Twitch and Tetanic Tension during Culture of Mature Xenopus laevis Single Muscle Fibres

R.T. Jaspers, Hiske Feenstra, M.B.E. Lee-de Groot, P.A.J.B.M. Huijing, W.J. van der Laarse

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Abstract

Investigation of the mechanisms of muscle adaptation requires independent control of the regulating factors. The aim of the present study was to develop a serum-free medium to culture mature single muscle fibres of Xenopus laevis. As an example, we used the culture system to study adaptation of twitch and tetanic force characteristics, number of sarcomeres in series and fibre cross-section. Fibres dissected from m. iliofibularis (n = 10) were kept in culture at a fibre mean sarcomere length of 2.3 µm in a culture medium without serum. Twitch and tetanic tension were determined daily. Before and after culture the number of sarcomeres was determined by laser diffraction and fibre cross-sectional area (CSA) was determined by microscopy. For five fibres twitch tension increased during culture and tetanic tension was stable for periods varying from 8 to 14 days (‘stable fibres’), after which fibres were removed from culture for analysis. Fibre CSA and the number of sarcomeres in series remained constant during culture. Five other fibres showed a substantial reduction in twitch and tetanic tension within the first five days of culture (‘unstable fibres’). After 7–9 days of culture, three of these fibres died. For two of the unstable fibres, after the substantial force reduction, twitch and tetanic tension increased again. Finally at day 14 and 18 of culture, respectively, the tensions attained values higher than their original values. For stable fibres, twitch contraction time, twitch half-relaxation time and tetanus 10%-relaxation time increased during culture. For unstable fibres these parameters fluctuated. For all fibres the stimulus threshold fluctuated during the first two days, and then remained constant, even for the fibres that were cultured for at least two weeks. It is concluded that the present culture system for mature muscle fibres allows long-term studies within a well-defined medium. Unfortunately, initial tetanic and twitch force are poor predictors of the long-term behaviour of the fibres.
Original languageEnglish
Pages (from-to)410-417
Number of pages20
JournalArchives of physiology and biochemistry
Volume109
Issue number5
DOIs
Publication statusPublished - 2001

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Sarcomeres
Xenopus laevis
Muscles
Serum-Free Culture Media
Tetanus
Culture Media
Microscopy
Lasers
Serum

Keywords

  • IR-103956
  • METIS-204055

Cite this

Jaspers, R. T., Feenstra, H., Lee-de Groot, M. B. E., Huijing, P. A. J. B. M., & van der Laarse, W. J. (2001). Twitch and Tetanic Tension during Culture of Mature Xenopus laevis Single Muscle Fibres. Archives of physiology and biochemistry, 109(5), 410-417. https://doi.org/10.1076/apab.109.5.410.11825
Jaspers, R.T. ; Feenstra, Hiske ; Lee-de Groot, M.B.E. ; Huijing, P.A.J.B.M. ; van der Laarse, W.J. / Twitch and Tetanic Tension during Culture of Mature Xenopus laevis Single Muscle Fibres. In: Archives of physiology and biochemistry. 2001 ; Vol. 109, No. 5. pp. 410-417.
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Jaspers, RT, Feenstra, H, Lee-de Groot, MBE, Huijing, PAJBM & van der Laarse, WJ 2001, 'Twitch and Tetanic Tension during Culture of Mature Xenopus laevis Single Muscle Fibres' Archives of physiology and biochemistry, vol. 109, no. 5, pp. 410-417. https://doi.org/10.1076/apab.109.5.410.11825

Twitch and Tetanic Tension during Culture of Mature Xenopus laevis Single Muscle Fibres. / Jaspers, R.T.; Feenstra, Hiske; Lee-de Groot, M.B.E.; Huijing, P.A.J.B.M.; van der Laarse, W.J.

In: Archives of physiology and biochemistry, Vol. 109, No. 5, 2001, p. 410-417.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Twitch and Tetanic Tension during Culture of Mature Xenopus laevis Single Muscle Fibres

AU - Jaspers, R.T.

