Human Genetics

, Volume 117, Issue 6, pp 545–557 | Cite as

Complex HTR2C linkage disequilibrium and promoter associations with body mass index and serum leptin

  • Shane McCarthy
  • Salim Mottagui-Tabar
  • Yumi Mizuno
  • Bengt Sennblad
  • Johan Hoffstedt
  • Peter Arner
  • Claes Wahlestedt
  • Björn Andersson
Original Investigation


The occurrence of obesity, eating disorders, and related diseases has increased in many parts of the world. Given that few strong genetic factors have been found, it is clear that these are complex multi-factorial diseases. The serotonin receptor 2C, a member of the 5-HTergic system, has been implicated in the control of phagia and obesity. We report a detailed investigation of linkage disequilibrium (LD) within and between the HTR2C promoter and the flanking sequences around a commonly utilized marker in the second coding exon of HTR2C. We suggest that inconsistent associations between HTR2C and several phenotypes, including obesity, may be due to the LD pattern across the gene in which recombination and gene conversion have been influential. The nucleotide and haplotype distribution is consistent with that of the neutral mutation model. The number of haplotypes suggests demographic influences or over dominant selection that may have a function in HTR2C expression. Using the fine LD pattern, we describe a possible association with promoter haplotypes and diplotypes, including a GT microsatellite, and body mass index (BMI) ≥30 kgm−2 (P<0.0001). SNP −995G>A heterozygotes, as well as promoter diplotypes, were found to marginally influence higher serum leptin corrected for percentage body fat (P=0.01), which might suggest that these subjects are leptin resistant. Our results complement previous studies of HTR2C in both mice and humans, and suggest the importance of genetic variation and elucidating the fine LD structure in uncovering the genetic factors of obesity.

