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The ascorbic acid transporter SVCT2 is expressed in slow-twitch skeletal muscle fibres

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Abstract

Ascorbic acid, the reduced form of vitamin C, functions as a potent antioxidant as well as in cell differentiation. Ascorbate is taken up by mammalian cells through the specific sodium/ascorbate co-transporters SVCT1 and SVCT2. Although skeletal muscle contains about 50% of the whole-body vitamin C, the expression of SVCT transporters has not been clearly addressed in this tissue. In this work, we analysed the expression pattern of SVCT2 during embryonic myogenesis using the chick as model system. We cloned the chick orthologue of SVCT2 (cSVCT2) that shares 93% identity with the mouse transporter. cSVCT2 mRNA and protein are expressed during chick embryonic muscle development. Immunohistochemical analyses showed that SVCT2 is preferentially expressed by type I slow-twitch muscle fibres throughout chick myogenesis as well as in post-natal skeletal muscles of several species, including human. Our results suggest that SVCT2-mediated uptake of ascorbate is relevant to the oxidative nature of type I muscle fibres.

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Acknowledegments

The authors are indebted to Sylvain Marcellini, Nelson Osses, Maria de los Angeles García, Hugo Olguin and members of our laboratory for useful discussion and comments on the manuscript. This work was supported by grants Anillo PBCT-CONICYT ACT-02 and ACI-12, DIUC-UdeC 204.031.098-1.0 and Fundación Andes C-13960/28.

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Authors and Affiliations

  1. Research Ring for the Study of the Nervous System (PBCT-CONICYT), Laboratory of Developmental Neurobiology, Department of Cell Biology, Faculty of Biological Sciences, Universidad de Concepción, Casilla 160-C, Concepción, Chile

    Marcela Low, Daniel Sandoval, Evelyn Avilés, Francisco Nualart & Juan Pablo Henríquez

  2. Hospital Guillermo Grant Benavente, Concepción, Chile

    Fernando Pérez

Authors
  1. Marcela Low
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  2. Daniel Sandoval
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  3. Evelyn Avilés
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  4. Fernando Pérez
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  5. Francisco Nualart
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  6. Juan Pablo Henríquez
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Corresponding author

Correspondence to Juan Pablo Henríquez.

Additional information

M. Low and D. Sandoval have contributed equally.

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Fig8

Fig. 1 Cloning and comparison of cSVCT2 mRNA sequence with cloned mammalian orthologues. (a) Total RNA was obtained from stage HH42 chick brain (lane 2) and hindlimb skeletal muscles (lane 3) for RT-PCR analysis to detect cSVCT2 (2,126bp). Lane 1 corresponds to a 1Kb DNA ladder standard. (b) Partial alignment of cSVCT2 mRNA sequence with that of mouse (GenBank accession AY004874) and human (GenBank accession EF032501) mRNAs. Sequences of the primers used are highlighted by black shading. The coding sequence of cSVCT2 is underlined. (JPG 2.69 MB)

Fig9

Fig. 2 A goat anti rat SVCT2 antibody specifically detects cSVCT2. Hindlimbs from chick embryos at stage HH40 were stained with a goat anti SVCT2 antibody (first panel). Control immunostaining experiments including those performed in the absence of any primary antibody (second panel), by using an irrelevant goat anti human MAP-1B antibody (third panel) and by coincubation of anti SVCT2 antibody along with its corresponding inhibitory peptide (IP, fourth panel), gave negative results. All sections were double stained with a mouse anti slow myosin heavy chain antibody (lower panels). Alexa488-conjugated anti goat and alexa633 conjugated anti mouse immunoglobulins were used as secondary antibodies. Bar, 20μm. (JPG 503 KB)

Fig10

Fig. 3 SVCT2 is expressed in post-natal type I skeletal muscle fibres from different mammalian species. Skeletal muscle cryosections from post-natal guinea-pig (soleus), rabbit (tibialis anterior) and rat (soleus) were double stained with a goat anti SVCT2 antibody (upper panel) together with a mouse anti slow myosin heavy chain antibody (middle panel). Alexa488-conjugated anti goat and alexa546-conjugated anti mouse immunoglobulins were used as secondary antibodies. Merge images (lower panel) indicate the co-localisation of anti SVCT2 with anti slow myosin staining. Bar, 100μm. (JPG 1.39 MB)

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Low, M., Sandoval, D., Avilés, E. et al. The ascorbic acid transporter SVCT2 is expressed in slow-twitch skeletal muscle fibres. Histochem Cell Biol 131, 565–574 (2009). https://doi.org/10.1007/s00418-008-0552-2

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