Transcranial magnetic stimulation (TMS) is a promising tool for human brain science, clinical application, and animal model research because of its non-/low invasiveness. During the early stage of TMS applications, measurement of the cortical effect in response to single- and paired-pulse TMS in humans and animals was restricted to the motor cortex; easily measurable output was limited to motor evoked potentials and induced myoelectric potentials involving the motor cortex¹^,². To expand the brain regions that can be measured by TMS modulation, electroencephalographic (EEG) recording was integrated with single- and paired-pulse TMS as a useful method to directly examine the excitability, connectivity, and spatiotemporal dynamics of areas throughout the whole brain³^,⁴^,⁵. Thus, the simultaneous application of TMS and EEG recording (TMS-EEG) to the brain has been used to probe various superficial cortical brain areas of humans and animals to investigate intracortical neural circuits (see Tremblay et al.⁶). Moreover, TMS-EEG systems can be used to examine additional cortical spatiotemporal characteristics, including the propagation of signals to other cortical areas and the generation of oscillatory activity⁷^,⁸.

However, the mechanism of action of TMS in the brain remains speculative because of the non-invasiveness of TMS, which limits our knowledge of how the brain functions during TMS applications. Therefore, invasive translational studies in animals ranging from rodents to humans are of crucial importance to understand the mechanism of the effects of TMS on neural circuits and their activity. In particular, for combined TMS-EEG experiments in animals, a simultaneous stimulation and measurement system has not been intensively developed for small animals. Therefore, experimentalists are required to construct such a system by trial and error according to their specific experimental requirements. In addition, mouse models are useful among other in vivo animal species models because many transgenic and strain-isolated mice strains are available as biological resources. Thus, a convenient method to build a TMS-EEG-combined measurement system for mice would be desirable for many neuroscience researchers.

This study proposes a TMS-EEG-combined method that can be applied for simultaneous stimulation and recording of the mouse brain, which is the main type of transgenic animal used in research, and that can easily be constructed in typical neuroscience laboratories. First, a low-cost EEG recording system is described using conventional screw electrodes and a flexible substrate to reproducibly assign an electrode-array position in each experiment. Second, a magnetic stimulation system is constructed using a millimeter-sized coil, which can easily be custom-made in typical laboratories. Third, the TMS-EEG-combined system records neural activity in response to sound and magnetic stimulation. The method presented in this study can reveal the mechanisms that generate specific disorders in small animals, and the results obtained in the animal models can be translated to understand the corresponding human disorders.

Electroencephalographic (EEG) Signal Recording System and Procedure