{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:44:35Z","timestamp":1760060675784,"version":"build-2065373602"},"reference-count":52,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,9,22]],"date-time":"2025-09-22T00:00:00Z","timestamp":1758499200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"NIH","award":["EY01869","EY002162"],"award-info":[{"award-number":["EY01869","EY002162"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Brain Sciences"],"abstract":"<jats:p>Background\/Objectives: Glutamatergic neurotransmission is essential for the normal functioning of the retina. Photoreceptor to bipolar and bipolar to ganglion cell signaling is mediated by L-glutamate, which is stored in and released from vesicular glutamate transporter 1 (VGLUT1) containing synaptic vesicles. VGLUT1 is expressed postnatally, P2 onwards, and is required for the glutamatergic retinal wave observed between P10 and P12 in the developing mouse retina. P9\u2013P13 postnatal age is critical for retinal development as VGLUT1 expressing ribbon synapses activate in the outer and inner plexiform layers, and rod\/cone mediated visual signaling commences in that period. Although it has been hypothesized that glutamatergic extrinsic signaling drives cell cycle exit and initiates cellular differentiation in the developing retina, it is not clear whether intracellular, synaptic, or extrasynaptic vesicular glutamate release contributes to this process. Recent studies have attempted to decipher VGLUT\u2019s role in retinal development. Here, we investigate the potential effect of genetic loss of VGLUT1 on early postnatal histogenesis and development of retinal neural circuitry. Methods: We employed immunohistochemistry and electrophysiology to ascertain the density of glutamatergic, cholinergic, and dopaminergic cells, spontaneous retinal activity, and light responses in VGLUT1 null retina, and contrasted them with wildtype (WT) and melanopsin null retina. Results: We have demonstrated here that VGLUT1 null retina shows signs of age dependent retinal degeneration, similar to other transgenic mice models with dysfunctional photoreceptor to bipolar cell synapses. The loss of VGLUT1 specifically alters glutamatergic cell density and morphological maturation of retinal ganglion cells. Moreover, VGLUT2 expression is lost in the majority of VGLUT2 cones in the absence of VGLUT1 coexpression, except when VGLUT2 coexpresses transiently with VGLUT3 in these cones, or when VGLUT1 null mice are dark reared. Conclusions: We present the first evidence that synaptic or extrasynaptic postnatal glutamate release from VGLUT1 containing vesicles impacts histogenesis of glutamatergic cells, pruning of retinal ganglion cell dendrites and VGLUT2 expression in cones.<\/jats:p>","DOI":"10.3390\/brainsci15091024","type":"journal-article","created":{"date-parts":[[2025,9,22]],"date-time":"2025-09-22T17:08:17Z","timestamp":1758560897000},"page":"1024","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Genetic Loss of VGLUT1 Alters Histogenesis of Retinal Glutamatergic Cells and Reveals Dynamic Expression of VGLUT2 in Cones"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1195-7137","authenticated-orcid":false,"given":"Sriparna","family":"Majumdar","sequence":"first","affiliation":[{"name":"Department of Biology, Stanford University School of Medicine, Stanford, CA 94305, USA"},{"name":"Computer Science Department, City College of San Francisco, San Francisco, CA 94112, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vincent","family":"Wu","sequence":"additional","affiliation":[{"name":"Kaiser Permanente Redwood City Medical Center, Redwood City, CA 94063, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Majumdar, S. (2023). Role of glutamate in the development of visual pathways. Front. Ophthanlmol., 3.","DOI":"10.3389\/fopht.2023.1147769"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Ichinose, T., and Habib, S. (2022). ON and OFF Signaling Pathways in the Retina and the Visual System. Front. Ophthalmol., 2.","DOI":"10.3389\/fopht.2022.989002"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2058","DOI":"10.1016\/j.celrep.2017.01.075","article-title":"Diverse Central Projection Patterns of Retinal Ganglion Cells","volume":"18","author":"Martersteck","year":"2017","journal-title":"Cell Rep."},{"key":"ref_4","unstructured":"Purves, D., Augustine, G.J., Fitzpatrick, D., Katz, L.C., LaMantia, A.S., McNamara, J.O., and Williams, S.M. (2001). Central Projections of Retinal Ganglion Cells. Neuroscience, Sinauer Associates. [2nd ed.]."