Skip to main content

Main menu

  • Home
  • Content
    • Published Ahead of Print
    • Current Issue
    • JASN Podcasts
    • Article Collections
    • Archives
    • Kidney Week Abstracts
    • Saved Searches
  • Authors
    • Submit a Manuscript
    • Author Resources
  • Editorial Team
  • Editorial Fellowship
    • Editorial Fellowship Team
    • Editorial Fellowship Application Process
  • More
    • About JASN
    • Advertising
    • Alerts
    • Feedback
    • Impact Factor
    • Reprints
    • Subscriptions
  • ASN Kidney News
  • Other
    • ASN Publications
    • CJASN
    • Kidney360
    • Kidney News Online
    • American Society of Nephrology

User menu

  • Subscribe
  • My alerts
  • Log in
  • My Cart

Search

  • Advanced search
American Society of Nephrology
  • Other
    • ASN Publications
    • CJASN
    • Kidney360
    • Kidney News Online
    • American Society of Nephrology
  • Subscribe
  • My alerts
  • Log in
  • My Cart
Advertisement
American Society of Nephrology

Advanced Search

  • Home
  • Content
    • Published Ahead of Print
    • Current Issue
    • JASN Podcasts
    • Article Collections
    • Archives
    • Kidney Week Abstracts
    • Saved Searches
  • Authors
    • Submit a Manuscript
    • Author Resources
  • Editorial Team
  • Editorial Fellowship
    • Editorial Fellowship Team
    • Editorial Fellowship Application Process
  • More
    • About JASN
    • Advertising
    • Alerts
    • Feedback
    • Impact Factor
    • Reprints
    • Subscriptions
  • ASN Kidney News
  • Follow JASN on Twitter
  • Visit ASN on Facebook
  • Follow JASN on RSS
  • Community Forum
Cell and Transport Physiology
You have accessRestricted Access

Distal Renal Tubular Acidosis in Mice Lacking the AE1 (Band3) Cl−/HCO3− Exchanger (slc4a1)

Paul A. Stehberger, Boris E. Shmukler, Alan K. Stuart-Tilley, Luanne L. Peters, Seth L. Alper and Carsten A. Wagner
JASN May 2007, 18 (5) 1408-1418; DOI: https://doi.org/10.1681/ASN.2006101072
Paul A. Stehberger
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Boris E. Shmukler
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Alan K. Stuart-Tilley
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Luanne L. Peters
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Seth L. Alper
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Carsten A. Wagner
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Article
  • Figures & Data Supps
  • Info & Metrics
  • View PDF
Loading

Article Figures & Data

Figures

  • Tables
  • Additional Files
  • Figure 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 1.

    Urine and blood gas analysis. Urinary pH and blood gas values of wild-type (Ae1+/+), heterozygous (Ae1+/−) and AE1-deficient (Ae1−/−) mice during metabolic cage experiments under control conditions for 2 d and after oral acid loading with 0.28 M NH4Cl for a third day. (A) Urinary pH in AE1-deficient mice was more alkaline than for other genotypes in all conditions but did acidify in response to the oral NH4Cl load. Blood pH (B) and [HCO3−] concentration (C) in wild-type and heterozygous mice were indistinguishable in all conditions. However, blood pH and [HCO3−] in Ae1−/− mice under control conditions revealed metabolic acidosis. This acidosis was severely exacerbated after NH4Cl loading (n = 8 mice for each genotype and time point). *P < 0.001 versus control conditions in the same genotype; #P < 0.001 versus wild-type.

  • Figure 2.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 2.

