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Trophic and proliferative effects of Shh on motor neurons in embryonic spinal cord culture from wildtype and G93A SOD1 mice.

Ma X, Turnbull P, Peterson R, Turnbull J - BMC Neurosci (2013)

Bottom Line: Shh increased the percentage of ciliated motor neurons, especially in G93A SOD1 culture.Moreover, Shh enhanced cell survival and differentiation of motor neuron precursors in WT culture.Shh is neurotrophic to motor neurons and has mitogenic effects in WT and mSOD1 G93A culture in vitro.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Medicine, McMaster University, 1200 Main St West, Hamilton, ON L8N 3Z5, Canada. turnbull@mcmaster.ca.

ABSTRACT

Background: The developmental morphogen sonic hedgehog (Shh) may continue to play a trophic role in the support of terminally-differentiated motor neurons, of potential relevance to motor neuron disease. In addition, it may support the proliferation and differentiation of endogenous stem cells along motor neuronal lineages. As such, we have examined the trophic and proliferative effects of Shh supplementation or Shh antagonism in embryonic spinal cord cell cultures derived from wildtype or G93A SOD1 mice, a mouse model of amyotrophic lateral sclerosis.

Results: Shh supported survival, and stimulated growth of motor neurons, neurite outgrowth, and neurosphere formation in primary culture derived from both G93A SOD1 and WT mice. Shh increased the percentage of ciliated motor neurons, especially in G93A SOD1 culture. Shh-treated cultures showed increased neuronal proliferation compared to controls and especially cyclopamine treated cultures, from G93A SOD1 and WT mice. Moreover, Shh enhanced cell survival and differentiation of motor neuron precursors in WT culture.

Conclusions: Shh is neurotrophic to motor neurons and has mitogenic effects in WT and mSOD1 G93A culture in vitro.

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Related in: MedlinePlus

The effect of Shh on cell survival and differentiation. Representative microscopic images show BrdU labeled surviving cells (green), Olig2 labeled motor neuron precursor cells (red), and doubling labeled BrdU and Olig2 cells in control (A) and in 500 ng/ml shh treated cell culture (B) from WT mice. The number (D) but not percentage (C) of BrdU labeled surviving cells was significantly higher in Shh treated cultures than in controls. Both the percentage and the number of cells double-labeled with Olig2 and BrdU were significantly higher in Shh treated cultures than in controls (Shh 39.2% ± 4.6% vs control 17.0% ± 3.0%, ** p = 0.0003; Shh 5 ± 1 vs control 2 ± 0, ** p = 0.0001, E, F). Scale bar =10 μm (A, B).
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Figure 5: The effect of Shh on cell survival and differentiation. Representative microscopic images show BrdU labeled surviving cells (green), Olig2 labeled motor neuron precursor cells (red), and doubling labeled BrdU and Olig2 cells in control (A) and in 500 ng/ml shh treated cell culture (B) from WT mice. The number (D) but not percentage (C) of BrdU labeled surviving cells was significantly higher in Shh treated cultures than in controls. Both the percentage and the number of cells double-labeled with Olig2 and BrdU were significantly higher in Shh treated cultures than in controls (Shh 39.2% ± 4.6% vs control 17.0% ± 3.0%, ** p = 0.0003; Shh 5 ± 1 vs control 2 ± 0, ** p = 0.0001, E, F). Scale bar =10 μm (A, B).

Mentions: Neuronal progenitors differentiate and mature into motor neurons, during which they express a progression of markers [16]. Commonly employed markers include (in order) Islet1, Olig2, Hb9, and SMI32. We first examined the effect of Shh on cell survival in WT culture by counting BrdU labeled cells 2 weeks after BrdU was removed (Figure 5A, 5B). Shh administration led to a significant increase in the number of BrdU+ cells (Figure 5D). The effect was seen mainly for cell counts rather than percentage of total cells, Figure 5C.


