Futuristic Animal Resource & Research Center (FARRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Chungcheongbuk-do, Republic of Korea.
Division of Animal Sciences, University of Missouri, Columbia, MO, USA.
J Alzheimers Dis. 2024;101(2):445-461. doi: 10.3233/JAD-231297.
Presenilin 1 (PSEN1) is one of the genes linked to the prevalence of early onset Alzheimer's disease. In mice, inactivation of Psen1 leads to developmental defects, including vertebral malformation and neural development. However, little is known about the role of PSEN1 during the development in other species.
To investigate the role of PSEN1 in vertebral development and the pathogenic mechanism of neurodegeneration using a pig model.
CRISPR/Cas9 system was used to generate pigs with different mutations flanking exon 9 of PSEN1, including those with a deleted exon 9 (Δexon9). Vertebral malformations in PSEN1 mutant pigs were examined by X-ray, micro-CT and micro-MRI. Neuronal cells from the brains of PSEN1 mutant pigs were analyzed by immunoflourescence, followed by image analysis including morphometric evaluation via image J and 3D reconstruction.
Pigs with a PSEN1 null mutation (Δexon9-12) died shortly after birth and had significant axial skeletal defects, whereas pigs carrying at least one Δexon9 allele developed normally and remained healthy. Effects of the null mutation on abnormal skeletal development were also observed in fetuses at day 40 of gestation. Abnormal distribution of astrocytes and microglia in the brain was detected in two PSEN1 mutant pigs examined compared to age-matched control pigs. The founder pigs were bred to establish and age PSEN1ΔE9/+ pigs to study their relevance to clinical Alzheimer's diseases.
PSEN1 has a critical role for normal vertebral development and PSEN1 mutant pigs serves as novel resources to study Alzheimer's disease.
早发型阿尔茨海默病的发病与早老素 1（PSEN1）基因有关。在小鼠中，PSEN1 的失活会导致发育缺陷，包括脊椎畸形和神经发育。然而，对于其他物种中 PSEN1 在发育过程中的作用知之甚少。
利用猪模型研究 PSEN1 在脊椎发育中的作用和神经退行性病变的发病机制。
使用 CRISPR/Cas9 系统生成 PSEN1 外显子 9 侧翼带有不同突变的猪，包括缺失外显子 9（Δexon9）的猪。通过 X 射线、微 CT 和微 MRI 检查 PSEN1 突变猪的脊椎畸形。通过免疫荧光法分析 PSEN1 突变猪的神经元细胞，然后通过图像分析包括使用 image J 进行形态计量评估和 3D 重建。
PSEN1 无功能突变（Δexon9-12）的猪在出生后不久死亡，且有明显的轴性骨骼缺陷，而携带至少一个 Δexon9 等位基因的猪则正常发育且保持健康。在妊娠第 40 天的胎儿中也观察到了该无功能突变对异常骨骼发育的影响。与年龄匹配的对照猪相比，在两只 PSEN1 突变猪中检测到大脑中星形胶质细胞和小胶质细胞的异常分布。创始人猪被繁殖以建立和增龄 PSEN1ΔE9/+猪，以研究它们与临床阿尔茨海默病的相关性。
PSEN1 对正常的脊椎发育至关重要，PSEN1 突变猪为研究阿尔茨海默病提供了新的资源。
