Background/Aims: Progress accomplished by complete genomes and cDNA-sequencing projects calls for methods that fully use these resources to study gene expression patterns in characterized cell populations. However, since the number of functional genes cannot be readily inferred from the genomic sequence, it is highly desirable to make use of methods enabling to study both known and unknown genes. Methods: The method of serial analysis of gene expression provides short diagnostic cDNA tags without bias towards known genes. In addition, the frequency of each tag in the library conveys quantitative information on gene expression. A microassay was set-up to perform serial analysis of gene expression in minute samples such as those obtained by microdissecting nephron segments. Results: Studies carried out in the thick ascending limb of Henle’s loop and the collecting duct of the mouse kidney provided expression data for several thousand genes. Known markers were found appropriately enriched, and several of the thick ascending limb or collecting duct specific transcripts had no database match. Conclusions: The microassay for serial analysis of gene expression makes possible large-scale quantitative measurements of mRNA levels in nephron segments. The comprehensive picture generated by analyzing both known and unknown transcripts in defined cell populations should help to discover genes with dedicated functions.

1.
Sanger F, Air GM, Barrell BG, Brown NL, Coulson AR, Fiddes CA, Hutchison CA, Slocombe PM, Smith M: Nucleotide sequence of bacteriophage Φ X174 DNA. Nature 1977;265:687–695.
2.
Anderson S, Bankier AR, Barrell BG, de Bruijn MHL, Coulson AR, Drouin J, Eperon IC, Nierlich DP, Roe BA, Sanger F, Schreier PH, Smith AJH, Staden R, Young IG: Sequence and organization of the human mitochondrial genome. Nature 1981;290:457–465.
3.
Bibb MJ, Van Etten RA, Wright CT, Walberg MW, Clayton DA: Sequence and gene organization of mouse mitochondrial DNA. Cell 1981;26:167–180.
4.
Fleischmann RD, Adams MD, White O, et al: Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science 1995;269:496–512.
5.
Goffeau A, Barrell BG, Bussey H, Davis RW, Dujon B, Feldmann H, Galibert F, Hoheisel JD, Jacq C, Johnston M, Louis EJ, Mewes HW, Murakami Y, Philippsen P, Tettelin H, Oliver SG: Life with 6,000 genes. Science 1996;274:563–567.
6.
The C. elegans Sequencing Consortium: Sequence and analysis of the genome of C. elegans. Science 1998;282:2012–2018.
7.
Adams MD, Celniker SE, Holt RA, et al: The genome sequence of Drosophila melanogaster. Science 2000;287:2185–2195.
8.
The Arabidopsis Genome Initiative: Analysis of the genome sequence of the flowering plant Arabidopsis thaliana. Nature 2000;408:796–805.
9.
International Human Genome Sequencing Consortium: Initial sequencing and analysis of the human genome. Nature 2001;409:860–921.
10.
Venter JC, Adams MD, Myers EW, et al: The sequence of the human genome. Science 2001;291:1304–1351.
11.
Schena M, Shalon D, Davis RW, Brown PO: Quantitative monitoring of gene expression patterns with a complementary DNA microarray. Science 1995;270:467–470.
12.
Velculescu VE, Zhang L, Vogelstein B, Kinzler KW: Serial analysis of gene expression. Science 1995;270:484–487.
13.
Wodicka L, Dong H, Mittmann M, Mh H, Lockhardt DJ: Genome-wide expression monitoring in Saccharomyces cerevisiae. Nat Biotechnol 1997;15:1359–1367.
14.
Velculescu VE, Zhang L, Zhou W, Vogelstein J, Basrai MA, Bassett DE, Hieter P, Vogelstein B, Kinzler KW: Characterization of the yeast transcriptome. Cell 1997;88:243–251.
15.
Ewing B, Green P: Analysis of expressed sequence tags indicates 35,000 human genes. Nat Genet 2000;25:232–234.
16.
Roest Crollius H, Jaillon O, Bernot O, Dasilva C, Bouneau L, Fischer C, Fizames C, Wincker P, Brottier P, Quétier F, Saurin W, Weissenbach J: Estimate of human gene number provided by genome-wide analysis using Tetraodon nigroviridis DNA sequence. Nat Genet 2000;25:235–238.
17.
Liang F, Holt I, Pertea G, Karamycheva S, Salzberg L, Quackenbush J: Gene index analysis of the human genome estimates approximately 120,000 genes. Nat Genet 2000;25:239–240.
18.
Hastie ND, Bishop JO: The expression of three abundance classes of messenger RNA in mouse tissues. Cell 1976;9:761–774.
19.
Adams MA, Kelley JM, Gocayne JD, Dubnick M, Polymeropoulos MH, Xiao H, Merril CR, Wu A, Olde B, Moreno RF, Kerlavage AR, McCombie WR, Venter JC: Complementary DNA sequencing: Expressed sequence tags and human genome project. Science 1991;252:1651–1656.
20.
Okubo K, Hori N, Matoba R, Niiyama T, Fukushima A, Kojima Y, Matsubara K: Large-scale cDNA sequencing for analysis of quantitative and qualitative aspects of gene expression. Nat Genet 1992;2:173–179.
21.
Adams MD, Kerlavage AR, Fleischmann RD, Fuldner RA, Bult CJ, Lee NH, Kirkness EF, Weinstock KG, Gocayne JD, White O, Sutton G, Blake JA, Brandon RC, Chin MW, Clayton RA: Initial assessment of human gene diversity and expression patterns based upon 83 million nucleotides of cDNA sequence. Nature 1995;377:3–174.
22.
