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In humans, homozygous mutations in\n                    <jats:italic>CLN5<\/jats:italic>\n                    and\n                    <jats:italic>CTSD<\/jats:italic>\n                    cause CLN5 disease and CLN10 disease, respectively, which are two subtypes of neuronal ceroid lipofuscinosis (commonly known as Batten disease). The mechanisms regulating the intracellular trafficking of CLN5 and CTSD and their release from cells are not well understood. Here, we used the social amoeba\n                    <jats:italic>Dictyostelium discoideum<\/jats:italic>\n                    as a model system to examine the pathways and cellular components that regulate the intracellular trafficking and release of the\n                    <jats:italic>D. discoideum<\/jats:italic>\n                    homologs of human CLN5 (Cln5) and CTSD (CtsD). We show that both Cln5 and CtsD contain signal peptides for secretion that facilitate their release from cells. Like Cln5, extracellular CtsD is glycosylated. In addition, Cln5 release is regulated by the amount of extracellular CtsD. Autophagy induction promotes the release of Cln5, and to a lesser extent CtsD. Release of Cln5 requires the autophagy proteins Atg1, Atg5, and Atg9, as well as autophagosomal\u2010lysosomal fusion. Atg1 and Atg5 are required for the release of CtsD. Together, these data support a model where Cln5 and CtsD are actively released from cells via their signal peptides for secretion and pathways linked to autophagy. The release of Cln5 and CtsD from cells also requires microfilaments and the\n                    <jats:italic>D. discoideum<\/jats:italic>\n                    homologs of human AP\u20103 complex mu subunit, the lysosomal\u2010trafficking regulator LYST, mucopilin\u20101, and the Wiskott\u2013Aldrich syndrome\u2010associated protein WASH, which all regulate lysosomal exocytosis in this model organism. These findings suggest that lysosomal exocytosis also facilitates the release of Cln5 and CtsD from cells. In addition, we report the roles of ABC transporters, microtubules, osmotic stress, and the putative\n                    <jats:italic>D. discoideum<\/jats:italic>\n                    homologs of human sortilin and cation\u2010independent mannose\u20106\u2010phosphate receptor in regulating the intracellular\/extracellular distribution of Cln5 and CtsD. In total, this study identifies the cellular mechanisms regulating the release of Cln5 and CtsD from\n                    <jats:italic>D. discoideum<\/jats:italic>\n                    cells and provides insight into how altered trafficking of CLN5 and CTSD causes disease in humans.\n                  <\/jats:p>","DOI":"10.1111\/tra.12925","type":"journal-article","created":{"date-parts":[[2024,1,21]],"date-time":"2024-01-21T23:30:00Z","timestamp":1705879800000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Mechanisms regulating the intracellular trafficking and release of\n                    <scp>CLN5<\/scp>\n                    and\n                    <scp>CTSD<\/scp>"],"prefix":"10.1111","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2606-6983","authenticated-orcid":false,"given":"Robert J.","family":"Huber","sequence":"first","affiliation":[{"name":"Department of Biology Trent University  Peterborough Ontario Canada"},{"name":"Environmental and Life Sciences Graduate Program Trent University  Peterborough Ontario Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0085-2188","authenticated-orcid":false,"given":"William D.","family":"Kim","sequence":"additional","affiliation":[{"name":"Environmental and Life Sciences Graduate Program Trent University  Peterborough Ontario Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Morgan L. 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