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High Sensitivity Method to Estimate Distribution of Hyaluronan Molecular Sizes in Small Biological Samples Using Gas-Phase Electrophoretic Mobility Molecular Analysis.

Do L, Dahl CP, Kerje S, Hansell P, Mörner S, Lindqvist U, Engström-Laurent A, Larsson G, Hellman U - Int J Cell Biol (2015)

Bottom Line: In this paper, we demonstrate that gas-phase electrophoretic molecular mobility analyzer (GEMMA) can be used to estimate the distribution of hyaluronan molecular sizes in biological samples with a limited amount of hyaluronan.The low detection level of the GEMMA method allows for estimation of hyaluronan molecular sizes from different parts of small organs.Hence, the GEMMA method opens opportunity to attain a profile over the distribution of hyaluronan molecular sizes and estimate changes caused by disease or experimental conditions that has not been possible to obtain before.

View Article: PubMed Central - PubMed

Affiliation: Cardiology, Heart Centre and Department of Public Health and Clinical Medicine, Division of Medicine, Umeå University, 901 85 Umeå, Sweden ; Department of Medical Biochemistry and Biophysics-Unit of Research Education and Development Östersund, Umeå University, 831 31 Umeå, Sweden.

ABSTRACT
Hyaluronan is a negatively charged polydisperse polysaccharide where both its size and tissue concentration play an important role in many physiological and pathological processes. The various functions of hyaluronan depend on its molecular size. Up to now, it has been difficult to study the role of hyaluronan in diseases with pathological changes in the extracellular matrix where availability is low or tissue samples are small. Difficulty to obtain large enough biopsies from human diseased tissue or tissue from animal models has also restricted the study of hyaluronan. In this paper, we demonstrate that gas-phase electrophoretic molecular mobility analyzer (GEMMA) can be used to estimate the distribution of hyaluronan molecular sizes in biological samples with a limited amount of hyaluronan. The low detection level of the GEMMA method allows for estimation of hyaluronan molecular sizes from different parts of small organs. Hence, the GEMMA method opens opportunity to attain a profile over the distribution of hyaluronan molecular sizes and estimate changes caused by disease or experimental conditions that has not been possible to obtain before.

No MeSH data available.


Related in: MedlinePlus

GEMMA data profile. Molecular size distribution of HA extracted from four different tissues and species, rat kidney (orange), chicken comb (green), human heart (blue), and moose heart (red), one of each. The electroforetic molecule diameter analyzed in the GEMMA was converted to molecular weight by analyzing HA standards from Hyalose L.L.C. ranging from 30 kDa to 2400 kDa samples [21]. HA from moose heart and chicken comb both showed uniform HMW HA peaks around ca 5000 kDa. HA from rat kidney showed the HMW peak and an additional broad peak with a center around 30 kDa. Human heart also showed presence of LMW HA in the same range as in rat kidney but the HMW HA content was of apparent greater size than the other three examples shown.
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fig1: GEMMA data profile. Molecular size distribution of HA extracted from four different tissues and species, rat kidney (orange), chicken comb (green), human heart (blue), and moose heart (red), one of each. The electroforetic molecule diameter analyzed in the GEMMA was converted to molecular weight by analyzing HA standards from Hyalose L.L.C. ranging from 30 kDa to 2400 kDa samples [21]. HA from moose heart and chicken comb both showed uniform HMW HA peaks around ca 5000 kDa. HA from rat kidney showed the HMW peak and an additional broad peak with a center around 30 kDa. Human heart also showed presence of LMW HA in the same range as in rat kidney but the HMW HA content was of apparent greater size than the other three examples shown.

Mentions: Weight needed for analysis was about 50 mg wet weight for heart and kidney and ca 20 mg for chicken comb samples. Purified HA extract of the heart and kidney were not necessary to dilute whereas the chicken comb HA required a 100-fold dilution prior to analysis to prevent overload of the instrument (Table 1). Differences in HA molecular size distribution were observed between all samples. The raw data from GEMMA analysis are reported as the electrophoretic mobility diameter (EMD) of the different samples. The EMD was converted to molecular weight by analyzing HA standards from Hyalose L.L.C. (Figure 1) [21]. HA from moose heart and chicken comb both showed uniform peaks around EMD 7.2 nm, which is consistent with HMW HA of ca 5000 kDa. In contrast, HA from rat kidney both showed the HMW peak seen in the moose and chicken samples, and in addition a broad peak with a center around EMD 5.2 nm, corresponding to a low molecular weight of ca 30 kDa [21]. Human heart also showed presence of LMW HA in the same range as in rat kidney but the HMW HA content was of apparent greater size than the other three examples shown.


