Desipramine inhibits salivary Ca<sup>2+</sup> signaling and aquaporin translocation

Lee, Kyung Hwa; Choi, Seongjin; Lm, Choi; J, Lee; Kim, Jae-Hoon; Chung, Grace H.; G, Lee; Sy, Choi; Park, K

doi:10.1111/odi.12317

Cited by 3 publications

(6 citation statements)

References 26 publications

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“…To determine the effect of MβCD on aquaporins, we analyzed the expression level of AQP5 via GFP intensity in the membrane fraction. In a previous study, we confirmed that 20 min of carbachol treatment was optimal for confirming activity-dependent AQP5 translocation [ 28 ]. We prepared the membrane fraction of HSG cells and analyzed the level of AQP5 in comparison with Na + /K + exchanger, a membrane-enriched protein.…”

Section: Resultssupporting

confidence: 78%

“…The cytosolic free Ca 2+ concentration ([Ca 2+ ] i ) was determined using the fluorescent Ca 2+ indicator fura-2/AM, as previously described [ 27 , 28 ]. Briefly, the cell suspension was incubated in fresh medium containing fura-2/AM (4 µM) for 60 min at 37 °C with continuous stirring.…”

Section: Methodsmentioning

confidence: 99%

“…AQP-5-transfected HSG cells were grown in a 60-mm dish, pre-incubated with MβCD for 3 min, and treated with carbachol or thapsigargin for 20 min at 37 °C. The membrane fraction was obtained as previously described [ 28 ] and sonicated in ice-cold 20 mM HEPES solution containing 1 mM MgCl 2 , 100 mM NaCl, 1 mM dithiothreitol, and 0.3 mM phenylmethylsulfonyl fluoride (3 × 30 s; Sigma) at pH 7.4. The samples were centrifuged at 600× g at 4 °C, and the supernatants were centrifuged further at 20,000× g at 4 °C.…”

Section: Methodsmentioning

confidence: 99%

“…The HSG cells were transiently transfected with the pEGFP-C1-AQP-5 plasmid using Lipofectamine 2000 [27]. AQP-5-transfected HSG cells were grown in a 60-mm dish, pre-incubated with MβCD for 3 min, and treated with carbachol or thapsigargin for 20 min at 37 • C. The membrane fraction was obtained as previously described [28] and sonicated in ice-cold 20 mM HEPES solution containing 1 mM MgCl 2 , 100 mM NaCl, 1 mM dithiothreitol, and 0.3 mM phenylmethylsulfonyl fluoride (3 × 30 s; Sigma) at pH 7.4. The samples were centrifuged at 600× g at 4 • C, and the supernatants were centrifuged further at 20,000× g at 4 • C. The pellets (P2 membrane fractions) were separated via SDS-PAGE, and the proteins were transferred onto a PVDF membrane for immunoblotting.…”

Section: Preparation Of Membrane Fractions and Western Blottingmentioning

confidence: 99%

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Muscarinic Receptors and BK Channels Are Affected by Lipid Raft Disruption of Salivary Gland Cells

Lee

Kim

Choi

et al. 2021

IJMS

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Activity-dependent fluid secretion is the most important physiological function of salivary glands and is regulated via muscarinic receptor signaling. Lipid rafts are important for G-protein coupled receptor (GPCR) signaling and ion channels in plasma membranes. However, it is not well understood whether lipid raft disruption affects all membrane events or only specific functions in muscarinic receptor-mediated water secretion in salivary gland cells. We investigated the effects of lipid raft disruption on the major membrane events of muscarinic transcellular water movement in human salivary gland (HSG) cells. We found that incubation with methyl-β-cyclodextrin (MβCD), which depletes lipid rafts, inhibited muscarinic receptor-mediated Ca2+ signaling in HSG cells and isolated mouse submandibular acinar cells. However, MβCD did not inhibit a Ca2+ increase induced by thapsigargin, which activates store-operated Ca2+ entry (SOCE). Interestingly, MβCD increased the activity of the large-conductance Ca2+-activated K+ channel (BK channel). Finally, we found that MβCD did not directly affect the translocation of aquaporin-5 (AQP5) into the plasma membrane. Our results suggest that lipid rafts maintain muscarinic Ca2+ signaling at the receptor level without directly affecting the activation of SOCE induced by intracellular Ca2+ pool depletion or the translocation of AQP5 into the plasma membrane.

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Section: Resultssupporting

confidence: 78%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Preparation Of Membrane Fractions and Western Blottingmentioning

confidence: 99%

See 2 more Smart Citations

Muscarinic Receptors and BK Channels Are Affected by Lipid Raft Disruption of Salivary Gland Cells

Lee

Kim

Choi

et al. 2021

IJMS

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“…The translocation of aquaporin‐5 into the plasma membrane is one of the cellular mechanisms of water secretion in salivary gland cells, HSG cells transfected with GFP‐tagged aquaporin‐5 have been used to analyze the amount of aquaporin‐5 translocated with Na + /K + ‐ATPase as a membrane protein marker . We monitored the bradykinin‐mediated modulation of aquaporin‐5 translocation in HSG cells transfected with GFP‐tagged aquaporin‐5 using the membrane protein marker, Na + /K + ‐ATPase.…”

