Skip to main content

Cell-specific expression of Epac2 in the subventricular and subgranular zones



cAMP signal transduction cascade activation is important in regulating neurogenesis in adult rodents by increasing the proliferation of newborn cells. Although the ventricular-subventricular zone (V-SVZ) and subgranular zone (SGZ) both contain large populations of neural stem/precursor cells; it remains unclear whether an alternative target of cAMP, the exchange protein directly activated by cAMP (Epac2), is involved in adult neurogenesis in the V-SVZ and SGZ. Here, we investigated the cell-specific expression of Epac2 protein in the V-SVZ and SGZ of the adult mouse brain.


Immunohistochemical analyses were performed using antibodies against Epac2, glial fibrillary acidic protein (GFAP), doublecortin (DCX), and beta-catenin, to examine the co-localization of Epac2 protein and neural stem/precursor cells in the V-SVZ and SGZ in three 8-week-old male mice.


In the V-SVZ of the lateral ventricle, most GFAP-positive adult neural stem cells (NSC, defined as type B cells) and 75% of DCX-positive migrating neuroblasts (type A cells) expressed Epac2 proteins. Ninety-three percent of beta-catenin-positive ependymal cells (type E cells), which are in direct contact with NSCs and the ventricles, also expressed Epac2 protein. Similarly, in the SGZ of the hippocampus, Epac2-immunopositive signals were shown by 83% of GFAP-positive radial-glia-like NSCs (type 1 cells), 86% of DCX-positive transiently amplifying cells (type 2 and type 3 cells), and 71% of DCX-positive immature neurons. The present data suggest that a PKA-independent cAMP signaling pathway via Epac2 may be party to adult neurogenesis in the V-SVZ and the SGZ.

Main text

In the adult mammalian brain, neural stem cells (NSCs) are retained in two regions, the ventricular-subventricular zone (V-SVZ) of the forebrain and the subgranular zone (SGZ) of the hippocampus (Fig. 1a, left) [1, 2]. The V-SVZ in the walls of the lateral ventricles contains a subpopulation of cells with astroglial properties (type B cells) that express glial fibrillary acidic protein (GFAP). These cells function as NSCs, giving rise to intermediate progenitor cells (type C cells), which in rodents, predominantly produce the migrating neuroblasts (type A cells) that mature into new neurons destined for the olfactory bulb (Fig. 1a, middle) [3]. In the SGZ of the dentate gyrus, NSCs also correspond to astroglial cells, which have a radial process that traverses the granule cell layer. These cells, known as type 1 progenitors [4] or radial glia-like cells [5], generate transiently amplifying progenitor cells (TAPs) including type 2 cells and migratory neuroblasts (type 3 cells), which amplify but subsequently exit the cell cycle ahead of maturation into granule neurons (Fig. 1a, right) [6]. Among the different intracellular signal transduction cascades, the PKA-dependent cAMP-cAMP response element-binding protein pathway is involved in hippocampal neurogenesis [7]. Interestingly, a recent study suggested that a PKA-independent alternative cAMP signal transduction pathway via an exchange protein directly activated by cAMP 2 (Epac2) also affects hippocampal adult neurogenesis by regulating progenitor cell proliferation [8]. Another study using Epac2 knockout mice [9] showed that Epac2 contributes to astrocytic differentiation in neural precursor cells [10]. However, there is little direct evidence of cell-specific expression of Epac2 in the V-SVZ and SGZ.

Fig. 1
figure 1

Colocalization of Epac2-immunopositive signals with NSCs, migrating neuroblasts, or ependymal cells in the V-SVZ of adult mice; and with NSCs, TAPs, or immature neurons in the SGZ. a. Overview of the V-SVZ and SGZ. Left panel, sagittal section through a mouse brain. Middle panel, cellular composition of the V-SVZ. A, type A cell; B, type B cell; C, type C cell; E, type E cell. Right panel, cellular composition of the SGZ. 1, type 1 cell (NSCs); 2, type 2 cell; 3, type 3 cell; imN, immature neurons. b-d. Representative immunofluorescence images of the V-SVZ. Epac2-immunoreactive signals are observed in the GFAP-positive NSCs (b, green boxes), DCX-positive neuroblasts (c, pink boxes), and beta-catenin-positive ependymal cells (d, yellow boxes). The white box shown in “c” indicates a neuroblast showing non-colocalization. e-f. Representative immunofluorescence images of the SGZ. GFAP-positive radial glia-like stem cells (e, green box), DCX-positive TAPs (f, pink boxes), and a DCX-positive immature neuron with one strong branching (f, yellow box) express Epac2 protein. Scale bars, 25 μm. g. Colocalization rates of Epac2-immunopositive cells with each of the cell marker protein-positive cells in the V-SVZ. H. Colocalization rates of Epac2-immunopositive cells with each type of cell marker protein-positive cell in the SGZ. g and h, Values in tables are presented as mean ± SEM.; n = cell numbers (See Materials and Methods)

