Continuous and intermittent transcranial magnetic theta burst stimulation modify tactile learning performance and cortical protein expression in the rat differently
Abstract
Repetitive transcranial magnetic stimulation (rTMS) can modulate cortical excitability in a stimulus-frequency-dependent manner. Two kinds of theta burst stimulation (TBS) [intermittent TBS (iTBS) and continuous TBS (cTBS)] modulate human cortical excitability differently, with iTBS increasing it and cTBS decreasing it. In rats, we recently showed that this is accompanied by changes in the cortical expression of proteins related to the activity of inhibitory neurons. Expression levels of the calcium-binding protein parvalbumin (PV) and of the 67-kDa isoform of glutamic acid decarboxylase (GAD67) were strongly reduced following iTBS, but not cTBS, whereas both increased expression of the 65-kDa isoform of glutamic acid decarboxylase. In the present study, to investigate possible functional consequences, we applied iTBS and cTBS to rats learning a tactile discrimination task. Conscious rats received either verum or sham rTMS prior to the task. Finally, to investigate how rTMS and learning effects interact, protein expression was determined for cortical areas directly involved in the task and for those either not, or indirectly, involved. We found that iTBS, but not cTBS, improved learning and strongly reduced cortical PV and GAD67 expression. However, the combination of learning and iTBS prevented this effect in those cortical areas involved in the task, but not in unrelated areas. We conclude that the improved learning found following iTBS is a result of the interaction of two effects, possibly in a homeostatic manner: a general weakening of inhibition mediated by the fast-spiking interneurons, and re-established activity in those neurons specifically involved in the learning task, leading to enhanced contrast between learning-induced and background activity.
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Experimentally induced theta bursts reduce PV expression in other brain regions.
“Along these lines, it is worth noting that experimentally induced theta bursts reduce PV expression in other brain regions (70; 71), raising the possibility that the changes we observed in PV intensity are the result of, as opposed to being responsible for, the altered electrophysiological profile observed with MSEW.”
From Perineuronal nets, inhibitory interneurons and anxiety-related ventral hippocampal neuronal oscillations are altered by early life adversity (2019) · cites this paper as [71]
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Ten years of theta burst stimulation in humans: Established knowledge, unknowns and prospects
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“In 2010, Mix et al. [Mix et al., 2010; 117] demonstrated that iTBS, but not cTBS, improved the ability of rats to learn a tactile discrimination task in darkness (Figure 4F1).”
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“However, theta burst stimulation (a variation of high frequency rTMS) can induce either depression or facilitation of cortical excitability, depending on burst-train duration, such that intermittent theta burst stimulation increases cortical excitability and continuous theta burst stimulation decreases cortical excita…”
BackgroundTheta-Burst Transcranial Magnetic Stimulation Alters Cortical Inhibition
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InfluentialTranscranial Magnetic Stimulation in Alzheimer’s Disease: Are We Ready?
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BackgroundMethodTargeted transcranial theta-burst stimulation alters fronto-insular network and prefrontal GABA
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“Moreover, rTMS could improve learning performance on a tactile discrimination task in normal rats (Mix et al., 2010).”
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BackgroundLifting the veil on the dynamics of neuronal activities evoked by transcranial magnetic stimulation
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BackgroundInfluentialGABA—from Inhibition to Cognition: Emerging Concepts
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“…while calbindin-expressing interneurons mediate dendritic inhibition (Mátyás et al., 2004), these findings imply that distinct rTMS-protocols may affect specific aspects of inhibition and therefore network activity and function (Mix et al., 2010; Funke and Benali, 2011; Volz et al., 2013).”
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“One of the most prominent finding of our previous rTMS studies in rats was a pronounced reduction in cortical PV expression following iTBS (Mix et al., 2010; Benali et al., 2011; Hoppenrath and Funke, 2013; Mix et al., 2013; Volz et al., 2013).”
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“…of cortical neurons expressing the 67 kDa isoform of the GABA-synthesizing enzyme glutamic acid decarboxylase (GAD67) and that of the calcium-binding proteins PV and calbindin (CB), indicative of reduced activity of inhibitory neurons (Trippe et al. 2009; Mix et al. 2010, 2013; Benali et al. 2011).”