Section: Resultsmentioning

confidence: 99%

Human salivary gland cells express bradykinin receptors that modulate the expression of proinflammatory cytokines

Lee

Kim

Choi

et al. 2016

European J Oral Sciences

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Bradykinin is an important peptide modulator that affects the function of neurons and immune cells. However, there is no evidence of the bradykinin receptors and their functions in human salivary glands. Here we have identified and characterized bradykinin receptors on human submandibular gland cells. Both bradykinin B1 and B2 receptors are expressed on human submandibular gland cells, A253 cells, and HSG cells. Bradykinin increased the intracellular Ca concentration ([Ca ] ) in a concentration-dependent manner. Interestingly, a specific agonist of the B1 receptor did not have any effect on [Ca ] in HSG cells, whereas specific agonists of the B2 receptor had a Ca mobilizing effect. Furthermore, application of the B1 receptor antagonist, R715, did not alter the bradykinin-mediated increase in cytosolic Ca , whereas the B2 receptor antagonist, HOE140, showed a strong inhibitory effect, which implies that bradykinin B2 receptors are functional in modulating the concentration of cytosolic Ca . Bradykinin did not affect a carbachol-induced rise of [Ca ] and did not modulate translocation of aquaporin-5. However, bradykinin did promote the expression of proinflammatory cytokines, including tumor necrosis factor-α (TNF-α), implying the role of bradykinin in salivary gland inflammation. These data suggest that bradykinin receptors are involved in Ca signaling in human submandibular gland cells and serve a unique role, which is separate from that of other salivary gland G protein-coupled receptors.

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Desipramine induces eryptosis in human erythrocytes, an effect blunted by nitric oxide donor sodium nitroprusside and N-acetyl-L-cysteine but enhanced by Calcium depletion

et al. 2023

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Background: Desipramine a representative of tricyclic antidepressants (TCAs) promotes recovery of depressed patients by inhibition of reuptake of neurotransmitters serotonin (SER) and norepinephrine (NE) in the presynaptic membrane by directly blocking their respective transporters SERT and NET. Aims: To study the effect of desipramine on programmed erythrocyte death (eryptosis) and explore the underlying mechanisms. Methods: Phosphatidylserine (PS) exposure on the cell surface as marker of cell death was estimated from annexin-V-binding, cell volume from forward scatter in flow cytometry. Hemolysis was determined photometrically, and intracellular glutathione [GSH] i from high performance liquid chromatography. Results: Desipramine dose-dependently significantly enhanced the percentage of annexin-V-binding cells and didn´t impact glutathione (GSH) synthesis. Desipramine-induced eryptosis was significantly reversed by pre-treatment of erythrocytes with either nitric oxide (NO) donor sodium nitroprusside (SNP) or N-acetyl-L-cysteine (NAC). The highest inhibitory effect was obtained by using both inhibitors together. Calcium (Ca 2+ ) depletion aggravated desipramine-induced eryptosis. Changing the order of treatment, i.e. desipramine first followed by inhibitors, could not influence the inhibitory effect of SNP or NAC. Conclusion: Antidepressants-caused intoxication can be treated by SNP and NAC, respectively. B) Patients with chronic hypocalcemia should not be treated with tricyclic anti-depressants or their dose should be noticeably reduced.

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Desipramine inhibits salivary Ca²⁺ signaling and aquaporin translocation

Abstract: These results imply that desipramine blocks muscarinic and histamine receptor-mediated Ca(2+) signaling and the subsequent translocation of aquaporin-5 in human salivary gland cells, suggesting a novel mechanism for the xerogenic effects of desipramine.

Cited by 3 publications

References 26 publications

Muscarinic Receptors and BK Channels Are Affected by Lipid Raft Disruption of Salivary Gland Cells

Muscarinic Receptors and BK Channels Are Affected by Lipid Raft Disruption of Salivary Gland Cells

Human salivary gland cells express bradykinin receptors that modulate the expression of proinflammatory cytokines

Desipramine induces eryptosis in human erythrocytes, an effect blunted by nitric oxide donor sodium nitroprusside and N-acetyl-L-cysteine but enhanced by Calcium depletion

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Desipramine inhibits salivary Ca2+ signaling and aquaporin translocation

Abstract: These results imply that desipramine blocks muscarinic and histamine receptor-mediated Ca(2+) signaling and the subsequent translocation of aquaporin-5 in human salivary gland cells, suggesting a novel mechanism for the xerogenic effects of desipramine.

Cited by 3 publications

References 26 publications

Muscarinic Receptors and BK Channels Are Affected by Lipid Raft Disruption of Salivary Gland Cells

Muscarinic Receptors and BK Channels Are Affected by Lipid Raft Disruption of Salivary Gland Cells

Human salivary gland cells express bradykinin receptors that modulate the expression of proinflammatory cytokines

Desipramine induces eryptosis in human erythrocytes, an effect blunted by nitric oxide donor sodium nitroprusside and N-acetyl-L-cysteine but enhanced by Calcium depletion

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Desipramine inhibits salivary Ca²⁺ signaling and aquaporin translocation