Supplementary material

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  1. Andolfatto P, Nordborg M (1998) The effect of gene conversion on intralocus associations. Genetics 148:1397–1399Google Scholar
  2. Ardlie KG, Kruglyak L, Seielstad M (2002) Patterns of linkage disequilibrium in the human genome. Nat Rev Genet 3:299–309Google Scholar
  3. Atmaca M, Kuloglu M, Tezcan E, Ustundag B (2003) Serum leptin and triglyceride levels in patients on treatment with atypical antipsychotics. J Clin Psychiatr 64:598–604Google Scholar
  4. Buckland PR, Hoogendoorn B, Guy CA, Smith SK, Coleman SL, O’Donovan M C (2005) Low gene expression conferred by association of an allele of the 5-HT2C receptor gene with antipsychotic-induced weight gain. Am J Psychiatry 162:613–615Google Scholar
  5. Burnet PW, Harrison PJ, Goodwin GM, Battersby S, Ogilvie AD, Olesen J, Russell MB (1997) Allelic variation in the serotonin 5-HT2C receptor gene and migraine. Neuroreport 8:2651–2653Google Scholar
  6. Burnet PW, Smith KA, Cowen PJ, Fairburn CG, Harrison PJ (1999) Allelic variation of the 5-HT2C receptor (HTR2C) in bulimia nervosa and binge eating disorder. Psychiatr Genet 9:101–104Google Scholar
  7. Cardon LR, Bell JI (2001) Association study designs for complex diseases. Nat Rev Genet 2:91–99Google Scholar
  8. Clark AG, Weiss KM, Nickerson DA, Taylor SL, Buchanan A, Stengard J, Salomaa V, Vartiainen E, Perola M, Boerwinkle E, Sing CF (1998) Haplotype structure and population genetic inferences from nucleotide-sequence variation in human lipoprotein lipase. Am J Hum Genet 63:595–612Google Scholar
  9. Deckert J, Meyer J, Catalano M, Bosi M, Sand P, DiBella D, Ortega G, Stober G, Franke P, Nothen MM, Fritze J, Maier W, Beckmann H, Propping P, Bellodi L, Lesch KP (2000) Novel 5’-regulatory region polymorphisms of the 5-HT2C receptor gene:association study with panic disorder. Int J Neuropsychopharmacol 3:321–325Google Scholar
  10. Drysdale CM, McGraw DW, Stack CB, Stephens JC, Judson RS, Nandabalan K, Arnold K, Ruano G, Liggett SB (2000) Complex promoter and coding region beta 2-adrenergic receptor haplotypes alter receptor expression and predict in vivo responsiveness. Proc Natl Acad Sci USA 97:10483–10488Google Scholar
  11. Ewens WJ (1972) The sampling theory of selectively neutral alleles. Theor Popul Biol 3:87–112Google Scholar
  12. Ewing B, Green P (1998) Base-calling of automated sequencer traces using phred. II. Error probabilities. Genome Res 8:186–194Google Scholar
  13. Ewing B, Hillier L, Wendl MC, Green P (1998) Base-calling of automated sequencer traces using phred I accuracy assessment. Genome Res 8: 175–185Google Scholar
  14. Finn PD, Cunningham MJ, Rickard DG, Clifton DK, Steiner RA (2001) Serotonergic neurons are targets for leptin in the monkey. J Clin Endocrinol Metab 86:422–426Google Scholar
  15. Frisse L, Hudson RR, Bartoszewicz A, Wall JD, Donfack J, Di Rienzo A (2001) Gene conversion and different population histories may explain the contrast between polymorphism and linkage disequilibrium levels. Am J Hum Genet 69:831–843Google Scholar
  16. Fu YX (1997) Statistical tests of neutrality of mutations against population growth, hitchhiking and background selection. Genetics 147: 915–925Google Scholar
  17. Fu YX, Li WH (1993) Statistical tests of neutrality of mutations. Genetics 133:693–709Google Scholar
  18. Gauderman WJ (2002) Sample size requirements for matched case-control studies of gene-environment interaction. Stat Med 21:35–50Google Scholar
  19. Glatt CE, DeYoung JA, Delgado S, Service SK, Giacomini KM, Edwards RH, Risch N, Freimer NB (2001) Screening a large reference sample to identify very low frequency sequence variants:comparisons between two genes. Nat Genet 27:435–438Google Scholar
  20. Glatt CE, Tampilic M, Christie C, DeYoung J, Freimer NB (2004) Re-screening serotonin receptors for genetic variants identifies population and molecular genetic complexity. Am J Med Genet 124B: 92–100Google Scholar
  21. Gordon D, Abajian C, Green P (1998) Consed: a graphical tool for sequence finishing. Genome Res 8:195–202Google Scholar
  22. Gutierrez B, Fananas L, Arranz MJ, Valles V, Guillamat R, van Os J, Collier D (1996) Allelic association analysis of the 5-HT2C receptor gene in bipolar affective disorder. Neurosci Lett 212:65–67Google Scholar
  23. Hartl DL, Clark AG (1997) Principles of population genetics, 3rd edn. Sinauer, SunderlandGoogle Scholar
  24. Hill W, Robertson A (1968) Linkage disequilibrium in finite populations. Theor Appl Genet 38:226–239Google Scholar
  25. Howell WM, Jobs M, Gyllensten U, Brookes AJ (1999) Dynamic allele-specific hybridization A new method for scoring single nucleotide polymorphisms. Nat Biotechnol 17:87–88Google Scholar
