Sarcoplasmic reticulum Ca2+ depletion in adult skeletal muscle fibres measured with the biosensor D1ER

  • Ramón Jiménez-Moreno
  • Zhong-Ming Wang
  • María Laura Messi
  • Osvaldo DelbonoEmail author
Muscle Physiology


The endoplasmic/sarcoplasmic reticulum (ER/SR) plays a crucial role in cytoplasmic signalling in a variety of cells. It is particularly relevant to skeletal muscle fibres, where this organelle constitutes the main Ca2+ store for essential functions, such as contraction. In this work, we expressed the cameleon biosensor D1ER by in vivo electroporation in the mouse flexor digitorum brevis (FDB) muscle to directly assess SR Ca2+ depletion in response to electrical and pharmacological stimulation. The main conclusions are: (1) D1ER is expressed in the SR of FDB fibres according to both di-8-(amino naphthyl ethenyl pyridinium) staining experiments and reductions in the Förster resonance energy transfer signal consequent to SR Ca2+ release; (2) the amplitude of D1ER citrine/cyan fluorescent protein (CFP) ratio evoked by either 4-chloro-m-cresol (4-CmC) or electrical stimulation is directly proportional to the basal citrine/CFP ratio, which indicates that SR Ca2+ modulates ryanodine-receptor-isoform-1-mediated SR Ca2+ release in the intact muscle fibre; (3) SR Ca2+ release, measured as D1ER citrine/CFP signal, is voltage-dependent and follows a Boltzmann function; and (4) average SR Ca2+ depletion is 20% in response to 4-CmC and 6.4% in response to prolonged sarcolemmal depolarization. These results indicate that significantly depleting SR Ca2+ content under physiological conditions is difficult.


Skeletal muscle Calcium Sarcoplasmic reticulum 



Endoplasmic/sarcoplasmic reticulum


Flexor digitorum brevis


Store-operated Ca2+ entry


Förster resonance energy transfer


Freund virus B


Cyan fluorescent protein


Yellow fluorescent protein


Ryanodine receptor isoform 1






Di-8-amino naphthyl ethenyl pyridinium



The present study was supported by grants from the National Institutes of Health/National Institute on Ageing (AG07157, AG33385 and AG15820) and the Muscular Dystrophy Association (MDA33149) to Osvaldo Delbono, and the Wake Forest University Claude D. Pepper Older Americans Independence Centre (P30-AG21332).


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Ramón Jiménez-Moreno
    • 1
    • 3
  • Zhong-Ming Wang
    • 1
  • María Laura Messi
    • 1
  • Osvaldo Delbono
    • 1
    • 2
    • 4
    Email author
  1. 1.Department of Internal Medicine, Section on Gerontology and Geriatric MedicineWake Forest University School of MedicineWinston-SalemUSA
  2. 2.Molecular Medicine and Neuroscience ProgramsWake Forest University School of MedicineWinston-SalemUSA
  3. 3.Department of Neurobiology and AnatomyWake Forest University School of MedicineWinston-SalemUSA
  4. 4.Wake Forest University School of MedicineWinston-SalemUSA

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