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FoxP1 marks medium spiny neurons from precursors to maturity and is required for their differentiation.

Precious SV, Kelly CM, Reddington AE, Vinh NN, Stickland RC, Pekarik V, Scherf C, Jeyasingham R, Glasbey J, Holeiter M, Jones L, Taylor MV, Rosser AE - Exp. Neurol. (2016)

Bottom Line: There was no co-localisation of FoxP1 with any interneuron markers.In summary, we show that FoxP1 labels MSN precursors prior to the expression of DARPP-32 during normal development, and in addition suggest that FoxP1 labels a sub-population of MSNs that are not co-labelled by DARPP-32.We demonstrate the utility of FoxP1 to label MSNs in vitro and following neural transplantation, and show that FoxP1 is required for DARPP-32 positive MSN differentiation in vitro.

View Article: PubMed Central - PubMed

Affiliation: Brain Repair Group, Sir Martin Evans Building, School of Biosciences, Cardiff University, Museum Avenue, Cardiff CF10 3AX, United Kingdom.

No MeSH data available.


Related in: MedlinePlus

Characterisation of FoxP1 expression in mouse cell culture.Mouse neural tissue from different brain regions was differentiated for 7 days and immunostained for FoxP1 (A–C). FoxP1 expression (green) was seen in cells from the WGE (A) and the cortex (B), but no expression was seen in cells form the ventral mesencephalon (C). Mouse WGE differentiated for 7 days in vitro, labelled for FoxP1 (green), which co-expressed the immature neuronal marker β-III-tubulin (red) (D) and the more mature neuronal marker Map2ab (red) (E). There was no co-expression of FoxP1 with the glial marker GFAP (red) (F). FoxP1 positive cells (green) co-expressed the mature striatal MSN marker DARPP-32 (red) (G), the GABAergic neuronal marker GAD-65/67 (red) (H) and the striatal neuronal marker met-enkephalin (red) (I). Nuclei were labelled with Hoechst (blue). Scale bar = 20 μm.
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f0015: Characterisation of FoxP1 expression in mouse cell culture.Mouse neural tissue from different brain regions was differentiated for 7 days and immunostained for FoxP1 (A–C). FoxP1 expression (green) was seen in cells from the WGE (A) and the cortex (B), but no expression was seen in cells form the ventral mesencephalon (C). Mouse WGE differentiated for 7 days in vitro, labelled for FoxP1 (green), which co-expressed the immature neuronal marker β-III-tubulin (red) (D) and the more mature neuronal marker Map2ab (red) (E). There was no co-expression of FoxP1 with the glial marker GFAP (red) (F). FoxP1 positive cells (green) co-expressed the mature striatal MSN marker DARPP-32 (red) (G), the GABAergic neuronal marker GAD-65/67 (red) (H) and the striatal neuronal marker met-enkephalin (red) (I). Nuclei were labelled with Hoechst (blue). Scale bar = 20 μm.

Mentions: A critical element of neural transplantation protocols is that donor cells are dissociated and suspended in medium in order to be injected into the CNS. Having seen FoxP1 expression in the intact developing brain, we wished to analyse FoxP1 expression in cells that have been dissected, dissociated, and placed in cell culture. To investigate whether the expression of FoxP1 is maintained in differentiating cell cultures, mouse neural cells from wild type embryos at E14 were assessed after seven days differentiation in vitro (Fig. 3). FoxP1 positive cells were detected in dissociated cultured WGE (Fig. 3A) and cortex (Fig. 3B), but not in ventral mesencephalon (Fig. 3C) or dorsal mesencephalon (data not shown). The FoxP1 immunopositive WGE cells were identified as neuronal because they co-labelled at early time points in culture with the neuronal marker β-III-tubulin (which is expressed from the start of neuronal maturation), and also with the more mature neuronal marker MAP2ab, but not the astrocytic marker GFAP (Fig. 3D-F). Furthermore, these neurons were identified as the MSN population of cells in these primary fetal mouse cultures through co-localisation of FoxP1 with the striatal MSN marker DARPP-32, the GABAergic neuronal marker GAD-65/67 and the striatal projection neuron marker met-enkephalin (Fig. 3G–I). Notably, as we found in the intact adult brain, although all DARPP-32 immunopositive cells co-expressed FoxP1 there were more FoxP1 immunopositive cells than DARPP-32 immunopositive cells (FoxP1: 40.2 ± 4.0% (mean ± SEM) as a percentage of total cells, and DARPP-32: 8.8 ± 1.0% as a percentage of FoxP1). As mentioned above, the relatively low percentage of DARPP-32 expression in cultured WGE is a common finding that compromises its use as an MSN marker in vitro. This finding probably reflects the relative immaturity of the cells in vitro. Our interpretation is that FoxP1 labels MSN precursors thus emphasising its utility as a marker of these cells in culture.


