Skip to main content

Advertisement

Log in

Effects of Ginko biloba leaf extract on the neurogenesis of the hippocampal dentate gyrus in the elderly mice

  • Original Article
  • Published:
Anatomical Science International Aims and scope Submit manuscript

Abstract

Aging is associated with reduced hippocampal neurogenesis, which may in turn contribute to cognitive impairment. We assessed the effect of Ginkgo biloba (Gb) on hippocampal neurogenesis in elderly male mice using immunohistochemistry. We used anti-caspase-3 as a marker of apoptosis, anti-GFAP as a marker of neural stem cells, anti-Ki-67 as a specific marker for cellular proliferation and anti-doublecortin (DCX) to detect newly born neurons in the hippocampal dentate gyrus (DG) of aged male mice. The 24-month-old male mice were divided into two groups: a control group treated with distilled water and a group fed with Gb at a dose of 100 mg/kg once daily for 28 days. A sharp decrease in apoptotic cells in Gb-treated compared to nontreated mice was observed by anti-csapase-3 immunostaining. A large number of GFAP+ve cells was found in the subgranular zone of the DG of Gb-treated mice, suggesting an increase in the pool of neural stem cells by Gb treatment. There was also an increase in Ki-67 immunoreactive cells, indicating increased cell proliferation in the DG in the Gb-treated compared to nontreated group. A significant increase in newborn DCX+ve neurons with well-developed tertiary dendrites was also found in the Gb-treated compared to nontreated group. Using Western blot analysis, the expression of DCX protein in the Gb group was also significantly increased compared to the control. The results support a beneficial role of Gb on hippocampal neurogenesis in the context of brain aging.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  • Alonso G (2001) Proliferation of progenitor cells in the adult rat brain correlates with the presence of vimentin-expressing astrocytes. Glia. 34:253–266

    Article  CAS  PubMed  Google Scholar 

  • Bolognin S, Buffelli M, Puolivali J, Iqbal K (2014) Rescue of cognitive-aging by administration of a neurogenic and/or neurotrophic compound Neurobiol. Aging 35:2134–2146

    CAS  Google Scholar 

  • Brinton RD, Wang JM (2006) Therapeutic potential of neurogenesis for prevention and recovery from Alzheimer’s disease: allopregnanolone as a proof of concept neurogenic agent. Curr Alzheimer Res 3:185–190

    Article  CAS  PubMed  Google Scholar 

  • Brown J, Cooper-Kuhn CM, Kempermann G, Van Praag H, Winkler J, Gage FH, Kuhn HG (2003) Enriched environment and physical activity stimulate hippocampal but not olfactory bulb neurogenesis. Eur J Neurosci 17:2042–2046

    Article  PubMed  Google Scholar 

  • Cameron HA, McKay RD (1999) Restoring production of hippocampal neurons in old age. Nat Neurosci 2:894–897

    Article  CAS  PubMed  Google Scholar 

  • Couillard-Despres S, Winner B, Schaubeck S, Aigner R, Vroemen M, Weidner N, Bogdahn U, Winkler J, Kuhn HG, Aigner L (2005) Doublecortin expression levels in adult brain reflect neurogenesis. Eur J Neurosci 21:1–14

    Article  PubMed  Google Scholar 

  • Dayer AG, Ford AA, Cleaver KM, Yassaee M, Cameron HA (2003) Short-term and long-term survival of new neurons in the rat dentate gyrus. J Comp Neurol 460:563–572

    Article  PubMed  Google Scholar 

  • Diamond BJ, Shiflett SC, Feiwel N, Matheis RJ, Noskin O, Richards JA, Schoenberger NE (2000) Ginkgo biloba extract: mechanisms and clinical indications. Arch Phys Med Rehabil 81:668–678

    CAS  PubMed  Google Scholar 

  • Drew MR, Hen R (2007) Adult hippocampal neurogenesis as target for the treatment of depression. CNS Neurol Disord Drug Targets 6:205–218

    Article  CAS  PubMed  Google Scholar 

  • Dupret D, Fabre A, Dobrossy MD, Panatier A, Rodriguez JJ, Lamarque S, Lemaire V, Oliet SH, Piazza PV, Abrous DN (2007) Spatial learning depends on both the addition and removal of new hippocampal neurons. PLoS Biol 5:e214

    Article  PubMed  PubMed Central  Google Scholar 

  • Encinas JM, Sierra A (2012) Neural stem cell deforestation as the main force driving the age-related decline in adult hippocampal neurogenesis. Behav Brain Res 227:433–439

    Article  PubMed  Google Scholar 

  • Encinas JM, Vaahtokari A, Enikolopov G 2006: Fluoxetine targets early progenitor cells in the adult brain. Proc Natl Acad Sci USA 103:8233–8238

