{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T05:03:51Z","timestamp":1789707831812,"version":"4.0.1"},"posted":{"date-parts":[[2026,9,14]]},"group-title":"Neuroscience","reference-count":8,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T00:00:00Z","timestamp":1789344000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"award":["1R01NS142268-01A1"],"award-info":[{"award-number":["1R01NS142268-01A1"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,14]]},"abstract":"Summary<\/jats:title>\n \n Following nervous system injury, surviving bystander neurons may rewire and form synapses with new postsynaptic targets to restore circuit function. This protocol quantifies compensatory motor neuron sprouting at the\n Drosophila<\/jats:italic>\n larval neuromuscular junction. We describe larval dissection, immunostaining, Airyscan imaging, image masking, muscle-specific region-of-interest selection, and Fiji-based quantification of Dlg-positive area, bouton number, mean Dlg-positive area per bouton, NMJ arbor length, axonal versus NMJ branch points, ectopic muscle acquisition, and sprouting penetrance.\n <\/jats:p>\n \n For complete details on the use and execution of this protocol, please refer to Olsen et al.\n 1<\/jats:sup>\n <\/jats:p>\n \n Graphical abstract<\/jats:title>\n \n \n <\/jats:fig>\n <\/jats:sec>\n \n Highlights<\/jats:title>\n \n \n Steps for larval dissection, immunostaining, Airyscan imaging, and preprocessing<\/jats:p>\n <\/jats:list-item>\n \n Instructions for Fiji-based masking to isolate single-neuron postsynaptic sites<\/jats:p>\n <\/jats:list-item>\n \n Guidance on muscle-specific quantification of synaptic area, boutons, and arbor length<\/jats:p>\n <\/jats:list-item>\n \n Steps for branch analysis to distinguish axonal sprouting from synaptic remodeling<\/jats:p>\n <\/jats:list-item>\n <\/jats:list>\n <\/jats:sec>\n \n eTOC blurb<\/jats:title>\n \n Olsen and Zarin present a workflow for quantifying compensatory sprouting by a single motor neuron at the\n Drosophila<\/jats:italic>\n larval neuromuscular junction. The protocol integrates Airyscan imaging, Fiji-based image masking, muscle-specific measurements, and branch classification to distinguish new axonal growth from remodeling of established synapses.\n <\/jats:p>\n <\/jats:sec>","DOI":"10.64898\/2026.09.06.749747","type":"posted-content","created":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T17:20:33Z","timestamp":1789406433000},"source":"Crossref","is-referenced-by-count":0,"title":["Quantification of compensatory motor neuron sprouting at the\n Drosophila<\/i>\n larval neuromuscular junction"],"prefix":"10.64898","author":[{"given":"Lizzy","family":"Olsen","sequence":"first","affiliation":[{"name":"Biology Graduate Program, Texas A&M University, College Station, TX 77843, USA"},{"name":"Department of Biology, Texas A&M University, College Station, TX 77843, USA"},{"name":"Texas A&M Institute for Neuroscience, Texas A&M University, College Station, TX 77843, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0484-3622","authenticated-orcid":false,"given":"Aref","family":"Zarin","sequence":"additional","affiliation":[{"name":"Department of Biology, Texas A&M University, College Station, TX 77843, USA"},{"name":"Texas A&M Institute for Neuroscience, Texas A&M University, College Station, TX 77843, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026091721150623000_2026.09.06.749747v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.isci.2026.116099"},{"key":"2026091721150623000_2026.09.06.749747v1.2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1004823"},{"key":"2026091721150623000_2026.09.06.749747v1.3","doi-asserted-by":"publisher","DOI":"10.3791\/55395"},{"key":"2026091721150623000_2026.09.06.749747v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2023.07.019"},{"key":"2026091721150623000_2026.09.06.749747v1.5","doi-asserted-by":"publisher","DOI":"10.1534\/g3.118.200809"},{"key":"2026091721150623000_2026.09.06.749747v1.6","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2303641120"},{"key":"2026091721150623000_2026.09.06.749747v1.7","doi-asserted-by":"crossref","unstructured":"Ankura Sitaula , Lizzy Olsen , Yuhan Huang , Dulanjana M Perera , Parinita Mitchelle Mandhyan , Nathan Byrd , Sara Arredondo , Lauren Carlisle , Isuru S Godage , and Zarin, A . (2026). Reconfiguration of Premotor Excitation and Inhibition Drives Behavior-Specific Protopodium Dynamics in Drosophila Larvae. bioRxiv.","DOI":"10.64898\/2026.01.23.701396"},{"key":"2026091721150623000_2026.09.06.749747v1.8","doi-asserted-by":"publisher","DOI":"10.1080\/01677060490894522"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.09.06.749747","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T04:15:15Z","timestamp":1789704915000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.09.06.749747"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,14]]},"references-count":8,"URL":"https:\/\/doi.org\/10.64898\/2026.09.06.749747","relation":{},"subject":[],"published":{"date-parts":[[2026,9,14]]},"subtype":"preprint"}}