FoxP1 marks medium spiny neurons from precursors to maturity and is required for their differentiation.

Precious SV, Kelly CM, Reddington AE, Vinh NN, Stickland RC, Pekarik V, Scherf C, Jeyasingham R, Glasbey J, Holeiter M, Jones L, Taylor MV, Rosser AE - Exp. Neurol. (2016)

Characterisation of FoxP1 expression in mouse cell culture.Mouse neural tissue from different brain regions was differentiated for 7 days and immunostained for FoxP1 (A–C). FoxP1 expression (green) was seen in cells from the WGE (A) and the cortex (B), but no expression was seen in cells form the ventral mesencephalon (C). Mouse WGE differentiated for 7 days in vitro, labelled for FoxP1 (green), which co-expressed the immature neuronal marker β-III-tubulin (red) (D) and the more mature neuronal marker Map2ab (red) (E). There was no co-expression of FoxP1 with the glial marker GFAP (red) (F). FoxP1 positive cells (green) co-expressed the mature striatal MSN marker DARPP-32 (red) (G), the GABAergic neuronal marker GAD-65/67 (red) (H) and the striatal neuronal marker met-enkephalin (red) (I). Nuclei were labelled with Hoechst (blue). Scale bar = 20 μm.
© Copyright Policy - CC BY
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4920670&req=5

f0015: Characterisation of FoxP1 expression in mouse cell culture.Mouse neural tissue from different brain regions was differentiated for 7 days and immunostained for FoxP1 (A–C). FoxP1 expression (green) was seen in cells from the WGE (A) and the cortex (B), but no expression was seen in cells form the ventral mesencephalon (C). Mouse WGE differentiated for 7 days in vitro, labelled for FoxP1 (green), which co-expressed the immature neuronal marker β-III-tubulin (red) (D) and the more mature neuronal marker Map2ab (red) (E). There was no co-expression of FoxP1 with the glial marker GFAP (red) (F). FoxP1 positive cells (green) co-expressed the mature striatal MSN marker DARPP-32 (red) (G), the GABAergic neuronal marker GAD-65/67 (red) (H) and the striatal neuronal marker met-enkephalin (red) (I). Nuclei were labelled with Hoechst (blue). Scale bar = 20 μm.
Mentions: A critical element of neural transplantation protocols is that donor cells are dissociated and suspended in medium in order to be injected into the CNS. Having seen FoxP1 expression in the intact developing brain, we wished to analyse FoxP1 expression in cells that have been dissected, dissociated, and placed in cell culture. To investigate whether the expression of FoxP1 is maintained in differentiating cell cultures, mouse neural cells from wild type embryos at E14 were assessed after seven days differentiation in vitro (Fig. 3). FoxP1 positive cells were detected in dissociated cultured WGE (Fig. 3A) and cortex (Fig. 3B), but not in ventral mesencephalon (Fig. 3C) or dorsal mesencephalon (data not shown). The FoxP1 immunopositive WGE cells were identified as neuronal because they co-labelled at early time points in culture with the neuronal marker β-III-tubulin (which is expressed from the start of neuronal maturation), and also with the more mature neuronal marker MAP2ab, but not the astrocytic marker GFAP (Fig. 3D-F). Furthermore, these neurons were identified as the MSN population of cells in these primary fetal mouse cultures through co-localisation of FoxP1 with the striatal MSN marker DARPP-32, the GABAergic neuronal marker GAD-65/67 and the striatal projection neuron marker met-enkephalin (Fig. 3G–I). Notably, as we found in the intact adult brain, although all DARPP-32 immunopositive cells co-expressed FoxP1 there were more FoxP1 immunopositive cells than DARPP-32 immunopositive cells (FoxP1: 40.2 ± 4.0% (mean ± SEM) as a percentage of total cells, and DARPP-32: 8.8 ± 1.0% as a percentage of FoxP1). As mentioned above, the relatively low percentage of DARPP-32 expression in cultured WGE is a common finding that compromises its use as an MSN marker in vitro. This finding probably reflects the relative immaturity of the cells in vitro. Our interpretation is that FoxP1 labels MSN precursors thus emphasising its utility as a marker of these cells in culture.

Bottom Line: There was no co-localisation of FoxP1 with any interneuron markers.In summary, we show that FoxP1 labels MSN precursors prior to the expression of DARPP-32 during normal development, and in addition suggest that FoxP1 labels a sub-population of MSNs that are not co-labelled by DARPP-32.We demonstrate the utility of FoxP1 to label MSNs in vitro and following neural transplantation, and show that FoxP1 is required for DARPP-32 positive MSN differentiation in vitro.

View Article: PubMed Central - PubMed

Affiliation: Brain Repair Group, Sir Martin Evans Building, School of Biosciences, Cardiff University, Museum Avenue, Cardiff CF10 3AX, United Kingdom.

No MeSH data available.


Related in: MedlinePlus