{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T02:11:09Z","timestamp":1787623869519,"version":"build-2736575974"},"reference-count":48,"publisher":"MDPI AG","issue":"23","license":[{"start":{"date-parts":[[2022,11,27]],"date-time":"2022-11-27T00:00:00Z","timestamp":1669507200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003246","name":"NWO-ALW VIDI","doi-asserted-by":"publisher","award":["016.121.346"],"award-info":[{"award-number":["016.121.346"]}],"id":[{"id":"10.13039\/501100003246","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003246","name":"NWO-ALW VIDI","doi-asserted-by":"publisher","award":["91210067"],"award-info":[{"award-number":["91210067"]}],"id":[{"id":"10.13039\/501100003246","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003246","name":"NWO-ALW VIDI","doi-asserted-by":"publisher","award":["865.14.004"],"award-info":[{"award-number":["865.14.004"]}],"id":[{"id":"10.13039\/501100003246","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001826","name":"Zon-MW TOP GO","doi-asserted-by":"publisher","award":["016.121.346"],"award-info":[{"award-number":["016.121.346"]}],"id":[{"id":"10.13039\/501100001826","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001826","name":"Zon-MW TOP GO","doi-asserted-by":"publisher","award":["91210067"],"award-info":[{"award-number":["91210067"]}],"id":[{"id":"10.13039\/501100001826","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001826","name":"Zon-MW TOP GO","doi-asserted-by":"publisher","award":["865.14.004"],"award-info":[{"award-number":["865.14.004"]}],"id":[{"id":"10.13039\/501100001826","id-type":"DOI","asserted-by":"publisher"}]},{"name":"C.J. 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Apart from its role in coordinating sensorimotor integration in the adult brain, the cerebello-thalamic tract (CbT) has also been implicated in developmental disorders, such as autism spectrum disorders. Although the development of the cerebellum, thalamus and cerebral cortex have been studied, there is no detailed description of the ontogeny of the mammalian CbT. Here we investigated the development of the CbT at embryonic stages using transgenic Ntsr1-Cre\/Ai14 mice and in utero electroporation of wild type mice. Wide-field, confocal and 3D light-sheet microscopy of immunohistochemical stainings showed that CbT fibers arrive in the prethalamus between E14.5 and E15.5, but only invade the thalamus after E16.5. We quantified the spread of CbT fibers throughout the various thalamic nuclei and found that at E17.5 and E18.5 the ventrolateral, ventromedial and parafascicular nuclei, but also the mediodorsal and posterior complex, become increasingly innervated. Several CbT fiber varicosities express vesicular glutamate transporter type 2 at E18.5, indicating cerebello-thalamic synapses. Our results provide the first quantitative data on the developing murine CbT, which provides guidance for future investigations of the impact that cerebellum has on thalamo-cortical networks during development.<\/jats:p>","DOI":"10.3390\/cells11233800","type":"journal-article","created":{"date-parts":[[2022,11,28]],"date-time":"2022-11-28T04:56:25Z","timestamp":1669611385000},"page":"3800","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Anatomical Development of the Cerebellothalamic Tract in Embryonic Mice"],"prefix":"10.3390","volume":"11","author":[{"given":"Dani\u00ebl B.","family":"Dumas","sequence":"first","affiliation":[{"name":"Department of Neuroscience, Erasmus Medical Center, 3015 AA Rotterdam, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Simona V.","family":"Gornati","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Erasmus Medical Center, 3015 AA Rotterdam, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Youri","family":"Adolfs","sequence":"additional","affiliation":[{"name":"Department of Translational Neuroscience, MIND Facility, University Medical Center Utrecht Brain Center, University of Utrecht, 3584 CX Utrecht, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomomi","family":"Shimogori","sequence":"additional","affiliation":[{"name":"RIKEN Center for Brain Science, Laboratory for Molecular Mechanisms of Brain Development, Wako City 351-0198, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"R. Jeroen","family":"Pasterkamp","sequence":"additional","affiliation":[{"name":"Department of Translational Neuroscience, MIND Facility, University Medical Center Utrecht Brain Center, University of Utrecht, 3584 CX