bioRxivNeurosciencePreprint
A significant enrichment that is not: spatial nulls, co-expression, and the imaging transcriptomics of EEG alpha-power genetics
Abstract
Background. Imaging transcriptomics routinely asks whether a trait-associated gene set is over-expressed in a region of interest, and the field standard is to guard that inference with a spatial-autocorrelation-preserving spin test. Electroencephalographic (EEG) oscillatory power is among the most heritable human neurophysiological traits, and the cortical generators of the alpha rhythm have been characterised independently from resting-state magnetoencephalography - making this a natural test bed both for asking whether trait genetics is regionally organised, and for asking what such a test actually establishes. Objective. To test whether alpha-associated genetic signal is spatially enriched in the cortical generators of the alpha rhythm, and to evaluate that inference against complementary null models. Methods. MAGMA gene-based analysis of ENIGMA-EEG summary statistics for six phenotypes: central and occipital alpha power, occipital alpha peak frequency, and theta, beta and delta power. Regional transcription was obtained from the Allen Human Brain Atlas (AHBA) with abagen in the Glasser HCP-MMP1.0 atlas, with Schaefer-100 and Yan-600 as sensitivity analyses. Enrichment in the 41 cortical alpha-source regions was quantified with a threshold-free continuous score and a top-100 gene-set composite, and assessed against three complementary nulls: a spin test (10,000 rotations, cross-checked against brainsmash surrogates), a co-expression-aware gene-set null (10,000 matched random gene sets), and a positive control on a known expression gradient. Results. Judged by the field-standard spin test alone, this study would have reported a positive, biologically coherent finding: alpha-power genes are enriched in the cortical alpha generators (continuous p_spin = 0.022; top-100 p_spin = 0.030), with the spin result corroborated by an independent surrogate model (p = 0.018) and the pipeline validated by a positive control (p_spin = 2 x 10^-4). Three further tests dissolve that conclusion. First, it is not band-specific: theta, beta and delta enrich comparably or more strongly (top-100 beta p_spin = 0.011; delta 0.042; continuous theta 0.042). Second, it does not replicate across alpha phenotypes (occipital alpha continuous p_spin = 0.20; alpha peak frequency non-significant, p_spin >= 0.066). Third, against random gene sets of matched size the alpha set is unremarkable (p_geneset = 0.33) - the apparent enrichment is a generic property of arbitrary gene sets in this cortical territory, and is invisible to a spatial-only null. No test survived false-discovery-rate correction across the 24-cell phenotype x score x region-set grid (minimum q = 0.127), and nominal significance did not survive a change of parcellation. Conclusion. EEG alpha-power genetics shows no regionally specific transcriptomic signature in the cortical generators of the rhythm; the weak tendency that is present is shared across frequency bands, consistent with their known genetic correlation. Methodologically, this is a worked demonstration that correcting for spatial autocorrelation is necessary but not sufficient: a spin-significant, surrogate-corroborated, mechanistically plausible enrichment can be fully accounted for by gene-set co-expression. Enrichment claims in imaging transcriptomics should report a gene-set null alongside the spatial null.
- organism: human
- assay: GWAS summary statistics
- assay: imaging
- assay: bulk RNA-seq
- research focus: GWAS / association mapping
- research focus: benchmarking
- approach: statistical model
In-depth summary
Claude Opus 5.5 · xhigh effort · 8 Nov 2026 · from the full text of version 4 (19 Sept 2026) · CC BY 4.0
EEG alpha-power genes appear enriched in cortical alpha generators under a spin test, but the effect is not band-specific and is matched by random gene sets (p_geneset = 0.33).
Imaging-transcriptomics enrichment tests are usually guarded by spin tests, which preserve spatial autocorrelation but never permute genes, so co-expression within a gene set can inflate significance unseen. This is, per the author, the first imaging-transcriptomics analysis of an EEG-trait GWAS, and it gives a real case where an enrichment that passes spin, surrogate and positive-control checks is fully explained by co-expression.
Key results
ShownWith the conventional spin test as the only check, genes linked to central alpha power (alphaCz) appear enriched in the 41 MEG-defined alpha-source regions. brainsmash surrogates corroborate this, and the pipeline passes a positive control.
Glasser parcellation: continuous score z = 1.98, p_spin = 0.022; top-100 score z = 1.77, p_spin = 0.030 (10,000 rotations). brainsmash surrogates p = 0.018 (1,000 maps). PC1 positive control: target-versus-rest difference 0.74, p_spin = 2 × 10^−4. Prefrontal-hub subset (8C, 8Av, 9a) not significant (continuous p_spin = 0.11; top-100 p_spin = 0.73). · 2.5 Alpha transcriptomic score and enrichment…; 2.6 Validation and complementary nulls; 3.2 The enrichment test is valid and the custom…; 3.3 By the spin test alone, alpha genes are…
ShownAgainst a co-expression-aware null built from random gene sets, the alpha top-100 set is unremarkable. Random sets of the same size also tend to score higher in this occipito-parieto-frontal territory, so the spin-test enrichment says nothing specific about alpha genetics.
Observed top-100 target-versus-rest difference 0.060 vs a mean of 0.043 across 10,000 random gene sets of matched size; p_geneset = 0.33. · 2.6 Validation and complementary nulls; 3.5 Second test: the alpha gene set is…
ShownThe enrichment is not specific to the alpha band and does not replicate across the other alpha phenotypes. No test in the 24-test grid survives FDR correction.
