Septo-dentate gyrus cholinergic circuits modulate function and morphogenesis of adult neural stem cells through granule cell intermediaries
Edited by Arturo Alvarez-Buylla, University of California San Francisco, San Francisco, CA; received March 15, 2024; accepted July 17, 2024
Significance
Radial neural stem cells (rNSCs) in the adult dentate gyrus (DG) can proliferate and generate new adult-born neurons throughout life contributing to hippocampus-dependent learning and memory. How this process is regulated by distinct neural circuits remains elusive. In this study, we reveal how septo-DG cholinergic circuits orchestrate the key niche cells to support neurogenic function and morphogenesis of rNSCs. Furthermore, using single-nucleus RNA sequencing, we identify cell-specific transcriptional changes in the adult DG in response to cholinergic circuit activity. Our study bridges a long-standing gap in understanding combined circuit and molecular mechanisms underlying activity-dependent regulation of rNSCs.
Abstract
Cholinergic neurons in the basal forebrain play a crucial role in regulating adult hippocampal neurogenesis (AHN). However, the circuit and molecular mechanisms underlying cholinergic modulation of AHN, especially the initial stages of this process related to the generation of newborn progeny from quiescent radial neural stem cells (rNSCs), remain unclear. Here, we report that stimulation of the cholinergic circuits projected from the diagonal band of Broca (DB) to the dentate gyrus (DG) neurogenic niche promotes proliferation and morphological development of rNSCs, resulting in increased neural stem/progenitor pool and rNSCs with longer radial processes and larger busy heads. Interestingly, DG granule cells (GCs) are required for DB-DG cholinergic circuit–dependent modulation of proliferation and morphogenesis of rNSCs. Furthermore, single-nucleus RNA sequencing of DG reveals cell type–specific transcriptional changes in response to cholinergic circuit stimulation, with GCs (among all the DG niche cells) exhibiting the most extensive transcriptional changes. Our findings shed light on how the DB-DG cholinergic circuits orchestrate the key niche components to support neurogenic function and morphogenesis of rNSCs at the circuit and molecular levels.
Data, Materials, and Software Availability
Single-nucleus RNA-seq data have been deposited to the Gene Expression Omnibus (GEO) under accession GSE202481 (56). Code is available in a Song Lab Repo on GitHub at https://github.com/zekachen/Chen_Quintanilla_cholinergic_neurogenesis2024 (57).
Acknowledgments
We thank all the members of the Song lab for comments and discussions. This work was supported by grants from NIH (R01MH111773, R01MH122692, RF1AG058160, R01NS104530, and R01MH132222 to J. S., R35NS116843 to H. S., and R35NS097370 to G-L.M.) and Alzheimer’s Association to J.S., L.Q. was partially supported by NIH T32 training grant (T32NS007431), National Institute of Aging Ruth L. Kirschstein F31 predoctoral fellowship (AG067718), and a NIMH R01 Diversity Supplement (3R01MH111773-03S1). R.N.S. was partially supported by NIH T32 training grant and Thomas Collum Butler Award from Department of Pharmacology. Confocal microscopy was performed at the UNC Neuroscience Microscopy Core Facility (RRID: SCR_019060). The Neuroscience Microscopy Core was supported in part by funding from the NIH-NINDS Neuroscience Center Support Grant P30 NS045892 and the NIH-NICHD Intellectual and Developmental Disabilities Research Center Support Grant U54 HD079124.
Author contributions
Z.-K.C., L.Q., G.-L.M., H.S., and J.S. designed research; Z.-K.C., L.Q., Y.S., R.N.S., Y.-J.L., Y.-D.L., and B.A. performed research; Z.-K.C. and L.Q. contributed new reagents/analytic tools; Z.-K.C., L.Q., Y.S., R.N.S., J.M.S., Y.-J.L., Y.-D.L., Z.C., B.A., D.S.T., and W.T.F. analyzed data; J.S. conceived the project; J.S. directed the project; and Z.-K.C., L.Q., and J.S. wrote the paper.
Competing interests
The authors declare no competing interest.
Supporting Information
Appendix 01 (PDF)
- Download
- 2.49 MB
Dataset S01 (XLSX)
- Download
- 332.69 KB
Dataset S02 (XLSX)
- Download
- 30.53 KB
References
1
M. E. Hasselmo, The role of acetylcholine in learning and memory. Curr. Opin. Neurobiol. 16, 710–715 (2006).
2
J. Winkler, S. T. Suhr, F. H. Gage, L. J. Thal, L. J. Fisher, Essential role of neocortical acetylcholine in spatial memory. Nature 375, 484–487 (1995).