    Measurement of Cl−/HCO3− exchanger activity in single type A intercalated cells (A-IC) in isolated outer medullary collecting duct (OMCD). (A and B) Representative original tracings of intracellular pH measurements (BCECF ratio fluorimetry) in the presence of HCO3−/CO2 and the alternating presence or absence of extracellular chloride. Addition of chloride caused a rapid acidification as a result of activation of Cl−/HCO3− exchange activity. The intracellular acidification rate was reduced by 400 nM of the AE1 inhibitor diBA(5)C4. Intracellular acidification was partially reduced in A-IC from Ae1−/− mice, and the remaining activity was insensitive to diBA(5)C4. (C) Summarized data from all experiments using the inhibitors diBA(5)C4 (400 nM) or DIDS (50 μM; n = 4 to 5 mice with five to six OMCD for each condition and 100 to 150 A-IC each). *P < 0.001 versus wild-type.

  • Figure 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 3.

    Expression of anion exchangers in kidney. (A) reverse transcriptase–PCR with primers detecting specific isoforms of several anion exchanger mRNA as well as KCC4 and aquaporin-6 (AQP6), two transport proteins implicated in A-IC function, did not reveal consistent changes in mRNA abundance in kidneys from Ae1−/− mice. β-Actin was used as a reference transcript. (B) Immunoblot showed that abundance of AE4/Slc4a9 polypeptide was not changed in kidneys from Ae1−/− mice. The abundance of pendrin/Slc26a4 anion exchanger polypeptide was strongly reduced in kidneys from Ae1−/− mice.

  • Figure 4.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 4.

    Severe urinary concentrating defect in Ae1−/− mice. (A) Under control conditions, urine volume normalized to body weight was approximately three-fold elevated in AE1-deficient mice and remained two-fold elevated during the severe dehydration produced by acid loading (see serum osmolarity, Table 2). (B) Urine osmolarity was much lower in AE1-deficient mice and did not increase during acid load–induced dehydration. (C) The acid load–induced loss of body weight in Ae1−/− mice was greatly increased. *P < 0.001 versus wild-type.

  • Figure 5.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 5.

    Dysregulation of AQP2 water channel in kidney of AE1-deficient mice. Immunoblots reveal increased AQP2 protein abundance in kidney cortex (A) and medulla (B) of Ae1−/− mice in control conditions. (C) Confocal immunofluorescence microscopy of outer medulla (top) shows increased AQP2 immunostaining (green) with strong apical localization in AE1-deficient mice. In contrast, in the inner medulla of Ae1−/− kidneys (bottom), AQP2 staining was confined mainly to intracellular vesicular structures, whereas AQP2 in wild-type kidney was predominantly luminal. AE1 staining is in red. Magnification, ×600.

  • Figure 6.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 6.

    Nephrocalcinosis in kidneys of Ae1−/− mice. (A) The fractional excretion (FE %) of calcium and phosphate was increased in acid-loaded AE1-deficient mice, whereas citrate excretion was reduced. **P < 0.001. (B) Van Kossa staining for calcium phosphate deposits (brown-black) detected tubular calcification predominantly in the medulla of Ae1−/− mice (bottom middle and right). Magnifications: ×100 (left), ×200 (middle), and ×400 (right).

Tables

  • Figures
  • Additional Files
    • View popup
    Table 1.

