Artículo
The syncytiotrophoblast (SCT), a multinucleated epithelium forming the outer layer of chorionic villi, acts in human placenta as a transporting barrier regulating the transference of nutrients, solutes and water between maternal and fetal blood. Electrolyte homeostasis and extracellular fluid volume are maintained primarily by regulated Na+ transport. The present study was conducted to analyze the presence of the epithelial Na channel (ENaC) in placental tissue from normal and pre-eclamptic women and in BeWo cell, a model of a human SCT. Changes in the expression of these proteins during sodium transport across the placenta may be related to the pathogeny of pre-eclampsia. The role that ENaC and Na+ transport deregulation play on human placental tissues still remains unknown although in aldosterone-responsive epithelial cells (kidney, colon), abnormalities upregulating its activity lead to increased Na+ uptake and hypertension (i.e. Liddle´s syndrome) whereas a diminished channel activity can result in the pseudohypoaldosteronisn syndrome with salt loss and hypotension. Our results show that ENaC is expressed in the apical membrane of normal syncytiotrophoblast. The amplified fragment of α-ENaC was cloned and sequenced having a 100% identity with the sequence of α-ENaC obtained from GenBankTM (SCNN1A, accession number Z92981). We found that the transcription of the α-ENaC mRNA was not detectable in preeclamptic placentas and the protein was not observed with immunohistochemistry staining, probably indicating a low protein expression level. In BeWo cells ENac was found and its expression is regulated by aldosterone, vasopressin, progesterone and estradiol. With patch clamp techniques we studied the currents trough ENaC channels in Bewo cells. We observed currents that were blocked by 10 µM amiloride in cells incubated in 100 nM aldosterone for 12 hs. The amplitude of this current was 20-fold the basal current, a reversal potential of 3 mV and a conductance of 127 ± 26 pS/pF with pulses between -60 and -140 mV. These characteristics are similar to those reported in ENaC channels in several tissues. Although their roles in placenta are still poorly understood, the differences in the expression of ENaC in pre-eclamptic placentas may have consequences for ion transport and these data could lead to future studies concerning the mechanism involved in the pathophysiology of pre-eclampsia. El sinciciotrofoblasto (SCT) de placenta humana regula la transferencia de solutos y agua entre la sangre fetal y materna. En el presente trabajo observamos que el canal de sodio ENaC (asociado a cuadros como el síndrome de Liddle y pseudohipoaldosteronismo) está presente en la membrana apical del SCT y que la subunidad α del canal tiene una expresión reducida en placentas con hipertensión gestacional (preeclampsia). Realizamos estudios a nivel de expresión de ARN (RT-PCR) y a nivel proteico (western blot e inmunohistoquímica). En la línea celular BeWo (modelo de SCT humano) el canal se encuentra presente y la expresión del mismo es regulada por las hormonas aldosterona, vasopresina, estradiol y progesterona. Analizamos la actividad del ENaC por electrofisiología y observamos corrientes sensibles a amiloride (10 µM) cuando las células BeWo se cultivaron 12 horas con aldosterona (100 nM). Esta corriente presentó una magnitud 20 veces mayor que las corrientes basales, un potencial de reversión cercano a 3 mV y una conductancia de 127 ± 26 pS/ pF entre los pulsos de –60 y –140 mV aplicados. Las características de esta corriente son similares a las producidas por ENaC en otros tejidos y evidencian la presencia de un canal funcional. El papel del ENaC en el SCT es poco comprendido, aunque la diferencia de expresión en la preeclampsia podría tener consecuencias para el transporte placentario de agua y iones. Nuestros datos son un aporte para futuros estudios de los mecanismos involucrados en la patofisiología de la preeclampsia.
Characterization of the epithelial sodium channel in human preeclampsia syncytiotrophoblast
Título:
Characterization of the epithelial sodium channel in human pre-eclampsia syncytiotrophoblast
del Monaco, Silvana Maria
; Assef, Yanina Andrea
; Damiano, Alicia Ermelinda
; Zotta, Elsa; Ibarra, Cristina Adriana
; Kotsias, Basilio Aristides
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Fecha de publicación:
12/2006
Editorial:
Fundación Revista Medicina
Revista:
Medicina (Buenos Aires)
ISSN:
0025-7680
e-ISSN:
1669-9106
Idioma:
Español
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
Palabras clave:
ENaC
,
placenta humana
,
preeclampsia
,
western blot
,
inmunohistoquímica
Archivos asociados
Licencia
Identificadores
Colecciones
Articulos(OCA HOUSSAY)
Articulos de OFICINA DE COORDINACION ADMINISTRATIVA HOUSSAY
Articulos de OFICINA DE COORDINACION ADMINISTRATIVA HOUSSAY
Citación
del Monaco, Silvana Maria; Assef, Yanina Andrea; Damiano, Alicia Ermelinda; Zotta, Elsa; Ibarra, Cristina Adriana; et al.; Characterization of the epithelial sodium channel in human preeclampsia syncytiotrophoblast; Fundación Revista Medicina; Medicina (Buenos Aires); 66; 1; 12-2006; 31-35
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