{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,12]],"date-time":"2025-12-12T13:47:54Z","timestamp":1765547274179,"version":"build-2065373602"},"reference-count":52,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2024,7,23]],"date-time":"2024-07-23T00:00:00Z","timestamp":1721692800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31371106","91632110","31700934"],"award-info":[{"award-number":["31371106","91632110","31700934"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Microscopy Res &amp; Technique"],"published-print":{"date-parts":[[2024,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:label\/><jats:p>Serotonin (5\u2010hydroxytryptamine, 5\u2010HT) is a very important neurotransmitter emerging from the raphe nuclei to several brain regions. Serotonergic neuronal connectivity has multiple functions in the brain. In this study, several techniques were used to trace serotonergic neurons in the dorsal raphe (DR) and median raphe (MnR) that project toward the arcuate nucleus of the hypothalamus (Arc), dorsomedial hypothalamic nucleus (DM), lateral hypothalamic area (LH), paraventricular hypothalamic nucleus (PVH), ventromedial hypothalamic nucleus (VMH), fasciola cinereum (FC), and medial habenular nucleus (MHb). Cholera toxin subunit B (CTB), retro\u2010adeno\u2010associated virus (rAAV\u2010CMV\u2010mCherry), glycoprotein\u2010deleted rabies virus (RV\u2010\u0394G), and simultaneous microinjection of rAAV2\u2010retro\u2010Cre\u2010tagBFP with AAV\u2010dio\u2010mCherry in C57BL\/6 mice were used in this study. In addition, rAAV2\u2010retro\u2010Cre\u2010tagBFP was microinjected into Ai9 mice. Serotonin immunohistochemistry was used for the detection of retrogradely traced serotonergic neurons in the raphe nuclei. The results indicated that rAAV2\u2010retro\u2010Cre\u2010tagBFP microinjection in Ai9 mice was the best method for tracing serotonergic neuron circuits. All of the previously listed nuclei exhibited serotonergic neuronal projections from the DR and MnR, with the exception of the FC, which had very few projections from the DR. The serotonergic neuronal projections were directed toward the Arc by the subpeduncular tegmental (SPTg) nuclei. Moreover, the RV\u2010\u0394G tracer revealed monosynaptic non\u2010serotonergic neuronal projections from the DR that were directed toward the Arc. Furthermore, rAAV tracers revealed monosynaptic serotonergic neuronal connections from the raphe nuclei toward Arc. These findings validate the variations in neurotropism among several retrograde tracers. The continued discovery of several novel serotonergic neural circuits is crucial for the future discovery of the functions of these circuits.<\/jats:p><\/jats:sec><jats:sec><jats:title>Research Highlights<\/jats:title><jats:p><jats:list list-type=\"bullet\">\n<jats:list-item><jats:p>Various kinds of retrograde tracers were microinjected into C57BL\/6 and Ai9 mice.<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p>The optimum method for characterizing serotonergic neuronal circuits is rAAV2\u2010retro\u2010Cre\u2010tagBFP microinjection in Ai9 mice.<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p>The DR, MnR, and SPTg nuclei send monosynaptic serotonergic neuronal projections toward the arcuate nucleus of the hypothalamus.<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p>Whole\u2010brain quantification analysis of retrograde\u2010labeled neurons in different brain nuclei following rAAV2\u2010retro\u2010Cre\u2010tagBFP microinjection in the Arc, DM, LH, and VMH is shown.<\/jats:p><\/jats:list-item>\n<jats:list-item><jats:p>Differential quantitative analysis of median and dorsal raphe serotonergic neurons emerging toward the PVH, DM, LH, Arc, VMH, MHb, and FC is shown.<\/jats:p><\/jats:list-item>\n<\/jats:list><\/jats:p><\/jats:sec>","DOI":"10.1002\/jemt.24662","type":"journal-article","created":{"date-parts":[[2024,7,23]],"date-time":"2024-07-23T09:52:53Z","timestamp":1721728373000},"page":"2894-2914","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Labeling of the serotonergic neuronal circuits emerging from the raphe nuclei via some retrograde tracers"],"prefix":"10.1002","volume":"87","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7144-2477","authenticated-orcid":false,"given":"Mona N.","family":"Hussein","sequence":"first","affiliation":[{"name":"State Key Laboratory of Agricultural Microbiology Huazhong Agricultural University  Wuhan China"},{"name":"College of Veterinary Medicine Huazhong Agricultural University  Wuhan China"},{"name":"Histology and Cytology Department, Faculty of Veterinary Medicine Benha University  Benha