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ADULT AND INFANT
                     Neurogenesis


Inés Cordón & Ingrid Meucci
What is Neurogenesis ?
                             Proliferation & 
           Stem Cell
                               Migration

                                                      Progenitor Cells
New neurons generated
from neural stem cells and
progenitors cells




                               Oligodendrocyte   Astrocyte     Neuron
Where does Neurogenesis happen?




                          Lateral Ventricle

                          Hippocampus
The SVZ : SubVentricular Zone
            In Adult
The RMS : Rostral Migratory Stream
Evidence of Migration in Rodent
DOES NEUROGENESIS
   DISAPPEAR ?
Technique: Immunohistochemistry
         Stains                  Mark


          DAPI                 All Cells


    DCX : Doublecortin    Immature neuron


       PSA-NCAM           Migrating neuron


          Ki67           Cellular proliferation


     GFAP/Vimentine        Astrocytes/Glia
Neurogenesis in Human

                             Lateral ventricle


     1 Subventricular Zone

                                 Neuroblasts
 2    Rostral Migratory Stream

Olfactory Bulb
                         3 Olfactory Tract
1. Neurogenesis in the SVZ
Horizontal Section of the SVZ




            Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
Immunochemistry of the SVZ
      During the first 18 months of age

                                                      Staining
                                         Cells

                                        Immature Neuron

                                        Glia




                           Large number of DCX(+) neurons
                             populate the future gap layer

                            The glia cells processes fill the gap
                                            area
                  Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
Quantification of Immature Neuron in the SVZ




                            Staining
          Immature neuron                         Proliferating subependymal cell



Sharp decline in proliferating cells and immature neuron
                            Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
Conclusion
         During infant and adult age

Infant                                 Adult




                        Where these cells go ?
2. Neurogenesis in the RMS
Caracterization of the RMS
                             At 5 months

  Sagital and Coronal Reconstitution
                                                                                          Tributaries of
                                                                                              cells

                                                                                               Isolated
                                                                                              ependymal
                                                                                                 islets




Evidence of the RMS between the SVZ
    and the Olfactory Bulb (OB)                                                            Out of the
                                                                                          sagital plane


                              Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
Immunohistochemistry of neurons in the RMS
                            At 5 months

                 Staining
  Immature neuron
                                                                                         Tributaries of
  Migrating neuron                                                                           cells

                                                                                              Isolated
                                                                                             ependymal
                                                                                                islets




                                                                                          Out of the
                                                                                         sagital plane
Clusters of immature and migrating neurons
                             Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
Immunohistochemistry and Electron Microscopie the RMS
                                    At 1 week
                                     Label
              Immature neuron                                  A : Immature neuron

               Glia                                              B : Glia

Individual DCX(+) neuronal chains                Ultrastructural analysis of a neuronal
                                                          chain cross-section




     Evidence of a chain of immature and migrating neurons within the RMS
                               surrounded by glia
                                    Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
General Structure of the RMS
Serial Coronal Sections of the Adult RMS

                                    Staining
                              Cells
                              Glia




                         Zhengang Yang et al, Cell Research (2011) 21:1534-1550
Horizontal Section of the Adult RMS

                                             Staining
                                        Glia
                                        Immature neuron

                                        Cells




The RMS-like pathway exists in the adult human forebrain

                                  Zhengang Yang et al, Cell Research (2011) 21:1534-1550
3. Neurogenesis in the Olfactory
            Tract
Immunohistochemistry in the Olfactory Tract
                         At 1 day,8 months, 7 years

Cross-Sectional Reconstitutions                                              Staining
                                               Immature neuron                                    Cells
                                               Migrating neuron




Decline of an active RMS
        with age

                                  Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
Toluidine Blue Staining in a Longitudinal Section of the OT
                        At 6 months




Central chain of immature neurons surrounded by a matrix of
                         astrocytes

                         Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
Quantification of cells in the Proximal and Distal RMS
                      At 6 months
                                                         Proximal RMS




                                                                             Distal RMS

  There is a decline in the total amount of cells and in
immature neurons between the proximal and distal RMS
                       Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
Hypothesis for decline in cell number
                               At 6 Months
                                                                         Staining
                                                           Immature neuron
                                                           Migrating and immature neuron




    A medial migratory stream (MMS) of immature neurons branches from the
 proximal rostral migratory stream (RMS) in the infant human brain to supply the
                     ventromedial prefrontal cortex (VMPFC)
                                 Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
Summary

                           Lateral ventricle


     Subventricular Zone

                               Neuroblasts
    Rostral Migratory Stream

Olfactory Bulb

                           Olfactory Tract
Is really Neurogenesis over in the Human 
                     adult brain ?

