Characterization of Hippocampal Adult-borne Granule Cells in a Transient Cerebral Ischemia Model
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Author:
Date:
2021-01-20
[Abstract] Long-term consequences of stroke significantly impair the quality of life in a growing population of stroke survivors. Hippocampal adult neurogenesis has been hypothesized to play a role in the pathophysiology of cognitive and neuropsychiatric long-term sequelae of stroke. Reliable animal models of stroke are paramount to understanding their biomechanisms and to advancing therapeutic strategies. We present a detailed protocol of a transient cerebral ischemia model which does not cause direct ischemic damage in the hippocampus, allowing investigations into the pathophysiology of long-term neurocognitive deficits of stroke. Furthermore, we describe a protocol for obtaining acute hippocampal slices for the purpose of electrophysiological and morphological characterization of adult-borne ...
[摘要] [摘要]中风的长期后果严重损害了越来越多的中风幸存者的生活质量。假定海马成年神经发生在中风的认知和神经精神病学长期后遗症的病理生理学中起作用。可靠的中风动物模型对于理解其生物机制和推进治疗策略至关重要。我们提出 暂时性脑缺血模型的详细协议,该模型不会在海马体中引起直接缺血损伤,从而可以研究中风的长期神经认知缺陷的病理生理学。此外,我们描述了一种用于获取成人海马颗粒细胞的电生理学和形态学特征的急性海马切片的协议。还讨论了与从小细胞(例如未成熟的成年颗粒细胞)进行电生理记录有关的特殊性。本协议可以通过多模式研究(行为,形态结构,生化)加以补充,以期有希望促进研究和中风的长期后遗症的发展以及发现新的治疗机会。
[背景]中风是在发达国家发病率和死亡率的重要原因引起急性和延迟赤字。虽然针对缺血性中风恢复血流的干预策略在降低急性发病率和死亡率方面已经变得更加有效,但中风的长期后果(如中风后抑郁和中风后认知功能障碍和痴呆症)目前正在逃避临床治疗(Wang等。,2010; Loubinoux等人,2012; Mijajlovic ...
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D-serine Measurements in Brain Slices or Other Tissue Explants
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Author:
Date:
2018-01-20
[Abstract] D-serine is an atypical amino acid present in the mammalian body (most amino acids in the mammalian body are L-isomers) that is mostly known in neuroscience for its role as a co-agonist controlling the N-methyl D-aspartate receptor (NMDAR). D-serine levels are decreased in patients with schizophrenia and this is thought to mediate, at least in part, the hypofunction of NMDARs that is central to the glutamate hypothesis for the etiology of this neuropsychiatric disorder. D-serine detection was first established using high performance liquid chromatography, a costly and complex technique that requires high levels of expertise. But with the increasing interest in this unconventional amino acid, there is an increasing need for easier, cheaper and more accessible detection methods. Here we ...
[摘要] D-丝氨酸是存在于哺乳动物体内的非典型氨基酸(哺乳动物体内的大部分氨基酸是L-异构体),其在神经科学中主要是作为控制N-甲基D-天冬氨酸受体的共激动剂的作用而已知的NMDA受体)。精神分裂症患者的D-丝氨酸水平降低,这被认为至少部分地介导了NMDARs功能减退,这种谷氨酸假说对于这种神经精神障碍的病因至关重要。 D-丝氨酸检测首先使用高效液相色谱法建立,这是一种昂贵且复杂的技术,需要高水平的专业知识。但随着对这种非常规氨基酸的兴趣日益增加,人们越来越需要更容易,更便宜和更容易获得的检测方法。在这里,我们描述了我们在最近的出版物(Papouin等人,2017b)中采用的安培法,基于生物传感器的方法。它可以对新鲜组织的D-丝氨酸水平进行可靠的测量,如急性脑切片,浓度高于100 nM,技术要求最低。
【背景】N-甲基D-天冬氨酸受体(NMDAR)是脑,脊髓和周围神经系统如肠神经元中神经递质谷氨酸的受体。在肾小管细胞和软骨细胞中也有发现。除谷氨酸外,NMDAR的激活需要在专用结合位点上结合共激动剂(Johnson和Ascher,1987; Kleckner和Dingledine,1988)。非常规氨基酸D-丝氨酸是神经系统众多区域中NMDAR的内源性共激动剂(参见Papouin等人,2017a)。在肝脏和肾脏中也可以发现其丰富的降解和排泄(Montesinos ...
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Obtaining Acute Brain Slices
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Author:
Date:
2018-01-20
[Abstract] Obtaining acute brain slices for electrophysiology or amperometric recordings has become a routine procedure in most labs in the field of neuroscience. Yet, protocols describing the step by step process are scarce, in particular for routine acute preparations such as from the mouse hippocampus. Here we provide a detailed protocol for the dissection, extraction and acute slicing of the mouse brain, including tips and list of material required.
[摘要] 在神经科学领域的大多数实验室中,获得用于电生理学或电流记录的急性脑切片已经成为常规手术。 然而,描述逐步过程的方案是稀缺的,特别是对于例如来自小鼠海马的常规急性制剂。 在这里,我们提供了一个详细的协议,解剖,提取和急性切片的小鼠大脑,包括技巧和所需材料清单。
【背景】随着体外电生理和电流记录技术的民主化,从啮齿动物大脑获取急性切片已经成为神经科学研究中经典和关键的步骤。然而,实现这一程序所需的技术诀窍通常是口头传递的,大多数实验室已经开发了自制的食谱,并根据自己的需要或感兴趣的区域调整了最重要的步骤,从而缺乏描述如何逐步获得高质量的急性脑切片。虽然可以发现一些方案描述特别具有挑战性的准备,例如成年小鼠脊髓的急性切片(Garre等人,Bioprotocol 2017: https://doi.org/10.21769/BioProtoc.2102 ),描述了更多日常准备的基本程序( ...
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