{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,22]],"date-time":"2026-02-22T08:08:01Z","timestamp":1771747681015,"version":"3.50.1"},"reference-count":48,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,9,11]],"date-time":"2025-09-11T00:00:00Z","timestamp":1757548800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000865","name":"Bill and Melinda Gates Foundation","doi-asserted-by":"publisher","award":["OPP1182584"],"award-info":[{"award-number":["OPP1182584"]}],"id":[{"id":"10.13039\/100000865","id-type":"DOI","asserted-by":"publisher"}]},{"name":"University of Washington Department of Pediatrics","award":["OPP1182584"],"award-info":[{"award-number":["OPP1182584"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Life"],"abstract":"<jats:p>Gyrification, the folding of the cerebral cortex, plays a crucial role in brain development and function. Perinatal hypoxia\u2013ischemia (HI) is a leading cause of neonatal brain injury, affecting cortical folding that can be measured by the gyrification index (GI). Using a late-preterm ferret model, our objective was to explore the relationships between HI injury, GI changes, and behavior, as well as the potential moderating effects of sex and treatment. Animals received 3 mg\/kg E. coli lipopolysaccharide and underwent bilateral carotid artery ligation followed by alternating hypoxia and hyperoxia (HIH) and were randomized to saline vehicle (n = 25), erythropoietin (n = 20), therapeutic hypothermia (6 h at 33.5 \u00b0C, n = 20), and uridine monophosphate (n = 6), with n = 20 unexposed littermates serving as controls. Early reflex testing, CatWalk gait analysis, open-field behavior, and an open-water swim test were performed. Average, peak, motor, and somatosensory strip GIs were then assessed using ex vivo MRI. In control animals, males had lower GI than females; however, HIH exposure resulted in male GI being more similar to females, where HIH animals had significantly higher average GI than controls (p = 0.02). Adjusting for brain volume and injury, GIs in motor and somatosensory areas were associated with faster reflex outcomes in males but not females. In female controls, motor and somatosensory GIs were associated with increased anxiety-like behaviors, such as spending less time in open water during the swim test. By comparison, in male controls, higher GI was associated with decreased anxiety-like behaviors, including higher exploration index in the swim test. These sex-specific relationships between GI and behavior were lost with HIH injury. Treatment did not meaningfully restore the relationship between GI and behavior after HIH, but targeting this outcome may be an important measure for use in future neuroprotection studies in the ferret.<\/jats:p>","DOI":"10.3390\/life15091428","type":"journal-article","created":{"date-parts":[[2025,9,11]],"date-time":"2025-09-11T14:23:50Z","timestamp":1757600630000},"page":"1428","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Ferreting Out the Effects of Neonatal Hypoxia\u2013Ischemia and Sex on Ferret Cortical Gyrification"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-0752-9509","authenticated-orcid":false,"given":"Olivia C.","family":"Brandon","sequence":"first","affiliation":[{"name":"Division of Neonatology, University of Washington, Seattle, WA 98105, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-6680-1736","authenticated-orcid":false,"given":"Olivia R.","family":"White","sequence":"additional","affiliation":[{"name":"Division of Neonatology, University of Washington, Seattle, WA 98105, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2796-2184","authenticated-orcid":false,"given":"Kylie A.","family":"Corry","sequence":"additional","affiliation":[{"name":"Division of Neonatology, University of Washington, Seattle, WA 98105, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andreea","family":"Stanescu","sequence":"additional","affiliation":[{"name":"Division of Neonatology, University of Washington, Seattle, WA 98105, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-6366-1890","authenticated-orcid":false,"given":"Arian","family":"Ariaye","sequence":"additional","affiliation":[{"name":"Division of Neonatology, University of Washington, Seattle, WA 98105, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel H.","family":"Moralejo","sequence":"additional","affiliation":[{"name":"Division of Neonatology, University of Washington, Seattle, WA 98105, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6279-1528","authenticated-orcid":false,"given":"Janessa B.","family":"Law","sequence":"additional","affiliation":[{"name":"Division of Neonatology, University of Washington, Seattle, WA 98105, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1397-8430","authenticated-orcid":false,"given":"Sarah