To address this issue, we conducted immunohistochemistry using antibodies against Epac2, GFAP to label NSCs, doublecortin (DCX) to label neuroblasts in the V-SVZ or TAPs and immature neurons in the SGZ, and beta-catenin to label ependymal cells. Double immunofluorescence staining of coronal brain sections of three 8-week-old male mice with Epac2 and one of GFAP, DCX, or beta-catenin showed that Epac2 protein was expressed in germinal zones of both the V-SVZ and SGZ. In the V-SVZ, Epac2-immunopositive signals were observed in over 98% of GFAP-positive NSCs (Fig. 1b, green box and 1Gg) and 75% of DCX-positive migrating neuroblasts (type A cells; Fig. 1c, pink box and g). The Epac2 signals were also co-localized with 93% of the beta-catenin-positive ependymal cells (type E cells; Fig. 1d, yellow box and g) that were in immediate contact with GFAP-positive NSCs and cerebrospinal fluid (CSF), which contains soluble factors that could modulate the behavior of NSCs [11]. When viewed en face from the ventricular side, the small apical endings of NSCs are surrounded by a rosette of ependymal cells with large apical surfaces [12], and ependymal cells are known to help maintain the molecular composition of the apical compartment of NSCs by propelling the CSF with their multiple motile cilia [13]. Although we could not evaluate the expression of Epac2 in the type C cells that are represented by less than 10% of the cells in the V-SVZ [14], these findings suggest that the NSCs, neuroblasts, and ependymal cells related to neurogenesis express Epac2 protein in the V-SVZ. Similar to the V-SVZ, Epac2-immunoreactive signals were also colocalized with 83% of GFAP-positive radial glia-like stem cells (type 1 cells; Fig. 1e, green box and 1H), although the total number of GFAP-positive NSCs was much less in the SGZ than in the V-SVZ (see the Additional file 1; Materials and Methods). DCX-immunopositive signals were found in TAPs, including type 2 cells (Fig. 1f, left pink box) and migrating neuroblasts with short processes (type 3 cells; Fig. 1f, right pink box), as well as in postmitotic immature neurons with strong dendritic branching (imN; Fig. 1f, yellow box) in the SGZ at the hippocampus. In the SGZ, we observed Epac2-immunoreactive signals in 86% of DCX-positive TAPs and 71% of immature neurons (Fig. 1h). These findings are consistent with the previous data published by Zhou et al. suggesting that Epac2 gene deletion induced the most obvious decrease in the number of DCX-positive cells with short processes and DCX-positive cells with one strong dendrite branching [10]. Taken together, our findings indicate that Epac2 protein may play an important role in both proliferative and postmitotic stages during neurogenesis in the SGZ.

In conclusion, the Epac2 protein that mediates PKA-independent cAMP signaling may be involved in neurogenesis in the V-SVZ and SGZ of adult mice, and further study to address the specific role of Epac2 in adult neurogenesis is needed.

Availability of data and materials

All materials are available in the Additional file 1 (Materials and Methods).



cerebrospinal fluid




exchange protein directly activated by cAMP 2


glial fibrillary acidic protein


neural stem cells


subgranular zone


transiently amplifying progenitor cells


ventricular-subventricular zone


  1. Luskin MB. Restricted proliferation and migration of postnatally generated neurons derived from the forebrain subventricular zone. Neuron. 1993;11:173–89.

    Article  CAS  Google Scholar 

  2. Seri B, Garcia-Verdugo JM, McEwen BS, Alvarez-Buylla A. Astrocytes give rise to new neurons in the adult mammalian hippocampus. J Neurosci. 2001;21:7153–60.

