全球老龄化人口(>60岁的人)正在稳步增长,导致与年龄相关的神经退行性疾病的患病率增加。尽管在过去的几十年中进行了大量的研究,仍然没有可以停止的治疗方法,治愈,或预防这些疾病。诱导成功的神经再生(即,可以在功能上整合到现有神经回路中的新神经元的产生)可以代表一种治疗方法,以替代因衰老的中枢神经系统中的损伤或疾病而丢失的神经元。非洲绿松石飞鱼,它的寿命特别短,已经成为研究衰老如何影响神经再生的有用模型。这里,我们描述了一种稳健且可重复的刺伤方案,以研究非洲绿松石killifish的端脑再生。受伤后,通过进行BrdU脉冲追踪实验来追踪新生细胞。为了识别新生神经元,进行BrdU和HuCD的双重免疫组织化学染色。技术如溴脱氧尿苷(BrdU)标记,心内灌注,冷冻切片,和免疫荧光染色被描述为单独的部分。
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.