Histochemistry and Cell Biology

, Volume 121, Issue 3, pp 189–199 | Cite as

Cell type-specific and developmentally regulated expression of the AE1 anion exchanger in the chicken chorioallantoic membrane

  • M. Gabriella GabrielliEmail author
  • John V. Cox
  • Giovanni Materazzi
  • Giovanna Menghi
Original Paper


Antibodies specific for the chicken AE1 anion exchanger have been used to determine the cell-type specific pattern of expression of this electroneutral transporter in the chick chorioallantoic membrane (CAM) during embryonic development. Immunolocalisation analyses demonstrated that the AE1 anion exchanger accumulated in the basolateral membrane of a subset of cells in both the chorionic and allantoic epithelial layers. Double immunostaining indicated that the AE1-positive cells in the chorionic and allantoic epithelia were also positive for the carbonic anhydrase isoform, CAII, which serves as a marker for the villus cavity (VC) cells of the chorionic epithelium and the mitochondria-rich cells of the allantoic epithelium. Immunoelectron microscopy revealed that AE1 accumulated in extensive projections that extended from the lateral membrane of VC cells towards the adjacent capillary covering cells. These results represent the first demonstration of anion exchanger expression in the chick CAM, and they suggest a role for basolateral AE1 in bicarbonate reabsorption that is required in the embryo for maintaining acid-base balance during development.


AE1 anion exchangers Chorioallantoic membrane Carbonic anhydrase Immunochemistry TEM CLSM 



We are grateful to Dr. P. J. Linser for the monoclonal antibody against chicken carbonic anhydrase and to Garbini S.p.A. for the fertilised eggs. The skilful technical assistance of Stefano Riccioni and Simonetta Cammertoni is gratefully acknowledged. This study was supported by grants from the University of Camerino.


