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In addition, the brain can metabolise circulating steroids and synthesise steroids de novo. Songbirds show high levels of neurosteroid synthesis. In the present study, we developed and validated a liquid chromatography\u2010tandem mass spectrometry (LC\u2010MS\/MS) assay for the measurement of 10 steroids in whole blood, plasma and microdissected brain tissue (1\u20102\u00a0mg) of song sparrows. Our assay is highly accurate, precise, specific and sensitive. Moreover, the liquid\u2010liquid extraction is fast, simple and effective. We quantified steroids in the blood and brain of wild male song sparrows in both breeding and non\u2010breeding seasons. As expected, systemic androgen levels were higher in the breeding season than in the non\u2010breeding season. Brain androgens were detectable only in the breeding season; androstenedione and 5\u03b1\u2010dihydrotestosterone levels were up to 20\u2010fold higher in specific brain regions than in blood. Oestrogens were not detectable in blood in both seasons. Oestrone and 17\u03b2\u2010oestradiol were detectable in brain in the breeding season only (up to 1.4\u00a0ng\u00a0g\n                    <jats:sup>\u20101<\/jats:sup>\n                    combined). Progesterone levels in several regions were higher in the non\u2010breeding season than the breeding season, despite the lack of seasonal changes in systemic progesterone. Corticosterone levels in the blood were higher in the breeding season than in the non\u2010breeding season but showed few seasonal differences in the brain. In general, the steroid levels presented here are lower than those in previous reports using immunoassays, because of the higher specificity of mass spectrometry. We conclude that (i) brain steroid levels can differ greatly from circulating steroid levels and (ii) brain steroid levels show region\u2010specific seasonal patterns that are not a simple reflection of circulating steroid levels. This approach using ultrasensitive LC\u2010MS\/MS is broadly applicable to other species and allows steroid profiling in microdissected brain regions.\n                  <\/jats:p>","DOI":"10.1111\/jne.12922","type":"journal-article","created":{"date-parts":[[2020,11,20]],"date-time":"2020-11-20T00:07:44Z","timestamp":1605830864000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":43,"title":["Profiling of systemic and brain steroids in male songbirds: Seasonal changes in neurosteroids"],"prefix":"10.1111","volume":"33","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4165-7610","authenticated-orcid":false,"given":"Cecilia","family":"Jalabert","sequence":"first","affiliation":[{"name":"Department of Zoology University of British Columbia  Vancouver BC Canada"},{"name":"Djavad Mowafaghian Centre for Brain Health University of British Columbia  Vancouver BC 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