Polycystin-2 cation channel function in the human syncytiotrophoblast is regulated by microtubular structures.
Polycystin-2 (PC2), encoded by PKD2, which is one of the genes whose mutations cause polycystic kidney disease, is abundantly produced in the apical domain of the syncytiotrophoblast (hST) of term human placenta. PC2, a TRP-type (TRPP2) non-selective cation channel, is present in primary cilia of renal epithelial cells, a microtubule-based ancillary structure with sensory function. The hST has abundant cytoskeletal structures, and actin filament dynamics regulate PC2 channel function in this epithelium. However, it is expected that the apical hST excludes microtubular structures. Here, we demonstrated by Western blot and immunocytochemical analyses that hST apical vesicles indeed contain microtubule structural components, including tubulin isoforms, acetylated alpha-tubulin, and the kinesin motor proteins KIF3A and KIF3B. PC2 and tubulin were substantially colocalized in hST vesicles. Treatment of hST vesicles with either the microtubular disrupter colchicine (15 microM) or the microtubular stabilizer paclitaxel (taxol, 15 microM) resulted in distinct patterns of microtubular re-organization and PC2 redistribution. We also observed that changes in microtubular dynamics regulate PC2 channel function. Addition of colchicine rapidly inhibited PC2 channel activity in lipid-bilayer reconstituted hST membranes. Addition of either tubulin and GTP, or taxol, however, stimulated PC2 channel activity in control hST membranes. Interestingly, we found that the kinesin motor protein KIF3A was capable of increasing PC2 channel activity in hST. We believe that the data are the first to provide a direct demonstration of a microtubular interaction with PC2 in the hST. This interaction thus plays an important regulatory role in the control of ion transport in the human placenta.
Montalbetti
Nicolás
N
Laboratorio de Canales Iónicos, Departamento de Fisicoquímica y Química Analítica, Facultad de Farmacia y Bioquímica, Buenos Aires, Argentina.
Li
Qiang
Q
Wu
Yuliang
Y
Chen
Xing-Zhen
XZ
Cantiello
Horacio F
HF
D002412:Cations / Q000378:metabolism; D002451:Cell Compartmentation / Q000502:physiology; D016764:Cell Polarity / Q000502:physiology; D003078:Colchicine / Q000494:pharmacology; D022162:Cytoplasmic Vesicles / Q000502:physiology; D005260:Female; D006160:Guanosine Triphosphate / Q000494:pharmacology; D006801:Humans; D016547:Kinesins / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008870:Microtubules / Q000187:drug effects / Q000502:physiology*; D017239:Paclitaxel / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D050396:TRPP Cation Channels / Q000378:metabolism*; D014327:Trophoblasts / Q000502:physiology* / Q000648:ultrastructure; D014404:Tubulin / Q000494:pharmacology; D050257:Tubulin Modulators / Q000494:pharmacology
D002412:Cations; D003078:Colchicine; D006160:Guanosine Triphosphate; D016547:Kinesins; D017239:Paclitaxel; D050396:TRPP Cation Channels; D014404:Tubulin; D050257:Tubulin Modulators; C509805:KIF2A protein, human