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No role for estrogen receptor 1 gene intron 1 Pvu II and exon 4 C325G polymorphisms in migraine susceptibility
© Colson et al; licensee BioMed Central Ltd. 2006
Received: 21 October 2005
Accepted: 28 February 2006
Published: 28 February 2006
We have previously reported an association between the estrogen receptor 1 (ESR1) gene exon 8 G594A polymorphism and migraine susceptibility in two independent Australian cohorts. In this paper we report results of analysis of two further single nucleotide polymorphisms (SNPs) in the ESR1 gene in the same study group, the T/C Pvu II SNP in intron 1 and the C325G SNP in exon 4, as well as results of linkage disequilibrium (LD) analysis on these markers.
We investigated these variants by case-control association analysis in a cohort of 240 migraineurs and 240 matched controls. The SNPs were genotyped using specific restriction enzyme assays. Results were analysed using contingency table methods incorporating the chi-squared statistic. LD results are presented as D' statistics with associated P values.
We found no evidence for association of the Pvu II T/C polymorphism and the C325G polymorphism and migraine susceptibility and no evidence for LD between these two SNPs and the previously implicated exon 8 G594A marker.
We have found no role for the polymorphisms in intron 1 and exon 4 with migraine susceptibility. To further investigate our previously implicated exon 8 marker, we suggest the need for studies with a high density of polymorphisms be undertaken, with particular focus on markers in LD with the exon 8 marker.
Migraine is a frequent debilitating neurological disorder that affects a significant proportion of the population. The pathophysiology of migraine is not fully understood, although cortical hypersensitivity, vasospasm, neurotransmitters, especially serotonin (5-hydroxytryptamine, 5-HT), platelet activation and sympathetic hyperactivity all appear to play a part, whether as part of the primary triggering event, or as a response mechanism. In the absence of any biological marker, migraine diagnosis is currently based on subjective criteria alone. To further compound the problem, treatment efficacy is limited. Migraine imparts a significant burden on society, both socially and financially. The World Health Organization has identified migraine among the world's top 20 leading causes of disability, with an impact that extends far beyond individual suffering .
There is significant evidence from family and twin studies to indicate a strong genetic component to migraine. The current understanding of migraine is that it is a polygenic multifactorial disorder . It has been postulated that genetic factors set the individual migraine threshold, with environmental influences playing a modulating role . It is likely that many genes may provide an important although small contribution to an individual's migraine susceptibility . The identification of migraine susceptibility genes has been the focus of substantial research to date and could eventually lead to improved treatments and greater understanding of the disorder. Several loci have shown promise, although these need to be followed up by both replication and functional studies to determine a definitive causative role [5–15].
The estrogen receptor 1 (ESR1) gene is a potential migraine candidate due to the well-known hormonal influence on migraine susceptibility. Migraines in women frequently occur during the childbearing years and are often influenced by significant hormonal milestones. The fluctuating hormone levels of the menstrual cycle have been implicated in migraine but a definitive role is yet to be established . It has been suggested that factors additional to the circulating hormone levels may be at play . Thus, we considered that variation in the ESR1 gene may confer increased migraine risk. To investigate the potential role of ESR1 in migraine we conducted an association study of the ESR1 G594A polymorphism (rs2228480) and migraine in two independent case-control groups. These previously reported results showed that individuals who carried the 594A allele were twice as likely to suffer from migraine than those who carried the 594G allele . The G594A polymorphism is in exon 8 of ESR1, a gene of approximately 295 kilobases in size and consisting of 8 exons. It is a synonymous polymorphism with no associated amino acid change, consequently it is unlikely that this polymorphism is causative, but may be in linkage disequilibrium (LD) with an unknown causative variant. In this study we have analysed two further single nucleotide polymorphisms (SNPs) in ESR1 in the same study group, the Pvu II C/T SNP in intron 1 (rs2234693,) and the C325G SNP in exon 4 (rs1801132) which is located in the hormone binding region. The Pvu II locus has been associated with variation in estradiol levels in post menopausal women  and with an increased risk of stroke in men . Interestingly both estrogen withdrawal and high estrogen concentrations have been implicated in migraine susceptibility in women , and there is evidence for an increased risk of stroke in MA sufferers . It has been reported that the C325G SNP may play a role in calcium metabolism  and susceptibility to breast cancer [23, 24] a disease in which hormones play a role. The minor allele at each of these SNPs has been shown to have a frequency of >20% as determined in Australian and other Caucasian populations [25, 26].
