Endocrinol Metab.  2014 Sep;29(3):349-355. 10.3803/EnM.2014.29.3.349.

Polarized and Stage-Dependent Distribution of Immunoreactivity for Novel PDZ-Binding Protein Preso1 in Adult Neurogenic Regions

• Lee ES
• Kim WR
• Kim Y
• Lee HW
• Kim H
• Sun W

• KMID: 2169472
• DOI: http://doi.org/10.3803/EnM.2014.29.3.349

Abstract

BACKGROUND
Adult neural stem cells have the potential for self-renewal and differentiation into multiple cell lineages via symmetric or asymmetric cell division. Preso1 is a recently identified protein involved in the formation of dendritic spines and the promotion of axonal growth in developing neurons. Preso1 can also bind to cell polarity proteins, suggesting a potential role for Preso1 in asymmetric cell division.
METHODS
To investigate the distribution of Preso1, we performed immunohistochemistry with adult mouse brain slice. Also, polarized distribution of Preso1 was assessed by immunocytochemistry in cultured neural stem cells.
RESULTS
Immunoreactivity for Preso1 (Preso1-IR) was strong in the rostral migratory stream and subventricular zone, where proliferating transit-amplifying cells and neuroblasts are prevalent. In cultured neural stem cells, Preso1-IR was unequally distributed in the cell cytosol. We also observed the distribution of Preso1 in the subgranular zone of the hippocampal dentate gyrus, another neurogenic region in the adult brain. Interestingly, Preso1-IR was transiently observed in the nuclei of doublecortin-expressing neuroblasts immediately after asymmetric cell division.
CONCLUSION
Our study demonstrated that Preso1 is asymmetrically distributed in the cytosol and nuclei of neural stem/progenitor cells in the adult brain, and may play a significant role in cell differentiation via association with cell polarity machinery.

Keyword

Preso1; Frmpd4; Cell polarity; Subventricular zone; Dentate gyrus; Adult neural stem cells

MeSH Terms

Adult*
Animals
Asymmetric Cell Division
Axons
Brain
Cell Differentiation
Cell Lineage
Cell Polarity
Cytosol
Dendritic Spines
Dentate Gyrus
Humans
Immunohistochemistry
Mice
Neural Stem Cells
Neurons
Rivers

Figure

• 

  Fig. 1 Distribution of Preso1-IR in the adult forebrain. (A-E) In the parasagittal view, Nestin (red)-expressing neuroblasts in the rostral migratory stream (RMS) are colabeled with Preso1 (green). (F-H) In the coronal section of the RMS, doublecortin (DCX)-expressing neuroblasts (red) also exhibited immunoreactivity for Preso1 (Preso1-IR; green). (I-K) In the subependymal layer (SEL) of the olfactory bulb (OB), NeuN-negative migrating neuroblasts strongly exhibited Preso1-IR, but Preso1-IR was markedly reduced in mature NeuN-expressing neurons (red) in the granule cell layer (GCL). Nuclei were counterstained with Hoechst33342 (blue). The scale bar in B=500 µm, in E, H, K=20 µm. LV, lateral ventricle.

• 

  Fig. 2 Immunoreactivity for Preso1 (Preso1-IR) in the subventricular zone (SVZ) of the adult mouse. (A-C) The majority of glial fibrillary acidic protein (GFAP)-expressing astrocytes (red) and stem cells did not exhibit Preso1-IR (green). (D-F) A few Nestin-expressing transit amplifying stem cells (red) exhibited Preso1-IR (green). (G-I) The majority of doublecortin (DCX)-expressing neuroblasts (red) also exhibited strong Preso1-IR (green). (J) Schematic diagram of the Preso1-expressing stage (green) of adult neural stem cells (NSCs) in the SVZ. The scale bar in I=10 µm, enlarged inset in I=5 µm. TAC, transit-amplifying cell.

• 

  Fig. 3 Polarized distributions of immunoreactivity for Preso1 (Preso1-IR) in cultured neural stem cells (NSCs). (A-C) NSCs were labeled with Preso1IR (green), F-actin (red), and nuclei (blue) were counterstained with rhodamine-phalloidin and Hoechst33342, respectively. Preso1-IR was unequally distributed in the cell cytosol. (D-F) NSCs migrating out from the neurosphere cores were stained with Preso1 (green) and GM130 (red), a Golgi apparatus marker. Preso1-IR was evenly distributed regardless of the cell polarity in migrating NSCs.

• 

  Fig. 4 Distribution of immunoreactivity for Preso1 (Preso1-IR) in the subgranular zone (SGZ) of the hippocampal formation. (A-C) Preso1-IR (green) was found throughout the adult hippocampus, while Nestin (red) was expressed in the SGZ. (D-F) In the SGZ, Preso1-IR was strongly expressed in the nuclei of a subset of cells. Cells with nuclear Preso1-IR did not express Nestin (red). (G-L) Most doublecortin (DCX)-expressing neuroblasts (red) exhibited nuclear Preso1-IR (green). Nuclei were counter-stained with Hoechst33342 (blue). The scale bar in F=5 µm, I=50 µm.

• 

  Fig. 5 Polarized Preso1 expression during asymmetric cell division. (A-C) Tuj-1 (red) expressing neuronal daughter cell-expressed Trim32 (green) in cultured neural stem cells (NSCs). (D-G) In the subgranular zone (SGZ), Trim32 (red)-expressing daughter cells also expressed nuclear Preso1-IR (green). (H) Schematic diagram of the Preso1 expression stages of the adult NSCs of the SGZ. RGL, radial glia-like; IPCs, intermediate precursor cells; GFAP, glial fibrillary acidic protein; DCX, doublecortin.

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