Søkilde Rolf, Kaadt Erik, Kristensen Lasse Sommer, Venø Morten Trillingsgaard, Kjems Jørgen, Nyengaard Jens Randel, Nielsen Boye Schnack, Elfving Betina
Experimental and Molecular Psychiatry, Translational Neuropsychiatry Unit, Department of Clinical Medicine, Aarhus University, Palle Juul-Jensens Blvd 11, 8200, Aarhus, Denmark.
The Laboratory for Gene-Regulatory Mechanisms in Cancer, Department of Biomedicine, Aarhus University, Høegh-Guldbergs Gade 10, 8000, Aarhus, Denmark.
Mol Neurobiol. 2025 May 31. doi: 10.1007/s12035-025-05077-z.
Here, we report that long-term stored human brain samples, formalin-fixed paraffin-embedded (FFPE) from The Brain Collection, University of Southern Denmark, Denmark ( https://www.sdu.dk/en/forskning/bridge/the-brain-collection ), can be used for in situ hybridization (ISH) analysis of selected microRNAs (miRNAs). The Human Brain Collection consists of brains from 9479 subjects who died at a Danish State Mental Hospital in the period of 1945-1982. In the present study we included tissue specimens from prefrontal cortex (PFC) and hippocampus (HIP) from 163 patients diagnosed with schizophrenia, bipolar disorder, or major depressive disorder. Initially, the Nanostring nCounter platform was used to identify miRNA candidates for ISH analysis using the miRNAscope technology. Based on the Nanostring nCounter quantifications with bulk tissue, we identified and selected 10 miRNAs from PFC (miR-9-5p, miR-29b-3p, miR-30c-5p, miR-124-3p, miR-125b-5p, miR-138-5p, miR-181a-5p, miR-224-5p, miR-302d-3p, and miR-432-5p) and 6 miRNAs from HIP (let-7a-5p, miR-7-5p, miR-124-3p, miR-127-3p, miR-145-5p and miR-149-5p). miRNAscope ISH analysis was then performed with the respective probes on 30 PFC and 30 HIP samples, respectively. In the PFC six miRNAs (miR-9-5p, miR-29b-3p, miR-124-3p, miR-125b-5p, miR-138-5p, and miR-181a-5p) were detected and four (miR-145-5p, let-7a-5p, miR-124-3p and miR-7-5p) in the HIP samples. In both brain regions miR-124-3p was the most abundantly expressed. We conclude, that the combination of the Nanostring nCounter technology and the miRNAscope analysis is a valid approach to study spatial expression of specific miRNAs in these up to 76 years old FFPE blocks. This opens a new avenue of possibilities for studying the underlying epigenetic mechanisms in mental disorders.
在此，我们报告来自丹麦南丹麦大学大脑库（https://www.sdu.dk/en/forskning/bridge/the-brain-collection）的长期保存的人脑样本，即福尔马林固定石蜡包埋（FFPE）样本，可用于所选 microRNA（miRNA）的原位杂交（ISH）分析。人脑库包含1945年至1982年期间在丹麦一家国立精神病院去世的9479名受试者的大脑。在本研究中，我们纳入了163例被诊断为精神分裂症、双相情感障碍或重度抑郁症患者的前额叶皮质（PFC）和海马体（HIP）的组织标本。最初，使用Nanostring nCounter平台，利用miRNAscope技术鉴定用于ISH分析的miRNA候选物。基于对大块组织的Nanostring nCounter定量分析，我们从PFC中鉴定并选择了10种miRNA（miR-9-5p、miR-29b-3p、miR-30c-5p、miR-124-3p、miR-125b-5p、miR-138-5p、miR-181a-5p、miR-224-5p、miR-302d-3p和miR-432-5p）以及从HIP中选择了6种miRNA（let-7a-5p、miR-7-5p、miR-124-3p、miR-127-3p、miR-145-5p和miR-149-5p）。然后分别用各自的探针在30个PFC样本和30个HIP样本上进行miRNAscope ISH分析。在PFC样本中检测到6种miRNA（miR-9-5p、miR-29b-3p、miR-124-3p、miR-125b-5p、miR-138-5p和miR-181a-5p），在HIP样本中检测到4种（miR-145-5p、let-7a-5p、miR-124-3p和miR-7-5p）。在两个脑区中，miR-124-3p表达最为丰富。我们得出结论，Nanostring nCounter技术与miRNAscope分析相结合是研究这些保存长达76年的FFPE样本中特定miRNA空间表达的有效方法。这为研究精神障碍潜在的表观遗传机制开辟了一条新的途径。
