Summary
Morphologic findings of widely dilated intercellular spaces in fluid transporting epithelia have been claimed as evidence for the existence of an epithelial compartment in which the coupling between solute and water fluxes takes place. The validity of using epithelial geometry in sectioned material as an argument can be questioned. The present report describes the morphological appearance of frog gallbladder epithelium — normal and ouabain-treated — in the living state in vitro and after fixation, dehydration and embedding. Gallbladder segments were photographed in the living state and at the end of each step of the preparative procedure. Direct observations of whole-mounted gallbladder segments were carried out, taking advantage of the possibility of optical sectioning and high resolution by Nomarski-microscopy. The same specimens were then sectioned and examined by conventional light and electron microscopy. The observations were quantitated and showed that the epithelial cells of normal and ouabain-treated gallbladders experienced an average linear shrinkage down to 70% of their length in Ringer's solution, which corresponds to a volume shrinkage down to 35%. Moreover, dilated lateral intercellular spaces appeared during the dehydration and embedding procedure in normal but only very moderately or not at all in ouabain-treated gallbladder specimens.
Similar content being viewed by others
References
Allen RD, David GB, Nomarski G (1969) The Zeiss-Nomarski differential interference equipment for transmitted-light microscopy. Z Wiss Mikrosk 69:193–221
Bahr GF, Bloom G, Friberg U (1957) Volume changes of tissues in physiological fluids during fixation in osmium tetroxide or formaldehyde and during subsequent treatment. Exp Cell Res 12:342–355
Bindslev N, Tormey J McD, Wright EM (1974) The effects of electrical and osmotic gradients on lateral intercellular spaces and membrane conductance in a low resistance epithelium. J Membr Biol 19:357–380
Blom H, Helander HF (1977) Quantitative electron microscopical studies on in vitro incubated rabbit gallbladder epithelium. J Membr Biol 37:45–61
Bloom W, Fawcett DW (1975) A textbook of histology, 10th ed. W B Saunders Company, Philadelphia London Toronto, pp 1–1033
Bone Q, Denton EJ (1971) The osmotic effects of electron microscope fixatives. J Cell Biol 49:571–581
Bone Q, Ryan KP (1972) Osmolarity of osmium tetroxide and glutaraldehyde fixatives. Histochem J 4:331–347
Borysko E (1956) Recent developments in methacrylate embedding. I. A study of the polymerization damage phenomenon by phase contrast microscopy. J Biophys Biochem Cytol 2:3–14
Boyde A, Bailey E, Jones SJ, Tamarin A (1977) Dimensional changes during specimen preparation for scanning electron microscopy. Scanning Electron Microsc 1:507–518
Curran PF, MacIntosh JR (1962) Model system for biological water transport. Nature 193:347
Diamond JM (1962) The reabsorptive function of the gallbladder. J Physiol 161:442–473
Diamond JM (1964) Transport of salt and water in rabbit and guinea pig gallbladder. J Gen Physiol 48:1–14
Diamond JM (1968) Transport mechanisms in the gallbladder. In: Code CF (ed) Handbook of Physiology, section 6. Williams & Wilkins Company, Baltimore, pp 2451–2482
Diamond JM, Tormey J McD (1966) Role of long extracellular channels in fluid transport across epithelia. Nature 210:817–820
Diamond JM, Bossert WH (1967) Standing-gradient osmotic flow. A mechanism for coupling of water and solute transport in epithelia. J Gen Physiol 50:2061–2083
Eisenberg BR, Mobley BA (1975) Size changes in single muscle fibers during fixation and embedding. Tissue & Cell 7:383–387
Frederiksen O (1978) Functional distinction between two transport mechanisms in rabbit gallbladder epithelium by use of ouabain, ethacrynic acid and metabolic inhibitors. J Physiol 280:373–387
Frederiksen O, Leyssac PP (1969) Transcellular transport of isosmotic volumes by the rabbit gallbladder in vitro. J Physiol 201:201–224
Frederiksen O, Rostgaard J (1974) Absence of dilated lateral intercellular spaces in fluid-transporting frog gallbladder epithelium. Direct microscopy observations. J Cell Biol 61:830–834
Frederiksen O, Møllgård K, Rostgaard J (1979) Lack of correlation between transepithelial transport capacity and paracellular pathway ultrastructure in Alcian blue-treated rabbit gallbladders. J Cell Biol 83:383–393
