Abstract
The subcommissural organ (SCO) is a gland located at the entrance of the aqueduct of Sylvius in the brain. It exists in species as distantly related as amphioxus and humans, but its function is largely unknown. Here, to explore its function, we compared transcriptomes of SCO and non-SCO brain regions and found three genes, Sspo, Car3 and Spdef, that are highly expressed in the SCO. Mouse strains expressing Cre recombinase from endogenous promoter/enhancer elements of these genes were used to genetically ablate SCO cells during embryonic development, resulting in severe hydrocephalus and defects in neuronal migration and development of neuronal axons and dendrites. Unbiased peptidomic analysis revealed enrichment of three SCO-derived peptides, namely, thymosin beta 4, thymosin beta 10 and NP24, and their reintroduction into SCO-ablated brain ventricles substantially rescued developmental defects. Together, these data identify a critical role for the SCO in brain development.
This is a preview of subscription content, access via your institution
Access options
Access Nature and 54 other Nature Portfolio journals
Get Nature+, our best-value online-access subscription
$29.99 / 30 days
cancel any time
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Data availability
All research materials and data are publicly available. New Cre mouse strains are in the process of being donated to the Jackson Laboratory mouse repository for distribution. RNA-seq are publicly available from NCBI GEO (accession numbers GSE214744 and GSE226349) and peptidomic data from IPROX (accession numbers 15E4 and U0Jm). RNA-seq results: these data have been deposited in the publicly available NCBI GEO database and will be released to the public upon publication. The accession information is as follows: SCO bulk RNA-seq (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE214744), secure token ivatogiazvabpoj; RNA-seq of neurons with peptide incubation (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE226349), secure token yfkvcygwxbkrvyr; peptideomics data (https://www.iprox.cn/page/PSV023.html;?url=1713509741810ndGU), accession number 15E4; proteomics data (https://www.iprox.cn/page/PSV023.html;?url=1697721703737jgBn), accession number U0Jm.
References
Peruzzo, B. et al. Ultrastructural immunocytochemistry and lectin histochemistry of the subcommissural organ in the snake Natrix maura with particular emphasis on its vascular and leptomeningeal projections. Histochemistry 93, 269–277 (1990).
Rodríguez, E. M., Oksche, A. & Montecinos, H. Human subcommissural organ, with particular emphasis on its secretory activity during the fetal life. Microsc. Res. Tech. 52, 573–590 (2001).
Guerra, M. M. et al. Understanding how the subcommissural organ and other periventricular secretory structures contribute via the cerebrospinal fluid to neurogenesis. Front. Cell. Neurosci. 9, 480 (2015).
Ortloff, A. R. et al. Role of the subcommissural organ in the pathogenesis of congenital hydrocephalus in the HTx rat. Cell Tissue Res. 352, 707–725 (2013).
Ortega, E. et al. The value of early and comprehensive diagnoses in a human fetus with hydrocephalus and progressive obliteration of the aqueduct of Sylvius: case report. BMC Neurol. 16, 45 (2016).
Kohn, D., Chinookoswong, N. & Chou, S. A new model of congenital hydrocephalus in the rat. Acta Neuropathol. 54, 211–218 (1981).
Somera, K. C. & Jones, H. C. Reduced subcommissural organ glycoprotein immunoreactivity precedes aqueduct closure and ventricular dilatation in H-Tx rat hydrocephalus. Cell Tissue Res. 315, 361–373 (2004).
Perez-Figares, J. et al. Spontaneous congenital hydrocephalus in the mutant mouse hyh. Changes in the ventricular system and the subcommissural organ. J. Neuropathol. Exp. Neurol. 57, 188–202 (1998).
Caprile, T., Hein, S., Rodrı́guez, S., Montecinos, H. & Rodrı́guez, E. Reissner fiber binds and transports away monoamines present in the cerebrospinal fluid. Mol. Brain. Res. 110, 177–192 (2003).
Muñoz, R. I. et al. The subcommissural organ and the Reissner fiber: old friends revisited. Cell Tissue Res. 375, 507–529 (2019).
Jiménez, A. et al. A programmed ependymal denudation precedes congenital hydrocephalus in the hyh mutant mouse. J. Neuropathol. Exp. Neurol. 60, 1105–1119 (2001).
Troutwine, B. R. et al. The Reissner fiber is highly dynamic in vivo and controls morphogenesis of the spine. Curr. Biol. 30, 2353–2362 e2353 (2020).
Rose, C. D. et al. SCO-spondin defects and neuroinflammation are conserved mechanisms driving spinal deformity across genetic models of idiopathic scoliosis. Curr. Biol. 30, 2363–2373 e2366 (2020).
Orts-Del'Immagine, A. et al. Sensory neurons contacting the cerebrospinal fluid require the Reissner fiber to detect spinal curvature in vivo. Curr. Biol. 30, 827–839 e824 (2020).
Kousi, M. & Katsanis, N. The genetic basis of hydrocephalus. Annu. Rev. Neurosci. 39, 409–435 (2016).
