed no significant changes in protein abundances. Confirmation of proteome data via immunoblotting (1D): immunoblot analyses of HSP75, SNX1, CSTF2, Sec63, HNRA3, and Rab1A revealed same changes in protein abundances upon G56S Caveolin-3 expression like in the G56S Caveolin-3 in vitro model. Tubulin was used as loading control.
PRCA-11-0-g001.jpg
PMC5248598_02_PRCA-11-0-g001.jpg
prca1813-fig-0001
Histopathological analysis of a quadriceps muscle biopsy revealed moderate chronic myopathy with few atrophic and several hypertrophic muscle fibers and a moderately elevated number of nonsubsarcolemmal muscle fiber nuclei (Fig. 1A III). As in the above-described cases, electron microscopy revealed subsarcolemmal and perinuclear vacuolar and tubular structures up to 0.5 mum in size (Fig. 1B VII). Information Table 2). Immunoblots for beta-Spectrin, Calpain-3, Lamin A/C, Myotilin, alpha- and gamma-Sarcoglycan, beta-Dystroglycqan, and Emerin gave normal results (Fig. 1C).
PMC5248598_02
Analysis of patient material. Microscopical investigations (1A): (I) M. vastus lateralis biopsy of patient 1 showing a myopathic pattern with minor muscle fiber atrophy and hypertrophy as well as frequent nonsubsarcolemmal nuclei (arrows). Cryostat section, H&E. Scale bar: 80 mum. (II) Low-grade muscle fiber atrophy and solitary rimmed vacuole (arrow) in cryostat section of the rectus femoris muscle biopsy of patient 2. Cryostat section, H&E. Scale bar: 50 mum. (III) M. quadriceps femoris biopsy of patient 3 showing minor atrophic as well as hypertrophic muscle fibers. Cryostat section, H&E. Scale bar: 80 mum Electron microscopic studies (1B): (I) Subsarcolemmal membrane-bound vacuoles, presumably corresponding to enlarged caveolae (arrows) in the muscle biopsy of patient 1. Scale bar: 250 nm. (II-VI) Patient 2; (II) subsarcolemmal membrane-bound vacuoles (arrows) similar to the one depicted in Fig. 1A. Scale bar: 200 nm. (III) Further abnormal membrane bound subsarcolemmal vacuoles as well as tubular structures (arrows). Scale bar: 2 mum. (IV) Large vacuole (asterisk) associated with a myonucleus. Scale bar: 4 mum. (V) Large (asterisk) and small (arrowheads) vacuoles in the vicinity of a myonucleus. Scale bar: 2 mum. (VI) Large vacuole containing autophagic material (arrows) associated with a deformed, possibly degenerating myonucleus. Arrowheads: Membranous material, presumably corresponding to a focal outfolding of the sarcoplasmic reticulum. Scale bar: 4 mum. (VII) Patient 3: subsarcolemmal membrane-bound vacuoles (arrows). Scale bar: 800 nm. Immunoblots of myopathy-associated proteins (1C): comparison of beta-Spectrin, Calpain-3, Lamin A/C, Myotilin, alpha- and gamma-Sarcoglycan, beta-Dystroglycan and Emerin in G56S Caveolin-3 patient and control muscle revealed no significant changes in protein abundances. Confirmation of proteome data via immunoblotting (1D): immunoblot analyses of HSP75, SNX1, CSTF2, Sec63, HNRA3, and Rab1A revealed same changes in protein abundances upon G56S Caveolin-3 expression like in the G56S Caveolin-3 in vitro model. Tubulin was used as loading control.
PRCA-11-0-g001.jpg
PMC5248598_02_PRCA-11-0-g001.jpg
prca1813-fig-0001
Histopathological analysis of a quadriceps muscle biopsy revealed moderate chronic myopathy with few atrophic and several hypertrophic muscle fibers and a moderately elevated number of nonsubsarcolemmal muscle fiber nuclei (Fig. 1A III). As in the above-described cases, electron microscopy revealed subsarcolemmal and perinuclear vacuolar and tubular structures up to 0.5 mum in size (Fig. 1B VII). Information Table 2). Immunoblots for beta-Spectrin, Calpain-3, Lamin A/C, Myotilin, alpha- and gamma-Sarcoglycan, beta-Dystroglycqan, and Emerin gave normal results (Fig. 1C).