Prevention of estradiol 17β-<scp>d</scp>-glucuronide–induced canalicular transporter internalization by hormonal modulation of cAMP in rat hepatocytes

Zucchetti, Andrés E.; Barosso, Ismael Ricardo; Boaglio, Andrea Carolina; Pellegrino, J.; Ochoa, Elena J.; Roma, Marcelo G.; Crocenzi, Fernando A.; Pozzi, Enrique J. Sánchez

doi:10.1091/mbc.e11-01-0047

Cited by 26 publications

(41 citation statements)

References 48 publications

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“…The likely involvement of PKA in E17G‐induced secretory failure was evaluated by incubating IRHCs with the PKA inhibitors, KT5720 (KT; 250 nM), H89 (1μM), or Rp‐cAMPS (Rp; 10 μM), for 15 minutes. Activation of PKA was confirmed evaluating phospho‐PKA substrates by western blotting …”

Section: Methodsmentioning

confidence: 99%

G-protein-coupled receptor 30/adenylyl cyclase/protein kinase A pathway is involved in estradiol 17ß-d-glucuronide-induced cholestasis

et al. 2014

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Estradiol-17ß-D-glucuronide (E17G) activates different signaling pathways (e.g., Ca 21 -dependent protein kinase C, phosphoinositide 3-kinase/protein kinase B, mitogenactivated protein kinases [MAPKs] p38 and extracellular signal-related kinase 1/2, and estrogen receptor alpha) that lead to acute cholestasis in rat liver with retrieval of the canalicular transporters, bile salt export pump (Abcb11) and multidrug resistanceassociated protein 2 (Abcc2). E17G shares with nonconjugated estradiol the capacity to activate these pathways. G-protein-coupled receptor 30 (GPR30) is a receptor implicated in nongenomic effects of estradiol, and the aim of this study was to analyze the potential role of this receptor and its downstream effectors in E17G-induced cholestasis. In vitro, GPR30 inhibition by G15 or its knockdown with small interfering RNA strongly prevented E17G-induced impairment of canalicular transporter function and localization. E17G increased cyclic adenosine monophosphate (cAMP) levels, and this increase was blocked by G15, linking GPR30 to adenylyl cyclase (AC). Moreover, AC inhibition totally prevented E17G insult. E17G also increased protein kinase A (PKA) activity, which was blocked by G15 and AC inhibitors, connecting the links of the pathway, GPR30-AC-PKA. PKA inhibition prevented E17G-induced cholestasis, whereas exchange protein activated directly by cyclic nucleotide/MAPK kinase, another cAMP downstream effector, was not implicated in cAMP cholestatic action. In the perfused rat liver model, inhibition of the GPR30-AC-PKA pathway totally prevented E17G-induced alteration in Abcb11 and Abcc2 function and localization. Conclusion: Activation of GPR30-AC-PKA is a key factor in the alteration of canalicular transporter function and localization induced by E17G. Interaction of E17G with GPR30 may be the first event in the cascade of signaling activation. (HEPATOLOGY 2014;59:1016-1029 H epatocanalicular adenosine-triphosphatedependent transporters are essential for bile secretion 1 and alteration in their expression, localization, or activity results in secretory failure and cholestasis. 2,3 Two of the most relevant canalicular transporters are the bile salt export pump (Abcb11; also named Bsep), which transports monoanionic bile salts, and multidrug resistance-associated protein 2 (Abcc2;

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Section: Methodsmentioning

confidence: 99%

G-protein-coupled receptor 30/adenylyl cyclase/protein kinase A pathway is involved in estradiol 17ß-d-glucuronide-induced cholestasis

et al. 2014

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“…While the protective effect of glucagon is dependent on PKA, but not EPAC, salbutamol's effect is EPAC/MEK dependent and PKA independent. Mechanistically, salbutamol/EPAC promotes a microtubule-dependent and long-range trafficking of Abcb11/Abcc2-containing vesicles, whereas glucagon/ PKA modulates the microfilament-mediated fusion of these vesicles with the apical membrane (1217). In a follow-up study, the preventive effects of salbutamol and glucagon were further validated in more physiological settings using perfused rat liver and in vivo alanine administration, which induces pancreatic glucagon secretion (1218).…”

Section: Epac Proteins and Hepatic Functionsmentioning

confidence: 96%

Intracellular cAMP Sensor EPAC: Physiology, Pathophysiology, and Therapeutics Development