Cifuentes, M., Fernández-LLebrez, P., Perez, J., Perez-Figares, J. & Rodriguez, E. Distribution of intraventricularly injected horseradish peroxidase in cerebrospinal fluid compartments of the rat spinal cord. Cell Tissue Res. 270, 485–494 (1992).
Meiniel, A. et al. The subcommissural organ and Reissner’s fiber complex: an enigma in the central nervous system? Prog. Histochem. Cytochem. 30, 1–66 (1996).
Gobron, S. et al. Subcommissural organ/Reissner’s fiber complex: characterization of SCO‐spondin, a glycoprotein with potent activity on neurite outgrowth. Glia 32, 177–191 (2000).
Gobron, S. et al. SCO-spondin: a new member of the thrombospondin family secreted by the subcommissural organ is a candidate in the modulation of neuronal aggregation. J. Cell Sci. 109, 1053–1061 (1996).
Rodríguez, S. et al. Isograft and xenograft of the subcommissural organ into the lateral ventricle of the rat and the formation of Reissner’s fiber. Cell Tissue Res. 296, 457–469 (1999).
Meiniel, A. SCO‐spondin, a glycoprotein of the subcommissural organ/Reissner’s fiber complex: evidence of a potent activity on neuronal development in primary cell cultures. Microsc. Res. Tech. 52, 484–495 (2001).
Brown, D. D. & Afifi, A. K. Histological and ablation studies on the relation of the subcommissural organ and rostral midbrain to sodium and water metabolism. Anat. Rec. 153, 255–263 (1965).
Voehringer, D., Liang, H.-E. & Locksley, R. M. Homeostasis and effector function of lymphopenia-induced ‘memory-like’ T cells in constitutively T cell-depleted mice. J. Immunol. 180, 4742–4753 (2008).
Daigle, T. L. et al. A suite of transgenic driver and reporter mouse lines with enhanced brain-cell-type targeting and functionality. Cell 174, 465–480 e422 (2018).
Madisen, L. et al. A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Nat. Neurosci. 13, 133–140 (2010).
Yulis, C. R. & Muñoz, R. I. Vertebrate floor plate transiently expresses a compound recognized by antisera raised against subcommissural organ secretion. Microsc. Res. Tech. 52, 608–614 (2001).
Rakic, P. & Sidman, R. L. Subcommissural organ and adjacent ependyma: autoradiographic study of their origin in the mouse brain. Am. J. Anat. 122, 317–335 (1968).
Hoyo-Becerra, C. et al. The subcommissural organ and the development of the posterior commissure in chick embryos. Cell Tissue Res. 339, 383–395 (2010).
Roth, L. W., Bormann, P., Bonnet, A. & Reinhard, E. β-Thymosin is required for axonal tract formation in developing zebrafish brain. Development 126, 1365–1374 (1999).
Mantri, M. et al. Spatiotemporal single-cell RNA sequencing of developing chicken hearts identifies interplay between cellular differentiation and morphogenesis. Nat. Commun. 12, 1771 (2021).
Delétage, N., Le Douce, J., Callizot, N., Godfrin, Y. & Lemarchant, S. SCO-spondin-derived peptide protects neurons from glutamate-induced excitotoxicity. Neuroscience 463, 317–336 (2021).
Hannapel, E. & van Kampen, M. Determination of thymosin β4 in human blood cells and serum. J. Chromatogr. A 397, 279–285 (1987).
Frohm, M. et al. Biochemical and antibacterial analysis of human wound and blister fluid. Eur. J. Biochem. 237, 86–92 (1996).
Pérez, J. et al. Light-and electron-microscopic immunocytochemical investigation of the subcommissural organ using a set of monoclonal antibodies against the bovine Reissner’s fiber. Histochem. Cell Biol. 104, 221–232 (1995).
Furey, C. G. et al. De novo mutation in genes regulating neural stem cell fate in human congenital hydrocephalus. Neuron 99, 302–314. e304 (2018).
Jin, S. C. et al. Exome sequencing implicates genetic disruption of prenatal neuro-gliogenesis in sporadic congenital hydrocephalus. Nat. Med. 26, 1754–1765 (2020).
Hale, A. T. et al. Multi-omic analysis elucidates the genetic basis of hydrocephalus. Cell Rep. 35, 109085 (2021).
Duy, P. Q. et al. Impaired neurogenesis alters brain biomechanics in a neuroprogenitor-based genetic subtype of congenital hydrocephalus. Nat. Neurosci. 25, 458–473 (2022).
Vio, K. et al. Hydrocephalus induced by immunological blockage of the subcommissural organ-Reissner’s fiber (RF) complex by maternal transfer of anti-RF antibodies. Exp. Brain Res. 135, 41–52 (2000).
Kiefer, M. et al. The ependyma in chronic hydrocephalus. Child’s Nerv. Syst. 14, 263–270 (1998).
