Journal: bioRxiv

Article Title: Amyloid β induces cardiac dysfunction and neuro-signaling impairment in the heart of an Alzheimer’s disease model

doi: 10.1101/2023.07.11.548558

Figure Lengend Snippet: ( A - B ): Representative immunoblots ( A ) and densitometric quantitative analysis ( B ) showing levels of NGF, BDNF, GAP-43, and DβH, in total cardiac (top panels) and cerebral cortex (bottom panels) lysates from WT and Tg2576 mice. ( C-D ): digital images (C, scale bar 100μm) and quantifications (D) showing cardiac adrenergic nerve fibers, labeled with anti-tyrosine-hydroxylase (TH, in green), and cardiac regenerating nerve endings, labeled with anti-neuronal regeneration marker (GAP-43, in red) in heart sections from WT and Tg2576 mice. n=3– 5 mice/group. Data are presented as a mean±SEM. *P<0.05, **P<0.01 and vs WT. Student t-tests have been performed between the 2 groups.

Article Snippet: Total lysates were used to evaluate the protein levels of NGF (AN-240; Alomone labs; 1:1000), BDNF (ANT-010; Alomone labs; 1:1000), GAP-43 (Millipore AB552; 1:1000), Cleaved Caspase-3- Asp175 (Cl-Casp-3- Cell Signaling- #94530; 1:200), dopamine β hydroxylase (DβH; AB1536; Millipore), human Aβ [mouse monoclonal anti-Aβ antibodies mixture composed of 4G8 epitope (residues Aβ18–22; SIG-39320; Covance) and 6E10 epitope (residues Aβ3–8; SIG-39220; Covance)], and GAPDH (sc-32233,6C5; Santa Cruz Biotechnology; 1: 2000), the latter which was used as the loading control.

Techniques: Western Blot, Labeling, Marker

Journal: bioRxiv

Article Title: Amyloid β induces cardiac dysfunction and neuro-signaling impairment in the heart of an Alzheimer’s disease model

doi: 10.1101/2023.07.11.548558

Figure Lengend Snippet: ( A - C ): Representative immunoblots ( A ) and densitometric quantitative analysis ( B - C ) showing protein levels of BDNF ( B ), and NGF ( C ) in total protein lysates from human cardiomyocytes (AC16) stimulated with human Aβ-40 oligomers for 16 h. GAPDH levels were used as a loading control. n= 6–7 biological replicates. Data are presented as a mean±SEM. **P<0.01 and vs WT. Student t-tests have been performed between the groups. (NT= Not Treated).

Article Snippet: Total lysates were used to evaluate the protein levels of NGF (AN-240; Alomone labs; 1:1000), BDNF (ANT-010; Alomone labs; 1:1000), GAP-43 (Millipore AB552; 1:1000), Cleaved Caspase-3- Asp175 (Cl-Casp-3- Cell Signaling- #94530; 1:200), dopamine β hydroxylase (DβH; AB1536; Millipore), human Aβ [mouse monoclonal anti-Aβ antibodies mixture composed of 4G8 epitope (residues Aβ18–22; SIG-39320; Covance) and 6E10 epitope (residues Aβ3–8; SIG-39220; Covance)], and GAPDH (sc-32233,6C5; Santa Cruz Biotechnology; 1: 2000), the latter which was used as the loading control.

Techniques: Western Blot, Control

Journal: Molecular Neurodegeneration

Article Title: Neuron loss in the 5XFAD mouse model of Alzheimer’s disease correlates with intraneuronal Aβ 42 accumulation and Caspase-3 activation

doi: 10.1186/1750-1326-8-2

Figure Lengend Snippet: Intraneuronal Aβ42 is partially localized within both endosomes and lysosomes in 5XFAD neurons. Parasagittal brain sections of 5XFAD ( A , B, E-K ) and 5XFAD; BACE1 −/− ( C , D ) brains were co-stained with anti-Aβ 42 -selective antibody ( A-K ) and anti-APP (6E10: B , D ; Karen : E ) or anti-Transferrin receptor (TfR) ( F-H ) or anti-LAMP1 ( I-K ) antibody, and DAPI for nuclei ( A-K ), and imaged by confocal microscopy. All mice were 4 months old, except mice used for LAMP1 immunostaining were 2 months ( I-K ). Label colors indicate fluorescence color. Note that APP staining appears widely distributed in the soma and cell surface ( B, D, E ), while Aβ 42 labeling is confined to intraneuronal puncta in 5XFAD mice ( A, B, E, F, I ). Asterisk in E identifies Aβ 42 -positive plaque (red) above large pyramidal neuron that exhibits intraneuronal Aβ 42 puncta (arrow). lntraneuronal Aβ 42 signal is absent in 5XFAD; BACE1 −/− neurons ( C, D ), again demonstrating the Aβ 42 -selectivity of the antibody. In 5XFAD neurons, intraneuronal Aβ 42 co-localizes with Transferrin receptor in early endosomes ( F-H ) and with LAMP1 in late endosomes and early lysosomes ( I-K ), at least in part. Scale bar in D = 12μm ( A, C ); = 20 μm ( B, D ). Scale bar in K = 12 μm ( E ); = 20 μm ( F-K ).

Article Snippet: Sections were incubated at 4°C on a shaker overnight with the following primary antibodies: rabbit monoclonal anti-Aβ 42 (1:1000; Invitrogen; 700254), mouse monoclonal anti-Aβ 42 (1:1000; Covance; SIG-39142), rabbit monoclonal anti-cleaved Caspse-3 (1:1000; Cell Signaling; #9664), rat monoclonal anti-Transferrin receptor (1:500; Abcam; ab60344), rat monoclonal anti-LAMP1 (1:500; Abcam; ab25245), 6E10 mouse monoclonal anti-Aβ (1:1000; Chemicon; MAB1560), Karen goat polyclonal anti-N-terminal APP (1:500; [ ]), DAPI (Invitrogen).

Techniques: Staining, Confocal Microscopy, Immunostaining, Fluorescence, Labeling