Robichaux

Cheng

2018

Physiological Reviews

162

226

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This review focuses on one family of the known cAMP receptors, the exchange proteins directly activated by cAMP (EPACs), also known as the cAMP-regulated guanine nucleotide exchange factors (cAMP-GEFs). Although EPAC proteins are fairly new additions to the growing list of cAMP effectors, and relatively "young" in the cAMP discovery timeline, the significance of an EPAC presence in different cell systems is extraordinary. The study of EPACs has considerably expanded the diversity and adaptive nature of cAMP signaling associated with numerous physiological and pathophysiological responses. This review comprehensively covers EPAC protein functions at the molecular, cellular, physiological, and pathophysiological levels; and in turn, the applications of employing EPAC-based biosensors as detection tools for dissecting cAMP signaling and the implications for targeting EPAC proteins for therapeutic development are also discussed.

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“…Data are expressed as mean ± standard error of the mean (SEM; n = 5) ▸ Hayashi et al (2012) demonstrated that internalization of Abcb11, a canalicular bile salt transporter, is clathrinmediated. Though the information about the mechanism of Abcc2 internalization is lacking, a similar mechanism is possible since confocal images in previous works showed that E17G action produced similar pattern of endocytosis of Abcb11 and Abcc2 Boaglio et al 2010;Crocenzi et al 2008;Zucchetti et al 2011).…”

Section: Discussionmentioning

confidence: 92%

“…These proteins are organized in several signaling pathways. Up to now there are evidences that support a pathway that involves ERα, PKC, and p38-MAPK responsible for the initial endocytic internalization of canalicular transporters, and pathways initiated in the action of E17G on its receptor GPR30 that activate either adenylyl cyclase-PKA or PI3K-Akt-ERK1/2, being the latter pathway responsible for keeping transporters in a subapical space, restraining reinsertion Boaglio et al 2010Boaglio et al , 2012Crocenzi et al 2008;Zucchetti et al 2011Zucchetti et al , 2014.…”

Section: Introductionmentioning

confidence: 99%

EGFR participates downstream of ERα in estradiol-17β-d-glucuronide-induced impairment of Abcc2 function in isolated rat hepatocyte couplets

et al. 2015

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Estradiol-17β-D-glucuronide (E17G) induces acute endocytic internalization of canalicular transporters, including multidrug resistance-associated protein 2 (Abcc2) in rat, generating cholestasis. Several proteins organized in at least two different signaling pathways are involved in E17G cholestasis: one pathway involves estrogen receptor alpha (ERα), Ca(2+)-dependent protein kinase C and p38-mitogen activated protein kinase, and the other pathway involves GPR30, PKA, phosphoinositide 3-kinase/AKT and extracellular signal-related kinase 1/2. EGF receptor (EGFR) can potentially participate in both pathways since it interacts with GPR30 and ERα. Hence, the aim of this study was to analyze the potential role of this receptor and its downstream effectors, members of the Src family kinases in E17G-induced cholestasis. In vitro, EGFR inhibition by Tyrphostin (Tyr), Cl-387785 or its knockdown with siRNA strongly prevented E17G-induced impairment of Abcc2 function and localization. Activation of EGFR was necessary but not sufficient to impair the canalicular transporter function, whereas the simultaneous activation of EGFR and GPR30 could impair Abcc2 transport. The protection of Tyr was not additive to that produced by the ERα inhibitor ICI neither with that produced by Src kinase inhibitors, suggesting that EGFR shared the signaling pathway of ERα and Src. Further analysis of ERα, EGFR and Src activations induced by E17G, demonstrated that ERα activation precedes that of EGFR and EGFR activation precedes that of Src. In conclusion, activation of EGFR is a key factor in the alteration of canalicular transporter function and localization induced by E17G and it occurs before that of Src and after that of ERα.

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Prevention of estradiol 17β-d-glucuronide–induced canalicular transporter internalization by hormonal modulation of cAMP in rat hepatocytes

Cited by 26 publications

References 48 publications

G-protein-coupled receptor 30/adenylyl cyclase/protein kinase A pathway is involved in estradiol 17ß-d-glucuronide-induced cholestasis

G-protein-coupled receptor 30/adenylyl cyclase/protein kinase A pathway is involved in estradiol 17ß-d-glucuronide-induced cholestasis

Intracellular cAMP Sensor EPAC: Physiology, Pathophysiology, and Therapeutics Development

EGFR participates downstream of ERα in estradiol-17β-d-glucuronide-induced impairment of Abcc2 function in isolated rat hepatocyte couplets

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