Research was approved by the Griffith University Ethics Committee for experimentation on human subjects. Informed consent was obtained from all participants prior to commencement. All were of Caucasian origin, and were recruited from the east coast of Australia through the Genomics Research Centre's patient clinic whereby each participant was interviewed, and completed a detailed questionnaire on personal and family medical history, migraine symptoms, age of onset, frequency, severity, treatment and response, and migraine triggers as previously described [27, 28]. Migraine was diagnosed by a clinical neurologist as either migraine with aura (MA), or migraine without aura (MO) based strictly on the widely accepted criteria specified by the International Headache Society . The study population was comprised of 240 migraineurs and 240 unrelated control individuals. To minimize potential bias from population stratification, the control group was matched for sex, age (+/- 5 years), and ethnicity.
Genotype data and allele frequencies were compared between the migraine case and control groups using standard chi-square analysis. Due to multiple testing, the Bonferroni correction for 5 tests was applied, which set the level of significance at 0.01 (ie. 0.05/5).
Linkage disequilibrium between the ESR1 intron 1, the ESR1 exon 4 polymorphism and the previously reported exon 8 polymorphism that was tested in the same study group was analysed using the 2LD program . LD results are presented as D' and P values.
Results and Discussion
Case Control analysis
Distribution of ESR Intron 1 Pvu II Polymorphism frequencies in migraineurs and controls
MA & MO
Distribution of ESR Exon 4 Codon C325G Polymorphism frequencies in migraineurs and controls
MA & MO
Linkage Disequilibrium analysis
Linkage disequilibrium D' values (upper right hand side) and distance in bases between markers (lower left hand side).
D' = 0.268 P = 0.0001
D' = 0.016 P = 0.52
D' = 0.060 P = 0.71
The human ESR1 gene is located on chromosome 6q25.1 and contains 8 exons . It is widely expressed in a broad range of tissues including CNS areas such as the hypothalamus, limbic system, hippocampus, cortices of the temporal lobe and the brainstem . Numerous studies have demonstrated the multifunctional role of the ESR, particularly in the CNS. It is understood to play a role in neuroprotection via activation of the MAPK pathway , as well as in cognition, mood, and memory . ESR can be activated by neurotransmitters and growth factors, in particular, dopamine . Estrogens can induce Ca2+ mobilization, and activate several kinases including protein kinase C, and phosphatidylinositol-3-OH kinase [41, 42]. Estrogen deficiency has been implicated in pathological and degenerative processes in the CNS, while elevated levels have been involved in the development and progression of tumours . In view of the wide variety of mechanisms under control of estrogen and its cognate receptor, particularly in known migraine pathways in the CNS, as well as the well-known role for hormones in migraine, variation in function of the estrogen receptor gene may play a role in neurological conditions, such as migraine. We have previously reported a role for the ESR1 G594A polymorphism in migraine susceptibility. This study investigated two further synonymous polymorphisms in ESR1, the widely studied Pvu II T/C polymorphism in intron 1, and the C325G polymorphism at exon 4.
Results showed no association with migraine in the case-control groups for both the intron 1 Pvu II marker and the exon 4 C325G marker. There was no evidence for LD between the exon 4 marker and the previously studied exon 8 marker nor between the intron 1 and exon 8 markers. The physical distance between the loci is ~155 kb for exon 4 and 8 markers, and ~257 kb between intron 1 and exon 8 markers. Absence of linkage disequilibrium at the same exon 4 and 8 loci has previously been reported in a different case-control panel by Curran et al (2001) . Our study showed that there was evidence for linkage disequilibrium between the intron 1 and exon 4 markers which are ~102 kb apart. Similar pairwise LD results between these two loci have been previously reported .
