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Microdialysis sampling from skeletal muscle and adipose tissue with special reference to the effects of insulin on tissue blood flow and glucose metabolism
Inst. för fysiologi och farmakologi, Karolinska Institutet.ORCID iD: 0000-0001-8161-5610
1998 (English)Doctoral thesis, comprehensive summary (Other academic)
Place, publisher, year, edition, pages
Stockholm: Norstedts Tryckeri AB , 1998. , 56 p.
Keyword [en]
microdialysis, insulin, skeletal muscle, adipose tissue, glucose, amino acids
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:gih:diva-1082ISBN: 91-628-3050-3 (print)OAI: oai:DiVA.org:gih-1082DiVA: diva2:284703
Public defence
1998-06-12, Aulan, GIH, Lidingövägen 1, Stockholm, 09:00 (English)
Opponent
Supervisors
Available from: 2010-01-08 Created: 2010-01-08 Last updated: 2017-03-01Bibliographically approved
List of papers
1. The ethanol technique of monitoring local blood flow changes in rat skeletal muscle: implications for microdialysis.
Open this publication in new window or tab >>The ethanol technique of monitoring local blood flow changes in rat skeletal muscle: implications for microdialysis.
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1992 (English)In: Acta Physiologica Scandinavica, ISSN 0001-6772, E-ISSN 1365-201X, Vol. 146, no 1, 87-97 p.Article in journal (Refereed) Published
Abstract [en]

We have investigated the feasibility of monitoring local skeletal muscle blood flow in the rat by including ethanol in the perfusion medium passing through a microdialysis probe placed in muscle tissue. Ethanol at 5, 55, or 1100 mM did not directly influence local muscle metabolism, as measured by dialysate glucose, lactate, and glycerol concentrations. The clearance of ethanol from the perfusion medium can be described by the outflow/inflow ratio ([ethanol]collected dialysate/[ethanol]infused perfusion medium), which was found to be similar (between 0.36 and 0.38) at all ethanol perfusion concentrations studied. With probes inserted in a flow-chamber, this ratio changed in a flow-dependent way in the external flow range of 5-20 microliters min-1. The ethanol outflow/inflow ratio in vivo was significantly (P less than 0.001) increased (to a maximum of 127 +/- 2.8% and 144 +/- 7.4% of the baseline, mean +/- SEM) when blood flow was reduced by either leg constriction or local vasopressin administration, and significantly (P less than 0.001) reduced (to 62 +/- 6.4% and 43 +/- 4.4% of baseline) with increases in blood flow during external heating or local 2-chloroadenosine administration, respectively. Dialysate glucose concentrations correlated negatively with the ethanol outflow/inflow ratio (P less than 0.01) and consequently decreased (to 46 +/- 7.6% and 56 +/- 5.6% of baseline) with constriction and vasopressin administration and increased (to 169 +/- 32.5% and 262 +/- 16.7% of baseline) following heating and 2-chloroadenosine administration. Dialysate lactate concentrations were significantly increased (approximately 2-fold, P less than 0.001) during all perturbations of blood flow. In conclusion, this technique makes it possible to monitor changes in skeletal muscle blood flow; however, methods of quantification remain to be established. The fact that blood flow changes were found to significantly affect interstitial glucose and lactate concentrations as revealed by microdialysis indicates that this information is critical in microdialysis experiments.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:gih:diva-1069 (URN)1442130 (PubMedID)
Available from: 2010-01-07 Created: 2010-01-07 Last updated: 2017-03-01Bibliographically approved
2. Interstitial glucose and lactate balance in human skeletal muscle and adipose tissue studied by microdialysis.
Open this publication in new window or tab >>Interstitial glucose and lactate balance in human skeletal muscle and adipose tissue studied by microdialysis.
1993 (English)In: Journal of Physiology, ISSN 0022-3751, E-ISSN 1469-7793, Vol. 471, 637-57 p.Article in journal (Refereed) Published
Abstract [en]