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Francis F, Koulakoff A, Boucher D, Chafey P, Schaar B, Vinet MC, Friocourt G, McDonnell N, Reiner O, Kahn A, McConnell SK, Berwald-Netter Y, Denoulet P, Chelly J (1999) Doublecortin is a developmentally regulated, microtubule-associated protein expressed in migrating and differentiating neurons. Neuron 23:247–256

    Article  CAS  PubMed  Google Scholar 

  • Franklin KBJ, Paxinos G (1997) The mouse brain in stereotaxic coordinates. Academic Press, San Diego

    Google Scholar 

  • Gebara E, Sultan S, Kocher-Braissant J, Toni N (2013) Adult hippocampal neurogenesis inversely correlates with microglia in conditions of voluntary running and aging. Front Neurosci 7:145–151

    Article  PubMed  PubMed Central  Google Scholar 

  • Gil-Mohapel J, Brocardo PS, Choquette W, Gothard R, Simpson JM, Christie BR (2013) Hippocampal neurogenesis levels predict WATERMAZE search strategies in the aging brain. PLoS One 8:e75125

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gleeson JG, Lin PT, Flanagan LA, Walsh CA (1999) Doublecortin is a microtubule-associated protein and is expressed widely by migrating neurons. Neuron 23:257–271

    Article  CAS  PubMed  Google Scholar 

  • Hüttmann K, Sadgrove M, Wallraff A, Hinterkeuser S, Kirchhoff F, Steinhäuser C, Gray WP (2003) Seizures preferentially stimulate proliferation of radial glia-like astrocytes in the adult dentate gyrus: functional and immunocytochemical analysis. Eur J Neurosci 18:2769–2778

    Article  PubMed  Google Scholar 

  • Hwang IK, Yoo KY, Kim DW, Lee CH, Choi JH, Kwon YG, Kim YM, Choi SY, Won MH (2010) Changes in the expression of mitochondrial peroxiredoxin and thioredoxin in neurons and glia and their protective effects in experimental cerebral ischemic damage. Free Radic Biol Med 48:1242–1251

    Article  CAS  PubMed  Google Scholar 

  • Jin K, Sun Y, Xie L, Batteur S, Mao XO, Smelick C, Logvinova A, Greenberg DA (2003) Neurogenesis and aging: FGF-2 and HB-EGF restore neurogenesis in hippocampus and subventricular zone of aged mice. Aging Cell 2:175–183

    Article  CAS  PubMed  Google Scholar 

  • Jones SP, Rahimi O, O’Boyle MP, Diaz DL, Claiborne BJ (2003) Maturation of granule cell dendrites after mossy fiber arrival in hippocampal field CA3. Hippocampus 13:413–427

    Article  PubMed  Google Scholar 

  • Kee N, Sivalingam S, Boonstra R, Wojtowicz JM (2002) The utility of Ki-67 and BrdU as proliferative markers of adult neurogenesis. J Neurosci Methods 115:97–105

    Article  CAS  PubMed  Google Scholar 

  • Kee N, Teixeira CM, Wang AH, Frankland PW (2007) Preferential incorporation of adult-generated granule cells into spatial memory networks in the dentate gyrus. Nat Neurosci 10:355–362

    Article  CAS  PubMed  Google Scholar 

  • Kempermann G, Kuhn HG, Gage FH (1998) Experience-induced neurogenesis in the senescent dentate gyrus. J Neurosci 18:3206–3212

    CAS  PubMed  Google Scholar 

  • Kheirbek MA (2015) Finding the roots of adult neurogenesis. Cell 161:1500–1502

    Article  CAS  PubMed  Google Scholar 

  • Kotakadi VS, Jin Y, Hofseth AB, Ying L, Cui X, Volate S, Chumanevich A, Wood PA, Price RL, McNeal A, Singh UP, Singh NP, Nagarkatti M, Nagarkatti PS, Matesic LE, Auclair K, Wargovich MJ, Hofseth LJ (2008) Ginkgo biloba extract EGb 761 has anti-inflammatory properties and ameliorates colitis in mice by driving effector T cell apoptosis. Carcinogenesis 29:1799–1806

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kronenberg G, Reuter K, Steiner B, Brandt MD, Jessberger S, Yamaguchi M, Kempermann G (2003) Subpopulations of proliferating cells of the adult hippocampus respond differently to physiologic neurogenic stimuli. J Comp Neurol 467:455–463

    Article  PubMed  Google Scholar 

  • Kuhn HG, Dickinson-Anson H, Gage FH (1996) Neurogenesis in the dentate gyrus of the adult rat: age-related decrease of neuronal progenitor proliferation. J Neurosci 16:2027–2033

    CAS  PubMed  Google Scholar 

  • Lemaire V, Lamarque S, Le Moal M, Piazza PV, Abrous DN (2006) Postnatal stimulation of the pups counteracts prenatal stress-induced deficits in hippocampal neurogenesis. Biol Psychiatry 59:786–792

