Skip to main content

Characterization of Ayu17-449 gene expression and resultant kidney pathology in a knockout mouse model

Transgenic Research volume 17, pages 599–608 (2008)Cite this article

Abstract

The gene trap technique is a powerful approach for characterizing and mutating genes in the mouse. We used this method to identify a mouse gene of unknown function and to establish a mutant mouse line. We subsequently identified one gene, denoted Ayu17-449, on mouse chromosome 3 that comprised 14 exons encoding 1920 amino acids with a granin motif in its N-terminal sequence. In adult mice, this gene was highly expressed in the brain, heart, lung, muscle, stomach, and kidney. The insertion of a trap vector into the second intron of this gene resulted in the null mutation. Homozygous mice for these mutation died by 1 day after birth. Mutant mice showed a loss of acidic granules in the proximal convoluted tubules of the kidney. Our data demonstrates that Ayu17-449 is important for mouse survival.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

References

  • Araki K, Araki M, Yamamura K (2002) Site-directed integration of the cre gene mediated by Cre recombinase using a combination of mutant lox sites. Nucleic Acids Res 30:e103

    Google Scholar 

  • Banks P, Helle KB (1965) The release of protein from the stimulated adrenal medulla. Biochem J 97:40C-41C

    PubMed  CAS  Google Scholar 

  • Birn H, Vorum H, Verroust PJ, Moestrup SK, Christensen EI (2000) Receptor associated protein is important for normal processing of megalin in kidney proximal tubules. J Am Soc Nephrol 11:191–202

    PubMed  CAS  Google Scholar 

  • Carlson CM, Largaespada DA (2005) Insertional mutagenesis in mice: new perspectives and tools. Nat Rev Genet 6:568–580

    PubMed  Article  CAS  Google Scholar 

  • Chen Z, Saffitz JE, Latour A, Schonfeld G (1999) Truncated apoB-70.5-containing lipoproteins bind to megalin but not the LDL receptor. J Clin Invest 103:1419–1430

    PubMed  Article  CAS  Google Scholar 

  • Christensen EI (1976) Rapid protein uptake and digestion in proximal tubule lysosomes. Kidney Int 10:301–310

    PubMed  Article  CAS  Google Scholar 

  • Christensen EI, Nielsen S, Moestrup SK, Borre C, Maunsbach AB, de Heer E, Ronco P, Hammond TG, Verroust P (1995) Segmental distribution of the endocytosis receptor gp330 in renal proximal tubules. Eur J Cell Biol 66:349–364

    PubMed  CAS  Google Scholar 

  • Christensen EI, Birn H, Verroust P, Moestrup SK (1998) Megalin-mediated endocytosis in renal proximal tubule. Ren Fail 20:191–199

    PubMed  CAS  Article  Google Scholar 

  • Ericsson JLE (1965) Transport and digestion of hemoglobin in the proximal tubule. II. Electron microscopy. Lab Invest 14:16–39

    PubMed  CAS  Google Scholar 

  • Evans MJ, Carlton MB, Russ AP (1997) Gene trapping and functional genomics. Trends Genet 13:370–374

    PubMed  Article  CAS  Google Scholar 

  • Fischer-Colbrie R, Laslop A, Kirchmair R (1995) Secretogranin II: molecular properties, regulation of biosynthesis and processing to the neuropeptide secretoneurin. Prog Neurobiol 46:49–70

    PubMed  Article  CAS  Google Scholar 

  • Frohman MA, Dush MK, Martin GR (1988) Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci USA 85:8998–9002

    PubMed  Article  CAS  Google Scholar 

  • Gossler A, Joyner AL, Rossant J, Skarnes WC (1989) Mouse embryonic stem cells and reporter constructs to detect developmentally regulated genes. Science 244:63–465

    Article  Google Scholar 

  • Helle KB (1966) Some chemical and physical properties of the soluble protein fraction of bovine adrenal chromaffin granules. Mol Pharmacol 2:298–310

    PubMed  CAS  Google Scholar 

  • Huttner WB, Gerdes HH, Rosa P (1991) The granin (chromogranin/secretogranin) family. Trends Biochem Sci 16:27–30

    PubMed  Article  CAS  Google Scholar 

  • Ischia R, Lovisetti-Scamihorn P, Hogue-Angeletti R, Wolkersdorfer M, Winkler H, Fischer-Colbrie R (1997) Molecular cloning and characterization of NESP55, a novel chromogranin-like precursor of a peptide with 5-HT1B receptor antagonist activity. J Biol Chem 272:11657–11662

    PubMed  Article  CAS  Google Scholar 

  • Krisch K, Buxbaum P, Horvat G, Krisch I, Neuhold N, Ulrich W, Srikanta S (1986) Monoclonal antibody HISL-19 as an immunocytochemical probe for neuroendocrine differentiation: its application in diagnostic pathology. Am J Pathol 123:100–108

    PubMed  CAS  Google Scholar 

  • Lako M, Hole N (2000) Searching the unknown with gene trapping. Expert Rev Mol Med 6:1–11

    Google Scholar 

  • Laurent T, Kimberly LH, O’Connor DT (2003) The chromogranin-secrectogranin family. N Engl J Med 348:1134–1149

    Article  Google Scholar 

  • Lee RW, Huttner WB (1983) Tyrosine-O-sulfated proteins of PC12 pheochromocytoma cells and their sulfation by a tyrosylprotein sulfotransferase. J Biol Chem 258:11326–11334

