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First published online February 1, 2008
Journal of Experimental Biology 211, 568-576 (2008)
Published by The Company of Biologists 2008
doi: 10.1242/jeb.010207

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Characterisation of neurotransmitter-induced electrolyte transport in cockroach salivary glands by intracellular Ca²⁺, Na⁺ and pH measurements in duct cells

Carsten Hille^* and Bernd Walz

University of Potsdam, Institute of Biochemistry and Biology, Department of Animal Physiology, 14476 Potsdam-Golm, Germany

View larger version (18K): [in this window] [in a new window]   Fig. 1. Comparison of 1 µmol l–1 dopamine (DA)-induced effects in duct cells of the two types of preparation: lobes and isolated ducts. Changes in intracellular pH (pHi) in duct cells as measured with BCECF in lobes (A) and isolated ducts (B). Simultaneous measurement of [Ca2+]i and [Na+]i changes in duct cells of lobes (C) and isolated ducts (D). The SBFI ratio (solid line) indicates changes in [Na+]i, whereas the Fluo-3 fluorescence (dashed line) indicates changes in [Ca2+]i (a.u., arbitrary units). These original recordings are representative of at least five independent experiments.

View larger version (12K): [in this window] [in a new window]   Fig. 2. Comparison of 1 µmol l–1 DA-induced and 1 µmol l–1 serotonin (5-HT)-induced effects in duct cells of both types of preparation: lobes and isolated ducts. Changes in pHi in duct cells as measured with BCECF in lobes (A) and isolated ducts (B). Changes in [Ca2+]i in duct cells as measured with Fura-2 in lobes (C) and isolated ducts (D). These original recordings are representative of at least five independent experiments.

View larger version (25K): [in this window] [in a new window]   Fig. 3. Effects of various factors on intracellular acidification induced by 1 µmol l–1 DA in duct cells of lobe preparations in HCO3–-free saline. (A) The acidification is abolished in Cl––free saline (N=4). (B) The acidification is not influenced by bath application of 500 µmol l–1 DIDS (N=9). (C) The acidification is transient following the bath application of 10 µmol l–1 bumetanide (N=6). (D) The combined bath application of 10 µmol l–1 bumetanide and 500 µmol l–1 DIDS abolishes the acidification (N=6). (E) The acidification is not influenced by bath application of 500 µmol l–1 acetozolamide (N=4). (F) The combined bath application of 10 µmol l–1 bumetanide and 500 µmol l–1 acetazolamide reduces the acidification dramatically (N=8).

View larger version (24K): [in this window] [in a new window]   Fig. 4. Effects of various factors on intracellular acidification induced by 1 µmol l–1 DA in duct cells of lobe preparations in CO2/HCO3–-buffered saline. (A) The acidification is still present in CO2/HCO3–-buffered saline (N=5). (B) Bath application of 10 µmol l–1 bumetanide does not influence the acidification (N=5). (C) The combined bath application of 10 µmol l–1 bumetanide and 500 µmol l–1 DIDS strongly reduces the acidification (N=5). (D) The combined bath application of 10 µmol l–1 bumetanide and 50 µmol l–1 EIPA also reduces the acidification dramatically (N=5). (E) In addition, the combined bath application of 10 µmol l–1 bumetanide and 500 µmol l–1 acetozolamide decreases the acidification (N=5).

View larger version (19K): [in this window] [in a new window]   Fig. 5. Schematic diagram of proposed ion transporters in salivary glands of Periplaneta americana. Dopamine-induced saliva secretion involves the formation of isosmotic primary saliva in acinar peripheral cells and modification in duct cells resulting in hyposmotic final saliva. CA, carbonic anhydrase. For details, see text.