    Article  PubMed  Google Scholar 

  • Liu Y, Namba T, Liu J, Suzuki R, Shioda S, Seki T (2010) Glial fibrillary acidic protein-expressing neural progenitors give rise to immature neurons via early intermediate progenitors expressing both glial fibrillary acidic protein and neuronal markers in the adult hippocampus. Neuroscience 166:241–251

    Article  CAS  PubMed  Google Scholar 

  • Madgula VL, Avula B, Yu YB, Wang YH, Tchantchou F, Fisher S, Luo Y, Khan IA, Khan SI (2010) Intestinal and blood-brain barrier permeability of Ginkgolides and Bilobalide: in vitro and in vivo approaches. Planta Med 76:599–606

    Article  CAS  PubMed  Google Scholar 

  • Molofsky AV, Slutsky SG, Joseph NM, He S, Pardal R, Krishnamurthy J, Sharpless NE, Morrison SJ (2006) Increasing p16INK4a expression decreases forebrain progenitors and neurogenesis during ageing. Nature 443:448–452

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nacher J, Alonso-Llosa G, Rosell DR, McEwen BS (2003) NMDA receptor antagonist treatment increases the production of new neurons in the aged rat hippocampus. Neurobiol Aging 24:273–284

    Article  CAS  PubMed  Google Scholar 

  • Namba T, Maekawa M, Yuasa S, Kohsaka S, Uchino S (2009) The Alzheimer’s disease drug memantine increases the number of radial glia-like progenitor cells in adult hippocampus. Glia 57:1082–1090

    Article  PubMed  Google Scholar 

  • O’Hara M, Kiefer D, Farrell K, Kemper K (1998) A review of 12 commonly used medicinal herbs. Arch Fam Med 7:523–536

    Article  PubMed  Google Scholar 

  • Rai KS, Hattiangady B, Shetty AK (2007) Enhanced production and dendritic growth of new dentate granule cells in the middle-aged hippocampus following intracerebroventricular FGF-2 infusions. Eur J Neurosci 26:1765–1779

    Article  PubMed  Google Scholar 

  • Rao MS, Shetty AK (2004) Efficacy of doublecortin as a marker to analyse the absolute number and dendritic growth of newly generated neurons in the adult dentate gyrus. Eur J Neurosci 19:234–246

    Article  PubMed  Google Scholar 

  • Rao MS, Hattiangady B, Abdel-Rahman A, Stanley DP, Shetty AK (2005) Newly born cells in the ageing dentate gyrus display normal migration, survival and neuronal fate choice but endure retarded early maturation. Eur J Neurosci 21:464–476

    Article  PubMed  Google Scholar 

  • Rigney U, Kimber S, Hindmarch I (1999) The effects of acute doses of standardized Ginkgo biloba extract on memory and psychomotor performance in volunteers. Phytother Res 13:408–415

    Article  CAS  PubMed  Google Scholar 

  • Scholzen T, Gerdes J (2000) The Ki-67 protein: from the known and the unknown. J Cell Physiol 182:311–322

    Article  CAS  PubMed  Google Scholar 

  • Seki T, Arai Y (1995) Age-related production of new granule cells in the adult dentate gyrus. NeuroReport 6:2479–2482

    Article  CAS  PubMed  Google Scholar 

  • Seri B, Garcia-Verdugo JM, Collado-Morente L, McEwen BS, Alvarez-Buylla A (2004) Cell types, lineage, and architecture of the germinal zone in the adult dentate gyrus. J Comp Neurol 478:359–378

    Article  PubMed  Google Scholar 

  • Shi C, Yao Z, Xu J, Yew DT (2009) Effects of Gingko extract (EGb761) on oxidative damage under different conditions of serum supply. J Bioenerg Biomembr 41:61–69

    Article  CAS  PubMed  Google Scholar 

  • Shif O, Gillette K, Damkaoutis CM, Carrano C, Robbins SJ, Hoffman JR (2006) Effects of Ginkgo biloba administered after spatial learning on water maze and radial arm maze performance in young adult rats. Pharmacol Biochem Behav 84:17–25

    Article  CAS  PubMed  Google Scholar 

  • Stackman RW, Eckenstein F, Frei B, Kulhanek D, Nowlin J, Quinn JF (2003) Prevention of age-related spatial memory deficits in a transgenic mouse model of Alzheimer’s disease by chronic Ginkgo biloba treatment. Exp Neurol 184:510–520

    Article  PubMed  Google Scholar 

  • Suh H, Consiglio A, Ray J, Sawai T, D’Amour KA, Gage FH (2007) In vivo fate analysis reveals the multipotent and self-renewal capacities of Sox2+ neural stem cells in the adult hippocampus. Cell Stem Cell 1:515–528