The fact that alleles of the two SNPs tested in the present study showed no association with migraine and were not in LD with alleles at the exon 8 SNP highlights the need for further studies with a high density of polymorphisms spanning the estrogen receptor to further investigate our previously reported susceptibility locus at exon 8. In particular, such studies should focus on markers that are in LD blocks with the G594A polymorphism. Additionally, we believe further investigation of the exon 8 locus for a potential functional variant is clearly warranted, perhaps utilising allele specific gene expression methods. Also worthy of note is the recent report of an association of the exon 4 C325G polymorphism with migraine in women in a large Spanish cohort . Although our results did not demonstrate this, there was an interesting trend in the female subgroup which we believe warrants further investigation in a larger study group.
This work was supported by funding from NHMRC and the Australian Brain Foundation. NC was supported by a Griffith University Postgraduate Scholarship and RL by an NHMRC CJ Martin Postdoctoral Fellowship.
- WHO: The World Health Report. Mental Health: New Understanding, New Hope. [http://www.who.int/whr2001/]
- Montagna P: Molecular genetics of migraine headaches: a review. Cephalalgia. 2000, 20: 3-14. 10.1046/j.1468-2982.2000.00003.x.View ArticlePubMedGoogle Scholar
- Ophoff RA, van den Maagdenberg AM, Roon KI, Ferrari MD, Frants RR: The impact of pharmacogenetics for migraine. Eur J Pharmacol. 2001, 413: 1-10. 10.1016/S0014-2999(00)00949-3.View ArticlePubMedGoogle Scholar
- Peroutka SJ: Sympathetic look at genetic basis of migraine. Headache. 2002, 42: 378-381. 10.1046/j.1526-4610.2002.02111.x.View ArticlePubMedGoogle Scholar
- Kowa H, Fusayasu E, Ijiri T, Ishizaki K, Yasui K, Nakaso K, Kusumi M, Takeshima T, Nakashima K: Association of the insertion/deletion polymorphism of the angiotensin I-converting enzyme gene in patients of migraine with aura. Neurosci Lett. 2005, 374: 129-131. 10.1016/j.neulet.2004.10.041.View ArticlePubMedGoogle Scholar
- Colson NJ, Lea RA, Quinlan S, MacMillan J, Griffiths LR: Investigation of hormone receptor genes in migraine. Neurogenetics. 2005, 6: 17-23. 10.1007/s10048-004-0205-0.View ArticlePubMedGoogle Scholar
- Marziniak M, Mossner R, Schmitt A, Lesch KP, Sommer C: A functional serotonin transporter gene polymorphism is associated with migraine with aura. Neurology. 2005, 64: 157-159. 10.1159/000088791.View ArticlePubMedGoogle Scholar
- Lea RA, Ovcaric M, Sundholm J, Solyom L, Macmillan J, Griffiths LR: Genetic variants of angiotensin converting enzyme and methylenetetrahydrofolate reductase may act in combination to increase migraine susceptibility. Brain Res Mol Brain Res. 2005, 136: 112-117. 10.1016/j.molbrainres.2005.01.006.View ArticlePubMedGoogle Scholar