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"480","DOI":"10.1002\/cne.10838","article-title":"Expression of vesicular glutamate transporter 1 in the mouse retina reveals temporal ordering in development of rod vs. cone and ON vs. OFF circuits","volume":"465","author":"Sherry","year":"2003","journal-title":"J. Comp. Neurol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1815","DOI":"10.1126\/science.1097468","article-title":"Vesicular glutamate transporters 1 and 2 target to functionally distinct synaptic release sites","volume":"304","author":"Fremeau","year":"2004","journal-title":"Science"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"544","DOI":"10.1002\/cne.20942","article-title":"Expression of the vesicular glutamate transporter vGluT2 in a subset of cones of the mouse retina","volume":"496","author":"Haverkamp","year":"2006","journal-title":"J. Comp. Neurol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1002\/cne.20250","article-title":"Vesicular glutamate transporter 3 expression identifies glutamatergic amacrine cells in the rodent retina","volume":"477","author":"Johnson","year":"2004","journal-title":"J. Comp. Neurol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/j.tins.2003.11.005","article-title":"VGLUTs define subsets of excitatory neurons and suggest novel roles for glutamate","volume":"27","author":"Fremeau","year":"2004","journal-title":"Trends Neurosci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"11","DOI":"10.3109\/03009730903572073","article-title":"Genetic inactivation of the vesicular glutamate transporter 2 (VGLUT2) in the mouse: What have we learnt about functional glutamatergic neurotransmission?","volume":"115","author":"Wootz","year":"2010","journal-title":"Upsala J. Med. Sci."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"263","DOI":"10.1016\/j.neuron.2007.11.032","article-title":"Sensorineural deafness and seizures in mice lacking vesicular glutamate transporter 3","volume":"57","author":"Seal","year":"2008","journal-title":"Neuron"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"7245","DOI":"10.1523\/JNEUROSCI.0815-07.2007","article-title":"Vesicular glutamate transporter 1 is required for photoreceptor synaptic signaling but not for intrinsic visual functions","volume":"27","author":"Johnson","year":"2007","journal-title":"J. Neurosci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"518","DOI":"10.1523\/JNEUROSCI.23-02-00518.2003","article-title":"Vesicular neurotransmitter transporter expression in developing postnatal rodent retina: GABA and glycine precede glutamate","volume":"23","author":"Johnson","year":"2003","journal-title":"J. Neurosci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1002\/cne.20354","article-title":"Expression of the vesicular glutamate transporters during development indicates the widespread corelease of multiple neurotransmitters","volume":"480","author":"Boulland","year":"2004","journal-title":"J. Comp. Neurol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"108040","DOI":"10.1016\/j.celrep.2020.108040","article-title":"Vesicular Glutamate Transporter Expression Ensures High-Fidelity Synaptic Transmission at the Calyx of Held Synapses","volume":"32","author":"Nakakubo","year":"2020","journal-title":"Cell Rep."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1038\/nrn3767","article-title":"Intrinsically different retinal progenitor cells produce specific types of progeny","volume":"15","author":"Cepko","year":"2014","journal-title":"Nat. Rev. Neurosci."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Duarte-Silva, A.T., Ximenes, L.G.R., Guimar\u00e3es-Souza, M., Domith, I., and Paes-de-Carvalho, R. (2022). Chemical signaling in the developing avian retina: Focus on cyclic AMP and AKT- dependent pathways. Front. Cell Dev. Biol., 10.","DOI":"10.3389\/fcell.2022.1058925"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Delwig, A., Majumdar, S., Ahern, K., LaVail, M.M., Edwards, R., Hnasko, T.S., and Copenhagen, D.R. (2013). Glutamatergic neurotransmission from melanopsin retinal ganglion cells is required for neonatal photoaversion but not adult pupillary light reflex. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0083974"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Verweij, J., Chaney, S.Y., Bredl, D., Vemaraju, S., K\u00f6nig, G.M., Kostenis, E., Lang, R.A., and Copenhagen, D.R. (2019). Light responses of melanopsin-expressing ganglion cells in the foetal mammalian retina. bioRxiv.","DOI":"10.1101\/675702"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1016\/j.neuron.2009.03.015","article-title":"Synaptic and extrasynaptic factors governing glutamatergic retinal waves","volume":"62","author":"Blankenship","year":"2009","journal-title":"Neuron"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1978","DOI":"10.1016\/j.neuron.2024.03.011","article-title":"Glutamatergic neuronal activity regulates angiogenesis and blood-retinal barrier maturation via Norrin\/\u03b2-catenin signaling","volume":"112","author":"Biswas","year":"2024","journal-title":"Neuron"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1046\/j.0953-816x.2001.01624.x","article-title":"Purification of Purkinje cells by fluorescence-activated cell sorting from transgenic mice that express green fluorescent protein","volume":"14","author":"Tomomura","year":"2001","journal-title":"Eur. J. Neurosci."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1002\/(SICI)1096-9861(19980330)393:1<93::AID-CNE9>3.0.CO;2-L","article-title":"Expression of the somatostatin subtype 2A receptor in the rabbit retina","volume":"393","author":"Johnson","year":"1998","journal-title":"J. Comp. Neurol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1002\/1096-9861(20000814)424:1<1::AID-CNE1>3.0.CO;2-V","article-title":"Immunocytochemical analysis of the mouse retina","volume":"424","author":"Haverkamp","year":"2000","journal-title":"J. Comp. Neurol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1284","DOI":"10.1083\/jcb.100.4.1284","article-title":"Identification of a transmembrane glycoprotein specific for secretory vesicles of neural and endocrine cells","volume":"100","author":"Buckley","year":"1985","journal-title":"J. Cell Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2633","DOI":"10.1523\/JNEUROSCI.3770-03.2004","article-title":"Vesicular glutamate transporter- dependent glutamate release from astrocytes","volume":"24","author":"Montana","year":"2004","journal-title":"J. Neurosci."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1083\/jcb.200408157","article-title":"Molecular dissection of the photoreceptor ribbon synapse: Physical interaction of Bassoon and RIBEYE is essential for the assembly of the ribbon complex","volume":"168","author":"Altrock","year":"2005","journal-title":"J. Cell Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1523\/JNEUROSCI.4442-08.2009","article-title":"Cone contacts, mosaics, and territories of bipolar cells in the mouse retina","volume":"29","author":"Puller","year":"2009","journal-title":"J. Neurosci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1098","DOI":"10.1002\/cne.22266","article-title":"Ret-PCP2 colocalizes with protein kinase C in a subset of primate ON cone bipolar cells","volume":"518","author":"Sulaiman","year":"2010","journal-title":"J. Comp. Neurol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1167\/iovs.08-2758","article-title":"Effects of presynaptic mutations on a postsynaptic Cacna1s calcium channel colocalized with mGluR6 at mouse photoreceptor ribbon synapses","volume":"50","author":"Specht","year":"2009","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_31","first-page":"1545","article-title":"Long-term preservation of cone photoreceptors and visual acuity in rd10 mutant mice exposed to continuous environmental enrichment","volume":"20","author":"Barone","year":"2014","journal-title":"Mol. Vis."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1016\/0006-8993(90)91774-B","article-title":"Pontine cholinergic neurons simultaneously innervate two thalamic targets","volume":"532","author":"Shiromani","year":"1990","journal-title":"Brain Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2405","DOI":"10.1002\/cne.22381","article-title":"Morphology and mosaics of melanopsin- expressing retinal ganglion cell types in mice","volume":"518","author":"Berson","year":"2010","journal-title":"J. Comp. Neurol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"17374","DOI":"10.1073\/pnas.1008533107","article-title":"Melanopsin-dependent light avoidance in neonatal mice","volume":"107","author":"Johnson","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1016\/j.ydbio.2021.04.001","article-title":"Development and maintenance of vision\u2019s first synapse","volume":"476","author":"Burger","year":"2021","journal-title":"Dev. Biol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1016\/S1568-1637(02)00007-7","article-title":"Photoreceptor degeneration and dysfunction in aging and age-related maculopathy","volume":"1","author":"Jackson","year":"2002","journal-title":"Ageing Res. Rev."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"8936","DOI":"10.1523\/JNEUROSCI.18-21-08936.1998","article-title":"The major cell populations of the mouse retina","volume":"18","author":"Jeon","year":"1998","journal-title":"J. Neurosci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"eaba7232","DOI":"10.1126\/sciadv.aba7232","article-title":"Molecular and functional architecture of the mouse photoreceptor network","volume":"6","author":"Jin","year":"2020","journal-title":"Sci. Adv."