PKCα regulates vasopressin-induced aquaporin-2 trafficking in mouse kidney collecting duct cells in vitro via altering microtubule assembly

Zhao, Hong; Yao, Xi; Wang, Taoxia; Jin, Wen-min; Ji, Qiu; Yang, Xiao; Duan, Qiuhong; Yao, Lei

doi:10.1038/aps.2011.160

Cited by 10 publications

(10 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In outer medullary tissues, AQP2 abundance was decreased in response to lithium treatment in WT mice; however, lithium only reduced AQP2 expression by 30% in PKCα KO mice ( Figure 4D ). Because PKCα reportedly plays a role in vasopressin-induced AQP2 trafficking [32] , we wanted to determine if the preservation of AQP2 expression in lithium-treated PKCα KO was ineffective due to inability of the channel to reach the apical membrane. AQP2 was diffusely localized in the cytosol of inner medullary collecting duct cells in both untreated WT and PKCα KO mice ( Figure 5 ).…”

Section: Resultsmentioning

confidence: 99%

“…Studies indicate that the inhibitory action of PKC on vasopressin-stimulated water permeability is due to PKC-initiated endocytosis of AQP2 [46]. Although PKC does not directly interact with AQP2 [32], activation of PKC leads to the short-chain ubiquitination of AQP2 resulting in lysosomal degradation of the channel [16]. Reports show that the PKCα isoform is involved in α-tubulin assembly in renal cells [47] and microtubule-dependent trafficking of AQP2 regulates intracellular localization following internalization; however, microtubules are not involved in the exocytic transport of the channel [48].…”

Section: Discussionmentioning

confidence: 99%

“…Reports show that the PKCα isoform is involved in α-tubulin assembly in renal cells [47] and microtubule-dependent trafficking of AQP2 regulates intracellular localization following internalization; however, microtubules are not involved in the exocytic transport of the channel [48]. In fact, overexpression of a constitutively active PKCα construct in IMCD cells prevented translocation of AQP2 to the plasma membrane and resulted in AQP2 distribution throughout the cytoplasm [32]. Our data support these collective findings and indicate that in the absence of PKCα, AQP2 remains at the apical membrane despite a lithium-mediated reduction in cAMP concentration.…”

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Absence of PKC-Alpha Attenuates Lithium-Induced Nephrogenic Diabetes Insipidus

et al. 2014

View full text Add to dashboard Cite

Lithium, an effective antipsychotic, induces nephrogenic diabetes insipidus (NDI) in ∼40% of patients. The decreased capacity to concentrate urine is likely due to lithium acutely disrupting the cAMP pathway and chronically reducing urea transporter (UT-A1) and water channel (AQP2) expression in the inner medulla. Targeting an alternative signaling pathway, such as PKC-mediated signaling, may be an effective method of treating lithium-induced polyuria. PKC-alpha null mice (PKCα KO) and strain-matched wild type (WT) controls were treated with lithium for 0, 3 or 5 days. WT mice had increased urine output and lowered urine osmolality after 3 and 5 days of treatment whereas PKCα KO mice had no change in urine output or concentration. Western blot analysis revealed that AQP2 expression in medullary tissues was lowered after 3 and 5 days in WT mice; however, AQP2 was unchanged in PKCα KO. Similar results were observed with UT-A1 expression. Animals were also treated with lithium for 6 weeks. Lithium-treated WT mice had 19-fold increased urine output whereas treated PKCα KO animals had a 4-fold increase in output. AQP2 and UT-A1 expression was lowered in 6 week lithium-treated WT animals whereas in treated PKCα KO mice, AQP2 was only reduced by 2-fold and UT-A1 expression was unaffected. Urinary sodium, potassium and calcium were elevated in lithium-fed WT but not in lithium-fed PKCα KO mice. Our data show that ablation of PKCα preserves AQP2 and UT-A1 protein expression and localization in lithium-induced NDI, and prevents the development of the severe polyuria associated with lithium therapy.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Absence of PKC-Alpha Attenuates Lithium-Induced Nephrogenic Diabetes Insipidus

et al. 2014

View full text Add to dashboard Cite

show abstract

“…In addition to confirming these results using the same techniques in this study, we also visually observed the expression pattern of UT-A1 induced by hypertonicity by the successful stable transfection of mIMCD 3 -UT-A1-GFP cells. Unlike other membrane proteins such as AQP2, which presents as a bright ring around the cell [22,23], hypertonicity induced concentrated spot distribution of UT-A1 in the cell. Whether the pattern of distribution is related to specific molecular structure of UT-A1 and what is the functional proteins associated with UT-A1 need further investigation.…”

Section: Discussionmentioning

confidence: 99%

Glycogen Synthase Kinase-3 Modulates Hyperosmotic-Induced Urea Transporter A1 Relocation in the Inner Medullary Collecting Duct Cells