Other pathways like MMS

  Serial sections (sagital, coronal, horizontal) : DAPI

  If yes : Immunohistochemistry (DCX, Hud+/Sox10-, NeuN)

Apoptosis ? 

  Hybridation in-situ : caspase, P53 ?

Which are the transcription factors expressed by the astrocytes ?

  Transcriptome : microarray

  Hybridation in-situ for these transcription factor
Thank For Your Attention

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Infant and Adult Neurogenesis

  • 1. ADULT AND INFANT Neurogenesis Inés Cordón & Ingrid Meucci
  • 2. What is Neurogenesis ? Proliferation & Stem Cell Migration Progenitor Cells New neurons generated from neural stem cells and progenitors cells Oligodendrocyte Astrocyte Neuron
  • 3. Where does Neurogenesis happen? Lateral Ventricle Hippocampus
  • 4. The SVZ : SubVentricular Zone In Adult
  • 5. The RMS : Rostral Migratory Stream
  • 7. DOES NEUROGENESIS DISAPPEAR ?
  • 8. Technique: Immunohistochemistry Stains Mark DAPI All Cells DCX : Doublecortin Immature neuron PSA-NCAM Migrating neuron Ki67 Cellular proliferation GFAP/Vimentine Astrocytes/Glia
  • 9. Neurogenesis in Human Lateral ventricle 1 Subventricular Zone Neuroblasts 2 Rostral Migratory Stream Olfactory Bulb 3 Olfactory Tract
  • 11. Horizontal Section of the SVZ Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
  • 12. Immunochemistry of the SVZ During the first 18 months of age Staining Cells Immature Neuron Glia Large number of DCX(+) neurons populate the future gap layer The glia cells processes fill the gap area Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
  • 13. Quantification of Immature Neuron in the SVZ Staining Immature neuron Proliferating subependymal cell Sharp decline in proliferating cells and immature neuron Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
  • 14. Conclusion During infant and adult age Infant Adult Where these cells go ?
  • 16. Caracterization of the RMS At 5 months Sagital and Coronal Reconstitution Tributaries of cells Isolated ependymal islets Evidence of the RMS between the SVZ and the Olfactory Bulb (OB) Out of the sagital plane Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
  • 17. Immunohistochemistry of neurons in the RMS At 5 months Staining Immature neuron Tributaries of Migrating neuron cells Isolated ependymal islets Out of the sagital plane Clusters of immature and migrating neurons Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
  • 18. Immunohistochemistry and Electron Microscopie the RMS At 1 week Label Immature neuron A : Immature neuron Glia B : Glia Individual DCX(+) neuronal chains Ultrastructural analysis of a neuronal chain cross-section Evidence of a chain of immature and migrating neurons within the RMS surrounded by glia Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
  • 20. Serial Coronal Sections of the Adult RMS Staining Cells Glia Zhengang Yang et al, Cell Research (2011) 21:1534-1550
  • 21. Horizontal Section of the Adult RMS Staining Glia Immature neuron Cells The RMS-like pathway exists in the adult human forebrain Zhengang Yang et al, Cell Research (2011) 21:1534-1550
  • 22. 3. Neurogenesis in the Olfactory Tract
  • 23. Immunohistochemistry in the Olfactory Tract At 1 day,8 months, 7 years Cross-Sectional Reconstitutions Staining Immature neuron Cells Migrating neuron Decline of an active RMS with age Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
  • 24. Toluidine Blue Staining in a Longitudinal Section of the OT At 6 months Central chain of immature neurons surrounded by a matrix of astrocytes Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
  • 25. Quantification of cells in the Proximal and Distal RMS At 6 months Proximal RMS Distal RMS There is a decline in the total amount of cells and in immature neurons between the proximal and distal RMS Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
  • 26. Hypothesis for decline in cell number At 6 Months Staining Immature neuron Migrating and immature neuron A medial migratory stream (MMS) of immature neurons branches from the proximal rostral migratory stream (RMS) in the infant human brain to supply the ventromedial prefrontal cortex (VMPFC) Arturo Alvarez- Buylla et al, Nature. ; 478(7369): 382–386. doi:10.1038/nature10487
  • 27. Summary Lateral ventricle Subventricular Zone Neuroblasts Rostral Migratory Stream Olfactory Bulb Olfactory Tract
  • 28. Is really Neurogenesis over in the Human adult brain ? Other pathways like MMS Serial sections (sagital, coronal, horizontal) : DAPI If yes : Immunohistochemistry (DCX, Hud+/Sox10-, NeuN) Apoptosis ? Hybridation in-situ : caspase, P53 ? Which are the transcription factors expressed by the astrocytes ? Transcriptome : microarray Hybridation in-situ for these transcription factor
  • 29. Thank For Your Attention