Laboratory Investigation

Kidney International (1981) 19, 491–502; doi:10.1038/ki.1981.47

Surface characteristics of carbonic-anhydrase-rich cells in turtle urinary bladder

Russell F Husted1, Allan L Mueller1, Richard G Kessel1 and Philip R Steinmetz1

1Laboratory of Epithelial Transport and Kidney Physiology, Department of Medicine and Department of Zoology, University of Iowa, Iowa City, Iowa

Correspondence: Dr R F Husted, 317 Medical Laboratories, University of Iowa, Iowa City, Iowa 52242, USA

Received 24 March 1980; Revised 4 May 1980.

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Abstract

Surface characteristics of carbonic-anhydrase-rich cells in turtle urinary bladder. Addition of a disulfonic stilbene, 4-acetamido-4'-isothiocyano-2,2'-disulfonic stilbene (SITS), to the serosal side of the turtle bladder blocks the efflux of bicarbonate ions from the acidifying cells and thereby inhibits hydrogen ion secretion into the luminal solution. Because SITS has little effect on other transport systems, we used it to define the relationship between hydrogen ion secretion (JH) and the different cell types facing the luminal surface. Cells were identified by scanning and transmission electron microscopy (SEM and TEM) and by histochemical localization of carbonic anhydrase (CA). SEM revealed that SITS caused marked alterations in luminal surface characteristics of a cell population with prominent microplicae. Two hours after the serosal addition of SITS, cells with identifiable microplicae had decreased from 12.7 to 0.5% of total cells. TEM studies and CA histochemistry showed that the number of cells rich in CA (CA cells) remained the same, whereas the individual luminal surface areas of a subpopulation of CA cells had decreased markedly. A comparison of the distribution of individual surface areas of cells with microplicae and CA cells revealed that the CA cells with large surface areas corresponded to the cells with microplicae and that both were affected by the serosal addition of SITS. Acetazolamide, which also inhibits JH caused similar changes. The luminal addition of SITS and an inactive analogue of acetazolamide, which have no effect on JH did not alter surface morphology. These results indicate that the CA cell with microplicae represents the active state of the hydrogen ion secreting cell.

Caractéristiques de surface des cellules riches en anhydrase carbonique de la vessie urinaire de tortue. L'action du stilbène disulfonique, 4-acetamido-4'-isothiocyano-2,2'-disulfonic stilbene (SITS), sur la face séreuse de la vessie de tortue bloque la sécrétion d'ions hydrogène vers la solution luminale. Du fait que le SITS a peu d'effets sur les autres systèmes de transport il a été employé pour définir la relation entre la sécrétion d'ions hydrogène (JH) et les différents types cellulaires qui sont en regard de la surface luminale. Les cellules ont été identifiées en microsco-pie électronique à balayage et ` transmission (SEM et TEM) et par la localisation histoenzymologique de l'anhydrase carbonique (CA). La SEM a montré que le SITS détermine des modifications des caractéristiques de la surface luminale d'une population cellulaire à microplis proéminents. Deux heures après l'addition séreuse de SITS, les cellules ayant des microplis identifiables ont diminué de 12,7 à 0,5% du nombre total de cellules. Les études au TEM et l'histochimie de CA montre que le nombre des cellules riches en CA (cellules CA) est le même alors que la surface luminale d'une sous-population de cellules CA a considérablement diminué. Une comparaison de la distribution des surfaces individuelles des cellules à microplis et des cellules CA montre que les cellules CA à grande surface correspondent aux cellules à microplis et que les deux sont affectées par le SITS du côté séreux. L'acétazolamide, qui inhibe aussi JH, détermine des modifications semblables. Le SITS luminal et un analogue inactif de l'acétazolamide qui n'a pas d'effet sur JH ne modifient pas la morphologie de surface. Ces résultats indiquent que les cellules CA à microplis représentent l'état actif de la cellule sécrétant d'ions hydrogène.

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