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“Due to its modulatory action on human cortical excitability, repetitive transcranial magnetic stimulation (rTMS) has become a promising tool for the treatment of psychiatric and neurodegenerative disorders and cortical reorganization after stroke (Di Lazzaro et al., 2008a; Grefkes et al., 2010;…”
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“In rats, iTBS has previously been shown to induce changes in markers of inhibitory and excitatory neuronal activity, as well as increases in the learning rate in an associative tactile task (Mix et al., 2010; Hoppenrath and Funke, 2013; Labedi et al., 2014).”
BackgroundMechanisms and pathways underlying the therapeutic effect of transcranial magnetic stimulation
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“RTMS parameters can be adjusted to have different effects: high frequency ( ≥ 5 Hz) rTMS and intermittent theta-burst stimulation (iTBS) facilitate neural activity, while low frequency ( ≤ 1 Hz) rTMS and continuous theta-burst stimulation (cTBS) inhibit it [9,18].”
Interneurons containing somatostatin are affected by learning-induced cortical plasticity
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“Parallel changes in the density of GAD and Prv immunoreactive cells were found, for example, in schizophrenia and as an effect of transcranial magnetic stimulation (Lewis et al., 2005; Mix et al., 2010).”
Fu, W., Cao, L., Zhang, Y., Huo, S., Du, J., Zhu, L. L., & Song, W. (2017) · Topics in Stroke Rehabilitation · doi:10.1080/10749357.2016.1253139 · Cited by 30 · see citing works
Gonsalves, M. A., White, T. L., Barredo, J., Fukuda, A. M., Joyce, H. E., Harris, A. D., & Carpenter, L. L. (2022) · NeuroImage Clinical · doi:10.1016/j.nicl.2022.103049 · Cited by 29 · see citing works
Castillo-Padilla, D. V., & Funke, K. (2015) · Developmental Neurobiology · doi:10.1002/dneu.22296 · Cited by 29 · see citing works
“Our previous studies showed that neuronal activity marker expression is mainly affected in layers 2/3 this way (e.g. Mix et al., 2010).”
BackgroundMethodResultInfluentialOng, R. C. S., & Tang, A. D. (2025) · Science Advances · doi:10.1126/sciadv.ado6705 · Cited by 22 · see citing works
Metaplasticity in human primary somatosensory cortex: effects on physiology and tactile perception
Jones, C. B., Lulic-Kuryllo, T., Bailey, A. Z., Mackenzie, T. N., Mi, Y. Q., Tommerdahl, M., & Nelson, A. J. (2016) · Journal of Neurophysiology · doi:10.1152/jn.00630.2015 · Cited by 21 · see citing works
Neurostimulation for the treatment of functional neurological disorder: A systematic review
Gonsalvez, I., Spagnolo, P. A., Dworetzky, B. A., & Baslet, G. C. (2021) · Epilepsy & Behavior Reports · doi:10.1016/j.ebr.2021.100501 · Cited by 21 · see citing works
Continuous theta-burst stimulation modulates tactile synchronization
Lee, K. G. H., Jacobs, M. F., Asmussen, M. J., Zapallow, C. M., Tommerdahl, M., & Nelson, A. J. (2013) · BMC Neuroscience · doi:10.1186/1471-2202-14-89 · Cited by 20 · see citing works
“However, in rats, cTBS does not alter the learning of a tactile discrimination task [57].”
BackgroundWyckhuys, T., Geeter, N. D., Crevecoeur, G., Stroobants, S. G., & Staelens, S. G. (2012) · Brain stimulation · doi:10.1016/j.brs.2012.10.004 · Cited by 18 · see citing works
Wu, M.-K., Satogami, K., Liang, C.-S., Stubbs, B., Carvalho, A. F., Brunoni, A., Su, K., Tu, Y.-X., Wu, Y.-C., Chen, T.-Y., et al. (2022) · Psychiatry and Clinical Neurosciences · doi:10.1111/pcn.13452 · Cited by 15 · see citing works
Kloosterboer, E., & Funke, K. (2019) · The Journal of Physiology · doi:10.1113/jp277507 · Cited by 17 · see citing works
Spagnolo, P. A., Pérez, L. J. G., Terraneo, A., Gallimberti, L., & Bonci, A. (2018) · European Journal of Neuroscience · doi:10.1111/ejn.14313 · Cited by 16 · see citing works
“However, theta burst stimulation (a variation of high-frequency rTMS) can either depress (continuous) or facilitate (intermittent) cortical excitability, depending on burst-train duration (Mix et al., 2010).”