Continuous score: thetaCz p_spin = 0.042; occipital alpha p_spin = 0.20. Top-100 score: occipital alpha p_spin = 0.008 and betaCz 0.011 vs central alpha 0.030; deltaCz 0.042. Alpha peak frequency: continuous p_spin = 0.13, top-100 0.066. Benjamini-Hochberg FDR over the 24-cell (6 phenotype × 2 score × 2 region-set) grid: minimum q = 0.127. · 3.4 First test: the enrichment is not…; Tables
ShownFor central alpha, the continuous-score effect stays positive in all three parcellations but reaches nominal significance only in the native Glasser atlas. Coarsening the parcellation toward MEG resolution does not recover it.
z between 1.5 and 2.0 across Glasser-360, Schaefer-100 and Yan-600; Glasser p_spin = 0.022, Schaefer-100 p_spin = 0.071, Yan-600 p_spin = 0.058; top-100 score significant only in Glasser. Target sets: 31/100 Schaefer and 180/600 Yan parcels (≥50% vertex overlap). · 2.4 Alpha-source region set; 3.3 By the spin test alone, alpha genes are…; 3.6 Third test: significance does not survive a…
ShownMAGMA gene-based analysis of central alpha power finds one genome-wide significant gene. It also finds sub-threshold signal at the chr3p21 locus, which the author uses as a sanity check on the gene-level pipeline.
18,053 genes tested; PRKG2 p = 4.0 × 10^−7, q = 0.007 (survives Bonferroni and FDR); chr3p21 genes GNL3, GLT8D1, NT5DC2, PBRM1 at p ≈ 4–5 × 10^−5. · 2.2 Gene-based analysis (MAGMA); 3.1 Gene-based analysis recovers credible signal
Methods, data and reproducibility
MAGMA gene-level z-statistics from ENIGMA-EEG summary statistics (six EEG phenotypes) were related to AHBA expression (abagen, Glasser HCP-MMP1.0), contrasting 41 MEG-defined alpha-source regions with other cortex. Tests used a 10,000-rotation custom spin test (checked by brainsmash; repeated for alphaCz in Schaefer-100 and Yan-600), 10,000 matched random gene sets (top-100 score) and a PC1 positive control.
- ENIGMA-EEG oscillatory-power GWAS summary statistics (2018-11-01 release, select_N6000_Hom100), six phenotypes, ≈4.96 million autosomal SNPs each (GRCh37) · human · n = N ≈ 6,000–7,700 per SNP
- Allen Human Brain Atlas microarray expression processed with abagen into 360 Glasser regions × 15,563 genes · human (Allen Human Brain Atlas) · n = 5 of 6 donors
- 1000 Genomes European reference panel used for linkage-disequilibrium modelling in MAGMA · human (1000 Genomes) · n = 503 individuals
- Cortical alpha-rhythm source regions from Tabarelli et al. (2022), derived from Cam-CAN resting-state MEG, applied bilaterally · human (alpha rhythm of human EEG/MEG; Cam-CAN cohort) · n = 41 regions (82 parcels)
- Methods & tools
- MAGMA v1.10, abagen (development build post-0.1.3), Allen Human Brain Atlas, brainsmash, custom spin test (10,000 rotations on fsaverage5 sphere), Glasser HCP-MMP1.0 atlas, Schaefer-100 parcellation, Yan-600 homotopic parcellation, fsaverage5, HCP resample_fsaverage registration spheres, 1000 Genomes European reference panel, NCBI build-37 gene locations
- Data & accessions
- https://doi.org/10.5281/zenodo.21514560
- Software & environment
- Python 3.12; MAGMA v1.10; abagen development build post-0.1.3 (needed for pandas ≥2); global seed 20260721; parameters, seeds and package versions logged to a machine-readable run log in the repository (MIT licence).
- Experimental conditions
- No hardware stated; the gene-set null is reported to take a few minutes of computation.
- Reproducibility gaps
- Raw ENIGMA-EEG summary statistics, AHBA data and 1000 Genomes/MAGMA reference files are not redistributed (download instructions only). AHBA donor 15496 excluded after HTTP 404 at source. Analysis grid fixed in advance but not formally pre-registered.
Limitations (6)
- Statistical power: the ENIGMA-EEG GWAS is modest and only PRKG2 passed genome-wide correction, so a small true effect could be masked. The author argues that the threshold-free score, positive control and gene-set null make a hidden alpha-specific signal unlikely. · stated by the authors
- Spatial and modality mismatch: the alpha sources come from MEG minimum-norm estimates with centimetre-scale point-spread, and MEG is relatively insensitive to radial sources, so the target regions are an imperfect proxy for where the genotyped scalp-EEG signal originates. · stated by the authors
- Band-power phenotypes mix periodic and aperiodic (1/f) spectral components, so part of the alpha-power association may be aperiodic. No GWAS of aperiodic parameters exists to separate them. · stated by the authors
- AHBA expression comes from five of six donors, and only two of the six have right-hemisphere tissue. The analysis covers only cortex and common variants tagged by the reference panel, and the 41-region set is a curated selection rather than an exhaustive generator map. · stated by the authors
- The gene-set null was run only for the top-100 score. The pre-declared primary continuous score was tested only against spatial nulls, so its robustness to co-expression is not directly assessed. · noted in this summary
- The positive control takes both the gene statistic (PC1 loadings) and the target regions (top-tertile PC1) from the same AHBA matrix. It therefore shows that the pipeline detects the dominant expression gradient, not that it can detect effects as small as GWAS-derived ones. · noted in this summary
Read this if you test gene-set enrichment against brain maps with abagen and spin tests, or study the genetics of EEG oscillations and want a worked example where a spin-significant result falls apart under a gene-set null.
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