Calcium absorption by Cav1.3 induces terminal web myosin II phosphorylation and apical GLUT2 insertion in rat intestine.
Glucose absorption in rat jejunum involves Ca(2+)- and PKC betaII-dependent insertion of GLUT2 into the apical membrane. Ca(2+)-induced rearrangement of the enterocyte cytoskeleton is thought to enhance paracellular flow. We have therefore investigated the relationships between myosin II regulatory light chain phosphorylation (RLC(20)), absorption of glucose, water and calcium, and mannitol clearance. ML-7, an inhibitor of myosin light chain kinase, diminished the phloretin-sensitive apical GLUT2 but not the phloretin-insensitive SGLT1 component of glucose absorption in rat jejunum perfused with 75 mM glucose. Western blotting and immunocytochemistry revealed marked decreases in RLC(20) phosphorylation in the terminal web and in the levels of apical GLUT2 and PKC betaII, but not SGLT1. Perfusion with phloridzin or 75 mM mannitol, removal of luminal Ca(2+), or inhibition of unidirectional (45)Ca(2+) absorption by nifedipine exerted similar effects. ML-7 had no effect on the absorption of 10 mM Ca(2+), nor clearance of [(14)C]-mannitol, which was less than 0.7% of the rate of glucose absorption. Water absorption did not correlate with (45)Ca(2+) absorption or mannitol clearance. We conclude that the Ca(2+) necessary for contraction of myosin II in the terminal web enters via an L-type channel, most likely Ca(v)1.3, and is dependent on SGLT1. Moreover, terminal web RLC(20) phosphorylation is necessary for apical GLUT2 insertion. The data confirm that glucose absorption by paracellular flow is negligible, and show further that paracellular flow makes no more than a minimal contribution to jejunal Ca(2+) absorption at luminal concentrations prevailing after a meal.
Mace
Oliver J
OJ
Department of Biology, The University of York, York YO10 5YW, UK.
Morgan
Emma L
EL
Affleck
Julie A
JA
Lister
Norma
N
Kellett
George L
GL
D000818:Animals; D001381:Azepines / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism*; D002132:Calcium Radioisotopes; D002250:Carbon Radioisotopes; D003599:Cytoskeleton / Q000378:metabolism; D020895:Enterocytes / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D051273:Glucose Transporter Type 2 / Q000378:metabolism*; D066298:In Vitro Techniques; D007583:Jejunum / Q000378:metabolism*; D008353:Mannitol / Q000378:metabolism; D024462:Myosin Type II / Q000378:metabolism*; D009219:Myosin-Light-Chain Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009281:Naphthalenes / Q000494:pharmacology; D010539:Permeability; D010766:Phosphorylation; D051381:Rats; D051269:Sodium-Glucose Transporter 1 / Q000378:metabolism; D050916:TRPV Cation Channels / Q000378:metabolism; D014867:Water / Q000378:metabolism
D001381:Azepines; D015220:Calcium Channels; D002132:Calcium Radioisotopes; D002250:Carbon Radioisotopes; D051273:Glucose Transporter Type 2; D009281:Naphthalenes; C495683:Slc2a2 protein, rat; C495676:Slc5a1 protein, rat; D051269:Sodium-Glucose Transporter 1; D050916:TRPV Cation Channels; D014867:Water; C070571:ML 7; C093712:Cacna1d protein, rat; D008353:Mannitol; D009219:Myosin-Light-Chain Kinase; D024462:Myosin Type II; D005947:Glucose; D002118:Calcium