Membrane-limited expression and regulation of Na+-H+ exchanger isoforms by P2 receptors in the rat submandibular gland duct

Min Goo Lee, Patrick J. Schultheis, Ming Yan, Gary E. Shull, Crescence Bookstein, Eugene Chang, Chung Ming Tse, Mark Donowitz, Keerang Park, Shmuel Muallem

Research output: Contribution to journalArticle

Abstract

1. Cell-specific reverse transcriptase-polymerase chain reaction (RT-PCR), immunolocalization and microspectrofluorometry were used to identify and localize the Na+-H+ exchanger (NHE) isoforms expressed in the submandibular gland (SMG) acinar and duct cells and their regulation by basolateral and luminal P2 receptors in the duct. 2. The molecular and immunofluorescence analysis showed that SMG acinar and duct cells expressed NHE1 in the basolateral membrane (BLM). Duct cells also expressed NHE2 and NHE3 in the luminal membrane (LM). 3. Expression of NHE3 was unequivocally established by the absence of staining in SMG from NHE3 knockout mice. NHE3 was expressed in the LM and in subluminal regions of the duct. 4. Measurement of the inhibition of NHE activity by the amiloride analogue HOE 694 (HOE) suggested expression of NHE1-like activity in the BLM and NKE2-like activity in the LM of the SMG duct. Several acute and chronic treatments tested failed to activate NHE activity with low affinity for HOE as expected for NHE3. Hence, the physiological function and role of NHE3 in the SMG duct is not clear at present. 5. Activation of P2 receptors resulted in activation of an NKE-independent, luminal H+ transport pathway that markedly and rapidly acidified the cells. This pathway could be blocked by luminal but not basolateral Ba2+. 6. Stimulation of P(2U) receptors expressed in the BLM activated largely NHE1-like activity, and stimulation of P(2Z) receptors expressed in the LM activated largely NHE2-like activity. 7. The interrelation between basolateral and luminal NHE activities and their respective regulation by P(2U) and P(2Z) receptors can be used to co-ordinate membrane transport events in the LM and BLM during active Na+ reabsorption by the SMG duct.

Original languageEnglish (US)
Pages (from-to)341-357
Number of pages17
JournalJournal of Physiology
Volume513
Issue number2
StatePublished - Dec 1 1998

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Sodium-Hydrogen Antiporter
Submandibular Gland
Protein Isoforms
Membranes
Acinar Cells
Amiloride
Reverse Transcriptase Polymerase Chain Reaction
Knockout Mice
Fluorescent Antibody Technique
Staining and Labeling

ASJC Scopus subject areas

  • Physiology

Cite this

Lee, M. G., Schultheis, P. J., Yan, M., Shull, G. E., Bookstein, C., Chang, E., ... Muallem, S. (1998). Membrane-limited expression and regulation of Na+-H+ exchanger isoforms by P2 receptors in the rat submandibular gland duct. Journal of Physiology, 513(2), 341-357.

Membrane-limited expression and regulation of Na+-H+ exchanger isoforms by P2 receptors in the rat submandibular gland duct. / Lee, Min Goo; Schultheis, Patrick J.; Yan, Ming; Shull, Gary E.; Bookstein, Crescence; Chang, Eugene; Tse, Chung Ming; Donowitz, Mark; Park, Keerang; Muallem, Shmuel.

In: Journal of Physiology, Vol. 513, No. 2, 01.12.1998, p. 341-357.

