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Human neurons lacking amyloid precursor protein exhibit cholesterol-associated developmental and presynaptic deficits.
- Source :
-
Journal of cellular physiology [J Cell Physiol] 2024 Jun; Vol. 239 (6), pp. e30999. Date of Electronic Publication: 2023 Mar 26. - Publication Year :
- 2024
-
Abstract
- Amyloid precursor protein (APP) produces aggregable β-amyloid peptides and its mutations are associated with familial Alzheimer's disease (AD), which makes it one of the most studied proteins. However, APP's role in the human brain remains unclear despite years of investigation. One problem is that most studies on APP have been carried out in cell lines or model organisms, which are physiologically different from human neurons in the brain. Recently, human-induced neurons (hiNs) derived from induced pluripotent stem cells (iPSCs) provide a practical platform for studying the human brain in vitro. Here, we generated APP-null iPSCs using CRISPR/Cas9 genome editing technology and differentiate them into matured human neurons with functional synapses using a two-step procedure. During hiN differentiation and maturation, APP-null cells exhibited less neurite growth and reduced synaptogenesis in serum-free but not serum-containing media. We have found that cholesterol (Chol) remedies those developmental defects in APP-null cells, consistent with Chol's role in neurodevelopment and synaptogenesis. The phenotypic rescue was also achieved by coculturing those cells with wild-type mouse astrocytes, suggesting that APP's developmental role is likely astrocytic. Next, we examined matured hiNs using patch-clamp recording and detected reduced synaptic transmission in APP-null cells. This change was largely due to decreased synaptic vesicle (SV) release and retrieval, which was confirmed by live-cell imaging using two SV-specific fluorescent reporters. Adding Chol shortly before stimulation mitigated the SV deficits in APP-null iNs, indicating that APP facilitates presynaptic membrane Chol turnover during the SV exo-/endocytosis cycle. Taken together, our study in hiNs supports the notion that APP contributes to neurodevelopment, synaptogenesis, and neurotransmission via maintaining brain Chol homeostasis. Given the vital role of Chol in the central nervous system, the functional connection between APP and Chol bears important implications in the pathogenesis of AD.<br /> (© 2023 Wiley Periodicals LLC.)
- Subjects :
- Animals
Humans
Mice
Alzheimer Disease metabolism
Alzheimer Disease genetics
Alzheimer Disease pathology
Astrocytes metabolism
Cell Differentiation
Neurogenesis
Presynaptic Terminals metabolism
Synapses metabolism
Female
Mice, Inbred C57BL
Amyloid beta-Protein Precursor metabolism
Amyloid beta-Protein Precursor genetics
Cholesterol metabolism
Induced Pluripotent Stem Cells metabolism
Neurons metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1097-4652
- Volume :
- 239
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Journal of cellular physiology
- Publication Type :
- Academic Journal
- Accession number :
- 36966431
- Full Text :
- https://doi.org/10.1002/jcp.30999