Advanced Neural Dynamics, Inc., Pennsylvania Biotechnology Center, 3805 Old Easton Road, Doylestown, PA, 18902, USA.
Sanford Children's Health Research Center, Sanford Research, Sioux Falls, SD, 57104, USA.
J Mol Neurosci. 2017 Sep;63(1):100-114. doi: 10.1007/s12031-017-0962-5. Epub 2017 Aug 15.
Juvenile Batten disease (JBD) is an inherited disorder that is characterized by the development of blindness, seizures, and progressive motor, psychiatric, and cognitive impairment. A model of JBD expressing the predominant human mutation (Cln3 ) has been explored. Dissociated brain cultures from Cln3 knock-in mice were compared to wild type (WT) for effects on granules of ceroid lipofuscin (CL) and neuronal structure. Utilizing high content image analysis of CL granules identified with antibodies to mitochondrial ATP synthase subunit c or tripeptidyl peptidase-1, significant increases in the areas for both immunoreactive granules were observed in Cln3 cultures in comparison to WT. CL granules also exhibit autofluorescence at 488 and 560 nm, and the areas of these autofluorescent spots were found to be significantly increased in Cln3 cultures in comparison to WT. Progressive increases in CL granule area in Cln3 cultures were observed during culture development. Because current therapies for JBD provide only symptomatic support, a therapeutic strategy has been explored based on the observations that JBD-related tissues are deficient in β-galactosyl ceramide. Treatment of cultures for 40 h with a potent analog of β-galactosyl ceramide (SNB-4050) produced significant decreases in CL granule area in the Cln3 cultures; whereas identical studies on WT cultures produced no detectible changes. Significant decreases in average neurite length and neurite branch point number were also observed in the Cln3 cultures that were attenuated by treatment with 1 nM SNB-4050. These studies indicate Cln3 brain cultures may be useful to screen therapeutic agents for treatment of JBD.
少年型 Batten 病（JBD）是一种遗传性疾病，其特征是失明、癫痫发作以及进行性运动、精神和认知障碍。已经探索了表达主要人类突变（Cln3）的 JBD 模型。将 Cln3 基因敲入小鼠的分离脑培养物与野生型（WT）进行比较，以研究其对神经细胞内的类脂褐素颗粒（CL）和神经元结构的影响。利用针对线粒体三磷酸腺苷合酶亚基 c 或三肽基肽酶-1 的抗体对 CL 颗粒进行高内涵图像分析，发现 Cln3 培养物中两种免疫反应性颗粒的面积均显著增加。CL 颗粒在 488nm 和 560nm 处也显示自发荧光，并且发现 Cln3 培养物中的这些自发荧光斑点的面积与 WT 相比显著增加。在培养过程中，观察到 Cln3 培养物中 CL 颗粒面积逐渐增加。由于目前治疗 JBD 的方法仅提供对症支持，因此已经探索了一种基于 JBD 相关组织中缺乏β-半乳糖基神经酰胺的治疗策略。用β-半乳糖基神经酰胺的有效类似物（SNB-4050）对培养物处理 40 小时，可使 Cln3 培养物中的 CL 颗粒面积显著减少；而对 WT 培养物进行相同的研究则未产生可检测到的变化。在 Cln3 培养物中还观察到平均神经突长度和神经突分支点数量的显著减少，用 1nM SNB-4050 处理可减轻这种减少。这些研究表明，Cln3 脑培养物可能可用于筛选治疗 JBD 的治疗剂。
