Budaszewski Pinto Charles, de Sá Couto-Pereira Natividade, Kawa Odorcyk Felipe, Cagliari Zenki Kamila, Dalmaz Carla, Losch de Oliveira Diogo, Calcagnotto Maria Elisa
Neurophysiology and Neurochemistry of Neuronal Excitability and Synaptic Plasticity Laboratory (NNNESP Lab.), Department of Biochemistry, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil; Graduate Program in Biological Sciences: Biochemistry, Department of Biochemistry, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
Neurophysiology and Neurochemistry of Neuronal Excitability and Synaptic Plasticity Laboratory (NNNESP Lab.), Department of Biochemistry, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil; Graduate Program in Neuroscience, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
Brain Res. 2021 Apr 1;1756:147334. doi: 10.1016/j.brainres.2021.147334. Epub 2021 Feb 2.
Acute seizures may cause permanent brain damage depending on the severity. The pilocarpine animal model has been broadly used to study the acute effects of seizures on neurogenesis and plasticity processes and the resulting epileptogenesis. Likewise, zebrafish is a good model to study neurogenesis and plasticity processes even in adulthood. Thus, the aim of this study is to evaluate the effects of pilocarpine-induced acute seizures-like behavior on neuroplasticity and long-term behavior in adult zebrafish. To address this issue, adult zebrafish were injected with Pilocarpine (350 mg/Kg, i.p; PILO group) or Saline (control group). Experiments were performed at 1, 2, 3, 10 or 30 days after injection. We evaluated behavior using the Light/Dark preference, Open Tank and aggressiveness tests. Flow cytometry and BrdU were carried out to detect changes in cell death and proliferation, while Western blotting was used to verify different proliferative, synaptic and neural markers in the adult zebrafish telencephalon. We identified an increased aggressive behavior and increase in cell death in the PILO group, with increased levels of cleaved caspase 3 and PARP1 1 day after seizure-like behavior induction. In addition, there were decreased levels of PSD95 and SNAP25 and increased BrdU positive cells 3 days after seizure-like behavior induction. Although most synaptic and cell death markers levels seemed normal by 30 days after seizures-like behavior, persistent aggressive and anxiolytic-like behaviors were still detected as long-term effects. These findings might indicate that acute severe seizures induce short-term biochemical alterations that ultimately reflects in a long-term altered phenotype.
急性癫痫发作可能会根据严重程度导致永久性脑损伤。毛果芸香碱动物模型已被广泛用于研究癫痫发作对神经发生和可塑性过程以及由此产生的癫痫发生的急性影响。同样，斑马鱼即使在成年期也是研究神经发生和可塑性过程的良好模型。因此，本研究的目的是评估毛果芸香碱诱导的急性癫痫样行为对成年斑马鱼神经可塑性和长期行为的影响。为了解决这个问题，对成年斑马鱼注射毛果芸香碱（350mg/Kg，腹腔注射；毛果芸香碱组）或生理盐水（对照组）。在注射后1、2、3、10或30天进行实验。我们使用明暗偏好、开放鱼缸和攻击性测试来评估行为。进行流式细胞术和BrdU检测以检测细胞死亡和增殖的变化，同时使用蛋白质印迹法验证成年斑马鱼端脑中不同的增殖、突触和神经标志物。我们发现毛果芸香碱组的攻击性行为增加且细胞死亡增加，在癫痫样行为诱导后1天，裂解的半胱天冬酶3和PARP1水平升高。此外，在癫痫样行为诱导后3天，PSD95和SNAP25水平降低，BrdU阳性细胞增加。尽管在癫痫样行为后30天大多数突触和细胞死亡标志物水平似乎正常，但仍检测到持续的攻击性行为和抗焦虑样行为作为长期影响。这些发现可能表明急性严重癫痫发作会诱导短期生化改变，最终反映为长期的表型改变。
