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Estimation of LINE-1 Copy Number in the Brain Tissue and Isolated Neuronal Nuclei

  • Miki BundoEmail author
  • Tadafumi Kato
  • Kazuya Iwamoto
Protocol
Part of the Neuromethods book series (NM, volume 131)

Abstract

The mammalian brain consists of heterogeneous cell types, including neurons and various glial cells. Because somatic mutations that include long interspersed element-1 (LINE-1) retrotranspositions are usually rare, the targeting of specific brain cell types in genomic analyses of these mutations is critical. We previously reported that isolated neuronal nuclei from the prefrontal cortex of patients with schizophrenia exhibit increased numbers of LINE-1 copies. In this chapter, we describe practical methods for isolating neuronal nuclei from frozen brain tissue and cultured cells, extracting genomic DNA, and estimating LINE-1 copy numbers with quantitative reverse transcription-polymerase chain reactions.

Key words

Psychiatric disorder Postmortem brain Retrotransposon Retrotransposition Neuronal nuclei (NeuN) 

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Department of Molecular Brain Science, Graduate School of Medical SciencesKumamoto UniversityChuo-kuJapan
  2. 2.PRESTO, Japan Science and Technology AgencyChiyodakuJapan
  3. 3.Laboratory for Molecular Dynamics of Mental Disorders, RIKEN Brain Science InstituteWakoJapan

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