Brain Case New Release｜Rapid, Precise Construction of PD/AD/AS Animal Models

Applications of AAV in Animal Modeling

Case Study

1. mGluR5 Protects Against α-Synuclein–Induced Microglial Inflammation and Neurotoxicity in a Parkinson’s Disease Model

Figure 1. Inflammatory response associated with progressive neurodegeneration in a rat Parkinson’s disease model induced by AAV-α-syn.
 

Atherosclerosis (AS)

Case Study

1. Induction of Atherosclerosis in Mice and Rats Without Germline Genetic Engineering

Figure 3. Atherosclerosis induced by rAAV8-D377Y-mPCSK9.
 

2. Ceramide Aggravates Atherosclerosis via CYSLTR2 and P2RY6 Sensing

🔺Experimental Animals: C57BL/6J mice
🔺Viral Vector: rAAV8-D377Y-hPCSK9
🔺Injection Protocol: Tail vein injection, 2.0 × 10¹¹ vg; 12-week expression period
🔺Diet Regimen: HFD — standard casein-based formula containing 1.25% cholesterol and 0.5% sodium cholate
🔺Experimental Findings: To explore whether CYSLTR2 and P2RY6 mediate ceramide-induced exacerbation of atherosclerosis, 8-week-old WT, Cysltr2⁻/⁻, P2ry6⁻/⁻, and double-knockout mice were injected with AAV8-hPCSK9-D377Y and fed a high-fat diet for 12 weeks. WT mice injected with AAV-Control and fed a normal diet served as blank controls. Starting at 10 weeks of age, “AAV-PCSK9 + high-fat” mice received intravenous C16:0 ceramide (5 mg/kg every 48 h). Results showed that C16:0 ceramide did not affect blood lipid levels but doubled plasma ceramide concentrations and significantly promoted plaque formation in WT mice. This effect was partially reduced in single knockouts and further attenuated in double knockouts, which exhibited less plaque formation, lipid accumulation, and macrophage infiltration. These findings demonstrate that CYSLTR2 and P2RY6 receptors synergistically mediate ceramide-induced aggravation of atherosclerosis.

Figure 4. Deletion of CYSLTR2/P2RY6 mitigates ceramide-aggravated atherosclerosis.
 

 

Alzheimer’s Disease (AD)

AD is a common age-related neurodegenerative disorder characterized by progressive cognitive decline and neuronal damage, with symptoms worsening over time. Neuropathologically, AD brains exhibit amyloid-β (Aβ) deposition forming senile plaques, hyperphosphorylated Tau protein aggregation forming neurofibrillary tangles (NFTs), as well as neuronal loss and synaptic damage.

In AD research, adeno-associated virus (AAV) vectors are frequently used to deliver AD-related pathogenic genes—such as mutant APP or mutant Tau—into specific brain regions (e.g., hippocampus, cortex) of experimental animals. This induces Aβ deposition, Tau aggregation, and cognitive impairment, creating animal models that closely replicate AD pathology and phenotype for mechanistic studies and drug screening.
 

Case Study

1. AAV-Mediated Tau Expression Induces Pyramidal Neuron Degeneration via Cell-Cycle Reentry Without Neurofibrillary Tangle Formation in Wild-Type Mice

💠Experimental Animals: Wild-type FVB/N mice
💠Viral Vector: rAAV1/2-hsyn-hTau (P301L)
💠Injection Protocol: Stereotaxic injection into the hippocampus, 1.0 × 10⁸ vg; 12-week expression period
💠Experimental Findings: Injection of AAV-Tau (P301L) into the mouse brain induced dose-dependent neurodegeneration. Low doses caused mild neuronal loss and thinning of the hippocampal CA1/2 region, with hTau levels decreasing as neuronal loss progressed due to reduced protein synthesis in degenerating neurons. Pathology first appeared in the CA2 region at 1.5 weeks post-injection, extended to CA1 by 3 weeks, and by 6–12 weeks nearly all CA pyramidal cells were lost, with degeneration spreading to the cortex. Fluoro-Jade B (FJB) staining confirmed degenerating neurons. Microglial activation emerged transiently and subsided by 12 weeks, while astrocyte activation persisted, indicating ongoing inflammation. Control mice injected with AAV-EGFP showed no neuronal loss or microglial activation. These results demonstrate that AAV-Tau (P301L) drives degeneration of hippocampal CA and cortical pyramidal neurons, with microglial activation specifically linked to Tau-mediated neurodegeneration rather than amyloid pathology or viral components.

Figure 5. Dose-dependent and temporal progression of neurodegeneration induced by AAV-Tau (P301L).

 

2. Hippocampal Overexpression of Human Tau to Establish a Non-Human Primate Model with Alzheimer’s-Like Pathology

🔸Animal Model: Adult rhesus monkeys (7–15 years old)
🔸Viral Vector: rAAV9-hTau (WT)
🔸Injection Protocol: Bilateral stereotaxic injections into the hippocampus, 10¹⁰–10¹¹ GC; expression for 6–12 weeks / up to 50 weeks
🔸Experimental Findings: To develop a non-human primate (NHP) model of Tauopathy with AD-like pathology, adult rhesus monkeys were injected bilaterally in the hippocampus with an AAV vector overexpressing human Tau. Immunostaining, PET, and MRI analyses revealed an AAV-mediated gene transduction efficiency of approximately 75%, with stable Tau expression maintained from 6 to 50 weeks. These results demonstrate that AAV enables efficient, widespread, and long-lasting Tau expression in the primate hippocampus, providing a robust NHP model for Alzheimer’s disease research.

Figure 6. Bilateral stereotaxic injection of AAV into the hippocampus achieves sustained high-level Tau expression in a non-human primate model.

 

Relevant AAV Modeling Tool Viruses Are All Available from Brain Case