JNK Isoforms Are Involved in the Control of Adult Hippocampal Neurogenesis in Mice, Both in Physiological Conditions and in an Experimental Model of Temporal Lobe Epilepsy

  • Rubén D. Castro-Torres
  • Jon Landa
  • Marina Rabaza
  • Oriol Busquets
  • Jordi Olloquequi
  • Miren Ettcheto
  • Carlos Beas-Zarate
  • Jaume Folch
  • Antoni Camins
  • Carme AuladellEmail author
  • Ester Verdaguer


Neurogenesis in the adult dentate gyrus (DG) of the hippocampus allows the continuous generation of new neurons. This cellular process can be disturbed under specific environmental conditions, such as epileptic seizures; however, the underlying mechanisms responsible for their control remain largely unknown. Although different studies have linked the JNK (c-Jun-N-terminal-kinase) activity with the regulation of cell proliferation and differentiation, the specific function of JNK in controlling adult hippocampal neurogenesis is not well known. The purpose of this study was to analyze the role of JNK isoforms (JNK1/JNK2/JNK3) in adult-hippocampal neurogenesis. To achieve this goal, we used JNK-knockout mice (Jnk1−/−, Jnk2−/−, and Jnk3−/−), untreated and treated with intraperitoneal injections of kainic acid (KA), as an experimental model of epilepsy. In each condition, we identified cell subpopulations at different stages of neuronal maturation by immunohistochemical specific markers. In physiological conditions, we evidenced that JNK1 and JNK3 control the levels of one subtype of early progenitor cells (GFAP+/Sox2+) but not the GFAP+/Nestin+ cell subtype. Moreover, the absence of JNK1 induces an increase of immature neurons (Doublecortin+; PSA-NCAM+ cells) compared with wild-type (WT). On the other hand, Jnk1−/− and Jnk3−/− mice showed an increased capacity to maintain hippocampal homeostasis, since calbindin immunoreactivity is higher than in WT. An important fact is that, after KA injection, Jnk1−/− and Jnk3−/− mice show no increase in the different neurogenic cell subpopulation analyzed, in contrast to what occurs in WT and Jnk2−/− mice. All these data support that JNK isoforms are involved in the adult neurogenesis control.


JNK isoforms Knockout mice Adult hippocampal neurogenesis Kainic acid 



avidin-biotin-peroxidase complex








Calcium-binding protein






dentate gyrus


fetal bovine serum


granular cell layer


granule cells


glial fibrillary acidic protein


intraperitoneal injection




c-Jun N-terminal kinases

JNK1, JNK2, and JNK3

JNK isoforms


Knockout mice for JNK1


knockout mice for JNK2


knockout mice for JNK3


kainic acid




neural stem cells




phosphate buffer


phosphate-buffered saline


polysialic acid neural cell adhesion molecule


room temperature


standard deviation


sodium dodecyl sulfate polyacrylamide gel electrophoresis.


sodium dodecyl sulfate


standard error of mean


subgranular zone


temporal lobe epilepsy


wild type



This work was supported by Ministerio Español de Ciencia e Innovación, SAF2017-84283-R; Centro de Investigación Biomédica en Red de Enfermedades Neurodegenerativas (CIBERNED) CB06/05/0024; Consejo Nacional de Ciéncia y Tecnologia, CONACYT, 177594; Generalitat de Catalunya, 2014SGR-525; Generalitat de Catalunya, 2017 SGR 625; Postdoctoral Fellowship CONACYT-MEXICO, 298337; Doctoral Program in Sciences in Molecular Biology in Medicine LGAC; and Molecular Bases of Chronic-Degenerative Diseases and its Applications 000091, PNPC, CONACYT-MEXICO.

Thanks to Kyra -Mae Leighton for her technical support.

Supplementary material

12035_2019_1476_Fig7_ESM.png (40 kb)
Figure S1

Supplement Quantification of the number of DCX/CR positive cells and the representative histogram. *P < 0.05, ****P < 0.0001, vs WT CT and; &P < 0.05 vs genotype control. (PNG 40 kb)

12035_2019_1476_MOESM1_ESM.tif (55 kb)
High resolution image (TIF 54 kb)
12035_2019_1476_Fig8_ESM.png (5.2 mb)
Figure S2

Supplement A. Representative DG hippocampal images of double immune-label NeuN (Green) and PSA-NCAM cells (Red), from control mice (A, C, E, G) and 24 h KA treated mice (B, D, F, H) of WT, jnk1−/, jnk2−/, jnk3−/. Arrows show the double immuno-positive cells against NeuN and PSA-NCAM. B. Quantification of the number of double NeuN/PSA-NCAM positive cells and the representative histogram is displayed. In all conditions the levels of double labeled cells is low. Only an icrease is observed after KA in WT. **P < 0.01, vs WT CT. ZSG: stratum granular; h: hilus. A-H: Scale bar 50 μm. (PNG 5361 kb)

12035_2019_1476_MOESM2_ESM.tif (18.4 mb)
High resolution image (TIF 18795 kb)


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Rubén D. Castro-Torres
    • 1
    • 2
    • 3
  • Jon Landa
    • 1
  • Marina Rabaza
    • 1
  • Oriol Busquets
    • 2
    • 4
    • 5
    • 6
  • Jordi Olloquequi
    • 7
  • Miren Ettcheto
    • 2
    • 4
    • 5
    • 6
  • Carlos Beas-Zarate
    • 3
  • Jaume Folch
    • 4
    • 5
  • Antoni Camins
    • 2
    • 5
    • 6
  • Carme Auladell
    • 1
    • 5
    • 6
    Email author
  • Ester Verdaguer
    • 1
    • 5
    • 6
  1. 1.Departament de Biologia Cellular, Fisiologia i Immunologia, Facultat de BiologiaUniversitat de BarcelonaBarcelonaSpain
  2. 2.Departament de Farmacologia, Toxicologia i Química Terapèutica, Facultat de Farmàcia i Ciències de l’AlimentacióUniversitat de BarcelonaBarcelonaSpain
  3. 3.Departamento de Biología Celular y Molecular, Laboratorio de Regeneración Neural, C.U.C.B.AUniversidad de GuadalajaraJaliscoMexico
  4. 4.Departament de Bioquímica i Biotecnologia, Facultat de Medicina i Ciències de la SalutUniversitat Rovira i VirgiliTarragonaSpain
  5. 5.Networking Research Center on Neurodegenerative Diseases (CIBERNED)Instituto de Salud Carlos IIIMadridSpain
  6. 6.Institut de NeurociènciesUniversitat de BarcelonaBarcelonaSpain
  7. 7.Facultad de Ciencias de la SaludUniversidad Autónoma de ChileTalcaChile

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