Virlon B, Cheval L, Buhler JM, Billon E, Doucet A, Elalouf JM: Serial microanalysis of renal transcriptomes. Proc Natl Acad Sci USA 1999;96:15286–15291.
23.
Cheval L, Virlon B, Elalouf JM: SADE: A microassay for serial analysis of gene expression; in Hunt SP, Livesey FJ (eds): Functional Genomics: A Practical Approach. Oxford, Oxford University Press, 2000, pp 139–163.
24.
Margulies EH, Innis JW: eSAGE: Managing and analysing data generated with serial analysis of gene expression (SAGE). Bioinformatics 2000;16:650–651.
25.
Van Kampen AHC, Van Schaik BDC, Pauws E, Michiels EMC, Ruijter JM, Caron HN, Versteeg R, Heisterkamp SH, Leunissen JAM, Baas F, Van der Mee M: USAGE: A web-based approach towards the analysis of SAGE data. Bioinformatics 2000;16:899–905.
26.
St Croix B, Rago C, Velculescu V, Traverso G, Romans KE, Montgomery E, Lal A, Riggins GJ, Lengauer C, Vogelstein B, Kinzler KW: Genes expressed in human tumor endothelium. Science 2000;289:1197–1202.
27.
Lash AE, Tolstoshev CM, Wagner L, Schuler GD, Strausberg RL, Riggins GJ, Altschul SF: SAGEmap: A public gene expression resource. Genome Res 2000;10:1051–1060.
28.
Knight J: When chips are down. Nature 2001;410:860–861.
29.
Zakin L, Reversade B, Virlon B, Rusniok C, Glaser P, Elalouf JM, Brûlet P: Gene expression profiles in normal and Otx2–/– early gastrulating mouse embryos. Proc Natl Acad Sci USA 2000;97:14388–14393.
30.
Fushimi K, Uchida S, Hara Y, Hirata Y, Marumo F, Sasaki S: Cloning and expression of apical membrane water channel of rat kidney collecting tubule. Nature 1993;361:549–552.
31.
Ishibashi K, Sasaki S, Fushimi K, Uchida S, Kuwahara M, Saito H, Furukawa T, Nakajima K, Yamaguchi Y, Gojobori T, Marumo F: Molecular cloning and expression of a member of the aquaporin family with permeability to glycerol and urea in addition to water expressed at the basolateral membrane of kidney collecting duct cells. Proc Natl Acad Sci USA 1994;91:6269–6273.
32.
Naray-Fejes-Toth A, Fejes-Toth G: Expression cloning of the aldosterone target cell-specific 11β-hydroxysteroid dehydrogenase from rabbit collecting duct cells. Endocrinology 1995:136;2579–2586.
33.
Thomson RB, Igarashi P, Biemesderfer D, Kim R, Abu-Alfa A, Soleimani M, Aronson PS: Isolation and cDNA cloning of Ksp-cadherin, a novel kidney-specific member of the cadherin multigene family. J Biol Chem 1995;270:17594–17601.
34.
Duc C, Farman N, Canessa CM, Bonvalet JP, Rossier BC: Cell-specific expression of epithelial sodium channel α, β, and γ subunits in aldosterone-responsive epithelia from the rat: Localization by in situ hybridization and immunocytochemistry. J Cell Biol 1994;127:1907–1921.
35.
Chen SY, Bhargava A, Mastroberardino L, Meijer OC, Wang J, Buse P, Firestone GL, Verrey F, Pearce D: Epithelial sodium channel regulated by aldosterone-induced protein sgk. Proc Natl Acad Sci USA 1999;96:2514–2519.
36.
Firsov D, Mandon B, Morel A, Merot J, Le Maout S, Bellanger AC, de Rouffignac C, Elalouf JM, Buhler JM: Molecular analysis of vasopressin receptors in the rat nephron: Evidence for alternative splicing of the V2 receptor. Pflügers Arch 1994;429:79–89.
37.
Young RM, Shull GE, Lingrel JB: Multiple mRNAs from rat kidney and brain encode a single Na+,K+-ATPase beta subunit protein. J Biol Chem 1987;262:4905–4910.
38.
Knepper MA, Wade JB, Terris J, Ecelbarger CA, Marples D, Mandon B, Chou CL, Kishore BK, Nielsen S: Renal aquaporins. Kidney Int 1996;49:1712–1717.
39.
Umenishi F, Verkman AS, Gropper MA: Quantitative analysis of aquaporin mRNA expression in rat tissues by RNase protection assay. DNA Cell Biol 1996;15:475–480.
40.
Hession C, Decker JM, Sherblom AP, Kumar S, Yue CC, Mattaliano RJ, Tizard R, Kawashima E, Schmeissner U, Heletky S, Chow EP, Burne CA, Shaw A, Muchmore AW: Uromodulin (Tamm-Horsfall glycoprotein): A renal ligand for lymphokines. Science 1987;237:1479–1484.
41.
Robert-Nicoud M, Flahaut M, Elalouf JM, Nicod M, Salinas M, Bens M, Doucet A, Wincker P, Artiguenave F, Horisberger JD, Vandewalle A, Rossier BC, Firsov D: Transcriptome of a mouse kidney cortical collecting duct cell line: Effects of aldosterone and vasopressin. Proc Natl Acad Sci USA 2001;98:2712–2716.
42.
Takenaka M, Imai E, Kaneko T, Ito T, Moriyama T, Yamauchi A, Hori M, Kawamoto S, Okubo K: Isolation of genes identified in mouse renal proximal tubule by comparing different gene expression profiles. Kidney Int 1998;53:562–572.
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