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"708","DOI":"10.1016\/j.neuron.2014.10.021","article-title":"An unconventional glutamatergic circuit in the retina formed by vGluT3 amacrine cells","volume":"84","author":"Lee","year":"2014","journal-title":"Neuron"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"7184","DOI":"10.1523\/JNEUROSCI.3500-15.2016","article-title":"M1 ipRGCs Influence Visual Function through Retrograde Signaling in the Retina","volume":"36","author":"Prigge","year":"2016","journal-title":"J. Neurosci."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e25591","DOI":"10.1002\/cne.25591","article-title":"Efficacy and specificity of melanopsin reporters for retinal ganglion cells","volume":"532","author":"Maloney","year":"2024","journal-title":"J. Comp. Neurol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1016\/j.neuron.2007.10.003","article-title":"Critical periods in the visual system: Changing views for a model of experience-dependent plasticity","volume":"56","author":"Hooks","year":"2007","journal-title":"Neuron"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/S0896-6273(03)00389-1","article-title":"Visual stimulation is required for refinement of ON and OFF pathways in postnatal retina","volume":"39","author":"Tian","year":"2003","journal-title":"Neuron"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"7256","DOI":"10.1523\/JNEUROSCI.0779-07.2007","article-title":"Brain-derived neurotrophic factor and TrkB modulate visual experience- dependent refinement of neuronal pathways in retina","volume":"27","author":"Liu","year":"2007","journal-title":"J. Neurosci."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1002\/dvdy.10376","article-title":"Major role of BAX in apoptosis during retinal development and in establishment of a functional postnatal retina","volume":"228","author":"Provost","year":"2003","journal-title":"Dev. Dynam"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1482","DOI":"10.1016\/j.exer.2006.08.007","article-title":"Analysis of apoptotic and survival mediators in the early post-natal and mature retina","volume":"83","author":"Donovan","year":"2006","journal-title":"Exp. Eye Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1059","DOI":"10.1016\/j.neuron.2018.10.011","article-title":"Light Prior to Eye Opening Promotes Retinal Waves and Eye-Specific Segregation","volume":"100","author":"Tiriac","year":"2018","journal-title":"Neuron"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"935","DOI":"10.1002\/dneu.22260","article-title":"Melanopsin ganglion cells extend dendrites into the outer retina during early postnatal development","volume":"75","author":"Renna","year":"2015","journal-title":"Dev. Neurobiol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"3124","DOI":"10.1016\/j.cell.2022.07.009","article-title":"Melanopsin retinal ganglion cells mediate light-promoted brain development","volume":"185","author":"Hu","year":"2022","journal-title":"Cell"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"2897","DOI":"10.1016\/j.devcel.2024.07.018","article-title":"Developmental control of rod number via a light- dependent retrograde pathway from intrinsically photosensitive retinal ganglion cells","volume":"59","author":"Upton","year":"2024","journal-title":"Dev. Cell"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1473","DOI":"10.1167\/iovs.04-0653","article-title":"Downregulation of cone-specific gene expression and degeneration of cone photoreceptors in the Rpe65-\/- mouse at early ages","volume":"46","author":"Znoiko","year":"2005","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Berry, C.T., Sceniak, M.P., Zhou, L., and Sabo, S.L. (2012). Developmental up-regulation of vesicular glutamate transporter-1 promotes neocortical presynaptic terminal development. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0050911"}],"container-title":["Brain Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-3425\/15\/9\/1024\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:47:18Z","timestamp":1760035638000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-3425\/15\/9\/1024"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,22]]},"references-count":52,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["brainsci15091024"],"URL":"https:\/\/doi.org\/10.3390\/brainsci15091024","relation":{},"ISSN":["2076-3425"],"issn-type":[{"type":"electronic","value":"2076-3425"}],"subject":[],"published":{"date-parts":[[2025,9,22]]}}}