Egypt"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,7,23]]},"reference":[{"key":"e_1_2_11_2_1","doi-asserted-by":"publisher","DOI":"10.1196\/annals.1296.005"},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/cne.23167"},{"key":"e_1_2_11_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molmet.2023.101676"},{"key":"e_1_2_11_5_1","doi-asserted-by":"publisher","DOI":"10.1021\/cn300224n"},{"key":"e_1_2_11_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2451"},{"key":"e_1_2_11_7_1","doi-asserted-by":"publisher","DOI":"10.1100\/2012\/823493"},{"key":"e_1_2_11_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.euroneuro.2015.12.032"},{"key":"e_1_2_11_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.yfrne.2019.04.003"},{"key":"e_1_2_11_10_1","doi-asserted-by":"publisher","DOI":"10.14814\/phy2.13807"},{"key":"e_1_2_11_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/nn.3039"},{"key":"e_1_2_11_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2014.07.002"},{"key":"e_1_2_11_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/nn.4442"},{"key":"e_1_2_11_14_1","doi-asserted-by":"publisher","DOI":"10.1002\/cne.22323"},{"key":"e_1_2_11_15_1","doi-asserted-by":"publisher","DOI":"10.1002\/cne.22283"},{"key":"e_1_2_11_16_1","doi-asserted-by":"publisher","DOI":"10.3389\/fncir.2013.00002"},{"key":"e_1_2_11_17_1","doi-asserted-by":"publisher","DOI":"10.1021\/acschemneuro.5b00045"},{"key":"e_1_2_11_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2019.06.051"},{"key":"e_1_2_11_19_1","doi-asserted-by":"publisher","DOI":"10.1038\/nn.4332"},{"key":"e_1_2_11_20_1","doi-asserted-by":"publisher","DOI":"10.3389\/fncom.2023.1189853"},{"key":"e_1_2_11_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molmet.2017.01.001"},{"key":"e_1_2_11_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.physbeh.2005.10.007"},{"key":"e_1_2_11_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molmet.2022.101604"},{"key":"e_1_2_11_24_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12264-019-00374-9"},{"key":"e_1_2_11_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.autneu.2010.05.006"},{"key":"e_1_2_11_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuropharm.2020.107993"},{"key":"e_1_2_11_27_1","doi-asserted-by":"publisher","DOI":"10.1002\/cne.24217"},{"key":"e_1_2_11_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.conb.2020.03.011"},{"key":"e_1_2_11_29_1","doi-asserted-by":"publisher","DOI":"10.1111\/jnc.14233"},{"key":"e_1_2_11_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2014.10.019"},{"key":"e_1_2_11_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pbb.2017.05.007"},{"key":"e_1_2_11_32_1","doi-asserted-by":"publisher","DOI":"10.1111\/ejn.14647"},{"key":"e_1_2_11_33_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00429-014-0924-4"},{"key":"e_1_2_11_34_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pbb.2017.05.001"},{"key":"e_1_2_11_35_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pneurobio.2022.102217"},{"key":"e_1_2_11_36_1","unstructured":"Park S. B. Yoo S. W. Jung H. S. Lim H. Y. Lee E. Sun W. &Lee I.(2019).The fasciola cinereum of the hippocampus is important in the acquisition but not the retrieval of visual contextual memory. Paper presented at the conference proceedings: Annual international conference of the IEEE engineering in medicine and biology society. IEEE Engineering in Medicine and Biology Society. Annual Conference."},{"key":"e_1_2_11_37_1","doi-asserted-by":"publisher","DOI":"10.1523\/ENEURO.0527-19.2020"},{"key":"e_1_2_11_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2018.07.043"},{"key":"e_1_2_11_39_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neulet.2015.11.035"},{"key":"e_1_2_11_40_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2019.05.048"},{"key":"e_1_2_11_41_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrmicro2260"},{"key":"e_1_2_11_42_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuropharm.2020.108155"},{"key":"e_1_2_11_43_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13024-019-0308-6"},{"key":"e_1_2_11_44_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbr.2016.11.026"},{"key":"e_1_2_11_45_1","doi-asserted-by":"publisher","DOI":"10.3389\/fnana.2015.00040"},{"key":"e_1_2_11_46_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2019.01.014"},{"key":"e_1_2_11_47_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12264-013-1327-x"},{"key":"e_1_2_11_48_1","doi-asserted-by":"publisher","DOI":"10.3389\/fnana.2022.886925"},{"key":"e_1_2_11_49_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2020.07.010"},{"key":"e_1_2_11_50_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10989-020-10120-9"},{"key":"e_1_2_11_51_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.neuron.2021.12.022"},{"key":"e_1_2_11_52_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2017.02.077"},{"key":"e_1_2_11_53_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.gt.3300938"}],"container-title":["Microscopy Research and Technique"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jemt.24662","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,16]],"date-time":"2025-10-16T14:24:42Z","timestamp":1760624682000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/jemt.24662"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,7,23]]},"references-count":52,"journal-issue":{"issue":"12","published-print":{"date-parts":[[2024,12]]}},"alternative-id":["10.1002\/jemt.24662"],"URL":"https:\/\/doi.org\/10.1002\/jemt.24662","archive":["Portico"],"relation":{},"ISSN":["1059-910X","1097-0029"],"issn-type":[{"type":"print","value":"1059-910X"},{"type":"electronic","value":"1097-0029"}],"subject":[],"published":{"date-parts":[[2024,7,23]]},"assertion":[{"value":"2024-02-13","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-07-11","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2024-07-23","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}