E.","family":"Kolnik","sequence":"additional","affiliation":[{"name":"Division of Neonatology, University of Washington, Seattle, WA 98105, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6861-6229","authenticated-orcid":false,"given":"Sandra E.","family":"Juul","sequence":"additional","affiliation":[{"name":"Division of Neonatology, University of Washington, Seattle, WA 98105, USA"},{"name":"Institute on Human Development and Disability, University of Washington, Seattle, WA 98195, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1130-2720","authenticated-orcid":false,"given":"Thomas R.","family":"Wood","sequence":"additional","affiliation":[{"name":"Division of Neonatology, University of Washington, Seattle, WA 98105, USA"},{"name":"Institute on Human Development and Disability, University of Washington, Seattle, WA 98195, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,11]]},"reference":[{"key":"ref_1","first-page":"631","article-title":"Research Progress of Treatment for Neonatal Hypoxic Ischemic Encephalopathy","volume":"24","author":"Yifan","year":"2021","journal-title":"Chin. J. Appl. Clin. Pediatr."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Millar, L.J., Shi, L., Hoerder-Suabedissen, A., and Moln\u00e1r, Z. (2017). Neonatal Hypoxia Ischaemia: Mechanisms, Models, and Therapeutic Challenges. Front. Cell. Neurosci., 11.","DOI":"10.3389\/fncel.2017.00078"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"396","DOI":"10.1001\/jamapediatrics.2024.6613","article-title":"Whole-Body Hypothermia for Neonatal Encephalopathy in Preterm Infants 33 to 35 Weeks\u2019 Gestation","volume":"179","author":"Faix","year":"2025","journal-title":"JAMA Pediatr."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"3553","DOI":"10.21037\/jtd.2017.09.64","article-title":"The Search Continues: Neuroprotection for All Neonates with Hypoxic-Ischemic Encephalopathy","volume":"9","author":"Lee","year":"2017","journal-title":"J. Thorac. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1001\/jama.2017.7218","article-title":"Effect of Depth and Duration of Cooling on Death or Disability at Age 18 Months Among Neonates With Hypoxic-Ischemic Encephalopathy","volume":"318","author":"Shankaran","year":"2017","journal-title":"JAMA"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"e1273","DOI":"10.1016\/S2214-109X(21)00264-3","article-title":"Hypothermia for Moderate or Severe Neonatal Encephalopathy in Low-Income and Middle-Income Countries (HELIX): A Randomised Controlled Trial in India, Sri Lanka, and Bangladesh","volume":"9","author":"Thayyil","year":"2021","journal-title":"Lancet Glob. Health"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Corry, K.A., White, O.R., Shearlock, A.E., Moralejo, D.H., Law, J.B., Snyder, J.M., Juul, S.E., and Wood, T.R. (2021). Evaluating Neuroprotective Effects of Uridine, Erythropoietin, and Therapeutic Hypothermia in a Ferret Model of Inflammation-Sensitized Hypoxic-Ischemic Encephalopathy. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22189841"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1159\/000498968","article-title":"A Ferret Model of Encephalopathy of Prematurity","volume":"40","author":"Wood","year":"2018","journal-title":"Dev. Neurosci."},{"key":"ref_9","first-page":"e60131","article-title":"A Ferret Model of Inflammation-Sensitized Late Preterm Hypoxic-Ischemic Brain Injury","volume":"153","author":"Wood","year":"2019","journal-title":"J. Vis. Exp."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Gilardi, C., and Kalebic, N. (2021). The Ferret as a Model System for Neocortex Development and Evolution. Front. Cell Dev. Biol., 9.","DOI":"10.3389\/fcell.2021.661759"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"80","DOI":"10.1203\/PDR.0b013e3181a291d9","article-title":"Characterization of Brain Development in the Ferret via MRI","volume":"66","author":"Barnette","year":"2009","journal-title":"Pediatr. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1016\/j.ijdevneu.2015.06.005","article-title":"Is the Ferret a Suitable Species for Studying Perinatal Brain Injury?","volume":"45","author":"Empie","year":"2015","journal-title":"Int. J. Dev. Neurosci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1444","DOI":"10.1093\/cercor\/bhm180","article-title":"Mapping the Early Cortical Folding Process in the Preterm Newborn Brain","volume":"18","author":"Dubois","year":"2008","journal-title":"Cereb. Cortex"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1002\/ana.24313","article-title":"Regional Impairments of Cortical Folding in Premature Infants","volume":"77","author":"Engelhardt","year":"2015","journal-title":"Ann. Neurol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.neuroscience.2017.09.015","article-title":"Biphasic Aspect of Sexually Dimorphic Ontogenetic Trajectory of Gyrification in the Ferret Cerebral Cortex","volume":"364","author":"Sawada","year":"2017","journal-title":"Neuroscience"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1136\/archdischild-2019-317466","article-title":"Early Cortical Maturation Predicts Neurodevelopment in Very Preterm Infants","volume":"105","author":"Kline","year":"2020","journal-title":"Arch. Dis. Child. Fetal Neonatal Ed."