{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,12]],"date-time":"2025-11-12T05:28:05Z","timestamp":1762925285822,"version":"3.45.0"},"reference-count":54,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2025,11,10]],"date-time":"2025-11-10T00:00:00Z","timestamp":1762732800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001807","name":"FAPESP","doi-asserted-by":"publisher","award":["2024\/05757-7"],"award-info":[{"award-number":["2024\/05757-7"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Calcineurin (CaN), a Ca2+-dependent phosphatase, plays key roles in eukaryotic cell signaling. We investigated whether Leishmania amazonensis\u2019 two infective forms\u2014promastigotes and amastigotes\u2014exhibit differences in CaN expression, localization, and functional impact, using two canonical inhibitors (cyclosporin A, CsA; tracolimus, FK506). At high 40 \u00b5M CsA, promastigotes showed reduced viability, whereas amastigotes remained resistant. FK506 had no effect on either form. At a sub-lethal 25 \u00b5M CsA, parasite proliferation remained unaffected. In parasite\u2013macrophage co-incubation assays, phosphorylation patterns differed: amastigotes\u2014but not promastigotes\u2014showed increased serine\/threonine phosphorylation upon CaN inhibition. Western blotting and in silico data revealed higher CaN catalytic (CaNA2) and regulatory (CaNB) subunit expression in amastigotes than promastigotes. Immunofluorescence localized CaNA prominently in both cytoplasm and nucleus of promastigotes, but predominantly cytoplasmic in amastigotes; CaNB was largely cytoplasmic in both. In silico localization predictions suggested strong membrane associations for CaNA in Leishmania, contrasting with mammalian models. Subcellular fractionation confirmed CaNA enrichment in membrane fractions, with CaNB in cytoplasmic and nuclear fractions. Collectively, these findings reveal form-specific differences in expression, subcellular distribution, and inhibitor responses of CaN in L. amazonensis, highlighting its potential as a stage-specific therapeutic target in leishmaniasis.<\/jats:p>","DOI":"10.3390\/pathogens14111139","type":"journal-article","created":{"date-parts":[[2025,11,10]],"date-time":"2025-11-10T15:07:31Z","timestamp":1762787251000},"page":"1139","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Stage-Specific Expression and Subcellular Localization of Calcineurin in Infective Forms of Leishmania amazonensis"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-5032-7744","authenticated-orcid":false,"given":"Deborah","family":"Brandt-Almeida","sequence":"first","affiliation":[{"name":"Department of Parasitology, Institute of Biomedical Sciences, University of S\u00e3o Paulo, S\u00e3o Paulo 05508000, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ismael Pretto","family":"Sauter","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Institute of Biomedical Sciences, University of S\u00e3o Paulo, S\u00e3o Paulo 05508000, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mario Costa","family":"Cruz","sequence":"additional","affiliation":[{"name":"Core Facilities to Support Research, Institute of Biomedical Sciences, University of S\u00e3o Paulo, S\u00e3o Paulo 05508000, SP, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1887-0808","authenticated-orcid":false,"given":"Cristian","family":"Cortez","sequence":"additional","affiliation":[{"name":"Escuela de Tecnolog\u00eda M\u00e9dica, Facultad de Ciencias, Universidad de San Sebasti\u00e1n, Santiago 8420000, Chile"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-2122-6660","authenticated-orcid":false,"given":"Patricio Reyes","family":"Orrego","sequence":"additional","affiliation":[{"name":"Departamento Biom\u00e9dico, Facultad de Ciencias de la Salud, Universidad de Antofagasta, Antofagasta 1270300, Chile"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6536-4647","authenticated-orcid":false,"given":"Mauro","family":"Cortez","sequence":"additional","affiliation":[{"name":"Department of Parasitology, Institute of Biomedical Sciences, University of S\u00e3o Paulo, S\u00e3o Paulo 05508000, SP, Brazil"},{"name":"Escuela de Tecnolog\u00eda M\u00e9dica, Facultad de Ciencias, Pontificia Universidad Cat\u00f3lica de Valpara\u00edso, Valpara\u00edso 2373223, Chile"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Soulat, D., and Bogdan, C. 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