  26. Huang XF, Huang X, Han M, Chen F, Storlien L, Lawrence AJ (2004) 5-HT(2A/2C) receptor and 5-HT transporter densities in mice prone or resistant to chronic high-fat diet-induced obesity: a quantitative autoradiography study. Brain Res 1018:227–235Google Scholar
  27. Hudson RR (1987) Estimating the recombination parameter of a finite population model without selection. Genet Res 50:245–250Google Scholar
  28. Hudson RR, Kaplan NL (1985) Statistical properties of the number of recombination events in the history of a sample of DNA sequences. Genetics 111:147–164Google Scholar
  29. Jaruzelska J, Zietkiewicz E, Batzer M, Cole DE, Moisan JP, Scozzari R, Tavare S, Labuda D (1999) Spatial and temporal distribution of the neutral polymorphisms in the last ZFX intron: analysis of the haplotype structure and genealogy. Genetics 152:1091–1101Google Scholar
  30. Lappalainen J, Long JC, Virkkunen M, Ozaki N, Goldman D, Linnoila M (1999) HTR2C Cys23Ser polymorphism in relation to CSF monoamine metabolite concentrations and DSM-III-R psychiatric diagnoses. Biol Psychiatr 46:821–826Google Scholar
  31. Lappalainen J, Zhang L, Dean M, Oz M, Ozaki N, Yu DH, Virkkunen M, Weight F, Linnoila M, Goldman D (1995) Identification, expression, and pharmacology of a Cys23-Ser23 substitution in the human 5-HT2c receptor gene (HTR2C). Genomics 27:274–279Google Scholar
  32. Lentes KU, Hinney A, Ziegler A, Rosenkranz K, Wurmser H, Barth N, Jacob K, Coners H, Mayer H, Grzeschik KH, Schafer H, Remschmidt H, Pirke KM, Hebebrand J (1997) Evaluation of a Cys23Ser mutation within the human 5-HT2C receptor gene: no evidence for an association of the mutant allele with obesity or underweight in children, adolescents and young adults. Life Sci 61:PL9–16Google Scholar
  33. Lewontin RC (1964) The interaction of selection and linkage. I. General considerations; heterotic models. Genetics 49:49–67Google Scholar
  34. Lindstrom M, Isacsson SO, Merlo J (2003) Increasing prevalence of overweight, obesity and physical inactivity: two population-based studies 1986 and 1994. Eur J Public Health 13:306–312Google Scholar
  35. Meyer J, Saam W, Mossner R, Cangir O, Ortega GR, Tatschner T, Riederer P, Wienker TF, Lesch KP (2002) Evolutionary conserved microsatellites in the promoter region of the 5-hydroxytryptamine receptor 2C gene (HTR2C) are not associated with bipolar disorder in females. J Neural Trans 109:939–946Google Scholar
  36. Miller SA, Dykes DD, Polesky HF (1988) A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 16:1215Google Scholar
  37. Nachman MW, Bauer VL, Crowell SL, Aquadro CF (1998) DNA variability and recombination rates at X-linked loci in humans. Genetics 150:1133–1141Google Scholar
  38. Nei M (1987) Molecular evolutionary genetics. Columbia University Press, New YorkGoogle Scholar
  39. Nickerson DA, Taylor SL, Weiss KM, Clark AG, Hutchinson RG, Stengard J, Salomaa V, Vartiainen E, Boerwinkle E, Sing CF (1998) DNA sequence diversity in a 9.7-kb region of the human lipoprotein lipase gene. Nat Genet 19:233–240Google Scholar
  40. Nonogaki K, Strack AM, Dallman MF, Tecott LH (1998) Leptin-independent hyperphagia and type 2 diabetes in mice with a mutated serotonin 5-HT2C receptor gene. Nat Med 4:1152–1156Google Scholar
  41. Parsian A, Cloninger CR (2001) Serotonergic pathway genes and subtypes of alcoholism: association studies. Psychiatr Genet 11:89–94Google Scholar
  42. Pooley EC, Fairburn CG, Cooper Z, Sodhi MS, Cowen PJ, Harrison PJ (2004) A 5-HT2C receptor promoter polymorphism (HTR2C-759C/T) is associated with obesity in women, and with resistance to weight loss in heterozygotes. Am J Med Genet 126B:124–127Google Scholar
  43. Quested DJ, Whale R, Sharpley AL, McGavin CL, Crossland N, Harrison PJ, Cowen PJ (1999) Allelic variation in the 5-HT2C receptor (HTR2C) and functional responses to the 5-HT2C receptor agonist, m-chlorophenylpiperazine. Psychopharmacology (Berl) 144:306–307Google Scholar
  44. Reynolds GP, Zhang Z, Zhang X (2003) Polymorphism of the promoter region of the serotonin 5-HT(2C) receptor gene and clozapine-induced weight gain. Am J Psychiatry 160:677–679Google Scholar
  45. Rieder MJ, Taylor SL, Clark AG, Nickerson DA (1999) Sequence variation in the human angiotensin converting enzyme. Nat Genet 22:59–62Google Scholar
  46. Rozas J, Gullaud M, Blandin G, Aguade M (2001) DNA variation at the rp49 gene region of Drosophila simulans:evolutionary inferences from an unusual haplotype structure. Genetics 158:1147–1155Google Scholar
  47. Rozas J, Rozas R (1999) DnaSP version 3: an integrated program for molecular population genetics and molecular evolution analysis. Bioinformatics 15:174–175Google Scholar