BackgroundLippmann, B., Barmashenko, G., & Funke, K. (2021) · European Journal of Neuroscience · doi:10.1111/ejn.15125 · Cited by 14 · see citing works
Transcriptional changes in the rat brain induced by repetitive transcranial magnetic stimulation
Weiler, M., Stieger, K. C., Shroff, K., Klein, J. P., Wood, W. H., Zhang, Y., Chandrasekaran, P., Lehrmann, E., Camandola, S., Long, J. M., et al. (2023) · Frontiers in Human Neuroscience · doi:10.3389/fnhum.2023.1215291 · Cited by 13 · see citing works
“Similar to procedures described in previous studies (Trippe et al., 2009; Mix et al., 2010, 2015; Benali et al., 2011; Hoppenrath et al., 2016), the coil was centered 8 mm above the rat’s skull, oriented with the handle to the left of the rat to produce a mediolaterally oriented electric field,…”
ResultLehmann, S. J., & Corneil, B. D. (2021) · Neuroscience & Biobehavioral Reviews · doi:10.1016/j.neubiorev.2021.10.040 · Cited by 11 · see citing works
Assessment and modulation of cortical inhibition using transcranial magnetic stimulation
Vlachos, A., Funke, K., & Ziemann, U. (2017) · e-Neuroforum · doi:10.1515/nf-2016-a103 · Cited by 13 · see citing works
“Consistent with these findings it was shown that rTMS promotes learning in animal experiments (Mix et al., 2010) and motor rehabilitation following ischemic stroke in humans (Brodie et al., 2014; Volz et al., 2016).”
ResultJazmati, D., Neubacher, U., & Funke, K. (2018) · Brain stimulation · doi:10.1016/j.brs.2018.02.017 · Cited by 13 · see citing works
Weiler, M., Moreno‐Castilla, P., Starnes, H. M., Melendez, E. T., Stieger, K. C., Long, J. M., & Rapp, P. R. (2021) · Brain stimulation · doi:10.1016/j.brs.2021.08.008 · Cited by 13 · see citing works
“The stimulation protocol 156 was adapted from earlier work in rats [42-44], using an intensity of stimulation just below 157 that expected to elicit an evoked motor response [45, 46] and adjusted to accommodate the 158 J urn al Pr e-p roo f”
MethodSafety of Special Waveform of Transcranial Electrical Stimulation (TES): In Vivo Assessment
Adeel, M., Chen, C.-C., Lin, B.-S., Chen, H., Liou, J., Li, Y., & Peng, C. (2022) · International Journal of Molecular Sciences · doi:10.3390/ijms23126850 · Cited by 10 · see citing works
“However, the frontal and barrel cortex, which are engaged in the learning process, exhibited a considerably smaller reduction in PV and CB expression than the visual cortex, which was not involved in the activity [53].”
BackgroundZhang, X., Yang, X.-H., Shi, Z.-M., Xu, R., Tan, J., Yang, J., Huang, X., Huang, X., & Zheng, W. (2023) · Journal of Personalized Medicine · doi:10.3390/jpm13030485 · Cited by 10 · see citing works
“An animal study found that iTBS rather than cTBS was associated with improvements in learning and strongly reduced cortical protein expression, such as parvalbumin [20].”
BackgroundZhang, T. R., Askari, B., Kesici, A., Guilherme, E. M., Vila‐Rodriguez, F., & Snyder, J. S. (2022) · European Journal of Neuroscience · doi:10.1111/ejn.15891 · Cited by 9 · see citing works
“…theta frequency stimulation reliably induces long-term potentiation, a synaptic model of memory, at hippocampal synapses (Cao & Harris, 2014; Larson & Munk acsy, 2015; Sheridan et al., 2014) and, iTBS enhances learning (Mix et al., 2010) and pattern discrimination (Castillo-Padilla & Funke, 2016).”