High Sensitivity Method to Estimate Distribution of Hyaluronan Molecular Sizes in Small Biological Samples Using Gas-Phase Electrophoretic Mobility Molecular Analysis.

Do L, Dahl CP, Kerje S, Hansell P, Mörner S, Lindqvist U, Engström-Laurent A, Larsson G, Hellman U - Int J Cell Biol (2015)

GEMMA data profile. Molecular size distribution of HA extracted from four different tissues and species, rat kidney (orange), chicken comb (green), human heart (blue), and moose heart (red), one of each. The electroforetic molecule diameter analyzed in the GEMMA was converted to molecular weight by analyzing HA standards from Hyalose L.L.C. ranging from 30 kDa to 2400 kDa samples [21]. HA from moose heart and chicken comb both showed uniform HMW HA peaks around ca 5000 kDa. HA from rat kidney showed the HMW peak and an additional broad peak with a center around 30 kDa. Human heart also showed presence of LMW HA in the same range as in rat kidney but the HMW HA content was of apparent greater size than the other three examples shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4581557&req=5

fig1: GEMMA data profile. Molecular size distribution of HA extracted from four different tissues and species, rat kidney (orange), chicken comb (green), human heart (blue), and moose heart (red), one of each. The electroforetic molecule diameter analyzed in the GEMMA was converted to molecular weight by analyzing HA standards from Hyalose L.L.C. ranging from 30 kDa to 2400 kDa samples [21]. HA from moose heart and chicken comb both showed uniform HMW HA peaks around ca 5000 kDa. HA from rat kidney showed the HMW peak and an additional broad peak with a center around 30 kDa. Human heart also showed presence of LMW HA in the same range as in rat kidney but the HMW HA content was of apparent greater size than the other three examples shown.
Mentions: Weight needed for analysis was about 50 mg wet weight for heart and kidney and ca 20 mg for chicken comb samples. Purified HA extract of the heart and kidney were not necessary to dilute whereas the chicken comb HA required a 100-fold dilution prior to analysis to prevent overload of the instrument (Table 1). Differences in HA molecular size distribution were observed between all samples. The raw data from GEMMA analysis are reported as the electrophoretic mobility diameter (EMD) of the different samples. The EMD was converted to molecular weight by analyzing HA standards from Hyalose L.L.C. (Figure 1) [21]. HA from moose heart and chicken comb both showed uniform peaks around EMD 7.2 nm, which is consistent with HMW HA of ca 5000 kDa. In contrast, HA from rat kidney both showed the HMW peak seen in the moose and chicken samples, and in addition a broad peak with a center around EMD 5.2 nm, corresponding to a low molecular weight of ca 30 kDa [21]. Human heart also showed presence of LMW HA in the same range as in rat kidney but the HMW HA content was of apparent greater size than the other three examples shown.

Bottom Line: In this paper, we demonstrate that gas-phase electrophoretic molecular mobility analyzer (GEMMA) can be used to estimate the distribution of hyaluronan molecular sizes in biological samples with a limited amount of hyaluronan.The low detection level of the GEMMA method allows for estimation of hyaluronan molecular sizes from different parts of small organs.Hence, the GEMMA method opens opportunity to attain a profile over the distribution of hyaluronan molecular sizes and estimate changes caused by disease or experimental conditions that has not been possible to obtain before.

View Article: PubMed Central - PubMed

Affiliation: Cardiology, Heart Centre and Department of Public Health and Clinical Medicine, Division of Medicine, Umeå University, 901 85 Umeå, Sweden ; Department of Medical Biochemistry and Biophysics-Unit of Research Education and Development Östersund, Umeå University, 831 31 Umeå, Sweden.

ABSTRACT
Hyaluronan is a negatively charged polydisperse polysaccharide where both its size and tissue concentration play an important role in many physiological and pathological processes. The various functions of hyaluronan depend on its molecular size. Up to now, it has been difficult to study the role of hyaluronan in diseases with pathological changes in the extracellular matrix where availability is low or tissue samples are small. Difficulty to obtain large enough biopsies from human diseased tissue or tissue from animal models has also restricted the study of hyaluronan. In this paper, we demonstrate that gas-phase electrophoretic molecular mobility analyzer (GEMMA) can be used to estimate the distribution of hyaluronan molecular sizes in biological samples with a limited amount of hyaluronan. The low detection level of the GEMMA method allows for estimation of hyaluronan molecular sizes from different parts of small organs. Hence, the GEMMA method opens opportunity to attain a profile over the distribution of hyaluronan molecular sizes and estimate changes caused by disease or experimental conditions that has not been possible to obtain before.

No MeSH data available.


Related in: MedlinePlus