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TRPC6 specifically interacts with APP to inhibit its cleavage by γ-secretase and reduce Aβ production.

Wang J, Lu R, Yang J, Li H, He Z, Jing N, Wang X, Wang Y - Nat Commun (2015)

Bottom Line: A fusion peptide derived from TRPC6 also reduces Aβ levels without effect on Notch cleavage.Crossing APP/PS1 mice with TRPC6 transgenic mice leads to a marked reduction in both plaque load and Aβ levels, and improvement in structural and behavioural impairment.Thus, TRPC6 specifically modulates γ-secretase cleavage of APP and preventing APP (C99) interaction with PS1 via TRPC6 could be a novel strategy to reduce Aβ formation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Neural Signal Transduction, Institute of Neuroscience, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, SIBS, CAS, 320 Yue-Yang Road, Shanghai 200031, China.

ABSTRACT
Generation of β-amyloid (Aβ) peptide in Alzheimer's disease involves cleavage of amyloid precursor protein (APP) by γ-secretase, a protease known to cleave several substrates, including Notch. Finding specific modulators for γ-secretase could be a potential avenue to treat the disease. Here, we report that transient receptor potential canonical (TRPC) 6 specifically interacts with APP leading to inhibition of its cleavage by γ-secretase and reduction in Aβ production. TRPC6 interacts with APP (C99), but not with Notch, and prevents C99 interaction with presenilin 1 (PS1). A fusion peptide derived from TRPC6 also reduces Aβ levels without effect on Notch cleavage. Crossing APP/PS1 mice with TRPC6 transgenic mice leads to a marked reduction in both plaque load and Aβ levels, and improvement in structural and behavioural impairment. Thus, TRPC6 specifically modulates γ-secretase cleavage of APP and preventing APP (C99) interaction with PS1 via TRPC6 could be a novel strategy to reduce Aβ formation.

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The Aβ levels were decreased in APP/PS1/TRPC6 mice.(a) Representative images of amyloid plaque immunostained with Aβ antibody (6E10) on brain sections from 6 months female APP/PS1 and APP/PS1/TRPC6 mice. Scale bar, 500 μm. (b) Quantification of the plaque load in indicated mouse brain sections shown in a (n=8). ELISA of total (c) TBST soluble (d) or insoluble (e) Aβ levels in the forebrain lysates of 6 month female indicated mice (n=8). Representative images (f) and quantification (g) of thioflavin S staining of amyloid plaque in the hippocampal region of 6 and 11 months APP/PS1 and APP/PS1/TRPC6 mice with B6C3 background (n=4–5 mice). Scale bar, 200 μm. Data were presented as means±s.e.m. Two-tailed Student's t-test was performed. *P<0.05, **P<0.01 versus APP/PS1.
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f5: The Aβ levels were decreased in APP/PS1/TRPC6 mice.(a) Representative images of amyloid plaque immunostained with Aβ antibody (6E10) on brain sections from 6 months female APP/PS1 and APP/PS1/TRPC6 mice. Scale bar, 500 μm. (b) Quantification of the plaque load in indicated mouse brain sections shown in a (n=8). ELISA of total (c) TBST soluble (d) or insoluble (e) Aβ levels in the forebrain lysates of 6 month female indicated mice (n=8). Representative images (f) and quantification (g) of thioflavin S staining of amyloid plaque in the hippocampal region of 6 and 11 months APP/PS1 and APP/PS1/TRPC6 mice with B6C3 background (n=4–5 mice). Scale bar, 200 μm. Data were presented as means±s.e.m. Two-tailed Student's t-test was performed. *P<0.05, **P<0.01 versus APP/PS1.

Mentions: To test whether TRPC6 could regulate Aβ levels in animals, we crossed TRPC6 transgenic mice, in which TRPC6 was overexpressed in the forebrain excitatory neurons19, with APP/PS1 mice, a mouse model commonly used to study Aβ formation32. Amyloid plaques were examined by Aβ immunostaining with 6E10 antibody on the brain sections of APP/PS1 and APP/PS1/TRPC6 mice (Fig. 5a; Supplementary Fig. 8a). The plaque load was markedly reduced in APP/PS1/TRPC6 male (Supplementary Fig. 8b) and female (Fig. 5b) mice compared with that in APP/PS1 mice. Plaque staining with Aβ specific antibody MOAB-2 (ref. 33) confirmed that the plaque load was greatly reduced in APP/PS1/TRPC6 mice compared with that in APP/PS1 mice (Supplementary Fig. 8f–i). Total Aβ levels in guanidine-HCl (GuHCl) extraction of APP/PS1/TRPC6 male (Supplementary Fig. 8c) and female (Fig. 5c) mouse forebrains were greatly reduced than those of APP/PS1 mouse forebrains. The TBST soluble and insoluble Aβ levels in APP/PS1/TRPC6 male (Supplementary Fig. 8d,e) and female (Fig. 5d,e) mice were also substantially reduced compared with those in APP/PS1 mice. To further confirm the effect of TRPC6, we examined whether TRPC6 could reduce amyloid plaque in APP/PS1 mice with another genetic background. The plaque load in the hippocampus of APP/PS1/TRPC6 mice in B6C3 background stained with thioflavin S (Fig. 5f) was also greatly reduced than that in APP/PS1 mice (Fig. 5g). Taken together, TRPC6 reduces Aβ accumulation in APP/PS1 mice.


TRPC6 specifically interacts with APP to inhibit its cleavage by γ-secretase and reduce Aβ production.