Research output: Contribution to journalArticle

Lee, MG, Schultheis, PJ, Yan, M, Shull, GE, Bookstein, C, Chang, E, Tse, CM, Donowitz, M, Park, K & Muallem, S 1998, 'Membrane-limited expression and regulation of Na+-H+ exchanger isoforms by P2 receptors in the rat submandibular gland duct', Journal of Physiology, vol. 513, no. 2, pp. 341-357.
Lee MG, Schultheis PJ, Yan M, Shull GE, Bookstein C, Chang E et al. Membrane-limited expression and regulation of Na+-H+ exchanger isoforms by P2 receptors in the rat submandibular gland duct. Journal of Physiology. 1998 Dec 1;513(2):341-357.
Lee, Min Goo ; Schultheis, Patrick J. ; Yan, Ming ; Shull, Gary E. ; Bookstein, Crescence ; Chang, Eugene ; Tse, Chung Ming ; Donowitz, Mark ; Park, Keerang ; Muallem, Shmuel. / Membrane-limited expression and regulation of Na+-H+ exchanger isoforms by P2 receptors in the rat submandibular gland duct. In: Journal of Physiology. 1998 ; Vol. 513, No. 2. pp. 341-357.
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abstract = "1. Cell-specific reverse transcriptase-polymerase chain reaction (RT-PCR), immunolocalization and microspectrofluorometry were used to identify and localize the Na+-H+ exchanger (NHE) isoforms expressed in the submandibular gland (SMG) acinar and duct cells and their regulation by basolateral and luminal P2 receptors in the duct. 2. The molecular and immunofluorescence analysis showed that SMG acinar and duct cells expressed NHE1 in the basolateral membrane (BLM). Duct cells also expressed NHE2 and NHE3 in the luminal membrane (LM). 3. Expression of NHE3 was unequivocally established by the absence of staining in SMG from NHE3 knockout mice. NHE3 was expressed in the LM and in subluminal regions of the duct. 4. Measurement of the inhibition of NHE activity by the amiloride analogue HOE 694 (HOE) suggested expression of NHE1-like activity in the BLM and NKE2-like activity in the LM of the SMG duct. Several acute and chronic treatments tested failed to activate NHE activity with low affinity for HOE as expected for NHE3. Hence, the physiological function and role of NHE3 in the SMG duct is not clear at present. 5. Activation of P2 receptors resulted in activation of an NKE-independent, luminal H+ transport pathway that markedly and rapidly acidified the cells. This pathway could be blocked by luminal but not basolateral Ba2+. 6. Stimulation of P(2U) receptors expressed in the BLM activated largely NHE1-like activity, and stimulation of P(2Z) receptors expressed in the LM activated largely NHE2-like activity. 7. The interrelation between basolateral and luminal NHE activities and their respective regulation by P(2U) and P(2Z) receptors can be used to co-ordinate membrane transport events in the LM and BLM during active Na+ reabsorption by the SMG duct.",
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AU - Lee, Min Goo

AU - Schultheis, Patrick J.

AU - Yan, Ming

AU - Shull, Gary E.

AU - Bookstein, Crescence

AU - Chang, Eugene

AU - Tse, Chung Ming

AU - Donowitz, Mark

AU - Park, Keerang

AU - Muallem, Shmuel

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N2 - 1. Cell-specific reverse transcriptase-polymerase chain reaction (RT-PCR), immunolocalization and microspectrofluorometry were used to identify and localize the Na+-H+ exchanger (NHE) isoforms expressed in the submandibular gland (SMG) acinar and duct cells and their regulation by basolateral and luminal P2 receptors in the duct. 2. The molecular and immunofluorescence analysis showed that SMG acinar and duct cells expressed NHE1 in the basolateral membrane (BLM). Duct cells also expressed NHE2 and NHE3 in the luminal membrane (LM). 3. Expression of NHE3 was unequivocally established by the absence of staining in SMG from NHE3 knockout mice. NHE3 was expressed in the LM and in subluminal regions of the duct. 4. Measurement of the inhibition of NHE activity by the amiloride analogue HOE 694 (HOE) suggested expression of NHE1-like activity in the BLM and NKE2-like activity in the LM of the SMG duct. Several acute and chronic treatments tested failed to activate NHE activity with low affinity for HOE as expected for NHE3. Hence, the physiological function and role of NHE3 in the SMG duct is not clear at present. 5. Activation of P2 receptors resulted in activation of an NKE-independent, luminal H+ transport pathway that markedly and rapidly acidified the cells. This pathway could be blocked by luminal but not basolateral Ba2+. 6. Stimulation of P(2U) receptors expressed in the BLM activated largely NHE1-like activity, and stimulation of P(2Z) receptors expressed in the LM activated largely NHE2-like activity. 7. The interrelation between basolateral and luminal NHE activities and their respective regulation by P(2U) and P(2Z) receptors can be used to co-ordinate membrane transport events in the LM and BLM during active Na+ reabsorption by the SMG duct.

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