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December 1, 1992Journal of Cell Science

Expression of naked plasmids by cultured myotubes and entry of plasmids into T tubules and caveolae of mammalian skeletal muscle

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Authors

JWJon A. WolffJohns Hopkins University MDMartin E. DowtyAbbVie (United States)SJShoushu JiaoAmgen (United States)

Discussion

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Implication

In vitro and in vivo investigation demonstrates specific uptake of naked plasmid DNA into T tubules and caveolae of skeletal muscle, suggesting specialized non-endocytic transport mechanisms.

Key Points

To determine the cellular and structural mechanisms governing naked plasmid DNA uptake and transgene expression in mammalian skeletal muscle cells.
Assessed in vitro transgene expression in primary rat muscle cells versus non-muscle primary and immortalized cell lines following naked plasmid DNA exposure.
Performed in vivo light and electron microscopy tracking of intramuscularly injected naked plasmid DNA and colloidal gold conjugates (DNA, polylysine, polyethylene glycol, or polyglutamate).
Primary rat myotubes expressed transgenes to a substantially greater extent than other immortalized or primary cell types upon exposure to naked plasmid DNA.
Colloidal gold-conjugated plasmid DNA crossed the external lamina and localized within T tubules and caveolae, whereas non-DNA polymer gold complexes remained in the extracellular space.
Plasmid uptake occurred without evidence of mechanical needle disruption, transient membrane tearing, or classical endocytosis, supporting transport via caveolar pathways such as potocytosis.

Cite This Study

Wolff et al. (1992) studied this question.

synapsesocial.com/papers/6a70abf631a3df824328ee9f https://doi.org/10.1242/jcs.103.4.1249

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