Wang J, Lu R, Yang J, Li H, He Z, Jing N, Wang X, Wang Y - Nat Commun (2015)

The Aβ levels were decreased in APP/PS1/TRPC6 mice.(a) Representative images of amyloid plaque immunostained with Aβ antibody (6E10) on brain sections from 6 months female APP/PS1 and APP/PS1/TRPC6 mice. Scale bar, 500 μm. (b) Quantification of the plaque load in indicated mouse brain sections shown in a (n=8). ELISA of total (c) TBST soluble (d) or insoluble (e) Aβ levels in the forebrain lysates of 6 month female indicated mice (n=8). Representative images (f) and quantification (g) of thioflavin S staining of amyloid plaque in the hippocampal region of 6 and 11 months APP/PS1 and APP/PS1/TRPC6 mice with B6C3 background (n=4–5 mice). Scale bar, 200 μm. Data were presented as means±s.e.m. Two-tailed Student's t-test was performed. *P<0.05, **P<0.01 versus APP/PS1.
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f5: The Aβ levels were decreased in APP/PS1/TRPC6 mice.(a) Representative images of amyloid plaque immunostained with Aβ antibody (6E10) on brain sections from 6 months female APP/PS1 and APP/PS1/TRPC6 mice. Scale bar, 500 μm. (b) Quantification of the plaque load in indicated mouse brain sections shown in a (n=8). ELISA of total (c) TBST soluble (d) or insoluble (e) Aβ levels in the forebrain lysates of 6 month female indicated mice (n=8). Representative images (f) and quantification (g) of thioflavin S staining of amyloid plaque in the hippocampal region of 6 and 11 months APP/PS1 and APP/PS1/TRPC6 mice with B6C3 background (n=4–5 mice). Scale bar, 200 μm. Data were presented as means±s.e.m. Two-tailed Student's t-test was performed. *P<0.05, **P<0.01 versus APP/PS1.
Mentions: To test whether TRPC6 could regulate Aβ levels in animals, we crossed TRPC6 transgenic mice, in which TRPC6 was overexpressed in the forebrain excitatory neurons19, with APP/PS1 mice, a mouse model commonly used to study Aβ formation32. Amyloid plaques were examined by Aβ immunostaining with 6E10 antibody on the brain sections of APP/PS1 and APP/PS1/TRPC6 mice (Fig. 5a; Supplementary Fig. 8a). The plaque load was markedly reduced in APP/PS1/TRPC6 male (Supplementary Fig. 8b) and female (Fig. 5b) mice compared with that in APP/PS1 mice. Plaque staining with Aβ specific antibody MOAB-2 (ref. 33) confirmed that the plaque load was greatly reduced in APP/PS1/TRPC6 mice compared with that in APP/PS1 mice (Supplementary Fig. 8f–i). Total Aβ levels in guanidine-HCl (GuHCl) extraction of APP/PS1/TRPC6 male (Supplementary Fig. 8c) and female (Fig. 5c) mouse forebrains were greatly reduced than those of APP/PS1 mouse forebrains. The TBST soluble and insoluble Aβ levels in APP/PS1/TRPC6 male (Supplementary Fig. 8d,e) and female (Fig. 5d,e) mice were also substantially reduced compared with those in APP/PS1 mice. To further confirm the effect of TRPC6, we examined whether TRPC6 could reduce amyloid plaque in APP/PS1 mice with another genetic background. The plaque load in the hippocampus of APP/PS1/TRPC6 mice in B6C3 background stained with thioflavin S (Fig. 5f) was also greatly reduced than that in APP/PS1 mice (Fig. 5g). Taken together, TRPC6 reduces Aβ accumulation in APP/PS1 mice.

Bottom Line: A fusion peptide derived from TRPC6 also reduces Aβ levels without effect on Notch cleavage.Crossing APP/PS1 mice with TRPC6 transgenic mice leads to a marked reduction in both plaque load and Aβ levels, and improvement in structural and behavioural impairment.Thus, TRPC6 specifically modulates γ-secretase cleavage of APP and preventing APP (C99) interaction with PS1 via TRPC6 could be a novel strategy to reduce Aβ formation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Neural Signal Transduction, Institute of Neuroscience, State Key Laboratory of Neuroscience, CAS Center for Excellence in Brain Science and Intelligence Technology, SIBS, CAS, 320 Yue-Yang Road, Shanghai 200031, China.

ABSTRACT
Generation of β-amyloid (Aβ) peptide in Alzheimer's disease involves cleavage of amyloid precursor protein (APP) by γ-secretase, a protease known to cleave several substrates, including Notch. Finding specific modulators for γ-secretase could be a potential avenue to treat the disease. Here, we report that transient receptor potential canonical (TRPC) 6 specifically interacts with APP leading to inhibition of its cleavage by γ-secretase and reduction in Aβ production. TRPC6 interacts with APP (C99), but not with Notch, and prevents C99 interaction with presenilin 1 (PS1). A fusion peptide derived from TRPC6 also reduces Aβ levels without effect on Notch cleavage. Crossing APP/PS1 mice with TRPC6 transgenic mice leads to a marked reduction in both plaque load and Aβ levels, and improvement in structural and behavioural impairment. Thus, TRPC6 specifically modulates γ-secretase cleavage of APP and preventing APP (C99) interaction with PS1 via TRPC6 could be a novel strategy to reduce Aβ formation.

Show MeSH
Related in: MedlinePlus