- McCarthy LC, Hosford DA, Riley JH, Bird MI, White NJ, Hewett DR, Peroutka SJ, Griffiths LR, Boyd PR, Lea RA, Bhatti SM, Hosking LK, Hood CM, Jones KW, Handley AR, Rallan R, Lewis KF, Yeo AJ, Williams PM, Priest RC, Khan P, Donnelly C, Lumsden SM, O'Sullivan J, See CG, Smart DH, Shaw-Hawkins S, Patel J, Langrish TC, Feniuk W, Knowles RG, Thomas M, Libri V, Montgomery DS, Manasco PK, Xu CF, Dykes C, Humphrey PP, Roses AD, Purvis IJ: Single-nucleotide polymorphism alleles in the insulin receptor gene are associated with typical migraine. Genomics. 2001, 78: 135-149. 10.1006/geno.2001.6647.View ArticlePubMedGoogle Scholar
- Kowa H, Yasui K, Takeshima T, Urakami K, Sakai F, Nakashima K: The homozygous C677T mutation in the methylenetetrahydrofolate reductase gene is a genetic risk factor for migraine. Am J Med Genet. 2000, 96: 762-764. 10.1002/1096-8628(20001204)96:6<762::AID-AJMG12>3.0.CO;2-X.View ArticlePubMedGoogle Scholar
- Lea RA, Dohy A, Jordan K, Quinlan S, Brimage PJ, Griffiths LR: Evidence for allelic association of the dopamine beta-hydroxylase gene (DBH) with susceptibility to typical migraine. Neurogenetics. 2000, 3: 35-40.PubMedGoogle Scholar
- Paterna S, Di Pasquale P, D'Angelo A, Seidita G, Tuttolomondo A, Cardinale A, Maniscalchi T, Follone G, Giubilato A, Tarantello M, Licata G: Angiotensin-converting enzyme gene deletion polymorphism determines an increase in frequency of migraine attacks in patients suffering from migraine without aura. Eur Neurol. 2000, 43: 133-136. 10.1159/000008151.View ArticlePubMedGoogle Scholar
- Del Zompo M, Cherchi A, Palmas MA, Ponti M, Bocchetta A, Gessa GL, Piccardi MP: Association between dopamine receptor genes and migraine without aura in a Sardinian sample. Neurology. 1998, 51: 781-786.View ArticlePubMedGoogle Scholar
- Ogilvie AD, Russell MB, Dhall P, Battersby S, Ulrich V, Smith CA, Goodwin GM, Harmar AJ, Olesen J: Altered allelic distributions of the serotonin transporter gene in migraine without aura and migraine with aura. Cephalalgia. 1998, 18: 23-26. 10.1046/j.1468-2982.1998.1801023.x.View ArticlePubMedGoogle Scholar
- Peroutka SJ, Wilhoit T, Jones K: Clinical susceptibility to migraine with aura is modified by dopamine D2 receptor (DRD2) NcoI alleles. Neurology. 1997, 49: 201-206.View ArticlePubMedGoogle Scholar
- MacGregor EA: Oestrogen and attacks of migraine with and without aura. Lancet Neurol. 2004, 3: 354-361. 10.1016/S1474-4422(04)00768-9.View ArticlePubMedGoogle Scholar
- Epstein MT, Hockaday JM, Hockaday TD: Migraine and reporoductive hormones throughout the menstrual cycle. Lancet. 1975, 1: 543-548. 10.1016/S0140-6736(75)91558-5.View ArticlePubMedGoogle Scholar
- Colson NJ, Lea RA, Quinlan S, MacMillan J, Griffiths LR: The estrogen receptor 1 G594A polymorphism is associated with migraine susceptibility in two independent case/control groups. Neurogenetics. 2004, 5: 129-133. 10.1007/s10048-004-0181-4.View ArticlePubMedGoogle Scholar
- Schuit SC, de Jong FH, Stolk L, Koek WN, van Meurs JB, Schoofs MW, Zillikens MC, Hofman A, van Leeuwen JP, Pols HA, Uitterlinden AG: Estrogen receptor alpha gene polymorphisms are associated with estradiol levels in postmenopausal women. Eur J Endocrinol. 2005, 153: 327-334. 10.1530/eje.1.01973.View ArticlePubMedGoogle Scholar
- Shearman AM, Cooper JA, Kotwinski PJ, Humphries SE, Mendelsohn ME, Housman DE, Miller GJ: Estrogen receptor alpha gene variation and the risk of stroke. Stroke. 2005, 36: 2281-2282. 10.1161/01.STR.0000181088.76518.ec.View ArticlePubMedGoogle Scholar
