Effect of extraction time and acid concentration on the separation of proglycogen and macroglycogen in horse muscle samples.
Abstract: The objective of this study was to determine whether the concentrations of proglycogen (PG) and macroglycogen (MG) in biopsy samples of horse muscle are influenced by extraction time or perchloric acid (PCA) concentration. In study 1, individual muscle-biopsy samples from 10 horses were divided into 4 parts each and then randomly subjected to 4 periods of extraction (10, 20, 60, or 120 min) with 1.5 M PCA. In study 2, individual muscle-biopsy samples from 6 horses were divided into 24 pieces each and then randomly subjected to 12 combinations of extraction time (10, 20, 30, or 40 min) and PCA concentration (0.5, 1.5, or 3.0 M). The results from study 1 indicated that PG and MG concentrations are affected only after extraction for 120 min; the PG concentration decreased significantly (P < 0.05), and the MG concentration increased (not significantly). In study 2, extraction in 3.0 M PCA yielded significantly lower PG and higher MG concentrations (P < 0.05) than extraction in 0.5 or 1.5 M PCA with each of the extraction times. The results of this study further support the existence of 2 glycogen pools and demonstrate that they are not an extraction artifact. The study also suggests that the 2 pools are stable during extraction over a range of extraction times and acid concentrations. However, if the exposure to acid is very long and, or, the acid concentration is high, some of the insoluble PG appears to be hydrolyzed and to enter the MG pool.
Publication Date: 2002-07-31 PubMed ID: 12146893PubMed Central: PMC227005
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research paper investigates how the extraction time and the concentration of perchloric acid affect the levels of proglycogen and macroglycogen in horse muscle samples. The findings suggest that these factors can influence the proportions of these two forms of glycogen, particularly with long extraction times or high acid concentrations.
Study Design and Methodology
- The study was designed to test the influence of extraction time and perchloric acid (PCA) concentration on the amounts of two types of glycogen—proglycogen (PG) and macroglycogen (MG)—in muscle biopsy specimens taken from horses.
- In the first study, muscle samples from 10 horses were each divided into four parts, which were then randomly assigned to undergo extraction for one of four different time periods (10, 20, 60, or 120 minutes) using 1.5 M PCA.
- In the second study, muscle samples from 6 horses were each divided into 24 pieces. These were then randomly subjected to 12 different combinations of extraction time (10, 20, 30, or 40 minutes) and PCA concentration (0.5, 1.5, or 3.0 M).
Key Findings
- In the first study, the results indicated that the concentrations of PG and MG were only affected when the extraction was carried out for the longest time period of 120 minutes. During this time, there was a significant decrease in PG levels and a non-significant increase in MG levels.
- In the second study, it was found that when extraction was conducted with the highest concentration of PCA (3.0 M), there was a significant decrease in PG levels and a significant increase in MG levels compared to the levels observed when using lower concentrations of PCA (0.5 or 1.5 M).
- The researchers concluded that the existence of two separate glycogen pools (PG and MG) is not simply an artifact of the extraction process, as these pools were stable within a certain range of extraction times and acid concentrations.
Implications and Conclusions
- The research suggests that high concentrations of PCA and prolonged extraction times can result in some of the insoluble PG being hydrolyzed and entering into the MG pool.
- This finding underscores the importance of carefully controlling extraction conditions when assessing proglycogen and macroglycogen levels in horse muscle tissue, as variations in these conditions could potentially distort the measured proportions of the two glycogen forms.
Cite This Article
APA
Bröjer JT, Stämpfli HR, Graham TE.
(2002).
Effect of extraction time and acid concentration on the separation of proglycogen and macroglycogen in horse muscle samples.
Can J Vet Res, 66(3), 201-206.
Publication
Researcher Affiliations
- Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Ontario. johan.brojer@kirmed.slu.se
MeSH Terms
- Animals
- Biopsy, Needle / veterinary
- Female
- Fluorometry / veterinary
- Glycogen / metabolism
- Horses / metabolism
- Male
- Muscle, Skeletal / chemistry
- Muscle, Skeletal / metabolism
- Perchlorates / chemistry
- Random Allocation
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Citations
This article has been cited 2 times.- Mojibi N, Rasouli M. Comparison of Methods to Assay Liver Glycogen Fractions: The Effects of Starvation. J Clin Diagn Res 2017 Mar;11(3):BC17-BC20.
- Marchand I, Tarnopolsky M, Adamo KB, Bourgeois JM, Chorneyko K, Graham TE. Quantitative assessment of human muscle glycogen granules size and number in subcellular locations during recovery from prolonged exercise. J Physiol 2007 Apr 15;580(Pt. 2):617-28.
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