CD40 is a major regulator of dendrite growth from developing excitatory and inhibitory neurons
- Cardiff University, United Kingdom
Figures
Figure 1 with 2 supplements
Phenotypic changes in hippocampal pyramidal neurons and striatal MSNs in P10 Cd40-/- mice.
(A) Representative low-power images of Golgi preparations of the hippocampal CA regions and dentate gyrus of Cd40+/+ and Cd40-/- mice. Scale bar, 250 μm. (B) High-power images of CA1 and CA3 …
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Figure 1—source data 1
Data of individual Golgi-stained pyramidal neurons in Cd40+/+ and Cd40-/- mice.
- https://doi.org/10.7554/eLife.30442.005
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Figure 1—source data 2
Data of individual Golgi-stained medium spiny neurons in Cd40+/+ and Cd40-/- mice.
- https://doi.org/10.7554/eLife.30442.006
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Figure 1—source data 3
2-way ANOVA for pyramidal neurons and medium spiny neurons in Cd40+/+ and Cd40-/- mice.
- https://doi.org/10.7554/eLife.30442.007
Figure 1—figure supplement 1
Opposite effects of the Cd40 null mutation on dendrite branching in hippocampal pyramidal neurons and striatal MSNs.
(A) Quantification of the number of branch points in the apical dendrite compartment (Cd40+/+, n = 86 and Cd40-/-, n = 71) and basal dendrite compartment (Cd40+/+, n = 64 and Cd40-/-, n = 53) and …
Figure 1—figure supplement 2
Phenotypic changes are maintained in P30 and adult Cd40-/- mice.
(A and B) Representative Golgi preparation images of the hippocampus of Cd40+/+ and Cd40-/- P30 and adult mice. (A) Low-power images of the hippocampal CA regions and dentate gyrus. Scale bar, 250 …
Figure 2 with 1 supplement
CD40-activated CD40L reverse signalling enhances dendrite growth from cultured hippocampal pyramidal neurons.
(A) Representative photomicrographs of hippocampal neurons of Cd40+/+ and Cd40-/- E18 embryos cultured for 9 days and treated 24 hr after plating with either soluble CD40L (sCD40L), CD40-Fc or Fc …
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Figure 2—source data 1
Means, s.e.m. and n numbers for bar charts.
- https://doi.org/10.7554/eLife.30442.010
Figure 2—figure supplement 1
Number of primary dendrites per neuron in the neurite arbors of cultured hippocampal pyramidal neurons.
The neurons of E18 Cd40+/+ or Cd40-/- littermates were cultured for 9 days. 24 hr after plating, the cultures were treated with either soluble CD40L (sCD40L), CD40-Fc or Fc protein (each at 1 μg/ml) …
Figure 3
CD40-activated CD40L reverse signalling enhances axon growth from cultured hippocampal pyramidal neurons.
(A) Representative photomicrographs of neurons of Cd40+/+ and Cd40-/- E18 embryos cultured for 3 days and treated 24 hr after plating with either soluble CD40L (sCD40L), CD40-Fc or Fc protein (each …
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Figure 3—source data 1
Means, s.e.m. and n numbers for bar charts.
- https://doi.org/10.7554/eLife.30442.012
Figure 4
CD40-activated CD40L reverse signalling reduces neurite growth from cultured MSNs.
(A) Representative photomicrographs of MSNs of Cd40+/+ and Cd40-/- E14 embryos cultured for 10 days and treated with either soluble CD40L (sCD40L) at 0.5 μg/ml, CD40-Fc or Fc protein (each at 1 …
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Figure 4—source data 1
Means, s.e.m. and n numbers for bar charts.
- https://doi.org/10.7554/eLife.30442.014
Figure 5
PKC mediates the effect of CD40-activated CD40L reverse signalling on dendrite growth.
(A) Representative photomicrographs of hippocampal neurons of Cd40-/- E18 embryos cultured for 9 days and treated 24 hr after plating with either 1 μg/ml CD40-Fc, 1 μg/ml Fc protein, 500 nM Go6983, …
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Figure 5—source data 1
Means, s.e.m. and n numbers for pyramidal neuron bar charts.
- https://doi.org/10.7554/eLife.30442.016
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Figure 5—source data 2
Means, s.e.m. and n numbers for medium spiny neuron bar charts.
- https://doi.org/10.7554/eLife.30442.017
Figure 6 with 1 supplement
PKC-β and PKC-γ selectively participate in mediating the effects of CD40-activated CD40L reverse signalling on dendritic growth.
(A) Representative photomicrographs of hippocampal neurons of Cd40+/+ and Cd40-/- E18 embryos transfected with either control siRNA or PKC-β siRNA and cultured for 9 days. The Cd40-/- neurons were …
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Figure 6—source data 1
Means, s.e.m. and n numbers for Cd40+/+ pyramidal neuron bar charts.
- https://doi.org/10.7554/eLife.30442.020
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Figure 6—source data 2
Means, s.e.m. and n numbers for Cd40+/+ medium spiny neuron bar charts.
- https://doi.org/10.7554/eLife.30442.021
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Figure 6—source data 3
Means, s.e.m. and n numbers for Cd40-/- pyramidal neuron bar charts.
- https://doi.org/10.7554/eLife.30442.022
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Figure 6—source data 4
Means, s.e.m. and n numbers for Cd40-/- medium spiny neuron bar charts.
- https://doi.org/10.7554/eLife.30442.023
Figure 6—figure supplement 1
Expression of PKC isoforms in the hippocampus and striatum and siRNA knockdown of PKC-β and PKC-γ hippocampal and striatal cultures.
(A) Representative western blots of hippocampal and striatal lysates of hippocampal and striatal tissues probed with antibodies to PKC-α, PKC-β,PKC-γ, PKC-εand GAPDH loading controls. (B) …
Figure 7 with 1 supplement
Localization of CD40 and CD40L in the developing hippocampus.
Representative images of sections of the CA1 region of P9 hippocampi labelled with either anti-CD40 (A) or anti-CD40L (B) together with DAPI and either anti-MAP-2 or anti-GFAP (pl, pyramidal layer; …
Figure 7—figure supplement 1
Expression of CD40 and CD40L in the developing hippocampus and striatum.
Representative western blots of hippocampal lysates of Cd40+/+ and Cd40-/- mice (A) and striatal lysates of Cd40+/+ mice (B) from embryonic, postnatal and adult (ad) mice probed with antibodies to …
Figure 8
Localization of CD40 and CD40L in the developing striatum.
(A) Low-power images of the expression of striatal MSNs labelled with DARPP-32 and double labelled with DAPI. Scale bar, 250 μm. Representative higher power images of sections of the striatum …
Figure 9
Co-localization of CD40 and CD40L in the cultured hippocampal pyramidal neurons and striatal MSNs.
(A) Representative images of dissociated E18 hippocampal cultures after 9 days in vitro triple labelling with anti-CD40, anti-CD40L and DAPI. The upper panels show images of neuronal cell bodies and …
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.30442.028