    Control conditionsa

    GenotypeAe1+/+Ae1+/−Ae1−/−
    Blood
        pH7.27 ± 0.027.30 ± 0.027.12 ± 0.02b
        HCO3−22.0 ± 0.822.2 ± 0.317.3 ± 0.5b
        Pco248.8 ± 1.744.9 ± 1.055.8 ± 2.1
        K+3.7 ± 0.13.5 ± 0.04.4 ± 0.1c
        Na+147.3 ± 1.1145.2 ± 0.8149.1 ± 1.5
        Cl−113.3 ± 1.1112.2 ± 1.1116.7 ± 1.2c
        Pi1.6 ± 0.11.6 ± 0.12.4 ± 0.3c
        anion gap12.0 ± 0.810.8 ± 0.215.1 ± 1.1
        osmolarity315.1 ± 2.6310.2 ± 1.5336.0 ± 9.1c
        creatinine0.07 ± 0.01ND0.11 ± 0.02
        urea6.4 ± 1.6ND26.8 ± 3.8b
    Urine
        pH6.48 ± 0.166.38 ± 0.136.77 ± 0.12
        osmolarity2519.8 ± 176.32336 ± 279.11085.1 ± 103.1b
        creatinine68.9 ± 3.866.2 ± 6.627.1 ± 3.4b
        HCO3−/creatinine0.2 ± 0.10.1 ± 0.00.5 ± 0.2
        K+/creatinine7.4 ± 1.02.9 ± 0.4c6.9 ± 1.1
        Na+/creatinine3.5 ± 0.51.2 ± 0.2c3.3 ± 0.5
        Cl−/creatinine3.8 ± 0.33.7 ± 0.64.2 ± 0.3
        Pi/creatinine0.7 ± 0.10.7 ± 0.10.5 ± 0.1
        NH4+/creatinine1.2 ± 0.21.0 ± 0.21.4 ± 0.2
        Ca2+/creatinine0.05 ± 0.010.05 ± 0.010.11 ± 0.02c
        Mg2+/creatinine0.44 ± 0.090.19 ± 0.040.66 ± 0.19
        citrate/creatinine0.71 ± 0.09ND0.25 ± 0.04b
        creatinine clearance1.03 ± 0.17ND1.11 ± 0.22
        FE Na+ (%)0.26 ± 0.05ND0.68 ± 0.15c
        FE K+ (%)21.25 ± 4.11ND10.31 ± 2.23
        FE Cl− (%)0.31 ± 0.08ND0.31 ± 0.05
        FE Ca2+ (%)0.76 ± 0.09ND0.58 ± 0.08
        FE Mg2+ (%)5.62 ± 2.76ND6.88 ± 1.16
        FE Pi (%)2.09 ± 0.42ND1.81 ± 0.44
        urinary anion gap177.8 ± 89.321.4 ± 36.336.8 ± 24.2
        titratable acids38.7 ± 4.628.0 ± 6.15.5 ± 6.6b
        NEA120.0 ± 10.294.1 ± 14.649.1 ± 12.4b
        NEA/creatinine1.9 ± 0.21.4 ± 0.11.6 ± 0.4
    • ↵a Summary of basal blood and urine data. Urine was collected during 24 h in metabolic cages, and blood was sampled at the end of the collection period. Concentrations of electrolytes, titratable acids, and urea are expressed in units of mmol/L, creatinine in mg/dl, Pco2 in mmHg, osmolarity in mOsmol/L, creatinine clearance in ml/min; n = 8 to 10 animals per genotype. ND, not determined; NEA, net acid excretion.

    • ↵b P < 0.001 versus wild-type.

    • ↵c P < 0.05 versus wild-type.

    • View popup
    Table 2.

