Anat Cell Biol.  2011 Sep;44(3):218-225. 10.5115/acb.2011.44.3.218.

The high dosage of earthworm (Eisenia andrei) extract decreases cell proliferation and neuroblast differentiation in the mouse hippocampal dentate gyrus

Affiliations
  • 1Department of Neurobiology, Kangwon National University School of Medicine, Chuncheon, Korea. mhwon@kangwon.ac.kr
  • 2Department of Oral Anatomy, College of Dentistry, Gangneung-Wonju National University, Gangneung, Korea.
  • 3Laboratory of Veterinary Pharmacology, College of Veterinary Medicine, Seoul National University, Seoul, Korea.
  • 4Department of Anatomy, College of Veterinary Medicine, Kangwon National University, Chuncheon, Korea.

Abstract

Earthworm extract has shown anticancer characteristics. In the present study, we examined the effect of chronic treatment with a high dose of earthworm (Eisenia andrei) extract (EE) on cell proliferation and neuroblast differentiation in the hippocampal dentate gyrus (DG) of 3-week-old mice using 5-bromo-2'-deoxyuridine (BrdU) and Ki-67 immunohistochemistry for cell proliferation and doublecortin (DCX) immunohistochemistry for neuroblast differentiation, respectively. BrdU-, Ki-67-, and DCX-immunoreactive cells were easily detected in the subgranular zone of the DG in vehicle (saline)-treated mice. However, BrdU-, Ki-67-, and DCX-immunoreactive cells in the 500 mg/kg EE-treated mice decreased distinctively compared to those in the vehicle-treated mice. In addition, brain-derived neurotrophic factor (BDNF) immunoreactivity and its protein level decreased markedly in the DG of the EE-treated group compared to those in the vehicle-treated group. These results indicate that chronic treatment with high dose EE decreased cell proliferation and neuroblast differentiation, and that BDNF immunoreactivity decreased in the DG of EE-treated mice.

Keyword

Neurogenesis; Subgranular zone; Eisenia andrei extract; Neurotrophic factor

MeSH Terms

Animals
Brain-Derived Neurotrophic Factor
Bromodeoxyuridine
Cell Proliferation
Dentate Gyrus
Immunohistochemistry
Mice
Neurogenesis
Oligochaeta
Brain-Derived Neurotrophic Factor
Bromodeoxyuridine

Figure

  • Fig. 1 Immunohistochemistry for 5-bromo-2'-deoxyuridine (BrdU) in the hippocampal dentate gyrus (DG) of the vehicle- (A, B) and earthworm extract (EE)-treated (C, D) groups. BrdU+ cells (arrows) in the DG of the EE-treated group were lower than those in the vehicle-treated group. GCL, granule cell layer; ML, molecular layer; PL, polymorphic layer; SGZ, subgranular layer. Scale bar=100 µm. (E) Mean number of BrdU+ cells per section in the vehicle- and EE-treated groups (n=7/group; *P<0.05, significantly different from the vehicle-treated group).

  • Fig. 2 Immunohistochemistry for Ki-67 in the hippocampal dentate gyrus of the vehicle- (A, B) and earthworm extract (EE)-treated (C, D) groups. Fewer Ki-67+ cells (arrows) were evident in the EE-treated group than those in the vehicle-treated group. GCL, granule cell layer; ML, molecular layer; PL, polymorphic layer; SGZ, subgranular layer. Scale bar=100 µm. (E) Mean number of Ki-67+ cells per section in the vehicle- and EE-treated groups (n=7/group; *P<0.05, significantly different from the vehicle-treated group).

  • Fig. 3 Immunohistochemistry for doublecortin (DCX) in the hippocampal dentate gyrus of the vehicle- (A, B) and earthworm extract (EE)-treated (C, D) groups. DCX+ neuroblasts (arrows) were abundant, and DCX+ processes were well stained in the vehicle-treated group. However, DCX immunoreactivity in the EE-treated group decreased markedly. GCL, granule cell layer; ML, molecular layer; PL, polymorphic layer; SGZ, subgranular layer. Scale bar=100 µm. (E) Mean number of DCX+ cells per section in the vehicle- and EE-treated groups (n=7/group; *P<0.05, significantly different from the vehicle-treated group).

  • Fig. 4 Immunohistochemistry for brain derived neurotrophic factor (BDNF) in the hippocampal dentate gyrus (DG) of the vehicle- (A) and earthworm extract (EE)-treated (B) groups. BDNF immunoreactivity in the EE-treated group was weaker than that in the vehicle-treated group. GCL, granule cell layer; ML, molecular layer; PL, polymorphic layer; SGZ, subgranular layer. Scale bar=100 µm. (C) Western blot analysis of BDNF in the DG of the vehicle- and EE-treated groups. Relative optical density (ROD) as a percentage of the immunoblot band is presented (n=5/group; *P<0.05, significantly different from the vehicle-treated group). Bars indicate mean±SD.


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