Heart rate variability is an important risk factor for cardiovascular disease and all-cause mortality. The acetylcholine pathway plays a key role in explaining heart rate variability in humans. We assessed whether 443 genotyped and imputed common genetic variants in eight key genes (CHAT, SLC18A3, SLC5A7, CHRNB4, CHRNA3, CHRNA, CHRM2 and ACHE) of the acetylcholine pathway were associated with variation in an established measure of heart rate variability reflecting parasympathetic control of the heart rhythm, the root mean square of successive differences (RMSSD) of normal RR intervals. The association was studied in a two stage design in individuals of European descent. First, analyses were performed in a discovery sample of four cohorts (n = 3429, discovery stage). Second, findings were replicated in three independent cohorts (n = 3311, replication stage), and finally the two stages were combined in a meta-analysis (n = 6740). RMSSD data were obtained under resting conditions. After correction for multiple testing, none of the SNPs showed an association with RMSSD. In conclusion, no common genetic variants for heart rate variability were identified in the largest and most comprehensive candidate gene study on the acetylcholine pathway to date. Future gene finding efforts for RMSSD may want to focus on hypothesis free approaches such as the genome-wide association study.

Document Type


Publication Date


Notes/Citation Information

Published in PLOS One, v. 9, no. 11, article e112476, p. 1-9.

© 2014 Riese et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Digital Object Identifier (DOI)


Funding Information

This work was supported by a Marie Curie International Re-Integration Grant from the EU to HS (IRG grant 44949) and a grant from the National Heart Lung and Blood Institute to XW (HL0836530). Participating centers of the TRacking Adolescents’ Individual Lives Survey (TRAILS) include various departments of the University Medical Center and University of Groningen, the Erasmus University Medical Center Rotterdam, the University of Utrecht, the Radboud Medical Center Nijmegen, and the Parnassia Bavo group, all in the Netherlands. TRAILS has been financially supported by various grants from the Netherlands Organization for Scientific Research NWO (Medical Research Council program grant GB-MW 940-38-011; ZonMW Brainpower grant 100-001-004; ZonMw Risk Behavior and Dependence grants 60-60600-97-118; ZonMw Culture and Health grant 261-98-710; Social Sciences Council medium-sized investment grants GB-MaGW 480-01-006 and GB-MaGW 480-07-001; Social Sciences Council project grants GB-MaGW 452-04-314 and GB-MaGW 452-06-004; NWO large-sized investment grant 175.010.2003.005; NWO Longitudinal Survey and Panel Funding 481-08-013), the Dutch Ministry of Justice (WODC), the European Science Foundation (EuroSTRESS project FP-006), Biobanking and Biomolecular Resources Research Infrastructure BBMRI-NL (CP 32), the participating universities, and Accare Center for Child and Adolescent Psychiatry. The authors are grateful to all adolescents, their parents and teachers who participated in this research and to everyone who worked on this project and made it possible. TWINS was supported by the Netherlands Organisation for Health Research and Development (ZonMw 904-57-130) and the UK–Netherlands Partnership Program in Science (BR 56–481 and BR 96–229), which is jointly run and financed by the British Council and the Netherlands Organization for Scientific Research (NWO). The BP Stress Study and Georgia Cardiovascular Twin Study were supported by the National Institute of Health (HL086530 and HL69999, respectively). The genotyping conducted in this study was supported by the National Institute of Health (R21 HL086530) and American Heart Association (SDG0730156N). NESDA was supported by the Geestkracht program of ZonMW [grant 10-000-1002]; matching funds from universities and mental health care institutes involved in NESDA (GGZ Buitenamstel-Geestgronden, Rivierduinen, University Medical Center Groningen, GGZ Lentis, GGZ Friesland, GGZ Drenthe). NTR was supported by the Genetic-epidemiology of anxious depression: analyses of twin-family data in the framework of the NEtherlands Twin-family Study of Anxious Depression (NETSAD; ZonMW 940-37-024) and the Centre for Medical Systems Biology (CMSB) 1, WP Genetic epidemiology & gene discovery for cardiovascular risk factors (Netherlands Genomics Initiative/Netherlands Organisation for Scientific Research NWO). Genotyping for NESDA and NTR was funded by the Genetic Association Information Network (GAIN) of the Foundation for the US National Institutes of Health, and analysis was supported by grants from GAIN and the NIMH (MH081802). Genotype data were obtained from dbGaP (www.ncbi.nlm.nih.gov/dbgap, accession number phs000020.v1.p1). PREVEND genetics is supported by the Dutch Kidney Foundation (Grant E033), the National Institutes of Health (grant LM010098), The Netherlands organisation for health research and development (NWO VENI grant 916.761.70), and the Dutch Inter University Cardiology Institute Netherlands. Part of the statistical analyses were carried out on the Genetic Cluster Computer (http://www.geneticcluster.org) hosted by SURFsara and financially supported by the Netherlands Scientific Organization (NWO 480-05-003 PI: Posthuma) along with a supplement from the Dutch Brain Foundation and the VU University Amsterdam. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Included in

Biostatistics Commons