Lab Anim Res.  2018 Dec;34(4):203-210. 10.5625/lar.2018.34.4.203.

Repeated restraint stress promotes hippocampal neuronal cell ciliogenesis and proliferation in mice

  • 1College of Pharmacy, Dongguk University, Goyang, Korea.
  • 2Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea.


Stress severely disturbs physiological and mental homeostasis which includes adult neurogenesis in hippocampus. Neurogenesis in hippocampus is a key feature to adapt to environmental changes and highly regulated by multiple cellular signaling pathways. The primary cilium is a cellular organelle, which acts as a signaling center during development and neurogenesis in adult mice. However, it is not clear how the primary cilia are involved in the process of restraint (RST) stress response. Using a mouse model, we examined the role of primary cilia in repeated and acute RST stress response. Interestingly, RST stress increased the number of ciliated cells in the adult hippocampal dentate gyrus (DG). In our RST model, cell proliferation in the DG also increased in a time-dependent manner. Moreover, the analysis of ciliated cells in the hippocampal DG with cell type markers indicated that cells that were ciliated in response to acute RST stress are neurons. Taken together, these findings suggest that RST stress response is closely associated with an increase in the number of ciliated neurons and leads to an increase in cell proliferation.


Restraint stress; primary cilia; adult neurogenesis; hippocampus

MeSH Terms

Cell Proliferation
Dentate Gyrus


  • Figure 1 Restraint stress increases cilia frequency in the adult hippocampal dentate gyrus. (A) Experimental design for the RST stress. (B) Total immobility in the FST was measured for five minutes. The total immobilization time was significantly increased in mice subjected to restraint stress compared to control mice. (CTRL: 165.83±14.55 vs RST: 205.29±10.57, P=0.0422) (P<0.01 using the student's t-test) (C) Immunofluorescent staining for neuronal cilia with type 3 adenylate cyclase (AC) antibody and DAPI for nucleus (blue) in the left panel. An increase in ciliated cells stained with ACIII antibody was found in RST mice compared to control mice (CTRL). The cilia frequency in the hippocampal DG in RST stress mice was significantly increased 3, 7, and 14 days after RST stress. Quantification of primary cilia frequency. *P<0.01 using one-way ANOVA.

  • Figure 2 Cell proliferation in the hippocampal SGZ is upregulated by acute restraint stress. (A) To compare the extent of cell proliferation, sectioned brain samples were immunostained with Ki67 (green) and mature neuronal marker, NeuN, antibodies (red). (B) Quantification of Ki67-positive cells in acute RST stress mice and the control group. Although extended subjection to restraint stress did not significantly change the number of proliferating cells, RST stress upregulated the number of Ki67-positive cells in the SGZ. *P<0.01 using one-way ANOVA.

  • Figure 3 The number of ciliated neuronal progenitor cells is increased in acute restraint stress mice. (A) A schematic of different neuronal markers specific for the different stages of neurogenesis. (B) Immunostaining for AC (green), GFAP, Tuj1, and NeuN (red). For nucleus staining, DAPI (blue) was used. We found that the number of ciliated cells labeled with the immature neuronal cell markers, GFAP and Tuj1, increased in acute RST mice one day after stress, but that of mature NeuN-labeled neurons did not change. We observed that acute RST stress increased the cilia frequency among neurons during the early stage of differentiation in the hippocampal DG. *P<0.01 using one-way ANOVA.


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