Huang²,

Liu³

et al. 2016

Nephron

Self Cite

View full text Add to dashboard Cite

Aim: Glycogen synthase kinase 3 (GSK3) regulates urine concentration by mediating the vasopressin-induced aquaporin 2 expression and water permeability, although it is unknown whether GSK3 also mediates the accumulation of the urea transporter A1 (UT-A1). The aim of this study is to investigate the effect of GSK3 on UT-A1 distribution. Methods: Mouse inner medullary collecting duct 3 cells were transfected with UT-A1-GFP construct. The stable transfected cells were cultured under hypertonic conditions, treated with GSK3 inhibitor lithium chloride, GSK3 activator, lysosome or proteasome inhibitor. The expression levels of UT-A1, GSK3, and phospho-GSK3 were analyzed using western blot. The interaction between UT-A1 and the Golgi apparatus was examined using confocal immunofluorescence microscope. The UT-A1 trafficking was examined using the biotinylation of surface membranes. Results: UT-A1 dissociated away from the Golgi apparatus and translocated to the plasma membrane under hypertonic-NaCl and NaCl plus urea stimulation. This movement was accompanied by the increased phosphorylation of GSK3 and its localization on the cellular membrane. Moreover, these results were duplicated by treating the cells with the GSK3 inhibitor, and by contrast, were partially reversed by the GSK3 activator. Treating cells with a lysosome or proteasome inhibitor failed to attenuate the effects of hypertonic stimulus, indicating that the loss of UT-A1 from the Golgi was not due to degradation. Conclusion: Our results suggest that GSK3 may in part modulate the hypertonic-induced intracellular UT-A1 redistribution and its accumulation on the plasma membrane, which may constitute another mechanism by which GSK3 modulates urine concentration.

show abstract

“…Conversely, parasite protein kinase C (PKC) activation by phorbol ester displaced the T. cruzi and Leishmania plasma membrane from subpellicular microtubules, forming surface protrusions [53][54]. In this regard AA can modulate PKC [55] and PKC activity can regulate microtubule functioning by phosphorylation of microtubule-associated proteins [56][57][58][59], including kinesin and trypanosome kinesins preserve parasite morphology via subpellicular microtubules regulation [60][61].…”

Section: Discussionmentioning

confidence: 99%

Modes of Action of Arjunolic Acid and Derivatives on Trypanosoma cruzi Cells

Souza-Neta¹,

Menezes²,

Lima³

et al. 2014

CTMC

View full text Add to dashboard Cite

Chagas disease causes considerable morbimortality in the Americas, with circa 7 to 8 million infected people, causing at least 12,000 annual deaths and 100 million people at risk. Its chemotherapy is poorly selective and effective, associated to severe side effects and unresponsive cases. Thus, R&D on therapeutic alternatives is undoubtedly required. The Brazilian poorly studied biodiversity offers uncountable bioagents, which may be exploited for chemotherapy. The triterpene arjunolic acid (AA), reduced the Trypanosoma cruzi epimastigote in vitro proliferation with an apparent IC₅₀ of 171 µM. Electron microscopy analysis revealed remarkable effects on the parasite surface and architecture. AA-treated parasites displayed minutely corrugated plasma membranes devoid of subpellicular microtubules as well as biogenesis of multiple basal bodies. As the AA effects appeared mainly restricted or originated at the parasite peripheral cytoplasm, including the cytoskeleton membrane linkage, we inferred that the compound targeted primarily the lipid bilayer; therefore, we performed synthetic modification to increase the molecule lipophilicity and thus membrane permeability. The methyl ester (MeAA) and tri-acetylated derivatives (3AcAA) had potentiated trypanocidal activity, producing IC₅₀ values of 21.9 and 15.8 µM, respectively. Both derivatives were able to produce remarkable ultrastructural alterations in the parasites, including inner compartments such as Golgi apparatus and the endocytic/autophagic pathway. Parasites cultured with both derivatives displayed numerous and large autophagic vacuoles, altered flagellar length and cell body connection. These data indicate that synthetically-modified natural products comprise valuable tools in antiparasitic chemotherapy and that electron microscopy may be useful not only in determining the mechanisms of action but also in directing such modifications for rational drug design.

show abstract

PKCα regulates vasopressin-induced aquaporin-2 trafficking in mouse kidney collecting duct cells in vitro via altering microtubule assembly

Cited by 10 publications

References 40 publications

Absence of PKC-Alpha Attenuates Lithium-Induced Nephrogenic Diabetes Insipidus

Absence of PKC-Alpha Attenuates Lithium-Induced Nephrogenic Diabetes Insipidus

Glycogen Synthase Kinase-3 Modulates Hyperosmotic-Induced Urea Transporter A1 Relocation in the Inner Medullary Collecting Duct Cells

Modes of Action of Arjunolic Acid and Derivatives on Trypanosoma cruzi Cells

Contact Info

Product

Resources

About