Wagner, C. et al. Cellular mechanisms involved in the stenosis and obliteration of the cerebral aqueduct of hyh mutant mice developing congenital hydrocephalus. J. Neuropathol. Exp. Neurol. 62, 1019–1040 (2003).
Rodríguez, S. et al. Changes in the cerebrospinal-fluid monoamines in rats with an immunoneutralization of the subcommissural organ–Reissner’s fiber complex by maternal delivery of antibodies. Exp. Brain Res. 128, 278–290 (1999).
Rodríguez, S. & Caprile, T. Functional aspects of the subcommissural organ–Reissner’s fiber complex with emphasis in the clearance of brain monoamines. Microsc. Res. Tech. 52, 564–572 (2001).
Lehtinen, M. K. & Walsh, C. A. Neurogenesis at the brain–cerebrospinal fluid interface. Annu. Rev. Cell Dev. Biol. 27, 653–679 (2011).
Lehtinen, M. K. et al. The cerebrospinal fluid provides a proliferative niche for neural progenitor cells. Neuron 69, 893–905 (2011).
Carter, C. S. et al. Abnormal development of NG2+ PDGFR-α+ neural progenitor cells leads to neonatal hydrocephalus in a ciliopathy mouse model. Nat. Med. 18, 1797–1804 (2012).
Rodriguez, E. M. & Guerra, M. M. Neural stem cells and fetal-onset hydrocephalus. Pediatr. Neurosurg. 52, 446–461 (2017).
Wisniewski, J. R., Zougman, A., Nagaraj, N. & Mann, M. Universal sample preparation method for proteome analysis. Nat. Methods 6, 359–362 (2009).
Zhang, P. et al. A dynamic mouse peptidome landscape reveals probiotic modulation of the gut–brain axis. Sci. Signal. https://doi.org/10.1126/scisignal.abb0443 (2020).
DeLaney, K. & Li, L. Data independent acquisition mass spectrometry method for improved neuropeptidomic coverage in crustacean neural tissue extracts. Anal. Chem. 91, 5150–5158 (2019).
Acknowledgements
We thank lab members in Ge laboratories, B. Samuels and T. Taylor for their feedback and critical reading of the manuscript. We thank Q. Guo, X. Gao and M. Jia in the imaging core of CIBR for assistance with imaging and data analysis. We thank W. Li, S. Huang and R. Shen at the animal core facility for assistance with animal care and purchasing. We thank J. Chen and X. Zhang at the Genomics Center for sequencing and fluorescence-activated cell sorting. We thank the MS facility of the Phoenix Center for use of their instrumentation. We thank F. Huang for suggestions. This work was supported in part by grants from the STI2030-Major Projects (2022ZD0204700), the Natural Science Foundation of China and Beijing Scholars to W.G. (32170964); startup funds from CIBR, the National Key R&D Program of China to C.J. (no. 2021YFA1302601); and a grant from the US National Institutes of Health (NIH, R01DK071801) to L.L. The Orbitrap instruments were purchased through support from an NIH shared instrument grant (NIH-NCRR S10RR029531).
Author information
Authors and Affiliations
Contributions
W.G. conceived and supervised the project. T.Z., D.A., P.W., L.L., C.J., W.S. and W.G. designed experiments. T.Z. performed most experiments, including mouse genetics, cell ablation, neuronal culture, immunostaining, imaging, RNAscope mFISH, mouse breeding and genotyping, CSF collection and rescue experiments, among others. P.W., Y.X., F.M., Y.Z., T.Z. and C.J. performed peptidomics and proteomics and data analysis. W.G., Z.Z., T.T. and X. Zhang performed SCO bulk RNA-seq; C.X., D.A. and W.G. analyzed the RNA-seq data. W.G. and T.Z. determined the genes for genetic mouse strains. T.Z. and J.L. assisted in MRI data acquisition and analysis, T.T. assisted in MRI imaging, Z.B. and T.Z. performed AAV viral labeling and peptide injection and assisted in slice imaging. X.-J.C., J.-L.L., J.Y. and L. Zheng assisted in brain slice imaging and electrophysiology. F.L. assisted in neuronal culture. M.Y. assisted in RNA-seq. C.L. assisted in electron microscopy. X. Zou and Z.F. assisted in mouse breeding, genotyping and immunostaining. Z.G. assisted in CSF collection. W.G., W.S., L.L., B.L., C.J., T.H., Z.L., L. Zhang, H. Zhang and H. Zeng provided reagents. T.Z. and W.G. wrote the manuscript. All authors discussed, reviewed and edited the manuscript.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing interests.
Peer review
Peer review information
Nature Neuroscience thanks Montserrat Guerra and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.
Additional information
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary information
Supplementary Information
Supplementary Figs. 1–24 and Tables 1–3.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Zhang, T., Ai, D., Wei, P. et al. The subcommissural organ regulates brain development via secreted peptides. Nat Neurosci (2024). https://doi.org/10.1038/s41593-024-01639-x
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41593-024-01639-x