Micropatterned cocultures are a useful experimental tool for the study of cell-cell interactions. Patterning methods often rely on sequential seeding of different cell types or removal of a barrier separating two populations, but it is difficult to pattern sharp interfaces between pure populations with low cross-contamination when using these approaches. Patterning by the use of reconfigurable substrates can overcome these limitations, but such methods can be costly and challenging to employ in a typical biology laboratory. Here, we describe a low-cost and simple-to-use reconfigurable substrate comprised of a transparent elastic material that is partially cut to form a slit that opens when the device is stretched. The slit seals back up when released, allowing two initially separate, adherent cell populations to be brought together to form a contact interface. Fluorescent imaging of patterned cocultures demonstrates the early establishment of a sharp cellular interface. As a proof of principle, we demonstrate the use of this device to study competition at the interface of two stem cell populations.
Previous studies show that activation of purinergic P2X receptors and transient receptor potential vanilloid type 1 (TRPV1) located on muscle afferent nerve evokes a pressor response. P2X and TRPV1 receptors are also sensitive to changes in pH. The aim of this study was to examine the effect of decreasing interstitial pH on the pressor reflex mediated by those receptors in skeletal muscle. In decerebrated rats, interstitial pH in the triceps surae muscle was adjusted by infusing Ringer solutions at pH: 4.5, 5.5, 6.5 and 7.4 into the femoral artery. Interstitial samples were collected using microdialysis probes inserted into the muscles and pH was measured. Arterial injection ofα, β‐methylene ATP (0.25 mM), a P2X agonist, at pH values of 7.4, 6.5 and 5.5 increased mean arterial pressure (MAP) by 29±2, 24±3 and 21±3 mmHg (P<0.05 for 5.5 vs. pH 7.4) respectively. Capsaicin (1μg/kg TRPV1 agonist) injected into the artery at pH 7.4, 6.5, 5.5 and 4.5 elevated MAP by 29±4, 33±2, 35±3 and 40±3 mmHg (P<0.05 for 4.5 vs. pH 7.4) respectively. Our data indicate that skeletal muscle acidosis attenuates P2X receptor‐mediated responses; but enhances TRPV1 receptor‐mediated response. Exaggerated TRPV1 response requires lower pH in muscle. This study provides evidence that muscle pH may be important in modulating P2X and TRPV1 responsiveness in exercising muscle. Supported by R01 info:ddbj-emblgenbank/HL075533, R01 info:ddbj-emblgenbank/HL078866 (Li) and R01 info:ddbj-emblgenbank/HL060800 (Sinoway).
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