  48. Rozas J, Sanchez-DelBarrio JC, Messeguer X, Rozas R (2003) DnaSP, DNA polymorphism analyses by the coalescent and other methods. Bioinformatics 19:2496–2497Google Scholar
  49. Sambrook J, Russell David W (2001) Molecular cloning : a laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar
  50. Sandelin A, Alkema W, Engstrom P, Wasserman WW, Lenhard B (2004) JASPAR: an open-access database for eukaryotic transcription factor binding profiles. Nucleic Acids Res 32, database issue:D91–94Google Scholar
  51. Sargent PA, Sharpley AL, Williams C, Goodall EM, Cowen PJ (1997) 5-HT2C receptor activation decreases appetite and body weight in obese subjects. Psychopharmacology (Berl) 133:309–312Google Scholar
  52. Schaffhauser AO, Madiehe AM, Braymer HD, Bray GA, York DA (2002) Effects of a high-fat diet and strain on hypothalamic gene expression in rats. Obes Res 10:1188–1196Google Scholar
  53. Slatkin M, Excoffier L (1996) Testing for linkage disequilibrium in genotypic data using the expectation-maximization algorithm. Heredity 76:377–383Google Scholar
  54. Stephens M, Donnelly P (2003) A comparison of bayesian methods for haplotype reconstruction from population genotype data. Am J Hum Genet 73:1162–1169Google Scholar
  55. Stephens M, Smith NJ, Donnelly P (2001) A new statistical method for haplotype reconstruction from population data. Am J Hum Genet 68:978–989Google Scholar
  56. Strobeck C (1987) Average number of nucleotide differences in a sample from a single subpopulation: a test for population subdivision. Genetics 117:149–153Google Scholar
  57. Taillon-Miller P, Bauer-Sardina I, Saccone NL, Putzel J, Laitinen T, Cao A, Kere J, Pilia G, Rice JP, Kwok PY (2000) Juxtaposed regions of extensive and minimal linkage disequilibrium in human Xq25 and Xq28. Nat Genet 25:324–328Google Scholar
  58. Tajima F (1989) Statistical method for testing the neutral mutation hypothesis by DNA polymorphism. Genetics 123:585–595Google Scholar
  59. Tecott LH, Sun LM, Akana SF, Strack AM, Lowenstein DH, Dallman MF, Julius D (1995) Eating disorder and epilepsy in mice lacking 5-HT2c serotonin receptors. Nature 374:542–546Google Scholar
  60. Watterson GA (1975) On the number of segregating sites in genetical models without recombination. Theor Popul Biol 7:256-76Google Scholar
  61. Weiss KM, Clark AG (2002) Linkage disequilibrium and the mapping of complex human traits. Trends Genet 18:19–24Google Scholar
  62. Westberg L, Bah J, Rastam M, Gillberg C, Wentz E, Melke J, Hellstrand M, Eriksson E (2002) Association between a polymorphism of the 5-HT2C receptor and weight loss in teenage girls. Neuropsychopharmacology 26:789–793Google Scholar
  63. Vickers SP, Clifton PG, Dourish CT, Tecott LH (1999) Reduced satiating effect of d-fenfluramine in serotonin 5-HT(2C) receptor mutant mice. Psychopharmacology (Berl) 143:309–314Google Scholar
  64. Vickers SP, Dourish CT, Kennett GA (2001) Evidence that hypophagia induced by d-fenfluramine and d-norfenfluramine in the rat is mediated by 5-HT2C receptors. Neuropharmacology 41:200–209Google Scholar
  65. Vincent JB, Masellis M, Lawrence J, Choi V, Gurling HM, Parikh SV, Kennedy JL (1999) Genetic association analysis of serotonin system genes in bipolar affective disorder. Am J Psychiatry 156:136–138Google Scholar
  66. von Meyenburg C, Langhans W, Hrupka BJ (2003) Evidence for a role of the 5-HT2C receptor in central lipopolysaccharide-, interleukin-1 beta-, and leptin-induced anorexia. Pharmacol Biochem Behav 74:1025–1031Google Scholar
  67. Xie E, Zhu L, Zhao L, Chang LS (1996) The human serotonin 5-HT2C receptor:complete cDNA, genomic structure, and alternatively spliced variant. Genomics 35:551–561Google Scholar
  68. Yamada J, Sugimoto Y, Hirose H, Kajiwara Y (2003) Role of serotonergic mechanisms in leptin-induced suppression of milk intake in mice. Neurosci Lett 348:195–197Google Scholar
  69. Yuan X, Yamada K, Ishiyama-Shigemoto S, Koyama W, Nonaka K (2000) Identification of polymorphic loci in the promoter region of the serotonin 5-HT2C receptor gene and their association with obesity and type II diabetes. Diabetologia 43:373–376Google Scholar
  70. Zuker M (2003) Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res 31:3406–3415Google Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Shane McCarthy
    • 1
  • Salim Mottagui-Tabar
    • 1
  • Yumi Mizuno
    • 1
  • Bengt Sennblad
    • 1
    • 3
  • Johan Hoffstedt
    • 2
  • Peter Arner
    • 2
  • Claes Wahlestedt
    • 1
  • Björn Andersson
    • 1
  1. 1.Center for Genomics and BioinformaticsKarolinska InstitutetStockholmSweden
  2. 2.Department of MedicineKarolinska Instututet, Karolinska University Hospital at HuddingeStockholmSweden
  3. 3.Stockholm Bioinformatics CenterAlbaNova Stockholm UniversityStockholmSweden

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