  1. Adair-Kirk TL, Cox KH, Cox JV (1999) Intracellular trafficking of variant chicken kidney AE1 anion exchangers: role of alternative NH2 termini in polarized sorting and Golgi recycling. J Cell Biol 147:1237–1248CrossRefPubMedGoogle Scholar
  2. Akins RE, Tuan RS (1993) Transepithelial calcium transport in the chick chorioallantoic membrane. I. Isolation and characterization of chorionic ectoderm cells. J Cell Sci 105:369–379Google Scholar
  3. Alper SL (1994) The band 3-related AE anion exchanger gene family. Cell Physiol Biochem 4:265–281Google Scholar
  4. Alper SL, Natale J, Gluck S, Lodish HF, Brown D (1989) Subtypes of intercalated cells in rat kidney collecting duct defined by antibodies against erythroid band 3 and renal vacuolar H+-ATPase. Proc Natl Acad Sci U S A 86:5429–5433PubMedGoogle Scholar
  5. Breton S, Alper SL, Gluck SL, Sly WS, Barker JE, Brown D (1995) Depletion of intercalated cells from collecting ducts of carbonic anhydrase II-deficient (CAR 2 null) mice. Am J Physiol 269:F761–F774PubMedGoogle Scholar
  6. Brosius FC III, Nguyen K, Stuart-Tilley AK, Haller C, Briggs JP, Alper SL (1995) Regional and segmental localization of AE2 anion exchanger mRNA and protein in rat kidney. Am Phys Soc 269:F461–F468Google Scholar
  7. Brown D, Breton S (1996) Mitochondria-rich, proton-secreting epithelial cells. J Exp Biol 199:2345–2358PubMedGoogle Scholar
  8. Castillo JE, Martinez-Anso E, Malumbres R, De Alava E, Garcìa C, Medina JF, Prieto J (2000) In situ localization of anion exchanger-2 in the human kidney. Cell Tissue Res 299:281–287PubMedGoogle Scholar
  9. Choi I, Romero MF, Khandoudi N, Bril A, Boron WF (1999) Cloning and characterization of a human electrogenic Na+-HCO3 cotransporter isoform (hhNBC). Am J Physiol 276:C576–C584PubMedGoogle Scholar
  10. Coleman JR, Terepka AR (1972) Fine structural changes associated with the onset of calcium, sodium and water transport by the chicken chorioallantoic membrane. J Membr Biol 7:111–127Google Scholar
  11. Cox KH, Cox JV (1995) Variant chicken AE1 anion exchangers possess divergent NH2-terminal cytoplasm domains. Am J Physiol 268:F503–F513PubMedGoogle Scholar
  12. Cox KH, Adair-Kirk TL, Cox JV (1995) Four variant chicken erythroid AE1 anion exchangers. J Biol Chem 270:19752–19760CrossRefPubMedGoogle Scholar
  13. Cox KH, Adair-Kirk TL, Cox JV (1996) Variant AE2 anion exchanger transcripts accumulate in multiple cell types in the chicken gastric epithelium. J Biol Chem 271:8895–8902CrossRefPubMedGoogle Scholar
  14. Crooks RJ, Simkiss K (1974) Respiratory acidosis and eggshell reabsorption by the chick embryo. J Exp Biol 61:197–202PubMedGoogle Scholar
  15. Dawes CM, Simkiss K (1969) The acid-base status of the blood of the developing chick embryo. J Exp Biol 50:79–86Google Scholar
  16. Dawes CM, Simkiss K (1971) The effects of respiratory acidosis in the chick embryo. J Exp Biol 55:77–84PubMedGoogle Scholar
  17. Drenckhahn D, Schluter K, Allen DP, Bennett V (1985) Colocalization of band 3 with ankyrin and spectrin at the basal membrane of intercalated cells in the rat kidney. Science 230:1287–1289PubMedGoogle Scholar
  18. Freeman BM, Vince MA (1974) Development of the avian embryo. A behavioural and physiological study. Chapman and Hall, LondonGoogle Scholar
  19. Gabrielli MG, Materazzi G, Cox JV, Menghi G (2001) Specialised cell types in the chorioallantoic membrane express carbonic anhydrase during chick embryogenesis. J Anat 198:229–238CrossRefPubMedGoogle Scholar
  20. Gabrielli MG, Materazzi G, Bondi AM, Menghi G (2003) Developmental expression of glycocomponents in the chick chorioallantoic membrane. Anat Embryol 207:63–72CrossRefPubMedGoogle Scholar
  21. Garcìa C, Montuenga LM, Medina JF, Prieto J (1998) In situ detection of AE2 anion exchanger mRNA in the human liver. Cell Tissue Res 291:481–488CrossRefPubMedGoogle Scholar
  22. Giffard RG, Papadopoulos MC, van Hooft JA, Xu LJ, Giuffrida R, Monyer H (2000) The electrogenic sodium bicarbonate cotransporter: developmental expression in rat brain and possible role in acid vulnerability. J Neurosci 20:1001–1008PubMedGoogle Scholar
  23. Holappa K, Mustonen M, Parvinen M, Vihko P, Rajaniemi H, Kellokumpu S (1999) Primary structure of a sperm cell anion exchanger and its messenger ribonucleic acid expression during spermatogenesis. Biol Reprod 61:981–986PubMedGoogle Scholar
  24. Jennings ML (1992) Anion transport protein. In: Seldin DW, Giebischs G (eds) The kidney physiology and pathophysiology. Raven, New York, pp 503–535Google Scholar
  25. Jensen LJ, Stuart-Tilley AK, Peters LL, Lux SE, Alper SL, Breton S (1999) Immunolocalization of AE2 anion exchanger in rat and mouse epididymis. Biol Reprod 61:973–980PubMedGoogle Scholar
  26. Jons T, Warrings B, Jons A, Drenckhahn D (1994) Basolateral localization of anion exchanger 2 (AE2) and actin in acid-secreting (parietal) cells of the human stomach. Histochemistry 102:255–263PubMedGoogle Scholar