AU - Feenstra, Hiske

AU - Lee-de Groot, M.B.E.

AU - Huijing, P.A.J.B.M.

AU - van der Laarse, W.J.

PY - 2001

Y1 - 2001

N2 - Investigation of the mechanisms of muscle adaptation requires independent control of the regulating factors. The aim of the present study was to develop a serum-free medium to culture mature single muscle fibres of Xenopus laevis. As an example, we used the culture system to study adaptation of twitch and tetanic force characteristics, number of sarcomeres in series and fibre cross-section. Fibres dissected from m. iliofibularis (n = 10) were kept in culture at a fibre mean sarcomere length of 2.3 µm in a culture medium without serum. Twitch and tetanic tension were determined daily. Before and after culture the number of sarcomeres was determined by laser diffraction and fibre cross-sectional area (CSA) was determined by microscopy. For five fibres twitch tension increased during culture and tetanic tension was stable for periods varying from 8 to 14 days (‘stable fibres’), after which fibres were removed from culture for analysis. Fibre CSA and the number of sarcomeres in series remained constant during culture. Five other fibres showed a substantial reduction in twitch and tetanic tension within the first five days of culture (‘unstable fibres’). After 7–9 days of culture, three of these fibres died. For two of the unstable fibres, after the substantial force reduction, twitch and tetanic tension increased again. Finally at day 14 and 18 of culture, respectively, the tensions attained values higher than their original values. For stable fibres, twitch contraction time, twitch half-relaxation time and tetanus 10%-relaxation time increased during culture. For unstable fibres these parameters fluctuated. For all fibres the stimulus threshold fluctuated during the first two days, and then remained constant, even for the fibres that were cultured for at least two weeks. It is concluded that the present culture system for mature muscle fibres allows long-term studies within a well-defined medium. Unfortunately, initial tetanic and twitch force are poor predictors of the long-term behaviour of the fibres.

AB - Investigation of the mechanisms of muscle adaptation requires independent control of the regulating factors. The aim of the present study was to develop a serum-free medium to culture mature single muscle fibres of Xenopus laevis. As an example, we used the culture system to study adaptation of twitch and tetanic force characteristics, number of sarcomeres in series and fibre cross-section. Fibres dissected from m. iliofibularis (n = 10) were kept in culture at a fibre mean sarcomere length of 2.3 µm in a culture medium without serum. Twitch and tetanic tension were determined daily. Before and after culture the number of sarcomeres was determined by laser diffraction and fibre cross-sectional area (CSA) was determined by microscopy. For five fibres twitch tension increased during culture and tetanic tension was stable for periods varying from 8 to 14 days (‘stable fibres’), after which fibres were removed from culture for analysis. Fibre CSA and the number of sarcomeres in series remained constant during culture. Five other fibres showed a substantial reduction in twitch and tetanic tension within the first five days of culture (‘unstable fibres’). After 7–9 days of culture, three of these fibres died. For two of the unstable fibres, after the substantial force reduction, twitch and tetanic tension increased again. Finally at day 14 and 18 of culture, respectively, the tensions attained values higher than their original values. For stable fibres, twitch contraction time, twitch half-relaxation time and tetanus 10%-relaxation time increased during culture. For unstable fibres these parameters fluctuated. For all fibres the stimulus threshold fluctuated during the first two days, and then remained constant, even for the fibres that were cultured for at least two weeks. It is concluded that the present culture system for mature muscle fibres allows long-term studies within a well-defined medium. Unfortunately, initial tetanic and twitch force are poor predictors of the long-term behaviour of the fibres.

KW - IR-103956

KW - METIS-204055

U2 - 10.1076/apab.109.5.410.11825

DO - 10.1076/apab.109.5.410.11825

M3 - Article

VL - 109

SP - 410

EP - 417

JO - Archives of physiology and biochemistry

JF - Archives of physiology and biochemistry

SN - 1381-3455

IS - 5

ER -