    PubMed  CAS  Google Scholar 

  • Mbikay M, Seidah NG, Chretien M (2001) Neuroendocrine secretory protein 7B2: structure, expression and functions. Biochem J 357:329–342

    PubMed  Article  CAS  Google Scholar 

  • Natori S, Huttner WB (1996) Chromogranin B (secretogranin I) promotes sorting to the regulated secretory pathway of processing intermediates derived from a peptide hormone precursor. Proc Natl Acad Sci USA 93:4431–4436

    PubMed  Article  CAS  Google Scholar 

  • Nobels FR, Kwekkeboom DJ, Bouillon R, Lambergs SW (1998) Chromogranin A: its clinical value as marker of neuroendocrine tumours. Eur J Clin Invest 28:431–440

    PubMed  Article  CAS  Google Scholar 

  • O’Connor DT (1983) Chromogranin: widespread immunoreactivity in polypeptide. Hormone producing tissues and in serum. Regul Pept 6:263–280

    PubMed  Article  CAS  Google Scholar 

  • O’Connor DT, Deftos LJ (1986) Secretion of chromogranin A by peptide-producing endocrine neoplasms. N Engl J Med 314:1145–1151

    PubMed  CAS  Article  Google Scholar 

  • Ottiger HP, Battenberg EF, Tsou AP, Bloom FE, Sutcliffe JG (1990) A brain- and pituitary-specific mRNA that encodes a novel chromogranin/secretogranin-like component of intracellular vesicles. J Neurosci 10:3135–3147

    PubMed  CAS  Google Scholar 

  • Rosa P, Zanini A (1981) Characterization of adenohypophysial polypeptides by two dimensional gel electrophoresis. II. Sulfated and glycosylated polypeptides. Mol Cell Endocrinol 24:181–193

    PubMed  Article  CAS  Google Scholar 

  • Sadaki Y, Hiroshi T, Keitaro K (1986) Immunocytochemical localization of cathepsin bin rat kidney. I. Light microscopic study using the indirect immunoenzyme technique. J Histochem Cytochem 34:891–897

    Google Scholar 

  • Shaw E, Dean RT (1980) The inhibition of macrophage protein turnover by a selective inhibitor of thiol proteinases. Biochem J 186:385–390

    PubMed  CAS  Google Scholar 

  • Stridsberg M, Oberg K, Li Q, Engstrom U, Lundqvist G (1995) Measurements of chromogranin A, chromogranin B (secretogranin I), chromogranin C (secretogranin II) and pancreastatin in plasma and urine from patients with carcinoid tumours and endocrine pancreatic tumours. J Endocrinol 144:49–59

    PubMed  CAS  Google Scholar 

  • Taniwaki T, Haruna K, Nakamura H, Sekimoto T, Oike Y, Imaizumi T, Saito F, Muta M, Soejima Y, Utoh A, Nakagata N, Araki M, Yamamura K, Araki K (2005) Characterization of an exchangeable gene trap using pU-17 carrying a stop codon-β-geo cassette. Dev Growth Diff 47:163–172

    Article  CAS  Google Scholar 

  • Tooze SA, Martens GJM, Huttner WB (2001) Secretory granule biogenesis: rafting to the SNARE. Trends Cell Biol 11:116–122

    PubMed  Article  CAS  Google Scholar 

  • Yagi T, Tokunaga T, Furuta Y, Nada S, Yoshida M, Tsukada T, Saga Y, Takeda N, Ikawa Y, Aizawa S (1993) A novel ES cell line, TT2, with high germline-differentiating potency. Anal Biochem 214:70–76

    PubMed  Article  CAS  Google Scholar 

  • Yokota S, Tsuji H, Kato K (1985) Immunocytochemical localization of cathepsin D in lysosomes of cortical collecting tubule cells of the rat kidney. J Histochem Cytochem 33:191–200

    PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Mrs. Nakada for staining of all tissue sections. This study was supported in part by KAKENHI (Grant-in-Aid for Scientific Research) on Priority Areas “Integrative Research Toward the Conquest of Cancer” from the Ministry of Education, Culture, Sports, Science and Technology of Japan, a grant from the Osaka Foundation of Promotion of Clinical Immunology; and a grant from the National Nature Science Foundation of China (30671080).

Author information

Affiliations

  1. Key Laboratory of Molecular Biology on Infectious Diseases, Chongqing Medical University, Yixueyuanlu-1, Chongqing, 400016, China

    Hua Tang

  2. Department of Developmental Genetics, Institute of Molecular Embryology and Genetics, Kumamoto University, Honjo 2-2-1, Kumamoto, 860-0811, Japan

    Hua Tang, Kimi Araki, Zhenghua Li & Kenichi Yamamura

Authors
  1. Hua Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kimi Araki
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zhenghua Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kenichi Yamamura
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Hua Tang or Kenichi Yamamura.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Tang, H., Araki, K., Li, Z. et al. Characterization of Ayu17-449 gene expression and resultant kidney pathology in a knockout mouse model. Transgenic Res 17, 599–608 (2008). https://doi.org/10.1007/s11248-008-9170-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11248-008-9170-y

Keywords

  • Acidic granules
  • ES cells
  • Gene trapping
  • Proximal convoluted tubules