Utrecht, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Freek E.","family":"Hoebeek","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Erasmus Medical Center, 3015 AA Rotterdam, The Netherlands"},{"name":"Department for Developmental Origins of Disease, Wilhelmina Children\u2019s Hospital and University Medical Center Utrecht Brain Center, 3584 EA Utrecht, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,11,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"9817","DOI":"10.1523\/JNEUROSCI.2707-07.2007","article-title":"Targeting Cre Recombinase to Specific Neuron Populations with Bacterial Artificial Chromosome Constructs","volume":"27","author":"Gong","year":"2007","journal-title":"J. Neurosci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1038\/nn.2467","article-title":"A robust and high-throughput Cre reporting and characterization system for the whole mouse brain","volume":"13","author":"Madisen","year":"2010","journal-title":"Nat. Neurosci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1191","DOI":"10.1016\/j.celrep.2014.10.037","article-title":"A Simple Method for 3D Analysis of Immunolabeled Axonal Tracts in a Transparent Nervous System","volume":"9","author":"Belle","year":"2014","journal-title":"Cell Rep."},{"key":"ref_4","first-page":"3024","article-title":"Mouse in Utero Electroporation: Controlled Spatiotemporal Gene Transfection","volume":"54","author":"Matsui","year":"2011","journal-title":"J. Vis. Exp."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Jacobowitz, D.M., and Abbott, L.C. (1998). Chemoarchitectonic Atlas of the Developing Mouse Brain, CRC Press.","DOI":"10.1201\/9781466593411"},{"key":"ref_6","unstructured":"Schambra, U.B., Lauder, J.M., and Silver, J. (1992). Atlas of the Prenatal Mouse Brain, Academic Press. [1st ed.]."},{"key":"ref_7","unstructured":"Paxinos, G. (2007). Atlas of the Developing Mouse Brain: At E17.5, PO, and P6, Academic Press."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"309","DOI":"10.1016\/j.neuron.2014.05.033","article-title":"A High-Resolution Spatiotemporal Atlas of Gene Expression of the Developing Mouse Brain","volume":"83","author":"Thompson","year":"2014","journal-title":"Neuron"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"266","DOI":"10.1002\/cne.10654","article-title":"Characterization of Foxp2 and Foxp1 mRNA and Protein in the Developing and Mature Brain","volume":"460","author":"Ferland","year":"2003","journal-title":"J. Comp. Neurol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1038\/nrn1605","article-title":"FOXP2 and the neuroanatomy of speech and language","volume":"6","author":"Gadian","year":"2005","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2493","DOI":"10.1007\/s00429-015-1052-5","article-title":"Molecular anatomy of the thalamic complex and the underlying transcription factors","volume":"221","author":"Nagalski","year":"2016","journal-title":"Brain Struct. Funct."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3090","DOI":"10.1523\/JNEUROSCI.5266-07.2008","article-title":"Structural correlates of efficient GABAergic transmission in the basal ganglia-thalamus pathway","volume":"28","author":"Bodor","year":"2008","journal-title":"J. Neurosci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2690","DOI":"10.1016\/j.celrep.2018.04.098","article-title":"Differentiating Cerebellar Impact on Thalamic Nuclei","volume":"23","author":"Gornati","year":"2018","journal-title":"Cell Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2254","DOI":"10.1002\/cne.23787","article-title":"Cerebellar Premotor Output Neurons Collateralize to Innervate the Cerebellar Cortex","volume":"523","author":"Houck","year":"2015","journal-title":"J. Comp. Neurol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"789","DOI":"10.1007\/s12311-015-0724-2","article-title":"Consensus Paper: Cerebellar Development","volume":"15","author":"Leto","year":"2016","journal-title":"Cerebellum"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"9184","DOI":"10.1523\/JNEUROSCI.1610-06.2006","article-title":"Development\/Plasticity\/Repair Unipolar Brush Cells of the Cerebellum Are Produced in the Rhombic Lip and Migrate through Developing White Matter","volume":"26","author":"Englund","year":"2006","journal-title":"J. Neurosci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"999","DOI":"10.1002\/cne.23888","article-title":"Interstitial branch formation within the red nucleus by deep cerebellar nuclei-derived commissural axons during target