3
A. Green et al., Muscarinic and nicotinic receptor modulation of object and spatial n-back working memory in humans. Pharmacol. Biochem. Behav. 81, 575–584 (2005).
4
J. J. Buccafusco, S. R. Letchworth, M. Bencherif, P. M. Lippiello, Long-lasting cognitive improvement with nicotinic receptor agonists: Mechanisms of pharmacokinetic-pharmacodynamic discordance. Trends Pharmacol. Sci. 26, 352–360 (2005).
5
K. M. Christian, H. Song, G. L. Ming, Functions and dysfunctions of adult hippocampal neurogenesis. Annu. Rev. Neurosci. 37, 243–262 (2014).
6
H. Bao, J. Song, Treating brain disorders by targeting adult neural stem cells. Trends Mol. Med. 24, 991–1006 (2018).
7
Y. D. Li, Y. J. Luo, J. Song, Optimizing memory performance and emotional states: Multi-level enhancement of adult hippocampal neurogenesis. Curr. Opin. Neurobiol. 79, 102693 (2023).
8
L. I. Madrid, J. Jimenez-Martin, E. J. Coulson, D. J. Jhaveri, Cholinergic regulation of adult hippocampal neurogenesis and hippocampus-dependent functions. Int. J. Biochem. Cell Biol. 134, 105969 (2021).
9
M. A. Bonaguidi et al., In vivo clonal analysis reveals self-renewing and multipotent adult neural stem cell characteristics. Cell 145, 1142–1155 (2011).
10
J. Song et al., Neuronal circuitry mechanism regulating adult quiescent neural stem-cell fate decision. Nature 489, 150–154 (2012).
11
M. A. Bonaguidi, J. Song, G. L. Ming, H. Song, A unifying hypothesis on mammalian neural stem cell properties in the adult hippocampus. Curr. Opin. Neurobiol. 22, 754–761 (2012).
12
E. Gebara et al., Heterogeneity of radial glia-like cells in the adult hippocampus. Stem. Cells 34, 997–1010 (2016).
13
C. Y. Yeh et al., Mossy cells control adult neural stem cell quiescence and maintenance through a dynamic balance between direct and indirect pathways. Neuron 99, 493–510.e4 (2018).
14
M. C. Senut, D. Menetrey, Y. Lamour, Cholinergic and peptidergic projections from the medial septum and the nucleus of the diagonal band of Broca to dorsal hippocampus, cingulate cortex and olfactory bulb: A combined wheatgerm agglutinin-apohorseradish peroxidase-gold immunohistochemical study. Neuroscience 30, 385–403 (1989).
15
C. Nyakas, P. G. Luiten, D. G. Spencer, J. Traber, Detailed projection patterns of septal and diagonal band efferents to the hippocampus in the rat with emphasis on innervation of CA1 and dentate gyrus. Brain Res. Bull. 18, 533–545 (1987).
16
A. Sans-Dublanc et al., Septal GABAergic inputs to CA1 govern contextual memory retrieval. Sci. Adv. 6, eaba5003 (2020).
17
B. Kiraly et al., The medial septum controls hippocampal supra-theta oscillations. Nat. Commun. 14, 6159 (2023).
18
X. Li et al., Molecularly defined and functionally distinct cholinergic subnetworks. Neuron 110, 3774–3788.e7 (2022), https://doi.org/10.1016/j.neuron.2022.08.025.
19
Y. D. Li et al., Hypothalamic modulation of adult hippocampal neurogenesis in mice confers activity-dependent regulation of memory and anxiety-like behavior. Nat. Neurosci. 25, 630–645 (2022).
20
H. Bao et al., Long-range GABAergic inputs regulate neural stem cell quiescence and control adult hippocampal neurogenesis. Cell Stem. Cell 21, 604–617 e605 (2017).
21
B. Asrican et al., Neuropeptides modulate local astrocytes to regulate adult hippocampal neural stem cells. Neuron 108, 349–366.e6 (2020).
22
H. Wu, J. Williams, J. Nathans, Complete morphologies of basal forebrain cholinergic neurons in the mouse. Elife 3, e02444 (2014).
23
J. Rossi et al., Melanocortin-4 receptors expressed by cholinergic neurons regulate energy balance and glucose homeostasis. Cell Metab. 13, 195–204 (2011).
24
C. H. Vanderwolf, Hippocampal electrical activity and voluntary movement in the rat. Electroencephalogr. Clin. Neurophysiol. 26, 407–418 (1969).
25
L. L. Colgin, Rhythms of the hippocampal network. Nat. Rev. Neurosci. 17, 239–249 (2016).
26
H. Zhang, S. C. Lin, M. A. Nicolelis, Spatiotemporal coupling between hippocampal acetylcholine release and theta oscillations in vivo. J. Neurosci. 30, 13431–13440 (2010).