1. Microdialysis was used to gain insight into the substrate exchanges in the interstitial space of skeletal muscle and adipose tissue. Probes were inserted in the quadriceps femoris muscle and para-umbilical subcutaneous adipose tissue of thirteen subjects and microdialysis was performed at different flow rates (1-4 microliters min-1) and during changes in tissue blood flow. 2. When ethanol (5 mM) is included in the perfusion solution, the ethanol clearance from the probe is a measure of tissue blood flow. Blood flow changes induced by adenosine or vasopressin perfusion, by exercise or by circulatory occlusion resulted in ethanol clearance values of 69-139% of the basal level. The ethanol clearance was higher in skeletal muscle than in adipose tissue (32-62%, P < 0.001), a difference compatible with a higher blood flow in muscle tissue. 3. The fraction of the interstitial glucose concentration that was recovered with the microdialysis was similar in skeletal muscle (the absolute values being 1.70 +/- 0.14 mM at 1 microliter min-1 and 0.59 +/- 0.05 mM at 4 microliters min-1) and adipose tissue (1.89 +/- 0.20 mM at 1 microliter min-1; 0.54 +/- 0.05 mM at 4 microliters min-1) and correlated inversely with the tissue ethanol clearance, both in the basal state and during changes in tissue blood flow (muscle: r = -0.56 to -0.67; adipose tissue r = -0.72 to -0.95). Coefficients of variation were 6-8% (glucose) and 11-16% (lactate) and were similar during isometric exercise. The reproducibility of the technique (comparison of two contralateral probes; perfusion flow rate 4 microliters min-1) was 5.3-8.3% (ethanol) and 23.9-20.8% (glucose) in muscle (n = 6) and adipose tissue (n = 4) respectively. 4. The skeletal muscle dialysate lactate concentration (1 microliter min-1: 1.16 +/- 0.2 mM) was higher than in adipose tissue (0.76 +/- 0.08 mM, P < 0.05), where the absolute amount of lactate that could be removed from the tissue (at 4 microliters min-1) was only half of that in skeletal muscle (0.8 +/- 0.11 vs. 1.76 +/- 0.23 nmol min-1, P < 0.05). The dialysate lactate level was not affected in either tissue by large changes in the interstitial glucose concentration indicating that in neither tissue is blood glucose a significant source of lactate formation. 5. The blood flow effects on the dialysate glucose concentration are the likely consequence of probe glucose drainage artificially shifting the balance between the supply and consumption of interstitial glucose.(ABSTRACT TRUNCATED AT 400 WORDS)

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:gih:diva-1068 (URN)8120827 (PubMedID)
Available from: 2010-01-07 Created: 2010-01-07 Last updated: 2017-03-01Bibliographically approved
3. Influence of adrenergic agonists on the release of amino acids from rat skeletal muscle studied by microdialysis.
Open this publication in new window or tab >>Influence of adrenergic agonists on the release of amino acids from rat skeletal muscle studied by microdialysis.
1998 (English)In: Acta Physiologica Scandinavica, ISSN 0001-6772, E-ISSN 1365-201X, Vol. 163, no 4, 349-60 p.Article in journal (Refereed) Published
Abstract [en]

The microdialysis technique was used to study the effects of adrenergic agonists on the release of amino acids from rat skeletal muscle. The release was monitored indirectly by measurements of interstitial concentrations. To distinguish metabolic from vasoactive effects, the adrenaline and isoprenaline results were compared with those of vasopressin, alpha-agonists and adenosine. As determined by the microdialysis ethanol technique, adrenaline, alpha-agonists and vasopressin induced vasoconstriction, whereas isoprenaline and adenosine induced vasodilatation. The lactate-to-pyruvate ratio increased fourfold with adrenaline (P < 0.001) and by 54% with isoprenaline (P < 0.05), whereas no change was observed with alpha-agonists and adenosine. Vasopressin induced a fivefold increase in the lactate-to-pyruvate ratio (P < 0.001), but with an unchanged pyruvate concentration, indicating that the effect may have been secondary to ischaemia. Adrenaline induced a twofold and vasopressin a 34% increase in the concentration of alanine (P < 0.001), whereas isoprenaline, adenosine and alpha-agonists had no significant effect. Adrenaline-perfusion induced an initial anabolic effect as evidenced by a reduced concentration of tyrosine. A significant decrease in the glutamate-to-glutamine ratio was observed with adrenaline and isoprenaline (22 and 27%, P < 0.01) whereas alpha-agonists, vasopressin and adenosine were without effect. In conclusion, the present study showed that adrenaline, via a beta-adrenergically mediated activation of glycogenolysis, possibly further stimulated by ischaemia, induced an increased release of alanine from skeletal muscle. The study indicates a beta-adrenergic stimulation on the glutamine synthetase step and a short lasting anabolic effect of adrenaline. Differences in the magnitude of the effects of adrenaline and isoprenaline could be related to their different vasoactive properties.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:gih:diva-1064 (URN)9789578 (PubMedID)
Available from: 2010-01-07 Created: 2010-01-07 Last updated: 2017-03-01Bibliographically approved
4. Microdialysis of rat skeletal muscle and adipose tissue: dynamics of the interstitial glucose pool.
Open this publication in new window or tab >>Microdialysis of rat skeletal muscle and adipose tissue: dynamics of the interstitial glucose pool.
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1994 (English)In: Acta Physiologica Scandinavica, ISSN 0001-6772, E-ISSN 1365-201X, Vol. 151, no 2, 249-60 p.Article in journal (Refereed) Published
Abstract [en]

Microdialysis was evaluated as a method for studying glucose metabolism in skeletal muscle. Dialysis probes (0.5 x 10 mm) were perfused at 0.5 or 1.0 microliter min-1. Based upon perfusion with glucose, the muscle interstitial glucose concentration was estimated to be 6.9 +/- 0.3 mM (n = 14), which was not significantly different from the blood glucose level. With insulin infusion (1200 mU kg-1 body wt i.v.), the insulin-induced change in the glucose concentration of the interstitial space of muscle was of equal magnitude to that of blood and adipose tissue. In spite of this, when the perfusion medium was not supplemented with glucose, the glucose concentration decreased more in skeletal muscle dialysates (to 36.7 +/- 4.9% of the initial level) than in blood (to 29.7 +/- 5.0%) but less than in adipose tissue (to 17.7 +/- 4.9% of the initial level) (P < 0.05). The results indicate that these differences are due to tissue-specific differences in the dynamic balance between the supply to, and removal from, the interstitial glucose pool. This balance is revealed as a result of the constant glucose drainage by the microdialysis probe. The present results show that, in skeletal muscle, increases in glucose uptake occur with a concomitant increase in tissue blood flow as revealed by the microdialysis ethanol technique, whereas in adipose tissue the glucose uptake increases in the absence of a corresponding increase in blood flow.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:gih:diva-1079 (URN)7942061 (PubMedID)
Available from: 2010-01-08 Created: 2010-01-08 Last updated: 2017-03-01Bibliographically approved
5. Effect of physiological hyperinsulinemia on blood flow and interstitial glucose concentration in human skeletal muscle and adipose tissue studied by microdialysis.
Open this publication in new window or tab >>Effect of physiological hyperinsulinemia on blood flow and interstitial glucose concentration in human skeletal muscle and adipose tissue studied by microdialysis.
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1998 (English)In: Diabetes, ISSN 0012-1797, E-ISSN 1939-327X, Vol. 47, no 8, 1296-301 p.Article in journal (Refereed) Published
Abstract [en]

