Jaszczyk Aneta, Stankiewicz Adrian M, Juszczak Grzegorz R
Department of Animal Behavior and Welfare, Institute of Genetics and Animal Biotechnology, Polish Academy of Sciences, 05-552 Jastrzebiec, Poland.
Department of Molecular Biology, Institute of Genetics and Animal Biotechnology, Polish Academy of Sciences, 05-552 Jastrzebiec, Poland.
Brain Sci. 2022 Jun 18;12(6):799. doi: 10.3390/brainsci12060799.
Many research methods applied in molecular neuroscience require the collection of hippocampal samples, but a still poorly recognized problem is contamination with the choroid plexus during brain dissection. Because of a distinct pattern of gene expression, its inclusion in brain samples can obscure or even confound conclusions drawn from molecular studies. Therefore, we tested our dissection method designed for removal of tissue contamination using expression of the transthyretin gene () as a marker of the choroid plexus. Additionally, we also validated dissection of the entire hippocampus into its dorsal, intermediate and ventral subdivisions using the expression of and genes as molecular markers of anatomical subdivisions. The PCR analysis showed that is expressed at a residual level in hippocampal samples that display an mRNA level several hundred lower than the adjacent control tissue colocalized with the choroid plexus. This indicates that the applied method for dissecting the hippocampus from a fresh brain allows for replicable removal of the majority of choroid plexus from hippocampal samples. In turn, differences in expression of and confirmed the proper dissection of dorsal, intermediate and ventral subdivisions from fresh brain tissue. Therefore, a special emphasis on the removal of tissue contamination and avoidance of tissue distortions makes our protocol especially suitable for molecular experiments performed either on the entire hippocampus or its subdivisions.
分子神经科学中应用的许多研究方法都需要采集海马体样本，但在脑解剖过程中脉络丛污染这一问题仍未得到充分认识。由于基因表达模式独特，其混入脑样本中可能会模糊甚至混淆分子研究得出的结论。因此，我们使用转甲状腺素蛋白基因（）的表达作为脉络丛的标志物，测试了我们设计的用于去除组织污染的解剖方法。此外，我们还利用和基因的表达作为解剖亚区的分子标志物，验证了将整个海马体解剖为背侧、中间和腹侧亚区的方法。PCR分析表明，在海马体样本中，的表达处于残留水平，其mRNA水平比与脉络丛共定位的相邻对照组织低数百倍。这表明，从新鲜大脑中解剖海马体的应用方法能够可重复地从海马体样本中去除大部分脉络丛。反过来，和表达的差异证实了从新鲜脑组织中正确解剖出背侧、中间和腹侧亚区。因此，特别强调去除组织污染和避免组织变形使我们的方案特别适用于对整个海马体或其亚区进行的分子实验。
