Department of Biomedical Sciences, Quillen College of Medicine, East Tennessee State University, Johnson City, United States.
Hong Kong Institute, Asia Metropolitan University, Hong Kong, China.
ASN Neuro. 2021 Jan-Dec;13:17590914211009730. doi: 10.1177/17590914211009730.
Dysfunction of the central noradrenergic and dopaminergic systems is the primary neurobiological characteristic of Parkinson's disease (PD). Importantly, neuronal loss in the locus coeruleus (LC) that occurs in early stages of PD may accelerate progressive loss of dopaminergic neurons. Therefore, restoring the activity and function of the deficient noradrenergic system may be an important therapeutic strategy for early PD. In the present study, the lentiviral constructions of transcription factors Phox2a/2b, Hand2 and Gata3, either alone or in combination, were microinjected into the LC region of the PD model VMAT2 Lo mice at 12 and 18 month age. Biochemical analysis showed that microinjection of lentiviral expression cassettes into the LC significantly increased mRNA levels of Phox2a, and Phox2b, which were accompanied by parallel increases of mRNA and proteins of dopamine β-hydroxylase (DBH) and tyrosine hydroxylase (TH) in the LC. Furthermore, there was considerable enhancement of DBH protein levels in the frontal cortex and hippocampus, as well as enhanced TH protein levels in the striatum and substantia nigra. Moreover, these manipulations profoundly increased norepinephrine and dopamine concentrations in the striatum, which was followed by a remarkable improvement of the spatial memory and locomotor behavior. These results reveal that over-expression of these transcription factors in the LC improves noradrenergic and dopaminergic activities and functions in this rodent model of PD. It provides the necessary groundwork for the development of gene therapies of PD, and expands our understanding of the link between the LC-norepinephrine and dopamine systems during the progression of PD.
中枢去甲肾上腺素能和多巴胺能系统功能障碍是帕金森病(PD)的主要神经生物学特征。重要的是,PD 早期蓝斑核(LC)神经元的丢失可能加速多巴胺能神经元的进行性丧失。因此,恢复去甲肾上腺素能系统的活性和功能可能是早期 PD 的重要治疗策略。在本研究中,转录因子 Phox2a/2b、Hand2 和 Gata3 的慢病毒构建物单独或组合,在 12 和 18 月龄的 VMAT2 Lo PD 模型小鼠的 LC 区被微注射。生化分析表明,LC 内慢病毒表达盒的微注射显著增加了 Phox2a 和 Phox2b 的 mRNA 水平,同时 LC 中的多巴胺 β-羟化酶(DBH)和酪氨酸羟化酶(TH)的 mRNA 和蛋白水平也平行增加。此外,前额叶皮层和海马中的 DBH 蛋白水平显著升高,纹状体和黑质中的 TH 蛋白水平也显著升高。此外,这些操作极大地增加了纹状体中的去甲肾上腺素和多巴胺浓度,随后空间记忆和运动行为显著改善。这些结果表明,LC 中这些转录因子的过表达改善了 PD 啮齿动物模型中去甲肾上腺素能和多巴胺能的活性和功能。它为 PD 的基因治疗的发展提供了必要的基础,并扩展了我们对 LC-去甲肾上腺素能和多巴胺系统在 PD 进展过程中联系的理解。