- Diener HC, Kurth T: Is migraine a risk factor for stroke?. Neurology. 2005, 64: 1496-1497.View ArticlePubMedGoogle Scholar
- Hoshino S, Hosoi T, Miyao M, Shiraki M, Orimo H, Ouchi Y, Inoue S: Identification of a novel polymorphism of estrogen receptor-alpha gene that is associated with calcium excretion in urine. J Bone Miner Metab. 2000, 18: 153-157. 10.1007/s007740050106.View ArticlePubMedGoogle Scholar
- Vasconcelos A, Medeiros R, Veiga I, Pereira D, Carrilho S, Palmeira C, Azevedo C, Lopes CS: Analysis of estrogen receptor polymorphism in codon 325 by PCR-SSCP in breast cancer: association with lymph node metastasis. Breast J. 2002, 8: 226-229. 10.1046/j.1524-4741.2002.08407.x.View ArticlePubMedGoogle Scholar
- Hsiao WC, Young KC, Lin SL, Lin PW: Estrogen receptor-alpha polymorphism in a Taiwanese clinical breast cancer population: a case-control study. Breast Cancer Res. 2004, 6: R180-6. 10.1186/bcr770.View ArticlePubMedPubMed CentralGoogle Scholar
- Curran JE, Lea RA, Rutherford S, Weinstein SR, Griffiths LR: Association of estrogen receptor and glucocorticoid receptor gene polymorphisms with sporadic breast cancer. Int J Cancer. 2001, 95: 271-275. 10.1002/1097-0215(20010720)95:4<271::AID-IJC1046>3.0.CO;2-D.View ArticlePubMedGoogle Scholar
- Sasaki M, Tanaka Y, Sakuragi N, Dahiya R: Six polymorphisms on estrogen receptor 1 gene in Japanese, American and German populations. Eur J Clin Pharmacol. 2003, 59: 389-393. 10.1007/s00228-003-0609-z.View ArticlePubMedGoogle Scholar
- Johnson MP, Lea RA, Curtain RP, MacMillan JC, Griffiths LR: An investigation of the 5-HT2C receptor gene as a migraine candidate gene. Am J Med Genet. 2003, 117B: 86-89. 10.1002/ajmg.b.10007.View ArticlePubMedGoogle Scholar
- Lea RA, Curtain RP, Hutchins C, Brimage PJ, Griffiths LR: Investigation of the CACNA1A gene as a candidate for typical migraine susceptibility. Am J Med Genet. 2001, 105: 707-712. 10.1002/ajmg.1609.View ArticlePubMedGoogle Scholar
- HCCIHS: Headache Classification Committee for the International Headache Society. Classification and diagnostic criteria for headache disorders, cranial neuralgias and facial pain 1st edition. Cephalgia. 1988, 8 (Suppl 7): 1-96.Google Scholar
- Lai IC, Liao DL, Bai YM, Lin CC, Yu SC, Chen JY, Wang YC: Association study of the estrogen receptor polymorphisms with tardive dyskinesia in schizophrenia. Neuropsychobiology. 2002, 46: 173-175. 10.1159/000067808.View ArticlePubMedGoogle Scholar
- Iwase H, Greenman JM, Barnes DM, Hodgson S, Bobrow L, Mathew CG: Sequence variants of the estrogen receptor (ER) gene found in breast cancer patients with ER negative and progesterone receptor positive tumors. Cancer Lett. 1996, 108: 179-184. 10.1016/S0304-3835(96)04406-0.View ArticlePubMedGoogle Scholar
- Etminan M, Takkouche B, Isorna FC, Samii A: Risk of ischaemic stroke in people with migraine: systematic review and meta-analysis of observational studies. Bmj. 2005, 330: 63-10.1136/bmj.38302.504063.8F.View ArticlePubMedPubMed CentralGoogle Scholar