27
R. Mu et al., A cholinergic medial septum input to medial habenula mediates generalization formation and extinction of visual aversion. Cell Rep. 39, 110882 (2022).
28
M. B. Ogando et al., Cholinergic modulation of dentate gyrus processing through dynamic reconfiguration of inhibitory circuits. Cell Rep. 36, 109572 (2021).
29
K. Martinello et al., Cholinergic afferent stimulation induces axonal function plasticity in adult hippocampal granule cells. Neuron 85, 346–363 (2015).
30
M. Pabst et al., Astrocyte intermediaries of septal cholinergic modulation in the hippocampus. Neuron 90, 853–865 (2016).
31
E. A. Oyarzabal et al., Chemogenetic stimulation of tonic locus coeruleus activity strengthens the default mode network. Sci. Adv. 8, eabm9898 (2022).
32
N. Habib et al., Div-Seq: Single-nucleus RNA-Seq reveals dynamics of rare adult newborn neurons. Science 353, 925–928 (2016).
33
H. Hochgerner, A. Zeisel, P. Lonnerberg, S. Linnarsson, Conserved properties of dentate gyrus neurogenesis across postnatal development revealed by single-cell RNA sequencing. Nat. Neurosci. 21, 290–299 (2018).
34
D. Arneson et al., Single cell molecular alterations reveal target cells and pathways of concussive brain injury. Nat. Commun. 9, 3894 (2018).
35
B. Artegiani et al., A single-cell RNA sequencing study reveals cellular and molecular dynamics of the hippocampal neurogenic niche. Cell Rep. 21, 3271–3284 (2017).
36
S. L. Ding et al., Distinct transcriptomic cell types and neural circuits of the subiculum and prosubiculum along the dorsal-ventral axis. Cell Rep. 31, 107648 (2020).
37
J. Shin et al., Single-cell RNA-seq with waterfall reveals molecular cascades underlying adult neurogenesis. Cell Stem. Cell 17, 360–372 (2015).
38
B. P. Rubin, R. P. Tucker, D. Martin, R. Chiquet-Ehrismann, Teneurins: A novel family of neuronal cell surface proteins in vertebrates, homologous to the Drosophila pair-rule gene product Ten-m. Dev. Biol. 216, 195–209 (1999).
39
B. P. Rubin, R. P. Tucker, M. Brown-Luedi, D. Martin, R. Chiquet-Ehrismann, Teneurin 2 is expressed by the neurons of the thalamofugal visual system in situ and promotes homophilic cell-cell adhesion in vitro. Development 129, 4697–4705 (2002).
40
T. R. Young et al., Ten-m2 is required for the generation of binocular visual circuits. J. Neurosci. 33, 12490–12509 (2013).
41
O. Rivero et al., Cadherin-13, a risk gene for ADHD and comorbid disorders, impacts GABAergic function in hippocampus and cognition. Transl Psychiatry 5, e655 (2015).
42
K. Street et al., Slingshot: Cell lineage and pseudotime inference for single-cell transcriptomics. BMC Genomics 19, 477 (2018).
43
R. Ji et al., TAM receptors support neural stem cell survival, proliferation and neuronal differentiation. PLoS One 9, e115140 (2014).
44
K. Khodosevich, P. H. Seeburg, H. Monyer, Major signaling pathways in migrating neuroblasts. Front. Mol. Neurosci. 2, 7 (2009).
45
M. H. Jang et al., Secreted frizzled-related protein 3 regulates activity-dependent adult hippocampal neurogenesis. Cell Stem. Cell 12, 215–223 (2013).
46
J. Dong et al., A neuronal molecular switch through cell-cell contact that regulates quiescent neural stem cells. Sci. Adv. 5, eaav4416 (2019).
47
R. Browaeys, W. Saelens, Y. Saeys, NicheNet: Modeling intercellular communication by linking ligands to target genes. Nat. Methods 17, 159–162 (2020).
48
J. L. Mignone, V. Kukekov, A. S. Chiang, D. Steindler, G. Enikolopov, Neural stem and progenitor cells in nestin-GFP transgenic mice. J. Comp. Neurol. 469, 311–324 (2004).
49
L. J. Quintanilla, C. Y. Yeh, H. Bao, C. Catavero, J. Song, Assaying circuit specific regulation of adult hippocampal neural precursor cells. J. Vis. Exp. 149, e59237 (2019), https://doi.org/10.3791/59237.
50
Y. Li et al., Supramammillary nucleus synchronizes with dentate gyrus to regulate spatial memory retrieval through glutamate release. Elife 9, e53129 (2020).
