{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,22]],"date-time":"2026-06-22T14:47:25Z","timestamp":1782139645286,"version":"3.54.5"},"reference-count":44,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2025,4,18]],"date-time":"2025-04-18T00:00:00Z","timestamp":1744934400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000071","name":"NICHD","doi-asserted-by":"publisher","award":["R01HD088549"],"award-info":[{"award-number":["R01HD088549"]}],"id":[{"id":"10.13039\/100000071","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Understanding the timing of fetal brain vulnerability to inflammatory changes in pregnancy complications is crucial for predicting neurodevelopmental risks. Beyond the placenta, the developing brain\u2019s vascular system is believed to form a secondary defense, the blood\u2013brain barrier (BBB), which restricts harmful substances that could disrupt neurodevelopment. However, the precise timing and mechanisms underlying BBB development are poorly understood. In this study, we examined the spatiotemporal expression of key BBB components and fetal brain vascularization in mice from gestational days (GD) 10 to 18. Fetal brain sections were immunostained to identify BBB components, including CD31, Factor VIII, NG2, and claudin-5. Our results showed that endothelial precursor cells form the primitive vascular network in a caudal-to-rostral gradient by GD10, with pericyte recruitment stabilizing vessels by GD12 in a lateral-to-medial gradient that aligns with neurogenesis, despite some regional exceptions. However, Factor VIII was not detected until GD15, and claudin-5 until GD18, suggesting a significant delay in endothelial maturation and tight junction formation. These findings highlight the critical timing of structural developments in the fetal brain vasculature and its vulnerability to placental diseases, laying the groundwork for future research on the impact of placental disorders on fetal brain development and potential therapeutic interventions.<\/jats:p>","DOI":"10.3390\/ijms26083862","type":"journal-article","created":{"date-parts":[[2025,4,20]],"date-time":"2025-04-20T20:24:16Z","timestamp":1745180656000},"page":"3862","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Spatiotemporal Angiogenic Patterns in the Development of the Mouse Fetal Blood\u2013Brain Barrier System During Pregnancy"],"prefix":"10.3390","volume":"26","author":[{"given":"Samuel Nofsinger","family":"Brown","sequence":"first","affiliation":[{"name":"Department of Obstetrics and Gynecology, University of Missouri Kansas City School of Medicine, Kansas, MO 64108, USA"},{"name":"University of Missouri School of Medicine, Columbia, MO 65212, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9910-3584","authenticated-orcid":false,"given":"Philemon","family":"Shallie","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, University of Missouri Kansas City School of Medicine, Kansas, MO 64108, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0465-0356","authenticated-orcid":false,"given":"Connor A.","family":"Sierra","sequence":"additional","affiliation":[{"name":"University of Missouri School of Medicine, Columbia, MO 65212, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Neha","family":"Nayak","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, University of Missouri Kansas City School of Medicine, Kansas, MO 64108, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anthony O.","family":"Odibo","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, University of Missouri Kansas City School of Medicine, Kansas, MO 64108, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paula","family":"Monaghan-Nichols","sequence":"additional","affiliation":[{"name":"Department of Biomedical Sciences, University of Missouri Kansas City School of Medicine, Kansas, MO 64108, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nihar R.","family":"Nayak","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, University of Missouri Kansas City School of Medicine, Kansas, MO 64108, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,4,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/j.placenta.2016.12.005","article-title":"Review: The blood-brain barrier; protecting the developing fetal brain","volume":"54","author":"Miller","year":"2017","journal-title":"Placenta"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1016\/j.rbmo.2012.03.017","article-title":"Placental adaptations to the maternal-fetal environment: Implications for fetal growth and developmental programming","volume":"25","author":"Sandovici","year":"2012","journal-title":"Reprod. Biomed. Online"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Saunders, N.R., Dreifuss, J.J., Dziegielewska, K.M., Johansson, P.A., Habgood, M.D., M\u00f8llg\u00e5rd, K., and Bauer, H.C. (2014). The rights and wrongs of blood-brain barrier permeability studies: A walk through 100 years of history. Front. Neurosci., 8.","DOI":"10.3389\/fnins.2014.00404"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"562","DOI":"10.1038\/nature09513","article-title":"Pericytes are required for blood-brain barrier integrity during embryogenesis","volume":"468","author":"Daneman","year":"2010","journal-title":"Nature"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"478","DOI":"10.1038\/s41593-020-00788-z","article-title":"Oligodendrocyte precursor cell specification is regulated by bidirectional neural progenitor-endothelial cell crosstalk","volume":"24","author":"Paredes","year":"2021","journal-title":"Nat. Neurosci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.devcel.2018.01.023","article-title":"Neurovascular Communication during CNS Development","volume":"45","author":"Paredes","year":"2018","journal-title":"Dev. Cell"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"641","DOI":"10.1073\/pnas.0805165106","article-title":"Wnt\/beta-catenin signaling is required for CNS, but not non-CNS, angiogenesis","volume":"106","author":"Daneman","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Mamber, C., Kamphuis, W., Haring, N.L., Peprah, N., Middeldorp, J., and Hol, E.M. (2012). GFAP\u03b4 expression in glia of the developmental and adolescent mouse brain. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0052659"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e63253","DOI":"10.7554\/eLife.63253","article-title":"Nano-scale architecture of blood-brain barrier tight-junctions","volume":"10","author":"Sasson","year":"2021","journal-title":"eLife"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Steinemann, A., Galm, I., Chip, S., Nitsch, C., and Maly, I.P. (2016). Claudin-1, -2 and -3 Are Selectively Expressed in the Epithelia of the Choroid Plexus of the Mouse from Early Development and into Adulthood While Claudin-5 is Restricted to Endothelial Cells. Front. Neuroanat., 10.","DOI":"10.3389\/fnana.2016.00016"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"643","DOI":"10.1038\/nrneurol.2014.187","article-title":"Maternal immune activation and abnormal brain development across CNS disorders","volume":"10","author":"Knuesel","year":"2014","journal-title":"Nat. Rev. Neurol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"192","DOI":"10.1038\/nrneurol.2015.13","article-title":"The role of inflammation in perinatal brain injury","volume":"11","author":"Hagberg","year":"2015","journal-title":"Nat. Rev. Neurol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1186\/1742-2094-9-175","article-title":"JNK signaling is the shared pathway linking neuroinflammation, blood-brain barrier disruption, and oligodendroglial apoptosis in the white matter injury of the immature brain","volume":"9","author":"Wang","year":"2012","journal-title":"J. Neuroinflamm."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"823","DOI":"10.1001\/jamapsychiatry.2020.0306","article-title":"Association of Preeclampsia in Term Births with Neurodevelopmental Disorders in Offspring","volume":"77","author":"Sun","year":"2020","journal-title":"JAMA Psychiatry"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1111\/bcpt.13456","article-title":"Placental transfer of pesticides studied in human placental perfusion","volume":"127","author":"Mathiesen","year":"2020","journal-title":"Basic Clin. Pharmacol. Toxicol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1186\/s12974-017-0981-8","article-title":"Acute in utero exposure to lipopolysaccharide induces inflammation in the pre- and postnatal brain and alters the glial cytoarchitecture in the developing amygdala","volume":"14","author":"Pakan","year":"2017","journal-title":"J. Neuroinflamm."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1002\/cne.20885","article-title":"Functional effectiveness of the blood-brain barrier to small water-soluble molecules in developing and adult opossum (Monodelphis domestica)","volume":"496","author":"Ek","year":"2006","journal-title":"J. Comp. Neurol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"5723","DOI":"10.1113\/JP275376","article-title":"Physiology and molecular biology of barrier mechanisms in the fetal and neonatal brain","volume":"596","author":"Saunders","year":"2018","journal-title":"J. Physiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1083\/jcb.201410131","article-title":"LSR\/angulin-1 is a tricellular tight junction protein involved in blood-brain barrier formation","volume":"208","author":"Sohet","year":"2015","journal-title":"J. Cell Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"507","DOI":"10.1038\/nature13324","article-title":"Mfsd2a is critical for the formation and function of the blood-brain barrier","volume":"509","author":"Lacoste","year":"2014","journal-title":"Nature"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1111\/joim.13263","article-title":"Developmental regulation of barrier- and non-barrier blood vessels in the CNS","volume":"292","author":"Liebner","year":"2022","journal-title":"J. Intern. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1097\/nen.0b013e31815f65d9","article-title":"Fetal blood-brain barrier P-glycoprotein contributes to brain protection during human development","volume":"67","author":"Virgintino","year":"2008","journal-title":"J. Neuropathol. Exp. Neurol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.mod.2015.07.001","article-title":"Vascularisation of the central nervous system","volume":"138","author":"Tata","year":"2015","journal-title":"Mech. Dev."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1675","DOI":"10.1007\/s00018-013-1277-5","article-title":"Neurovascular development and links to disease","volume":"70","author":"Ruhrberg","year":"2013","journal-title":"Cell Mol. Life Sci."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Puelles, L., Mart\u00ednez-Marin, R., Melgarejo-Otalora, P., Ayad, A., Valavanis, A., and Ferran, J.L. (2019). Patterned Vascularization of Embryonic Mouse Forebrain, and Neuromeric Topology of Major Human Subarachnoidal Arterial Branches: A Prosomeric Mapping. Front. Neuroanat., 13.","DOI":"10.3389\/fnana.2019.00059"},{"key":"ref_26","unstructured":"(2025, February 13). Allen Reference Atlas\u2014Mouse Brain [Brain Atlas]. Available online: https:\/\/atlas.brain-map.org\/."