- Schwaag S, Nabavi DG, Frese A, Husstedt IW, Evers S: The association between migraine and juvenile stroke: a case-control study. Headache. 2003, 43: 90-95. 10.1046/j.1526-4610.2003.03023.x.View ArticlePubMedGoogle Scholar
- Merikangas KR, Fenton BT, Cheng SH, Stolar MJ, Risch N: Association between migraine and stroke in a large-scale epidemiological study of the United States. Arch Neurol. 1997, 54: 362-368.View ArticlePubMedGoogle Scholar
- Donaghy M, Chang CL, Poulter N: Duration, frequency, recency, and type of migraine and the risk of ischaemic stroke in women of childbearing age. J Neurol Neurosurg Psychiatry. 2002, 73: 747-750. 10.1136/jnnp.73.6.747.View ArticlePubMedPubMed CentralGoogle Scholar
- Hubbard T, Andrews D, Caccamo M, Cameron G, Chen Y, Clamp M, Clarke L, Coates G, Cox T, Cunningham F, Curwen V, Cutts T, Down T, Durbin R, Fernandez-Suarez XM, Gilbert J, Hammond M, Herrero J, Hotz H, Howe K, Iyer V, Jekosch K, Kahari A, Kasprzyk A, Keefe D, Keenan S, Kokocinsci F, London D, Longden I, McVicker G, Melsopp C, Meidl P, Potter S, Proctor G, Rae M, Rios D, Schuster M, Searle S, Severin J, Slater G, Smedley D, Smith J, Spooner W, Stabenau A, Stalker J, Storey R, Trevanion S, Ureta-Vidal A, Vogel J, White S, Woodwark C, Birney E: Ensembl 2005. Nucleic Acids Res. 2005, 33: D447-53. 10.1093/nar/gki138.View ArticlePubMedGoogle Scholar
- Osterlund MK, Grandien K, Keller E, Hurd YL: The human brain has distinct regional expression patterns of estrogen receptor alpha mRNA isoforms derived from alternative promoters. J Neurochem. 2000, 75: 1390-1397. 10.1046/j.1471-4159.2000.0751390.x.View ArticlePubMedGoogle Scholar
- Mize AL, Shapiro RA, Dorsa DM: Estrogen receptor-mediated neuroprotection from oxidative stress requires activation of the mitogen-activated protein kinase pathway. Endocrinology. 2003, 144: 306-312. 10.1210/en.2002-220698.View ArticlePubMedGoogle Scholar
- McEwen B: Estrogen actions throughout the brain. Recent Prog Horm Res. 2002, 57: 357-384. 10.1210/rp.57.1.357.View ArticlePubMedGoogle Scholar
- Power RF, Mani SK, Codina J, Conneely OM, O'Malley BW: Dopaminergic and ligand-independent activation of steroid hormone receptors. Science. 1991, 254: 1636-1639.View ArticlePubMedGoogle Scholar
- Kelly MJ, Levin ER: Rapid actions of plasma membrane estrogen receptors. Trends Endocrinol Metab. 2001, 12: 152-156. 10.1016/S1043-2760(01)00377-0.View ArticlePubMedGoogle Scholar
- Simoncini T, Hafezi-Moghadam A, Brazil D, Ley K, Chin W, Liao J: Interaction of estrogen receptor with regulatory subunit of phosphatidylinositol-OH kinase. Nature. 2000, 407:Google Scholar
- Diel P: Tissue-specific estrogenic response and molecular mechanisms. Toxicol Lett. 2002, 127: 217-224. 10.1016/S0378-4274(01)00503-3.View ArticlePubMedGoogle Scholar
- Wedren S, Lovmar L, Humphreys K, Magnusson C, Melhus H, Syvanen AC, Kindmark A, Landegren U, Fermer ML, Stiger F, Persson I, Baron J, Weiderpass E: Oestrogen receptor alpha gene haplotype and postmenopausal breast cancer risk: a case control study. Breast Cancer Res. 2004, 6: R437-49. 10.1186/bcr811.View ArticlePubMedPubMed CentralGoogle Scholar
- The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2350/7/12/prepub
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