    24-h NH4Cl loadinga

    GenotypeAe1+/+Ae1+/−Ae1−/−
    Blood
        pH7.21 ± 0.017.20 ± 0.016.82 ± 0.05b
        HCO3−20.6 ± 0.718.8 ± 0.77.1 ± 0.9b
        Pco253.3 ± 1.849.6 ± 1.544.4 ± 3.1c
        K+3.7 ± 0.24.0 ± 0.14.9 ± 0.3b
        Na+147.9 ± 1.0148.1 ± 1.0156.4 ± 2.8c
        Cl−117.0 ± 1.8119.9 ± 2.0133.0 ± 4.3b
        Pi2.0 ± 0.22.1 ± 0.23.4 ± 0.1b
        anion gap10.1 ± 1.89.5 ± 2.316.3 ± 2.4c
        osmolarity304.1 ± 1.2302.4 ± 1.5346.8 ± 3.4b
        creatinine0.08 ± 0.020.10 ± 0.020.15 ± 0.02c
        urea10.8 ± 1.6ND29.6 ± 5.3b
    Urine
        pH6.07 ± 0.066.10 ± 0.106.47 ± 0.12c
        osmolarity3302.5 ± 178.42388 ± 97.11176.5 ± 69.7b
        creatinine83.1 ± 5.886.1 ± 13.133.2 ± 3.3b
        HCO3−/creatinine0.0 ± 0.00.1 ± 0.00.1 ± 0.0
        K+/creatinine3.5 ± 0.63.1 ± 0.75.2 ± 1.0
        Na+/creatinine1.8 ± 0.41.4 ± 0.33.5 ± 0.7
        Cl−/creatinine7.6 ± 0.57.3 ± 1.06.7 ± 1.0
        Pi/creatinine0.6 ± 0.10.5 ± 0.10.8 ± 0.2
        NH4+/creatinine2.1 ± 0.22.2 ± 0.32.2 ± 0.3
        Ca2+/creatinine0.04 ± 0.010.05 ± 0.020.06 ± 0.01
        Mg2+/creatinine0.19 ± 0.040.20 ± 0.080.26 ± 0.07
        creatinine clearance1.46 ± 0.400.80 ± 0.310.47 ± 0.06b
        FE Na+ (%)0.10 ± 0.030.09 ± 0.050.34 ± 0.08c
        FE K+ (%)6.78 ± 2.063.99 ± 1.0516.72 ± 4.06
        FE Cl− (%)0.44 ± 0.100.58 ± 0.220.76 ± 0.15
        FE Ca2+ (%)0.15 ± 0.060.24 ± 0.110.31 ± 0.07
        FE Mg2+ (%)4.42 ± 1.692.41 ± 1.783.80 ± 1.08
        FE Pi (%)1.98 ± 0.433.02 ± 1.013.52 ± 0.62
        urinary anion gap−202.3 ± 69.7−223.2 ± 57.680.6 ± 61.8c
        titratable acids37.8 ± 3.220.0 ± 4.716.1 ± 4.4c
        NEA205.7 ± 9.5176.6 ± 10.285.2 ± 5.2b
        NEA/Crea2.6 ± 0.22.9 ± 0.52.7 ± 0.3
    • ↵a Summary of blood and urine data after 24 h of oral NH4Cl loading. Urine was collected during 24 h in metabolic cages, and blood was sampled at the end of the collection period. Concentrations of electrolytes, titratable acids, and urea are expressed in units of mmol/L, creatinine in mg/dl, Pco2 in mmHg, osmolarity in mOsmol/L, creatinine clearance in ml/min; n = 8 to 10 animals per genotype.

    • ↵b P < 0.001 versus wild-type.

    • ↵c P < 0.05 versus wild-type.

Additional Files

  • Figures
  • Tables
  • Distal renal tubular acidosis in mice lacking the AE1 (band3) Cl-/HCO3- exchanger (slc4a1)

    Files in this Data Supplement:

    • Supplemental Data - 1
    • Supplemental Data - 2
    • Supplemental Data - 3
    • Supplemental Data - 4
    • Supplemental Data - 5
PreviousNext
Back to top

In this issue

Journal of the American Society of Nephrology: 18 (5)
Journal of the American Society of Nephrology
Vol. 18, Issue 5
May 2007
  • Table of Contents
  • Table of Contents (PDF)
  • Index by author
View Selected Citations (0)
Print
Download PDF
Sign up for Alerts
Email Article
Thank you for your help in sharing the high-quality science in JASN.
Enter multiple addresses on separate lines or separate them with commas.
Distal Renal Tubular Acidosis in Mice Lacking the AE1 (Band3) Cl−/HCO3− Exchanger (slc4a1)
(Your Name) has sent you a message from American Society of Nephrology
(Your Name) thought you would like to see the American Society of Nephrology web site.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Distal Renal Tubular Acidosis in Mice Lacking the AE1 (Band3) Cl−/HCO3− Exchanger (slc4a1)
Paul A. Stehberger, Boris E. Shmukler, Alan K. Stuart-Tilley, Luanne L. Peters, Seth L. Alper, Carsten A. Wagner
JASN May 2007, 18 (5) 1408-1418; DOI: 10.1681/ASN.2006101072

