Introduction: Transcranial ultrasound stimulation (TUS) is a promising new form of non-invasive neuromodulation. As a nascent technique, replication of its effects on brain function is important. Of particular interest is offline 5Hz repetitive TUS (5Hz-rTUS), originally reported by Zeng and colleagues [1] to elicit lasting corticospinal excitability increases, with large effect sizes. Material and method: Here, we conducted a pre-registered (https://osf.io/p5n4q) replication of this protocol that benefitted from three additional features: double-blind application of TUS, neuronavigation for consistent TMS positioning, and acoustic simulations to assess M1 target exposure to TUS. Changes in resting motor thresholds (rMT), motor-evoked potential (MEP) amplitude, short-interval intracortical inhibition (SICI) and intracortical facilitation (ICF) in response to TUS (5Hz-rTUS vs. sham) were measured in the right first dorsal interosseous (FDI), abductor digiti minimi (ADM) and abductor pollicis brevis (APB) muscles, with unbiased selection of participants. Transducer location was determined by the TMS-hotspot for motor representations of the right FDI, as in the original work. Results: No significant effects of 5Hz-TUS (vs sham) were observed. Post-hoc simulations showed considerable variability of the acoustic focus, which was outside the anatomical M1-hand area in 67% of participants, in line with the known poor correspondence of TMS-hotspot location and M1-hand area. Conclusion: Our results indicate that the effect sizes of the neuromodulatory effects of 5Hz-rTUS on M1 may be more variable than previously appreciated. We suggest that double-blinding, neuronavigated TMS, individualised acoustic simulations for TUS targeting and pre-registration will aid reproducibility across studies.