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Sawada, K., Kamiya, S., and Aoki, I. (2021). Neonatal Valproic Acid Exposure Produces Altered Gyrification Related to Increased Parvalbumin-Immunopositive Neuron Density with Thickened Sulcal Floors. PLoS ONE, 16.","DOI":"10.1371\/journal.pone.0250262"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1159\/000430860","article-title":"Hypothermia Does Not Reverse Cellular Responses Caused by Lipopolysaccharide in Neonatal Hypoxic-Ischaemic Brain Injury","volume":"37","author":"Osredkar","year":"2015","journal-title":"Dev. Neurosci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1159\/000481980","article-title":"Neonatal Systemic Inflammation Induces Inflammatory Reactions and Brain Apoptosis in a Pathogen-Specific Manner","volume":"113","author":"Falck","year":"2018","journal-title":"Neonatology"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1038\/nmeth.2089","article-title":"NIH Image to ImageJ: 25 Years of Image Analysis","volume":"9","author":"Schneider","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"575","DOI":"10.1016\/j.neuroimage.2017.03.009","article-title":"Population Based MRI and DTI Templates of the Adult Ferret Brain and Tools for Voxelwise Analysis","volume":"152","author":"Hutchinson","year":"2017","journal-title":"Neuroimage"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4847","DOI":"10.21105\/joss.04847","article-title":"rigr: Regression, Inference, and General Data Analysis Tools in R","volume":"7","author":"Chen","year":"2022","journal-title":"J. Open Source Softw."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"74135","DOI":"10.7554\/eLife.74135","article-title":"Sex Difference Analyses under Scrutiny","volume":"10","author":"Vorland","year":"2021","journal-title":"eLife"},{"key":"ref_24","unstructured":"R Development Core Team (2022). R: A Language and Environment for Statistical Computing, R Foundation for Statistical Computing."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Kline, J.E., Illapani, V.S.P., He, L., Altaye, M., and Parikh, N.A. (2019). Retinopathy of Prematurity and Bronchopulmonary Dysplasia Are Independent Antecedents of Cortical Maturational Abnormalities in Very Preterm Infants. Sci. Rep., 9.","DOI":"10.1038\/s41598-019-56298-x"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1255","DOI":"10.1093\/brain\/awz071","article-title":"Aberrant Gyrification Contributes to the Link between Gestational Age and Adult IQ after Premature Birth","volume":"142","author":"Hedderich","year":"2019","journal-title":"Brain"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1016\/j.neuropsychologia.2005.05.015","article-title":"Increased Temporal Lobe Gyrification in Preterm Children","volume":"44","author":"Kesler","year":"2006","journal-title":"Neuropsychologia"},{"key":"ref_28","first-page":"640","article-title":"Altered Cortical Gyrification in Adults Who Were Born Very Preterm and Its Associations With Cognition and Mental Health","volume":"5","author":"Papini","year":"2020","journal-title":"Biol. Psychiatry Cogn. Neurosci. Neuroimaging"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"e227045","DOI":"10.1001\/jamanetworkopen.2022.7045","article-title":"Association of Prenatal Exposure to Early-Life Adversity With Neonatal Brain Volumes at Birth","volume":"5","author":"Triplett","year":"2022","journal-title":"JAMA Netw. Open"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2055","DOI":"10.1212\/WNL.0b013e31821f442d","article-title":"Predicting Motor Outcome and Death in Term Hypoxic-Ischemic Encephalopathy","volume":"76","author":"Kapellou","year":"2011","journal-title":"Neurology"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4228","DOI":"10.1523\/JNEUROSCI.3976-13.2014","article-title":"Mapping Longitudinal Development of Local Cortical Gyrification in Infants from Birth to 2 Years of Age","volume":"34","author":"Li","year":"2014","journal-title":"J. Neurosci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1016\/j.neuroimage.2016.07.010","article-title":"Relation between Clinical Risk Factors, Early Cortical Changes, and Neurodevelopmental Outcome in Preterm Infants","volume":"142","author":"Kersbergen","year":"2016","journal-title":"Neuroimage"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1186\/s13229-015-0035-y","article-title":"Sex