TY - JOUR
T1 - Characterization of purinergic receptor expression in ARPKD cystic epithelia
AU - Palygin, Oleg
AU - Ilatovskaya, Daria V.
AU - Levchenko, Vladislav
AU - Klemens, Christine A.
AU - Dissanayake, Lashodya
AU - Williams, Anna Marie
AU - Pavlov, Tengis S.
AU - Staruschenko, Alexander
N1 - Publisher Copyright:
© 2018, Springer Nature B.V.
PY - 2018/12/1
Y1 - 2018/12/1
N2 - Polycystic kidney diseases (PKDs) are a group of inherited nephropathies marked by formation of fluid-filled cysts along the nephron. Growing evidence suggests that in the kidney formation of cysts and alteration of cystic electrolyte transport are associated with purinergic signaling. PCK/CrljCrl-Pkhd1pck/CRL (PCK) rat, an established model of autosomal recessive polycystic kidney disease (ARPKD), was used here to test this hypothesis. Cystic fluid of PCK rats and their cortical tissues exhibited significantly higher levels of ATP compared to Sprague Dawley rat kidney cortical interstitium as assessed by highly sensitive ATP enzymatic biosensors. Confocal calcium imaging of the freshly isolated cystic monolayers revealed a stronger response to ATP in a higher range of concentrations (above 100 μM). The removal of extracellular calcium results in the profound reduction of the ATP evoked transient, which suggests calcium entry into the cyst-lining cells is occurring via the extracellular (ionotropic) P2X channels. Further use of pharmacological agents (α,β-methylene-ATP, 5-BDBD, NF449, isoPPADS, AZ10606120) and immunofluorescent labeling of isolated cystic epithelia allowed us to narrow down potential candidate receptors. In conclusion, our ex vivo study provides direct evidence that the profile of P2 receptors is shifted in ARPKD cystic epithelia in an age-related manner towards prevalence of P2X 4 and/or P2X 7 receptors, which opens new avenues for the treatment of this disease.
AB - Polycystic kidney diseases (PKDs) are a group of inherited nephropathies marked by formation of fluid-filled cysts along the nephron. Growing evidence suggests that in the kidney formation of cysts and alteration of cystic electrolyte transport are associated with purinergic signaling. PCK/CrljCrl-Pkhd1pck/CRL (PCK) rat, an established model of autosomal recessive polycystic kidney disease (ARPKD), was used here to test this hypothesis. Cystic fluid of PCK rats and their cortical tissues exhibited significantly higher levels of ATP compared to Sprague Dawley rat kidney cortical interstitium as assessed by highly sensitive ATP enzymatic biosensors. Confocal calcium imaging of the freshly isolated cystic monolayers revealed a stronger response to ATP in a higher range of concentrations (above 100 μM). The removal of extracellular calcium results in the profound reduction of the ATP evoked transient, which suggests calcium entry into the cyst-lining cells is occurring via the extracellular (ionotropic) P2X channels. Further use of pharmacological agents (α,β-methylene-ATP, 5-BDBD, NF449, isoPPADS, AZ10606120) and immunofluorescent labeling of isolated cystic epithelia allowed us to narrow down potential candidate receptors. In conclusion, our ex vivo study provides direct evidence that the profile of P2 receptors is shifted in ARPKD cystic epithelia in an age-related manner towards prevalence of P2X 4 and/or P2X 7 receptors, which opens new avenues for the treatment of this disease.
KW - ARPKD
KW - ATP
KW - Intracellular calcium flux
KW - Kidney
KW - P2X
KW - P2X
KW - P2X receptors
KW - P2rx4
KW - P2rx7
KW - PCK rat
KW - Polycystic kidney disease
KW - Purinergic receptor
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U2 - 10.1007/s11302-018-9632-5
DO - 10.1007/s11302-018-9632-5
M3 - Article
C2 - 30417216
AN - SCOPUS:85056334780
SN - 1573-9538
VL - 14
SP - 485
EP - 497
JO - Purinergic Signalling
JF - Purinergic Signalling
IS - 4
ER -