51
A. B. Rosenberg et al., Single-cell profiling of the developing mouse brain and spinal cord with split-pool barcoding. Science 360, 176–182 (2018).
52
Y. Su et al., Neuronal activity modifies the chromatin accessibility landscape in the adult brain. Nat. Neurosci. 20, 476–483 (2017).
53
X. Qian et al., Sliced human cortical organoids for modeling distinct cortical layer formation. Cell Stem. Cell 26, 766–781.e9 (2020).
54
S. Parekh, C. Ziegenhain, B. Vieth, W. Enard, I. Hellmann, zUMIs–A fast and flexible pipeline to process RNA sequencing data with UMIs. Gigascience 7, giy059 (2018).
55
J. Harrow et al., GENCODE: The reference human genome annotation for The ENCODE Project. Genome Res. 22, 1760–1774 (2012).
56
Z.-K. Chen et al., Data from “Septo-dentate gyrus cholinergic circuits modulate function and morphogenesis of adult neural stem cells through granule cell intermediaries.” NCBI GEO. https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE202481. Deposited 8 May 2022.
57
J. M. Simon et al., Code from “Chen_Quintanilla_cholinergic_neurogenesis 2024.” Github. https://github.com/zekachen/Chen_Quintanilla_cholinergic_neurogenesis2024. Deposited 31 July 2024.
Information & Authors
Information
Published in
Cover image: Pictured is green foxtail (Setaria viridis), an annual grass. Magda Garbowski et al. combined data from more than 75,000 vegetation plots across the United States with functional trait data for more than 5,500 plant species to uncover how naturalized species affect plant community traits. Compared with communities dominated by native species, communities dominated by naturalized species were shorter, more shallowly rooted, and exhibited traits supporting rapid resource acquisition and low reliance on mycorrhizal symbionts. According to the authors, community-level trait shifts likely underpin the functional changes observed in invaded ecosystems. See the article by Garbowski et al. e2403120121. Image credit: Magda Garbowski.
Classifications
Copyright
Copyright © 2024 the Author(s). Published by PNAS. This open access article is distributed under Creative Commons Attribution License 4.0 (CC BY).
Data, Materials, and Software Availability
Single-nucleus RNA-seq data have been deposited to the Gene Expression Omnibus (GEO) under accession GSE202481 (56). Code is available in a Song Lab Repo on GitHub at https://github.com/zekachen/Chen_Quintanilla_cholinergic_neurogenesis2024 (57).
Submission history
Received: March 15, 2024
Accepted: July 17, 2024
Published online: September 23, 2024
Published in issue: October 1, 2024
Keywords
Acknowledgments
We thank all the members of the Song lab for comments and discussions. This work was supported by grants from NIH (R01MH111773, R01MH122692, RF1AG058160, R01NS104530, and R01MH132222 to J. S., R35NS116843 to H. S., and R35NS097370 to G-L.M.) and Alzheimer’s Association to J.S., L.Q. was partially supported by NIH T32 training grant (T32NS007431), National Institute of Aging Ruth L. Kirschstein F31 predoctoral fellowship (AG067718), and a NIMH R01 Diversity Supplement (3R01MH111773-03S1). R.N.S. was partially supported by NIH T32 training grant and Thomas Collum Butler Award from Department of Pharmacology. Confocal microscopy was performed at the UNC Neuroscience Microscopy Core Facility (RRID: SCR_019060). The Neuroscience Microscopy Core was supported in part by funding from the NIH-NINDS Neuroscience Center Support Grant P30 NS045892 and the NIH-NICHD Intellectual and Developmental Disabilities Research Center Support Grant U54 HD079124.
Author Contributions
Z.-K.C., L.Q., G.-L.M., H.S., and J.S. designed research; Z.-K.C., L.Q., Y.S., R.N.S., Y.-J.L., Y.-D.L., and B.A. performed research; Z.-K.C. and L.Q. contributed new reagents/analytic tools; Z.-K.C., L.Q., Y.S., R.N.S., J.M.S., Y.-J.L., Y.-D.L., Z.C., B.A., D.S.T., and W.T.F. analyzed data; J.S. conceived the project; J.S. directed the project; and Z.-K.C., L.Q., and J.S. wrote the paper.
Competing Interests
The authors declare no competing interest.
Notes
This article is a PNAS Direct Submission.
Authors
Metrics & Citations
Metrics
Citation statements
Altmetrics
Citations
If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click Download.
View Options
View options
PDF format
Download this article as a PDF file
DOWNLOAD PDFGet Access
Login options
Check if you have access through your login credentials or your institution to get full access on this article.
Personal login Institutional LoginRecommend to a librarian
Recommend PNAS to a LibrarianPurchase options
Purchase this article to access the full text.