Electrogenic cation transport across leech caecal epithelium

Abstract

Electrogenic cation transport across the caecal epithelium of the leech Hirudo medicinalis was investigated using modified Ussing chambers. Transepithelial resistance (RT) and potential difference (VT) were 61.0 +/- 3.5 omega.cm2 and -1.1 +/- 0.2 mV (n = 149), respectively, indicating that leech caecal epithelium is a "leaky" epithelium. Under control conditions short circuit current (ISC) and transepithelial Na+ transport rate (INa) averaged at 22.1 +/- 1.5 microA.cm-2 and 49.7 +/- 2.6 microA.cm-2, respectively. Mucosal application of amiloride (100 mumol.l-1) or benzamil (50 mumol.l-1) influenced neither ISC nor INa. The transport system in the apical membrane showed no pronounced cation selectivity and a linear dependence on mucosal Na+ concentration. Removal of mucosal Ca2+ increased ISC by about 50% due to an increase of transepithelial Na+ transport. Trivalent cations (La3+ and Tb3+, 1 mmol.l-1 both) added to the mucosal Ringer solution reduced INa by more than 40%. Serosal ouabain (1 mmol.l-1) almost halved ISC and INa while 0.1% (= 5.4 mmol.l-1) DNP decreased INa to 11.8 +/- 5.1% of initial values. Serosal addition of cAMP increased both ISC and INa whereas the neurotransmitters. FMRFamide, acetylcholine, GABA, L-dopa, serotonin and dopamine failed to show any effects; octopamine, glycine and L-glutamate reduced INa markedly. On the basis of these results we conclude that in leech caecal epithelium apical uptake of monovalent cations is mediated by non-selective cation conductances which are sensitive to extracellular Ca2+ but insensitive to amiloride. Basolaterally Na+ is extruded via ouabain-sensitive and -insensitive ATPases. cAMP activates Na+ transport across leech caecal epithelium, although the physiological stimulus for cAMP-production remains unknown.