BackgroundTranscranial Magnetic Stimulation of Rodents
Funke, K. (2018) · Handbook of behavioral neuroscience · doi:10.1016/b978-0-12-812028-6.00020-3 · Cited by 8 · see citing works
Effects of iTBS-rTMS on the Behavioral Phenotype of a Rat Model of Maternal Immune Activation
Rittweger, N., Ishorst, T., Barmashenko, G., Aliane, V., Winter, C., & Funke, K. (2021) · Frontiers in Behavioral Neuroscience · doi:10.3389/fnbeh.2021.670699 · Cited by 7 · see citing works
Makale, M. T., Nybo, C., Blum, K., Dennen, C. A., Elman, I., & Murphy, K. T. (2024) · Journal of Personalized Medicine · doi:10.3390/jpm14080857 · Cited by 6 · see citing works
“On the other hand, low intensity PrTMS improves outcomes owing to the induction of beneficial forms of neuroplasticity [46–50].”
BackgroundCortical Modulation of the Transient Visual Response at Thalamic Level: A TMS Study
Espinosa, N., Mariño, J., de Labra, C., & Cudeiro, J. (2011) · PLoS ONE · doi:10.1371/journal.pone.0017041 · Cited by 5 · see citing works
Nouhaud, C., Sherrard, R. M., & Belmin, J. (2016) · La Revue de Médecine Interne · doi:10.1016/j.revmed.2016.06.005 · Cited by 5 · see citing works
James, J. C., & Funke, K. (2020) · Developmental Neurobiology · doi:10.1002/dneu.22785 · Cited by 5 · see citing works
“, 2013), indicative of a reduced activity of these neurons and less cortical inhibition (Funke & Benali, 2011) which was accompanied by increased sensory responses (Thimm & Funke, 2015) and improved learning performance (Mix et al., 2010) in rats.”
BackgroundMethodModulating motor cortex plasticity via cortical and peripheral somatosensory stimulation
Bao, S., & Lei, Y. (2025) · Journal of Neurophysiology · doi:10.1152/jn.00600.2024 · Cited by 4 · see citing works
Pecsok, M. K., Mordy, A. E., Cristancho, M. A., Oathes, D. J., & Roalf, D. R. (2024) · Psychopathology · doi:10.1159/000538690 · Cited by 3 · see citing works
Kanigowski, D., & Urban‐Ciećko, J. (2025) · Journal of Neural Transmission · doi:10.1007/s00702-025-02949-5 · Cited by 3 · see citing works
“Such magnetic stimulation of the cortex can also positively influence asso - ciative learning related to tactile stimuli recognized by vibrissae (Mix et al. 2010 ).”
Siebner, H. R. (2010) · European Journal of Neuroscience · doi:10.1111/j.1460-9568.2010.07476.x · Cited by 3 · see citing works
“In the article published in this issue of EJN, Annika Mix and collaborators (Mix et al., 2010) addressed the question how transcranial stimulation and subsequent learning influence the cortical expression of proteins linked to inhibitory neurotransmission.”
BackgroundGore, I. R., Brown, C. J., Waters, R. C., & Gould, E. (2024) · Learning & Memory · doi:10.1101/lm.053968.124 · Cited by 3 · see citing works
“Although it remains unknown whether the PV intensity decreaseresultsfromorisacauseofchangesinnetworkactivity,previous studies have shown that experimentally induced theta bursts reduce PV expression in other brain regions (Mix et al. 2010, 2015).”
Dastgheib, Z. A., Lithgow, B., & Moussavi, Z. (2025) · Cognitive Neurodynamics · doi:10.1007/s11571-025-10310-5 · Cited by 1 · see citing works
“It comprises an increase of GABAergic synthesizing enzymes and transporters after low-frequency rTMS stimulation (Trippe et al. 2009; Mix et al. 2010) and a decrease in the number of inhibitory cells after high frequency rTMS stimulation (Benali et al. 2011; Jazmati et al. 2018).”