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Request Permissions
Share
Distal Renal Tubular Acidosis in Mice Lacking the AE1 (Band3) Cl−/HCO3− Exchanger (slc4a1)
Paul A. Stehberger, Boris E. Shmukler, Alan K. Stuart-Tilley, Luanne L. Peters, Seth L. Alper, Carsten A. Wagner
JASN May 2007, 18 (5) 1408-1418; DOI: 10.1681/ASN.2006101072
del.icio.us logo Digg logo Reddit logo Twitter logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like

Jump to section

  • Article
    • Abstract
    • Materials and Methods
    • Results
    • Discussion
    • Conclusion
    • Disclosures
    • Acknowledgments
    • Footnotes
    • References
  • Figures & Data Supps
  • Info & Metrics
  • View PDF

More in this TOC Section

  • Acute Regulation of the Epithelial Na+ Channel by Phosphatidylinositide 3-OH Kinase Signaling in Native Collecting Duct Principal Cells
  • Role for TGF-β in Cyclosporine-Induced Modulation of Renal Epithelial Barrier Function
  • Increased Renal Responsiveness to Vasopressin and Enhanced V2 Receptor Signaling in RGS2−/− Mice
Show more Cell and Transport Physiology

Cited By...

  • Introduction of single nucleotide variations into the promoter region of the mouse kidney anion exchanger 1 gene downstream of a TATA box changes its transcriptional activity
  • A Ser725Arg mutation in Band 3 abolishes transport function and leads to anemia and renal tubular acidosis
  • Intercalated Cell Depletion and Vacuolar H+-ATPase Mistargeting in an Ae1 R607H Knockin Model
  • Regulation of breathing by CO2 requires the proton-activated receptor GPR4 in retrotrapezoid nucleus neurons
  • Physical and Functional Links between Anion Exchanger-1 and Sodium Pump
  • GATA2 Regulates Body Water Homeostasis through Maintaining Aquaporin 2 Expression in Renal Collecting Ducts
  • Overexpression of Pendrin in Intercalated Cells Produces Chloride-Sensitive Hypertension
  • A mouse model for distal renal tubular acidosis reveals a previously unrecognized role of the V-ATPase a4 subunit in the proximal tubule
  • Specification of ion transport cells in the Xenopus larval skin
  • Deletion of the pH Sensor GPR4 Decreases Renal Acid Excretion
  • Anion Exchanger 1 Interacts with Nephrin in Podocytes
  • Function of human Rh based on structure of RhCG at 2.1 A
  • Deletion of the Chloride Transporter Slc26a7 Causes Distal Renal Tubular Acidosis and Impairs Gastric Acid Secretion
  • The Calcium-Sensing Receptor Promotes Urinary Acidification to Prevent Nephrolithiasis
  • Molecular physiology and genetics of Na+-independent SLC4 anion exchangers
  • Bicarbonate transport in cell physiology and disease
  • Google Scholar

Similar Articles

Related Articles

  • This Month's Highlights
  • PubMed
  • Google Scholar

Articles

  • Current Issue
  • Early Access
  • Subject Collections
  • Article Archive
  • ASN Annual Meeting Abstracts

Information for Authors

  • Submit a Manuscript
  • Author Resources
  • Editorial Fellowship Program
  • ASN Journal Policies
  • Reuse/Reprint Policy

About

  • JASN
  • ASN
  • ASN Journals
  • ASN Kidney News

Journal Information

  • About JASN
  • JASN Email Alerts
  • JASN Key Impact Information
  • JASN Podcasts
  • JASN RSS Feeds
  • Editorial Board

More Information

  • Advertise
  • ASN Podcasts
  • ASN Publications
  • Become an ASN Member
  • Feedback
  • Follow on Twitter
  • Password/Email Address Changes
  • Subscribe to ASN Journals

© 2022 American Society of Nephrology

Print ISSN - 1046-6673 Online ISSN - 1533-3450

Powered by HighWire