Differences in Cortical Volume and Gyrification in Autism","volume":"6","author":"Schaer","year":"2015","journal-title":"Mol. Autism"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1016\/j.neuroimage.2013.05.076","article-title":"Sex Differences in Thickness, and Folding Developments throughout the Cortex","volume":"82","author":"Mutlu","year":"2013","journal-title":"Neuroimage"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1093\/brain\/awl277","article-title":"Perinatal Risk Factors Altering Regional Brain Structure in the Preterm Infant","volume":"130","author":"Thompson","year":"2007","journal-title":"Brain"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"845","DOI":"10.1097\/WNR.0000000000000844","article-title":"Early Detection of Neonatal Hypoxic\u2013Ischemic White Matter Injury","volume":"28","author":"Seo","year":"2017","journal-title":"Neuroreport"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1037\/0735-7044.100.4.563","article-title":"Sexual Differentiation of Play Behavior in the Ferret","volume":"100","author":"Stockman","year":"1986","journal-title":"Behav. Neurosci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1007\/s12144-009-9047-0","article-title":"Stress, Sex Differences, and Coping Strategies Among College Students","volume":"28","author":"Brougham","year":"2009","journal-title":"Curr. Psychol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3109\/08990220.2010.548465","article-title":"An Examination of Somatosensory Area SIII in Ferret Cortex","volume":"28","author":"Foxworthy","year":"2011","journal-title":"Somat. Mot. Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"161","DOI":"10.3109\/08990229309028830","article-title":"Cytoarchitecture of the Ferret Suprasylvian Gyrus Correlated with Areas Containing Multiunit Responses Elicited by Stimulation of the Face","volume":"10","author":"Rice","year":"1993","journal-title":"Somat. Mot. Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1080\/08990229870673","article-title":"Organization of the Forepaw Representation in Ferret Somatosensory Cortex","volume":"15","author":"McLaughlin","year":"1998","journal-title":"Somat. Mot. Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1016\/j.cub.2018.11.017","article-title":"Ferrets as a Model for Higher-Level Visual Motion Processing","volume":"29","author":"Lempel","year":"2019","journal-title":"Curr. Biol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1016\/j.clp.2015.04.004","article-title":"Erythropoietin and Neonatal Neuroprotection","volume":"42","author":"Juul","year":"2015","journal-title":"Clin. Perinatol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"fcab172","DOI":"10.1093\/braincomms\/fcab172","article-title":"Recombinant Erythropoietin Does Not Augment Hypothermic White Matter Protection after Global Cerebral Ischaemia in Near-Term Fetal Sheep","volume":"3","author":"Wassink","year":"2021","journal-title":"Brain Commun."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1159\/000486820","article-title":"High-Dose Erythropoietin for Asphyxia and Encephalopathy (HEAL): A Randomized Controlled Trial\u2014Background, Aims, and Study Protocol","volume":"113","author":"Juul","year":"2018","journal-title":"Neonatology"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.ijdevneu.2018.05.006","article-title":"Ontogeny of White Matter, Toll-like Receptor Expression, and Motor Skills in the Neonatal Ferret","volume":"70","author":"Snyder","year":"2018","journal-title":"Int. J. Dev. Neurosci."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"2039","DOI":"10.1093\/cercor\/bhr284","article-title":"Germinal Zones in the Developing Cerebral Cortex of Ferret: Ontogeny, Cell Cycle Kinetics, and Diversity of Progenitors","volume":"22","author":"Reillo","year":"2012","journal-title":"Cereb. Cortex"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"e2023598118","DOI":"10.1073\/pnas.2023598118","article-title":"Development of Human White Matter Pathways in Utero over the Second and Third Trimester","volume":"118","author":"Wilson","year":"2021","journal-title":"Proc. Natl. Acad. Sci. USA"}],"container-title":["Life"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-1729\/15\/9\/1428\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:44:11Z","timestamp":1760035451000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-1729\/15\/9\/1428"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,11]]},"references-count":48,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["life15091428"],"URL":"https:\/\/doi.org\/10.3390\/life15091428","relation":{},"ISSN":["2075-1729"],"issn-type":[{"value":"2075-1729","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,11]]}}}