Trophic and proliferative effects of Shh on motor neurons in embryonic spinal cord culture from wildtype and G93A SOD1 mice.

Ma X, Turnbull P, Peterson R, Turnbull J - BMC Neurosci (2013)

The effect of Shh on cell survival and differentiation. Representative microscopic images show BrdU labeled surviving cells (green), Olig2 labeled motor neuron precursor cells (red), and doubling labeled BrdU and Olig2 cells in control (A) and in 500 ng/ml shh treated cell culture (B) from WT mice. The number (D) but not percentage (C) of BrdU labeled surviving cells was significantly higher in Shh treated cultures than in controls. Both the percentage and the number of cells double-labeled with Olig2 and BrdU were significantly higher in Shh treated cultures than in controls (Shh 39.2% ± 4.6% vs control 17.0% ± 3.0%, ** p = 0.0003; Shh 5 ± 1 vs control 2 ± 0, ** p = 0.0001, E, F). Scale bar =10 μm (A, B).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3852546&req=5

Figure 5: The effect of Shh on cell survival and differentiation. Representative microscopic images show BrdU labeled surviving cells (green), Olig2 labeled motor neuron precursor cells (red), and doubling labeled BrdU and Olig2 cells in control (A) and in 500 ng/ml shh treated cell culture (B) from WT mice. The number (D) but not percentage (C) of BrdU labeled surviving cells was significantly higher in Shh treated cultures than in controls. Both the percentage and the number of cells double-labeled with Olig2 and BrdU were significantly higher in Shh treated cultures than in controls (Shh 39.2% ± 4.6% vs control 17.0% ± 3.0%, ** p = 0.0003; Shh 5 ± 1 vs control 2 ± 0, ** p = 0.0001, E, F). Scale bar =10 μm (A, B).
Mentions: Neuronal progenitors differentiate and mature into motor neurons, during which they express a progression of markers [16]. Commonly employed markers include (in order) Islet1, Olig2, Hb9, and SMI32. We first examined the effect of Shh on cell survival in WT culture by counting BrdU labeled cells 2 weeks after BrdU was removed (Figure 5A, 5B). Shh administration led to a significant increase in the number of BrdU+ cells (Figure 5D). The effect was seen mainly for cell counts rather than percentage of total cells, Figure 5C.

Bottom Line: Shh increased the percentage of ciliated motor neurons, especially in G93A SOD1 culture.Moreover, Shh enhanced cell survival and differentiation of motor neuron precursors in WT culture.Shh is neurotrophic to motor neurons and has mitogenic effects in WT and mSOD1 G93A culture in vitro.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Medicine, McMaster University, 1200 Main St West, Hamilton, ON L8N 3Z5, Canada. turnbull@mcmaster.ca.

ABSTRACT

Background: The developmental morphogen sonic hedgehog (Shh) may continue to play a trophic role in the support of terminally-differentiated motor neurons, of potential relevance to motor neuron disease. In addition, it may support the proliferation and differentiation of endogenous stem cells along motor neuronal lineages. As such, we have examined the trophic and proliferative effects of Shh supplementation or Shh antagonism in embryonic spinal cord cell cultures derived from wildtype or G93A SOD1 mice, a mouse model of amyotrophic lateral sclerosis.

Results: Shh supported survival, and stimulated growth of motor neurons, neurite outgrowth, and neurosphere formation in primary culture derived from both G93A SOD1 and WT mice. Shh increased the percentage of ciliated motor neurons, especially in G93A SOD1 culture. Shh-treated cultures showed increased neuronal proliferation compared to controls and especially cyclopamine treated cultures, from G93A SOD1 and WT mice. Moreover, Shh enhanced cell survival and differentiation of motor neuron precursors in WT culture.

Conclusions: Shh is neurotrophic to motor neurons and has mitogenic effects in WT and mSOD1 G93A culture in vitro.

Show MeSH
Related in: MedlinePlus