Long, J., Liao, X., Han, K., Chen, J., Tang, Z., Wang, W., Wang, X., Qianqian, C., Zhang, X., & Zhang, H. (2026) · Cellular and Molecular Life Sciences · doi:10.1007/s00018-026-06096-2 · Cited by 2 · see citing works
“Similarly, Mix et al. [38 ] reported that iTBS, but not cTBS, significantly enhanced learning performance in a tactile discrimination task in rats. iTBS also improved the performance of rodent models [48–50 ], the present study found no signifi - cant difference in gut microbial α-diversity but a…”
李, 查. (2022) · Advances in Clinical Medicine · doi:10.12677/acm.2022.12121579 · Cited by 1 · see citing works
Ceravolo, L., Moisa, M., Grandjean, D., Ruff, C. C., & Frühholz, S. (2021) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2021.12.15.472758 · Cited by 1 · see citing works
Jones, C. B. (2016) · MacSphere (McMaster University) · doi:10.71548/9403
Epidemiologie und Versorgungssituation von Zwangsstörungen
Roseneck, S. K. (2011)
Untersuchung und Modulation kortikaler Inhibition mittels transkranieller Magnetstimulation
Vlachos, A., Funke, K., & Ziemann, U. (2017) · e-Neuroforum · doi:10.1515/nf-2016-1103
Tang, A. D. (2016) · UWA Profiles and Research Repository (UWA)
Fulopova, B. (2015) · UTAS Research Repository · doi:10.25959/23238728
Kanigowski, D., & Urban‐Ciećko, J. (2024) · Research Square · doi:10.21203/rs.3.rs-5071377/v1
Ceravolo, L., Moisa, M., Grandjean, D., Ruff, C. C., & Frühholz, S. (2026) · Scientific Reports · doi:10.1038/s41598-026-39668-0 · Cited by 1 · see citing works
“Such assumptions could also be tested in the future by using intermittent—as opposed to continuous—TBS over the right and/or left IFG pars triangularis , known to enhance brain activity and not alter it 30,59 .”
Dufor, T. (2017) · theses.fr (ABES) · doi:10.70675/2b83812fzee8fz42ebz8376z569f5731c84e · Cited by 3 · see citing works
“In other studies from the group, modulation of inhibitory neuron activation induced by rTMS occurred with functional alterations such as tactile learning performance (Mix et al., 2010) and sensory responses in rat barrel cortex (Thimm and Funke, 2015).”
BackgroundLee, K. G. H. (2013) · MacSphere (McMaster University) · doi:10.71548/24834
- Authors
- Mix, A., Benali, A., Eysel, U. T., & Funke, K.
- Affiliations
- Ruhr University Bochum (DE)
- Corresponding author
- Funke, Klaus
- Venue
- European Journal of Neuroscience
- Publisher
- Wiley
- Published
- 2010-10-18
- Volume
- 32
- Issue
- 9
- Pages
- 1575-1586
- ISSN
- 0953-816X (ISSN-L)
- 1460-9568
- OpenAlex
- W1689956239
- Cited by
- 78
- References
- 62
- Citations by year
- 2026: 2 · 2025: 4 · 2024: 5 · 2023: 3 · 2022: 6 · 2021: 6 · 2020: 1 · 2019: 4 · 2018: 4 · 2017: 6 · 2016: 12 · 2015: 4 · 2014: 6 · 2013: 6 · 2012: 3
- FWCI
- 2.86
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- European Journal of Neuroscience (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Deutsche Forschungsgemeinschaft
- Topics
- Life Sciences › Neuroscience › Neurology › Transcranial Magnetic Stimulation Studies
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Neuroscience › Cognitive Neuroscience › Memory and Neural Mechanisms
- Keywords
- theta burst stimulation, intermittent theta burst stimulation, continuous theta burst stimulation, parvalbumin, GAD67, cortical inhibition, fast-spiking interneurons, cortical excitability, repetitive transcranial magnetic stimulation, learning-induced plasticity, homeostatic plasticity
- MeSH terms
- Animals, Behavior, Animal, Cerebral Cortex, Glutamate Decarboxylase, Humans, Learning, Male, Nerve Tissue Proteins, Parvalbumins, Psychomotor Performance, Theta Rhythm, Rats, Sprague-Dawley, Transcranial Magnetic Stimulation, Rats
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