  27. Kollert-Jons A, Wagner S, Hubner S, Appelhans H, Drenckhahn D (1993) Anion exchanger 1 in human kidney and oncocytoma differs from erythroid AE1 in its NH2 terminus. Am J Physiol 265:F813–F821PubMedGoogle Scholar
  28. Kopito RR (1990) Molecular biology of the anion exchanger gene family. Int Rev Cytol 123:177–199PubMedGoogle Scholar
  29. Kudrycki KE, Shull GE (1993) Rat kidney band 3 chloride/bicarbonate exchanger mRNA is transcribed from an alternative promoter. Am J Physiol 264:F540–F547PubMedGoogle Scholar
  30. Kudrycki KE, Newman PR, Shull GE (1990) cDNA cloning and tissue distribution of mRNAs for two proteins that are related to the band 3 Cl/HCO3 exchanger. J Biol Chem 265:462–471PubMedGoogle Scholar
  31. Linn SC, Kudrycki KE, Shull GE (1992) The predicted translation product of the cardiac AE3 mRNA contains an N terminus distinct from that of the brain AE3 Cl/HCO3 exchanger. J Biol Chem 267:7927–7935PubMedGoogle Scholar
  32. Linn SC, Askew G, Menon AG, Shull GE (1995) Conservation of an AE3 Cl/HCO3 exchanger cardiac-specific exon and promoter region and AE3 mRNA expression patterns in murine and human hearts. Circ Res 76:584–591PubMedGoogle Scholar
  33. Linser P, Perkins MS, Fitch FW, Moscona AA (1984) Comparative characterization of monoclonal antibodies to carbonic anhydrase. Biochem Biophys Res Commun 125:690–697PubMedGoogle Scholar
  34. Narbaitz R, Bastani B, Galvin NJ, Kapal VK, Levine DZ (1995) Ultrastructural and immunocytochemical evidence for the presence of polarized plasma membrane H+-ATPase in two specialised cell types in the chick chorioallantoic membrane. J Anat 186:245–252PubMedGoogle Scholar
  35. Park K, Hurley PT, Roussa E, Cooper GJ, Smith CP, Thévenod F, Steward MC, Case RM (2002) Expression of a sodium bicarbonate cotransporter in human parotid salivary glands. Arch Oral Biol 47:1–9CrossRefPubMedGoogle Scholar
  36. Reithmeier RAF (2001) A membrane metabolon linking carbonic anhydrase with chloride/bicarbonate anion exchangers. Blood Cells Mol Dis 27:85–89CrossRefPubMedGoogle Scholar
  37. Romero MF, Fong PY, Berger UV, Hediger MA, Boron WF (1998) Cloning and functional expression of rNBC, an electrogenic Na+-HCO3 cotransporter from rat kidney. Am J Physiol 274:F425–F432PubMedGoogle Scholar
  38. Roussa E (2001) H+ and HCO3 transporters in human salivary ducts. An immunohistochemical study. Histochem J 33:77–84Google Scholar
  39. Roussa E, Romero MF, Schmitt BM, Boron WF, Alper SL, Thévenod F (1999) Immunolocalization of anion exchanger AE2 and Na+-HCO3 cotransporter in rat parotid and submandibular glands. Am J Physiol 277:G1288–G1296PubMedGoogle Scholar
  40. Schwartz GJ, Barasch J, Al-Awqati Q (1985) Plasticity of functional epithelial polarity. Nature 318:368–371PubMedGoogle Scholar
  41. Sterling D, Reithmeier RA, Casey JR (2001) A transport metabolon. Functional interaction of carbonic anhydrase II and chloride/bicarbonate exchangers. J Biol Chem 276:47886–47894PubMedGoogle Scholar
  42. Stewart ME, Terepka AR (1969) Transport functions of the chick chorio-allantoic membrane. 1. Normal histology and evidence for active electrolyte transport from the allantoic fluid, in vivo. Exp Cell Res 58:93–106PubMedGoogle Scholar
  43. Stuart-Tilley AK, Shmukler BE, Brown D, Alper SL (1998) Immunolocalization and tissue-specific splicing of AE2 anion exchanger in mouse kidney. J Am Soc Nephrol 9:946–959PubMedGoogle Scholar
  44. Thévenod F, Roussa E, Schmitt BM, Romero MF (1999) Cloning and immunolocalization of a rat pancreatic Na+ bicarbonate cotransporter. Biochem Biophys Res Commun 264:291–298CrossRefPubMedGoogle Scholar
  45. Vince JW, Reithmeier RAF (1998) Carbonic anhydrase II binds to carboxyl terminus of human band 3, the erythrocyte Cl/HCO3 exchanger. J Biol Chem 273:28430–28437CrossRefPubMedGoogle Scholar
  46. Vince JW, Reithmeier RAF (2000) Identification of the carbonic anhydrase II binding site in the Cl/HCO3 anion exchanger AE1. Biochemistry 39:5527–5533CrossRefPubMedGoogle Scholar
  47. Wagner SR, Vogel R, Lietzke R, Koob R, Drenckhahn D (1987) Immunochemical characterization of the human band 3-like anion exchanger in collecting duct of human kidney. Am J Physiol 253:F213–F222PubMedGoogle Scholar
  48. Yannoukakos D, Stuart-Tilley AK, Fernandez HA, Fey P, Duyk G, Alper SL (1994) Molecular cloning, expression and chromosomal localization of two isoforms of the AE3 anion exchanger from human heart. Circ Res 75:603–614PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • M. Gabriella Gabrielli
    • 1
    Email author
  • John V. Cox
    • 2
  • Giovanni Materazzi
    • 1
  • Giovanna Menghi
    • 1
  1. 1.Department of Comparative Morphology and BiochemistryUniversity of CamerinoCamerino (MC)Italy
  2. 2.Department of Molecular SciencesUniversity of Tennessee Health Science CenterMemphisUSA

Personalised recommendations