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New method for determining total calcium content in tissue applied to skeletal muscle with and without calsequestrin.

Lamboley CR, Kake Guena SA, Touré F, Hébert C, Yaddaden L, Nadeau S, Bouchard P, Wei-LaPierre L, Lainé J, Rousseau EC, Frenette J, Protasi F, Dirksen RT, Pape PC - J. Gen. Physiol. (2015)

Bottom Line: In both fast-twitch (extensor digitorum longus, EDL) and slow-twitch (soleus) muscles from mice, [CaT]WM increased approximately linearly with decreasing muscle weight, increasing approximately twofold with a twofold decrease in muscle weight.Knocking out the high capacity Ca-binding protein calsequestrin (CSQ) did not significantly reduce [CaT]WM in mouse EDL or soleus muscle.Because greater reductions in [CaT]WM would be predicted in both muscle types, we hypothesize that there is a substantial increase in Ca bound to other sites in the CSQ knockout muscles.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Sport, Exercise and Active Living, Victoria University, Melbourne, Victoria 8001, Australia.

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Absorbance spectra corrected for stray light used to determine [CaT]WM with the BAPTA method. (A–D) The absorbance spectra in this figure are from the same experiment shown in Fig. 3, except that they are corrected for the stray light component. Details of this correction are given in the text associated with this figure and Section 3 of the supplemental text. The format of the figure is the same as that used for Fig. 3. For B, the best-fit ratios (A0 − A∞)/(A0 − AM) and (AM − AS)/(A0 − A∞) were, respectively, 3.809 and 0.2275. For D, the best-fit ratios (S0 − S∞)/(S0 − SM) and (SM − SS)/(S0 − S∞) were, respectively, 16.73 and 0.2368.
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fig4: Absorbance spectra corrected for stray light used to determine [CaT]WM with the BAPTA method. (A–D) The absorbance spectra in this figure are from the same experiment shown in Fig. 3, except that they are corrected for the stray light component. Details of this correction are given in the text associated with this figure and Section 3 of the supplemental text. The format of the figure is the same as that used for Fig. 3. For B, the best-fit ratios (A0 − A∞)/(A0 − AM) and (AM − AS)/(A0 − A∞) were, respectively, 3.809 and 0.2275. For D, the best-fit ratios (S0 − S∞)/(S0 − SM) and (SM − SS)/(S0 − S∞) were, respectively, 16.73 and 0.2368.

Mentions: Fig. 4 shows the same absorbance spectra from Fig. 3 corrected for stray light using the procedure described in Section 3 of the supplemental text. As in Fig. 3 B, the solid red curve shows the least-squares best fit of the A0 − AM spectrum to the A0 − A∞ spectrum over the wavelength range of 282 to 340 nm. In contrast to Fig. 3, the A0 − AM difference spectrum matches well with that of A0 − A∞ over the nonfitted region of the spectrum (250–282 nm). The solid and dashed black curves give the corresponding fit of A0 − A∞ to the AM − AS difference spectrum. These black curves are difficult to see because, again, the fit is excellent for the full wavelength range. Likewise, the corresponding fits without muscle present in Fig. 4 D are excellent for the full measured wavelength range. Similar good matches were obtained in other experiments with uncorrected spectra if the effects of stray light were greatly decreased by decreasing the magnitude of the absorbance signals. The lower absorbance values were achieved by either diluting the muscle and BAPTA concentrations by a factor of ∼3 or by using quartz cuvettes with shorter path lengths (1 or 3 mm) compared with that used for all of the data given in this article (path length of 1 cm).


New method for determining total calcium content in tissue applied to skeletal muscle with and without calsequestrin.

Lamboley CR, Kake Guena SA, Touré F, Hébert C, Yaddaden L, Nadeau S, Bouchard P, Wei-LaPierre L, Lainé J, Rousseau EC, Frenette J, Protasi F, Dirksen RT, Pape PC - J. Gen. Physiol. (2015)

Absorbance spectra corrected for stray light used to determine [CaT]WM with the BAPTA method. (A–D) The absorbance spectra in this figure are from the same experiment shown in Fig. 3, except that they are corrected for the stray light component. Details of this correction are given in the text associated with this figure and Section 3 of the supplemental text. The format of the figure is the same as that used for Fig. 3. For B, the best-fit ratios (A0 − A∞)/(A0 − AM) and (AM − AS)/(A0 − A∞) were, respectively, 3.809 and 0.2275. For D, the best-fit ratios (S0 − S∞)/(S0 − SM) and (SM − SS)/(S0 − S∞) were, respectively, 16.73 and 0.2368.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4306712&req=5

fig4: Absorbance spectra corrected for stray light used to determine [CaT]WM with the BAPTA method. (A–D) The absorbance spectra in this figure are from the same experiment shown in Fig. 3, except that they are corrected for the stray light component. Details of this correction are given in the text associated with this figure and Section 3 of the supplemental text. The format of the figure is the same as that used for Fig. 3. For B, the best-fit ratios (A0 − A∞)/(A0 − AM) and (AM − AS)/(A0 − A∞) were, respectively, 3.809 and 0.2275. For D, the best-fit ratios (S0 − S∞)/(S0 − SM) and (SM − SS)/(S0 − S∞) were, respectively, 16.73 and 0.2368.
Mentions: Fig. 4 shows the same absorbance spectra from Fig. 3 corrected for stray light using the procedure described in Section 3 of the supplemental text. As in Fig. 3 B, the solid red curve shows the least-squares best fit of the A0 − AM spectrum to the A0 − A∞ spectrum over the wavelength range of 282 to 340 nm. In contrast to Fig. 3, the A0 − AM difference spectrum matches well with that of A0 − A∞ over the nonfitted region of the spectrum (250–282 nm). The solid and dashed black curves give the corresponding fit of A0 − A∞ to the AM − AS difference spectrum. These black curves are difficult to see because, again, the fit is excellent for the full wavelength range. Likewise, the corresponding fits without muscle present in Fig. 4 D are excellent for the full measured wavelength range. Similar good matches were obtained in other experiments with uncorrected spectra if the effects of stray light were greatly decreased by decreasing the magnitude of the absorbance signals. The lower absorbance values were achieved by either diluting the muscle and BAPTA concentrations by a factor of ∼3 or by using quartz cuvettes with shorter path lengths (1 or 3 mm) compared with that used for all of the data given in this article (path length of 1 cm).

Bottom Line: In both fast-twitch (extensor digitorum longus, EDL) and slow-twitch (soleus) muscles from mice, [CaT]WM increased approximately linearly with decreasing muscle weight, increasing approximately twofold with a twofold decrease in muscle weight.Knocking out the high capacity Ca-binding protein calsequestrin (CSQ) did not significantly reduce [CaT]WM in mouse EDL or soleus muscle.Because greater reductions in [CaT]WM would be predicted in both muscle types, we hypothesize that there is a substantial increase in Ca bound to other sites in the CSQ knockout muscles.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Sport, Exercise and Active Living, Victoria University, Melbourne, Victoria 8001, Australia.

Show MeSH
Related in: MedlinePlus