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tada Y, Kagota S, Kubota Y, Nejime N, Nakamura K, Kunitomo M, Shinozuka K (2008) Long-term feeding of Ginkgo biloba extract impairs peripheral circulation and hepatic function in aged spontaneously hypertensive rats. Biol Pharm Bull 31:68–72

    Article  CAS  PubMed  Google Scholar 

  • Tang F, Nag S, Shiu SY, Pang SF (2002) The effects of melatonin and Ginkgo biloba extract on memory loss and choline acetyltransferase activities in the brain of rats infused intracerebroventricularly with β-amyloid 1–40. Life Sci 71:2625–2631

    Article  CAS  PubMed  Google Scholar 

  • Taupin P (2007) BrdU immunohistochemistry for studying adult neurogenesis: paradigms, pitfalls, limitations, and validation. Brain Res Rev 53:198–214

    Article  CAS  PubMed  Google Scholar 

  • Tchantchou F, Lacor PN, Cao Z, Lao L, Hou Y, Cui C, Klein WL, Luo Y (2009) Stimulation of neurogenesis and synaptogenesis by bilobalide and quercetin via common final pathway in hippocampal neurons. J Alzheimers Dis 18:787–798

    CAS  PubMed  Google Scholar 

  • Thuret S, Toni N, Aigner S, Yeo GW, Gage FH (2009) Hippocampus-dependent learning is associated with adult neurogenesis in MRL/MpJ mice. Hippocampus 19:658–669

    Article  CAS  PubMed  Google Scholar 

  • Toni N, Teng EM, Bushong EA, Aimone JB, Zhao C, Consiglio A, van Praag H, Martone ME, Ellisman MH, Gage FH (2007) Synapse formation on neurons born in the adult hippocampus. Nat Neurosci 10:727–734

    Article  CAS  PubMed  Google Scholar 

  • Tropepe V, Craig CG, Morshead CM, van Der Kooy D (1997) Transforming growth factor-alpha null and senescent mice show decreased neural progenitor cell proliferation in the forebrain subependyma. J Neurosci 17:7850–7859

    CAS  PubMed  Google Scholar 

  • Van Praag H, Shubert T, Zhao C, Gage FH (2005) Exercise enhances learning and hippocampal neurogenesis in aged mice. J Neurosci 25:8680–8685

    Article  PubMed  PubMed Central  Google Scholar 

  • Von Bohlen Und Halbach O (2007) Immunohistological markers for staging neurogenesis in adult hippocampus. Cell Tissue Res 329:409–420

    Article  CAS  PubMed  Google Scholar 

  • Wang Y, Wang L, Wu J, Cai J (2006) The in vivo synaptic plasticity mechanism of EGb 761-induced enhancement of spatial learning and memory in aged rats. Br J Pharmacol 148:147–153

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wesnes KA, Ward T, McGinty A, Petrini O (2000) The memory enhancing effects of a Ginkgo biloba/Panax ginseng combination in healthy middle-aged volunteers. Psychopharmacology 152:353–361

    Article  CAS  PubMed  Google Scholar 

  • Williams B, Watanabe CM, Schultz PG, Rimbach G, Krucker T (2004) Age-related effects of Ginkgo biloba extract on synaptic plasticity and excitability. Neurobiol Aging 25:955–962

    Article  PubMed  Google Scholar 

  • Winter JC (1998) The effects of an extract of Ginkgo biloba, EGb 761, on cognitive behavior and longevity in the rat. Physiol Behav 63:425–433

    Article  CAS  PubMed  Google Scholar 

  • Wojtowicz JM, Kee N (2006) BrdU assay for neurogenesis in rodents. Nat Protoc 1:1399–1405

    Article  CAS  PubMed  Google Scholar 

  • Wu YZ, Li SQ, Zu XG, Du J, Wang FF (2008) Ginkgo biloba extract improves coronary artery circulation in patients with coronary artery disease: contribution of plasma nitric oxide and endothelin-1. Phytother Res 22:734–739

    Article  PubMed  Google Scholar 

  • Yoo DY, Nam YY, Kim W, Yoo KY, Park J, Lee CH, Choi JH, Yoon YS, Kim DW, Won MH, Hwang IK (2011) Effects of Ginkgo biloba extract on promotion of neurogenesis in the hippocampal dentate gyrus in C57BL/6 mice. J Vet Med Sci 73:71–76

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Noura M. S. Osman.

Ethics declarations

Conflict of interest

None.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Osman, N.M.S., Amer, A.S. & Abdelwahab, S. Effects of Ginko biloba leaf extract on the neurogenesis of the hippocampal dentate gyrus in the elderly mice. Anat Sci Int 91, 280–289 (2016). https://doi.org/10.1007/s12565-015-0297-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12565-015-0297-7

Keywords

Navigation