The effect of an euglycemic-hyperinsulinemic glucose clamp (94 +/- 5 microU/ml) on blood flow and glucose extraction fraction in human skeletal muscle and adipose tissue was investigated. Limb blood flow was measured by venous occlusion pletysmography and tissue blood flow by the microdialysis ethanol technique. Insulin infusion resulted in an increased blood flow in the calf and forearm (64 and 36%, respectively; P < 0.01) but not in the studied muscles of these limbs (ethanol outflow-to-inflow ratio: m. gastrocnemius 0.144 +/- 0.009 to 0.140 +/- 0.011, NS; m. brachioradialis 0.159 +/- 0.025 to 0.168 +/- 0.027, NS). This was accompanied by an increased extraction fraction of glucose, as measured by an increased arteriovenous difference over the forearm (0.16 +/- 0.04 to 0.70 +/- 0.10 mmol/l; P < 0.001) and by an increase in the estimated arterial-interstitial glucose difference in the gastrocnemius (0.82-1.42 mmol/l) and brachioradialis muscle (0.82-1.97 mmol/l). The blood flow in adipose tissue was significantly increased during insulin infusion, as evidenced by a decreased ethanol outflow-to-inflow ratio (0.369 +/- 0.048 to 0.325 +/- 0.046; P < 0.01). This was accompanied by an unchanged concentration of glucose in the dialysate (-2.6%, NS). In summary, during physiological hyperinsulinemia 1) a blood flow increase was detected in the calf and forearm, but not in the studied muscles of these limbs; 2) the blood flow increased in the subcutaneous adipose tissue; and 3) the estimated arterial-interstitial glucose difference increased in both muscles studied and was larger in the forearm muscle than the arteriovenous glucose difference over the forearm. The present study shows that microdialysis is a useful tool to obtain tissue-specific information about the effect of insulin on blood flow and glucose extraction in human skeletal muscle and adipose tissue.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:gih:diva-1065 (URN)9703331 (PubMedID)
Available from: 2010-01-07 Created: 2010-01-07 Last updated: 2017-03-01Bibliographically approved
6. Microdialysis in human skeletal muscle and adipose tissue at low flow rates is possible if dextran-70 is added to prevent loss of perfusion fluid.
Open this publication in new window or tab >>Microdialysis in human skeletal muscle and adipose tissue at low flow rates is possible if dextran-70 is added to prevent loss of perfusion fluid.
1997 (English)In: Acta Physiologica Scandinavica, ISSN 0001-6772, E-ISSN 1365-201X, Vol. 159, no 3, 261-2 p.Article in journal (Refereed) Published
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:gih:diva-1067 (URN)9079158 (PubMedID)
Available from: 2010-01-07 Created: 2010-01-07 Last updated: 2017-03-01Bibliographically approved
7. Effect of adding a colloid to the perfusate on net fluid transport across the microdialysis membrane and on concentrations of metabolites in dialysate.
Open this publication in new window or tab >>Effect of adding a colloid to the perfusate on net fluid transport across the microdialysis membrane and on concentrations of metabolites in dialysate.
(English)Manuscript (preprint) (Other academic)
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:gih:diva-1080 (URN)
Note
Arbetet publicerades år 2002 i J. Appl. Physiol. med titeln Microdialysis in human skeletal muscle: effetcs of adding a colloid to the perfusateAvailable from: 2010-01-08 Created: 2010-01-08 Last updated: 2017-03-01Bibliographically approved
8. Metabolite levels in human skeletal muscle and adipose tissue studied with microdialysis at low perfusion flow.
Open this publication in new window or tab >>Metabolite levels in human skeletal muscle and adipose tissue studied with microdialysis at low perfusion flow.
1998 (English)In: The American journal of physiology, ISSN 0002-9513, Vol. 274, no 5 Pt 1, E936-45 p.Article in journal (Refereed) Published
Abstract [en]

To identify a perfusion flow at which the interstitial fluid completely equilibrates with the microdialysis perfusion fluid, a protocol with successively lower perfusion flows was used. A colloid was included in the perfusion fluid to make sampling possible at the lowest perfusion flows. At 0.16 microliter/min, the measured metabolites had reached a complete equilibration in both tissues, and the measured concentrations of glucose, glycerol, and urea were in good agreement with expected tissue-specific levels. The glucose concentration in adipose tissue (4.98 +/- 0.14 mM) was equal to that of plasma (5.07 +/- 0.07 mM), whereas the concentration in muscle (4.41 +/- 0.11 mM) was lower than in plasma and adipose tissue (P < 0.001). The concentration of lactate was higher (P < 0.001) in muscle (2.39 +/- 0.22 mM) than in adipose tissue (1.30 +/- 0.12 mM), whereas the glycerol concentration in adipose tissue (233 +/- 19.7 microM) was higher (P < 0.001) than in muscle (40.8 +/- 3.0 microM) and in plasma (68.7 +/- 3.97 microM). The concentration of urea was equal in the two tissues. Overall, the study indicates that microdialysis at a low perfusion flow may be a tool to continuously monitor tissue interstitial concentrations.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:gih:diva-1066 (URN)9612253 (PubMedID)
Available from: 2010-01-07 Created: 2010-01-07 Last updated: 2017-03-01Bibliographically approved
9. Insulin perfused through a microdialysis catheter with a 100 KDa dialysis membrane induces local metabolic effects in skeletal muscle.
Open this publication in new window or tab >>Insulin perfused through a microdialysis catheter with a 100 KDa dialysis membrane induces local metabolic effects in skeletal muscle.
(English)Manuscript (preprint) (Other academic)
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:gih:diva-1081 (URN)
Note
Arbetet publicerades år 2000 i Int. J. of Biological Macromolecules med titeln A microdialysis method for the in situ investigation of the action of large peptide molecules in human skeletal muscle:detection of local metabolic effects of insulinAvailable from: 2010-01-08 Created: 2010-01-08 Last updated: 2017-03-01Bibliographically approved

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