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"167","DOI":"10.4161\/cam.2.3.6485","article-title":"Angiogenesis in the embryonic CNS: A new twist on an old tale","volume":"2","author":"Vasudevan","year":"2008","journal-title":"Cell Adhes. Migr."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1182\/blood-2009-12-257832","article-title":"Tissue macrophages act as cellular chaperones for vascular anastomosis downstream of VEGF-mediated endothelial tip cell induction","volume":"116","author":"Fantin","year":"2010","journal-title":"Blood"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"4489","DOI":"10.1242\/dev.107193","article-title":"Excessive vascular sprouting underlies cerebral hemorrhage in mice lacking \u03b1V\u03b28-TGF\u03b2 signaling in the brain","volume":"141","author":"Arnold","year":"2014","journal-title":"Development"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Ruiz-Reig, N., and Studer, M. (2017). Rostro-Caudal and Caudo-Rostral Migrations in the Telencephalon: Going Forward or Backward?. Front. Neurosci., 11.","DOI":"10.3389\/fnins.2017.00692"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1038\/nature09522","article-title":"Pericytes regulate the blood-brain barrier","volume":"468","author":"Armulik","year":"2010","journal-title":"Nature"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"705","DOI":"10.1177\/0192623317728134","article-title":"Histology Atlas of the Developing Prenatal and Postnatal Mouse Central Nervous System, with Emphasis on Prenatal Days E7.5 to E18.5","volume":"45","author":"Chen","year":"2017","journal-title":"Toxicol. Pathol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1018","DOI":"10.1002\/dneu.20954","article-title":"Extracellular matrix and matrix receptors in blood-brain barrier formation and stroke","volume":"71","author":"Baeten","year":"2011","journal-title":"Dev. Neurobiol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Li, Y., Shen, M., Stockton, M.E., and Zhao, X. (2019). Hippocampal deficits in neurodevelopmental disorders. Neurobiol. Learn. Mem., 165.","DOI":"10.1016\/j.nlm.2018.10.001"},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Cie\u015blik, M., Gassowska-Dobrowolska, M., Zawadzka, A., Frontczak-Baniewicz, M., Gewartowska, M., Dominiak, A., Czapski, G.A., and Adamczyk, A. (2020). The Synaptic Dysregulation in Adolescent Rats Exposed to Maternal Immune Activation. Front. Mol. Neurosci., 13.","DOI":"10.3389\/fnmol.2020.555290"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2190","DOI":"10.1002\/jnr.21673","article-title":"Maternal immune activation in mice delays myelination and axonal development in the hippocampus of the offspring","volume":"86","author":"Makinodan","year":"2008","journal-title":"J. Neurosci. Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"892","DOI":"10.2353\/ajpath.2008.080001","article-title":"von-Willebrand factor influences blood brain barrier permeability and brain inflammation in experimental allergic encephalomyelitis","volume":"173","author":"Noubade","year":"2008","journal-title":"Am. J. Pathol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2112","DOI":"10.1161\/ATVBAHA.113.301362","article-title":"Endothelial Von Willebrand factor promotes blood-brain barrier flexibility and provides protection from hypoxia and seizures in mice","volume":"33","author":"Suidan","year":"2013","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"van Leeuwen, L.M., Evans, R.J., Jim, K.K., Verboom, T., Fang, X., Bojarczuk, A., Malicki, J., Johnston, S.A., and van der Sar, A.M. (2018). A transgenic zebrafish model for the in vivo study of the blood and choroid plexus brain barriers using claudin 5. Biol. Open, 7.","DOI":"10.1242\/bio.030494"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1016\/j.ajhg.2018.10.009","article-title":"Human Models Are Needed for Studying Human Neurodevelopmental Disorders","volume":"103","author":"Zhao","year":"2018","journal-title":"Am. J. Hum. Genet."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"103","DOI":"10.1016\/j.conb.2020.09.006","article-title":"Modeling brain development and diseases with human cerebral organoids","volume":"66","author":"Shi","year":"2021","journal-title":"Curr. Opin. Neurobiol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"4941","DOI":"10.1172\/JCI76864","article-title":"Endometrial VEGF induces placental sFLT1 and leads to pregnancy complications","volume":"124","author":"Fan","year":"2014","journal-title":"J. Clin. Investig."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Anamthathmakula, P., Shallie, P.D., Nayak, N., Dhal, S., Vivian, J.L., Mor, G., Soares, M.J., and Nayak, N.R. (2023). Variable Cre Recombination Efficiency in Placentas of Cyp19-Cre ROSAmT\/mG Transgenic Mice. Cells, 12.","DOI":"10.3390\/cells12162096"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"3571","DOI":"10.1096\/fj.08-111401","article-title":"VEGF blockade inhibits angiogenesis and reepithelialization of endometrium","volume":"22","author":"Fan","year":"2008","journal-title":"FASEB J."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/8\/3862\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:17:35Z","timestamp":1760030255000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/8\/3862"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,4,18]]},"references-count":44,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2025,4]]}},"alternative-id":["ijms26083862"],"URL":"https:\/\/doi.org\/10.3390\/ijms26083862","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,4,18]]}}}