SAMPLING INTERSTITIAL FLUID FROM RAT SKELETAL-MUSCLES BY INTERMUSCULAR WICKS

A modification of the implanted wick method (K. Aukland and H. O. Fadnes. Acta Physiol. Scand. 88:350-358, 1973) was devised to sample interstitial fluid from rat muscles. Dry nylon wicks were inserted postmortem into intermuscular spaces between leg muscles by means of a plastic catheter, which was subsequently withdrawn. Inserting the wicks postmortem avoids contaminating wick fluid with proteins extravasated as a result of local inflammatory reactions; placing them intermuscularly avoids contamination by fluid and proteins from damaged muscle cells. Wick fluid protein concentrations (mg/ml) averaged 24.1 +/- 1.1 and 28.5 +/- 1.5 (means +/- SE) in medial and lateral hindlimb muscles, respectively. The corresponding albumin concentrations were 13.0 +/- 0.7 and 13.9 +/- 0.7 mg/ml. Total protein and albumin concentrations in plasma were 54.1 +/- 0.8 and 22.5 +/- 0.3 mg/ml. Electrophoresis of wick fluid showed a pattern of peaks similar to that of plasma, with albumin relatively high and larger molecules relatively low. Proteins from muscle cells were not detected. Isotope studies (I-125-labeled albumin, Cr-51-EDTA) showed that < 2% of the albumin in wick fluid came directly from plasma and that wick fluid was not concentrated by cell swelling postmortem. Wick fluid from intermuscular wicks implanted in anesthetized rats in vivo had nearly the same total protein concentration as fluid from postmortem wicks, but albumin-to-globulin (A/G) ratios were slightly lower (1.22 +/- 0.07 vs. 1.53 +/- 0.21 measured by gel electrophoresis), and more significantly, nearly 50% of the albumin leaked to wick fluid from plasma as a result of wick implantation. Fluid recovered from wicks implanted intramuscularly postmortem had higher total protein concentrations and lower A/G ratios than intermuscular wick fluid; electrophoresis showed large contamination by muscle cell proteins. Among the methods tested here, we found that intermuscular wicks implanted postmortem provided the most satisfactory access to muscle interstitial fluid in rats.