recognition","volume":"524","author":"Hara","year":"2016","journal-title":"J. Comp. Neurol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1038\/nature10835","article-title":"Gain control by layer six in cortical circuits of vision","volume":"483","author":"Olsen","year":"2012","journal-title":"Nature"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1111\/j.1460-9568.2006.05258.x","article-title":"Visualization of corticofugal projections during early cortical development in a \u03c4-GFP-transgenic mouse","volume":"25","author":"Jacobs","year":"2007","journal-title":"Eur. J. Neurosci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1038\/s41586-018-0633-x","article-title":"A cortico-cerebellar loop for motor planning","volume":"563","author":"Gao","year":"2018","journal-title":"Nature"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/S0896-6273(01)00344-0","article-title":"The expression of vesicular glutamate transporters defines two classes of excitatory synapse","volume":"31","author":"Fremeau","year":"2001","journal-title":"Neuron"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1016\/S0169-328X(02)00442-4","article-title":"Expression of inorganic phosphate\/vesicular glutamate transporters (BNPI\/VGLUT1 and DNPI\/VGLUT2) in the cerebellum and precerebellar nuclei of the rat","volume":"107","author":"Hisano","year":"2002","journal-title":"Brain Res. Mol. Brain Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1016\/S0168-0102(02)00009-3","article-title":"Complementary distribution of vesicular glutamate transporters in the central nervous system","volume":"42","author":"Kaneko","year":"2002","journal-title":"Neurosci. Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"53","DOI":"10.3389\/fnins.2012.00053","article-title":"Development of the Corticothalamic Projections","volume":"6","author":"Grant","year":"2012","journal-title":"Front. Neurosci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1159\/000113567","article-title":"Brainstem Reticular Nuclei that Project to the Thalamus in Rats: A Retrograde Tracer Study","volume":"44","author":"Newman","year":"2008","journal-title":"Brain Behav. Evol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1002\/cne.903590107","article-title":"Ascending projections of the posterior nucleus of the hypothalamus: PHA-L analysis in the rat","volume":"359","author":"Vertes","year":"1995","journal-title":"J. Comp. Neurol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.neures.2014.10.016","article-title":"Efferent and afferent connections of the ventromedial hypothalamic nucleus determined by neural tracer analysis: Implications for lordosis regulation in female rats","volume":"91","author":"Shimogawa","year":"2015","journal-title":"Neurosci. Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1002\/cne.23183","article-title":"Development of the principal nucleus trigeminal lemniscal projections in the mouse","volume":"521","author":"Kivrak","year":"2013","journal-title":"J. Comp. Neurol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1016\/j.tcb.2017.01.005","article-title":"The interplay of axonal energy homeostasis and mitochondrial trafficking and anchoring","volume":"27","author":"Sheng","year":"2017","journal-title":"Trends Cell Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1002\/cne.902840405","article-title":"Development of the rat thalamus: IV. The intermediate lobule of the thalamic neuroepithelium, and the time and site of origin and settling pattern of neurons of the ventral nuclear complex","volume":"284","author":"Altman","year":"1989","journal-title":"J. Comp. Neurol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1016\/j.neuron.2012.11.031","article-title":"Pathfinding of Corticothalamic Axons Relies on a Rendezvous with Thalamic Projections","volume":"77","author":"Deck","year":"2013","journal-title":"Neuron"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3066","DOI":"10.1523\/JNEUROSCI.5203-05.2006","article-title":"Development of the deep cerebellar nuclei: Transcription factors and cell migration from the rhombic lip","volume":"26","author":"Fink","year":"2006","journal-title":"J. Neurosci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"12542","DOI":"10.1523\/JNEUROSCI.2521-09.2009","article-title":"Semaphorin 3F is a bifunctional guidance cue for dopaminergic axons and controls their fasciculation, channeling, rostral growth, and intracortical targeting","volume":"29","author":"Kolk","year":"2009","journal-title":"J. Neurosci."