Journal: Neural Regeneration Research

Article Title: Pathological axonal enlargement in connection with amyloidosis, lysosome destabilization, and bleeding is a major defect in Alzheimer’s disease

doi: 10.4103/NRR.NRR-D-24-01440

Figure Lengend Snippet: Blood and vascular proteins co-exist with intracellular Aβ in neural cells in AD frontal lobe tissues and may also be related to lysosome instability. (A) Representative images showing the co-expression of blood-related markers HBA (green, Alexa Fluor 488) and Aβ (red, Alexa Fluor 594). (B) Representative images showing the co-expression of vascular markers ACTA2 (green, Alexa Fluor 488) and Aβ (red, Alexa Fluor 594). (C) Representative images showing the co-expression of vascular markers ColIV (red, Alexa Fluor 594) and Aβ (green, Alexa Fluor 488). The colocalization of markers is indicated by yellow arrows. Two small blood vessels are indicated by asterisks in the ColIV staining panel. (D) Intracellular HBA (green, Alexa Fluor 488) expression domains overlapped with the region of lysosomal destabilization, which is indicated by enlarged and clustered lysosomal compartments and diffuse Cathepsin D staining (red, Alexa Fluor 594, marked with yellow dashed circles). Control HBA-unaffected lysosomes are indicated by white arrows. Scale bars: 25 μm. ACTA2: Smooth muscle actin alpha 2; AD: Alzheimer’s disease; Aβ: amyloid-β; ColIV: collagen type IV; HBA: hemoglobin.

Article Snippet: The following primary antibodies and dilutions were used: Aβ (1:200, Abcam, Cambridge, UK, Cat# ab201061, RRID: AB_2722492), Aβ/AβPP (1:200, CST, Danvers, MA, USA, Cat# 2450, RRID: AB_490857), phos-tau (1:200, Abcam, Cat# ab151559, RRID: AB_2893278), microtubule associated protein 2 (MAP2; 1:200, Proteintech, Rosemont, IL, USA, Cat# 17490-1-AP, RRID: AB_2137880), apolipoprotein E (ApoE; 1:200, Abcam, Cat# ab183597, RRID: AB_3331650), Sortilin1 (1:200, Abcam, Cat# ab263864, RRID: AB_2884942), alpha-hemoglobin (HBA; 1:200, Abcam, Cat# ab92492, RRID: AB_10561594), Cathepsin D (1:200, Abcam, Cat# ab75852, RRID: AB_1523267), lysosome-associated membrane protein 2 (Lamp2; 1:100, Proteintech, Cat# 66301-1-Ig, RRID: AB_2881684), Cathepsin D (1:100, Proteintech, Cat# 66534-1-Ig, RRID: AB_2881897), glycosylated hemoglobin type A1C (HbA1c; 1:100, OkayBio, Nanjing, Jiangsu, China, Cat# K5a2), hemin (1:100, Absolute Antibody, Redcar, UK, Cat# 1D3), advanced glycation end products (AGE; 1:200, Abcam, Cat# ab23722, RRID: AB_447638), collagen type IV (ColIV; 1:200, Abcam, Cat# ab236640), and smooth muscle actin alpha 2 (ACTA2; 1:400, Proteintech, Cat# 23081-1-AP, RRID: AB_2815024).

Techniques: Expressing, Staining, Control