Neuroplasticity and Neuroproteomics Research Group, Animal Physiology and Neurobiology Section, Department of Biology, KU Leuven, 3000, Leuven, Belgium.
Neuroplasticity and Neuroproteomics Research Group, Animal Physiology and Neurobiology Section, Department of Biology, KU Leuven, 3000, Leuven, Belgium
Cold Spring Harb Protoc. 2024 Feb 1;2024(2):107809. doi: 10.1101/pdb.prot107809.
The aging population (people >60 yr old) is steadily increasing worldwide, resulting in an increased prevalence of age-related neurodegenerative diseases. Despite intensive research efforts in the past decades, there are still no therapies available to stop, cure, or prevent these diseases. Induction of successful neuroregeneration (i.e., the production of new neurons that can functionally integrate into the existing neural circuitry) could represent a therapy to replace neurons lost by injury or disease in the aged central nervous system. The African turquoise killifish, with its particularly short life span, has emerged as a useful model to study how aging influences neuroregeneration. Here, we describe a robust and reproducible stab-injury protocol to study regeneration in the telencephalon of the African turquoise killifish. After the injury, newborn cells are traced by conducting a BrdU pulse-chase experiment. To identify newborn neurons, a double immunohistochemical staining for BrdU and HuCD is carried out. Techniques such as bromodeoxyuridine (BrdU) labeling, intracardial perfusion, cryosectioning, and immunofluorescence staining are described as separate sections.
人口老龄化（60 岁以上的人）在全球范围内稳步增加，导致与年龄相关的神经退行性疾病的患病率增加。尽管过去几十年进行了密集的研究，但仍然没有可用的疗法来阻止、治愈或预防这些疾病。成功的神经再生（即产生新的神经元，这些神经元可以在功能上整合到现有的神经回路中）可以作为一种治疗方法，以替代因衰老中枢神经系统损伤或疾病而丧失的神经元。非洲蓝珍珠鱼，其寿命特别短，已成为研究衰老如何影响神经再生的有用模型。在这里，我们描述了一种稳健且可重复的刺伤损伤方案，用于研究非洲蓝珍珠鱼端脑的再生。损伤后，通过进行 BrdU 脉冲追踪实验来追踪新生细胞。为了鉴定新生神经元，进行 BrdU 和 HuCD 的双重免疫组织化学染色。溴脱氧尿苷 (BrdU) 标记、心脏内灌注、冷冻切片和免疫荧光染色等技术分别作为单独的章节进行描述。