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Bibollet-Bahena, O., Okafuji, T., Hokamp, K., Tear, G., and Mitchell, K.J. (2017). A dual-strategy expression screen for candidate connectivity labels in the developing thalamus. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0177977"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1016\/S0079-6123(00)24014-4","article-title":"Topography of cerebellar nuclear projections to the brain stem in the rat","volume":"124","author":"Teune","year":"2000","journal-title":"Prog. Brain Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"298","DOI":"10.1038\/s41583-019-0152-2","article-title":"Emerging connections between cerebellar development, behaviour and complex brain disorders","volume":"20","author":"Sathyanesan","year":"2019","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"14172","DOI":"10.1038\/ncomms14172","article-title":"Prenatal thalamic waves regulate cortical area size prior to sensory processing","volume":"8","author":"Filipchuk","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_38","first-page":"575","article-title":"Development of the thalamus","volume":"Volume 1","author":"Jones","year":"2007","journal-title":"The Thalamus"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1016\/0361-9230(82)90165-4","article-title":"In vivo and in vitro development of serotonergic neurons","volume":"9","author":"Lauder","year":"1982","journal-title":"Brain Res. Res. Bull."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1002\/cne.902600204","article-title":"Development of brainstem and cerebellar projections to the diencephalon with notes on thalamocortical projections: Studies in the north american opossum","volume":"260","author":"Martin","year":"1987","journal-title":"J. Comp. Neurol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/0165-3806(85)90230-5","article-title":"Electrophysiological studies on the postnatal development of intracerebellar nuclei neurons in rat cerebellar slices maintained in vitro. I. Postsynaptic potentials","volume":"351","author":"Gardette","year":"1985","journal-title":"Brain Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"5215","DOI":"10.1523\/JNEUROSCI.4685-06.2007","article-title":"Interplay between laminar specificity and activity-dependent mechanisms of thalamocortical axon branching","volume":"27","author":"Uesaka","year":"2007","journal-title":"J. Neurosci."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1002\/dneu.22317","article-title":"Synapse-dependent and independent mechanisms of thalamocortical axon branching are regulated by neuronal activity","volume":"76","author":"Matsumoto","year":"2016","journal-title":"Dev. Neurobiol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"518","DOI":"10.1016\/j.neuron.2014.07.016","article-title":"The cerebellum, sensitive periods, and autism","volume":"83","author":"Wang","year":"2014","journal-title":"Neuron"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"549","DOI":"10.1146\/annurev.neuro.27.070203.144327","article-title":"Critical period regulation","volume":"27","author":"Hensch","year":"2004","journal-title":"Annu. Rev. Neurosci."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"181","DOI":"10.1159\/000515189","article-title":"Abnormal Cerebellar Development in Autism Spectrum Disorders","volume":"43","author":"Gill","year":"2021","journal-title":"Dev. Neurosci."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"659","DOI":"10.1038\/mp.2013.78","article-title":"The autism brain imaging data exchange: Towards a large-scale evaluation of the intrinsic brain architecture in autism","volume":"19","author":"Yan","year":"2014","journal-title":"Mol. Psychiatry"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"4497","DOI":"10.1002\/hbm.22938","article-title":"Regional specificity of aberrant thalamocortical connectivity in autism","volume":"36","author":"Nair","year":"2015","journal-title":"Hum. Brain Mapp."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/23\/3800\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:28:00Z","timestamp":1760146080000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/23\/3800"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,11,27]]},"references-count":48,"journal-issue":{"issue":"23","published-online":{"date-parts":[[2022,12]]}},"alternative-id":["cells11233800"],"URL":"https:\/\/doi.org\/10.3390\/cells11233800","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/731968","asserted-by":"object"}]},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,11,27]]}}}