Hertwig G (1931) Der Einfluss der Fixierung auf das Kern-und Zellvolumen. Z Mikrosk Anat Forsch 23:484–504
Hill AE (1975) Solute-solvent coupling in epithelia: contribution of the junctional pathway to fluid production. Proc R Soc London B 191:537–547
Hill BS, Hill AE (1978) Fluid transfer by Necturus gallbladder epithelium as a function of osmolarity. Proc R Soc London B 200: 151–162
Kaye GI, Wheeler HO, Whitlock RT, Lane N (1966) Fluid transport in the rabbit gallbladder. Acombined physiological and electron microscopic study. J Cell Biol 30:237–268
Kushida H (1962) A study of cellular swelling and shrinkage during fixation, dehydration and embedding in various standard media. J Electron Microsc 11:135–138
Loeschke K, Eisenbach GM, Bentzel CJ (1975) Water flow across Necturus gallbladder and small intestine. Excerpta Med Int Congr Ser 391:406–412
Luciano L (1972) Die Feinstruktur der Gallenblase und der Gallengänge. I. Das Epithel der Gallenblase der Maus. Z Zellforsch 135:87–102
Luft JH (1961) Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol 9:409–414
Luft JH (1973) Embedding media — old and new. In: Koehler JK (ed) Advanced Techniques in Biological Electron Microscopy. Springer, Berlin Heidelberg New York, pp 1–34
Machen TE, Diamond JM (1969) An estimate of the salt concentration in the lateral intercellular spaces of rabbit gall-bladder during maximal fluid transport. J Membr Biol 1:194–213
Mathieu O, Claassen H, Weibel ER (1978) Differential effect of glutaraldehyde and buffer osmolarity on cell dimensions: A study on lung tissue. J Ultrastruct Res 63:20–34
Maunsbach AB (1966) The influence of different fixatives and fixation methods on the ultrastructure of rad kidney proximal tubule cells. II. Effects of varying osmolality, ionic strength, buffer system and fixative concentration of glutaraldehyde solutions. J Ultrastruct Res 15:283–309
Millonig G, Marinozzi V (1968) Fixation and embedding in electron microscopy. In: Cosslett VE (ed) Advances in optical and electron microscopy. Vol 2. Academic Press, London New York, pp 282–341
Mueller JC, Jones AL, Long JA (1972) Topographic and subcellular anatomy of the Guinea pig gallbladder. Gastroenterology 63:856–868
Patlak CS, Goldstein DA, Hoffman JF (1963) The flow of solute and solvent across a two-membrane system. J Theoret Biol 5:426–442
Porter K, Prescott D, Frye J (1973) Changes in surface morphology of Chinese hamster ovary cells during the cell cycle. J Cell Biol 57:815–836
Poyton RO, Branton D (1970) A multipurpose microperfusion chamber. Exp Cell Res 60:109–114
Smulders AP, Tormey J McD, Wright EM (1972) The effect of osmotically induced water flows on the permeability and ultrastructure of the rabbit gallbladder. J Membr Biol 7:164–197
Spring KR (1979) Optical techniques for the evaluation of epithelial transport processes. Am J Physiol 237:F167-F174
Spring KR, Hope A (1978) Size and shape of the lateral intercellular spaces in a living epithelium. Science 200:54–58
Spring KR, Hope A (1979) Fluid transport and the dimensions of cells and interspaces of living Necturus gallbladder. J Gen Physiol 73:287–305
Tormey J McD, Diamond JM (1967) The ultrastructural route of fluid transport in rabbit gallbladder. J Gen Physiol 50:2031–2061
Van Os CH, Slegers JFG (1971) Correlation between (Na+-K+)-activated ATPase activities and the rate of isotonic fluid transport of gallbladder epithelium. Biochim Biophys Acta 241:89–96
Webster H deF, Ames A III, Nesbett FB (1969) A quantitative morphological study of osmotically induced swelling and shrinkage in nervous tissue. Tissue & Cell 1:201–216
Weibel ER, Knight BW (1964) A morphometric study on the thickness of the pulmonary air-blood barrier. J Cell Biol 21:367–384
Welling DJ, Welling LW, Hill JJ (1978) Phenomenological model relating cell shape to water reabsorption in proximal nephron. Am J Physiol 234:F308-F317
Whitlock RT, Wheeler HO (1964) Coupled transport of solute and water across rabbit gallbladder epithelium. J Clin Invest 43:2249–2265
Wüstenfeld E (1955) Experimentelle Beiträge zur Frage der Volumenänderungen und Eindringdauer in der histologischen Technik. Z Wiss Mikrosk 63:7–15
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Rostgaard, J., Frederiksen, O. Fluid transport and dimensions of epithelial cells and intercellular spaces in frog gallbladder. Cell Tissue Res. 215, 223–247 (1981). https://doi.org/10.1007/BF00239111
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00239111