    Article  CAS  Google Scholar 

  3. Arnold K, Arturo AB. The glial nature of embryonic and adult neural stem cells. Annu Rev Neurosci. 2009;32:149–84.

    Article  Google Scholar 

  4. Filippov V, Kronenberg G, Pivneva T, Reuter K, Steiner B, Wang LP, et al. Subpopulation of nestin-expressing progenitor cells in the adult murine hippocampus shows electrophysiological and morphological characteristics of astrocytes. Mol Cell Neurosci. 2003;23:373–82.

    Article  CAS  Google Scholar 

  5. Bonaguidi MA, Wheeler MA, Shapiro JS, Stadel RP, Sun GJ, Ming GL, Song H. In vivo clonal analysis reveals self-renewing and multipotent adult neural stem cell characteristics. Cell. 2011;145:1142–55.

    Article  CAS  Google Scholar 

  6. Kempermann G, Song H, Gage FH. Neurogenesis in the adult hippocampus. Cold Spring Harb Perspect Biol. 2015;7(9):a018812.

    Article  Google Scholar 

  7. Shin N, Ji-Eun K, Rena L, Jessica EM, Jingshan C, Cathy S, et al. Regulation of neurogenesis in adult mouse Hippocampus by cAMP and the cAMP response element-binding protein. J Neurosci. 2002;22(9):3673–82.

    Article  Google Scholar 

  8. Zhou L, Ma SL, Yeung PKK, Wong YH, Tsim KWK, So KF, Lam LCW, Chung SK. Anxiety and depression with neurogenesis defects in exchange protein directly activated by cAMP 2-deficient mice are ameliorated by a selective serotonin reuptake inhibitor. Prozac Transl Psychiatry. 2016;6:e881.

    Article  CAS  Google Scholar 

  9. Lee K, Kobayashi Y, Seo H, Kwak JH, Masuda A, Lim CS, Lee HR, Kang SJ, Park P, Sim SE, Kogo N, Kawasaki H, Kaang BK, Itohara S. Involvement of cAMP-guanine nucleotide exchange factor II in hippocampal long-term depression and behavioral flexibility. Mol Brain. 2015;8:38.

    Article  CAS  Google Scholar 

  10. Seo H, Lee K. Epac2 contributes to PACAP-induced astrocytic differentiation through calcium ion influx in neural precursor cells. BMB Rep. 2016;49(2):128–33.

    Article  CAS  Google Scholar 

  11. Lehtinen MK, Zappaterra MW, Chen X, Yang YJ, Hill AD, Lun M, Maynard T, Gonzalez D, Kim S, Ye P, et al. The cerebrospinal fluid provides a proliferative niche for neural progenitor cells. Neuron. 2011;69:893–905.

    Article  CAS  Google Scholar 

  12. Mirzadeh Z, Merkle FT, Soriano-Navarro M, Garcia-Verdugo JM, Alvarez-Buylla A. Neural stem cells confer unique pinwheel architecture to the ventricular surface in neurogenic regions of the adult brain. Cell Stem Cell. 2008;3:265–78.

    Article  CAS  Google Scholar 

  13. Sawamoto K, Wichterle H, Gonzalez-Perez O, Cholfin JA, Yamada M, Spassky N, et al. New neurons follow the flow of cerebrospinal fluid in the adult brain. Science. 2006;311:629–32.

    Article  CAS  Google Scholar 

  14. Doetsch F, Garcia-Verdugo JM, Alvarez-Buylla A. Cellular composition and three-dimensional organization of the subventricular germinal zone in the adult mammalian brain. J Neurosci. 1997;17(13):5046–61.

    Article  CAS  Google Scholar 

Download references


This research was supported by Kyungpook National University Research Fund, 2017.

Author information

Authors and Affiliations



HH Seo performed immunohistochemistry experiments and analyzed the data. K Lee designed the experiments and wrote the article. Both authors read and approved the final manuscript.

Corresponding author

Correspondence to Kyungmin Lee.

Ethics declarations

Ethics approval and consent to participate

The experimental procedures and animal housing conditions were approved by the Institutional Animal Care and Use Committee at Kyungpook National University (Daegu, Republic of Korea).

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Additional file 1.

Materials and Methods.

Rights and permissions

Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( applies to the data made available in this article, unless otherwise stated.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Seo, H., Lee, K. Cell-specific expression of Epac2 in the subventricular and subgranular zones. Mol Brain 12, 113 (2019).

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: