SHIGA TOXIN TYPE 2 (STX2) INCREASES CELL VOLUME OF HUMAN GLOMERULAR ENDOTHELIAL CELLS (HGEC) THAT EXPRESS AQUAPORIN 1 AND AQUAPORIN 4 (AQP1 AND AQP4)

FERNANDO DANIEL GOMEZ; JULIETA REPETTI; CLAUDIO DANIEL GIRÓN REYES; FLAVIA SACERDOTI; CRISTINA IBARRA; NORA MARTINEZ ; ALICIA DAMIANO ; GISELA DI GIUSTO; AMARAL MARÍA MARTA

Mar del Plata

Congreso; Reunion Anual de Sociedades de Investigación Clinica; 2022

Sociedad de Investigación clinica

Hemolytic Uremic Syndrome associated to Shiga toxin–producing Escherichia coli (STEC-HUS) infection is the major cause ofacute renal injury in pediatric age groups in Argentina. Previously,we demonstrated that Stx2 inhibited the net water transport (Jw)across HGEC monolayers, possibly as a consequence of direct alterations in the Jw mechanisms. In addition, we showed that Stx2increased the cell area of HGEC and AQPs inhibitors were able toreverse this effect. Therefore, in this work we evaluate changes incell volume of monolayers of HGEC treated with Stx2 as well aswe analyze the expression of both AQP1 and AQP4 by these cells.Immunohistochemical and immunofluorescence studies were performed in HGEC that were grown on gelatinized glass coverslips (12mm) and then treated or not with Stx2 for 40 minutes. Cell imageswere acquired and digitized using a confocal microscope OlympusFluoView (FV1000). A z-stack consisting of a series of 0.5 µm optical 2D slices was taken from top to bottom of the cells and then 3Dreconstructed. The measure of total cell volume and the analysis ofAQP1 or AQP4 expression and localization in HGEC cells imageswere processed using the Surface tool of Imaris 7.1.0 software. Ourresults show that HGEC treated with Stx2 exhibited an increase inthe cell volume (Stx2: 9374 ± 992 µm3 vs CTRL 4411.15 ± 470 µm3 n=6, p