Literature DB >> 32202523

Combining Laser Capture Microdissection and Microfluidic qPCR to Analyze Transcriptional Profiles of Single Cells: A Systems Biology Approach to Opioid Dependence.

Sean J O'Sullivan1, Beverly A S Reyes2, Rajanikanth Vadigepalli3, Elisabeth J Van Bockstaele2, James S Schwaber3.   

Abstract

Profound transcriptional heterogeneity in anatomically adjacent single cells suggests that robust tissue functionality may be achieved by cellular phenotype diversity. Single-cell experiments investigating the network dynamics of biological systems demonstrate cellular and tissue responses to various conditions at biologically meaningful resolution. Herein, we explain our methods for gathering single cells from anatomically specific locations and accurately measuring a subset of their gene expression profiles. We combine laser capture microdissection (LCM) with microfluidic reverse transcription quantitative polymerase chain reactions (RT-qPCR). We also use this microfluidic RT-qPCR platform to measure the microbial abundance of gut contents.

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Year:  2020        PMID: 32202523      PMCID: PMC8015684          DOI: 10.3791/60612

Source DB:  PubMed          Journal:  J Vis Exp        ISSN: 1940-087X            Impact factor:   1.355


  9 in total

1.  Computational analysis of cell-to-cell heterogeneity in single-cell RNA-sequencing data reveals hidden subpopulations of cells.

Authors:  Florian Buettner; Kedar N Natarajan; F Paolo Casale; Valentina Proserpio; Antonio Scialdone; Fabian J Theis; Sarah A Teichmann; John C Marioni; Oliver Stegle
Journal:  Nat Biotechnol       Date:  2015-01-19       Impact factor: 54.908

Review 2.  Single-cell RNA sequencing to explore immune cell heterogeneity.

Authors:  Efthymia Papalexi; Rahul Satija
Journal:  Nat Rev Immunol       Date:  2017-08-07       Impact factor: 53.106

3.  Identifying functional gene regulatory network phenotypes underlying single cell transcriptional variability.

Authors:  James Park; Babatunde Ogunnaike; James Schwaber; Rajanikanth Vadigepalli
Journal:  Prog Biophys Mol Biol       Date:  2014-11-27       Impact factor: 3.667

4.  Dysbiosis in inflammatory bowel disease.

Authors:  C P Tamboli; C Neut; P Desreumaux; J F Colombel
Journal:  Gut       Date:  2004-01       Impact factor: 23.059

5.  Inputs drive cell phenotype variability.

Authors:  James Park; Anthony Brureau; Kate Kernan; Alexandria Starks; Sonali Gulati; Babatunde Ogunnaike; James Schwaber; Rajanikanth Vadigepalli
Journal:  Genome Res       Date:  2014-03-26       Impact factor: 9.043

6.  A comprehensive assessment of RNA-seq accuracy, reproducibility and information content by the Sequencing Quality Control Consortium.

Authors: 
Journal:  Nat Biotechnol       Date:  2014-08-24       Impact factor: 54.908

7.  Gut inflammation and dysbiosis in human motor neuron disease.

Authors:  Julie Rowin; Yinglin Xia; Barbara Jung; Jun Sun
Journal:  Physiol Rep       Date:  2017-09

8.  Single-Cell Glia and Neuron Gene Expression in the Central Amygdala in Opioid Withdrawal Suggests Inflammation With Correlated Gut Dysbiosis.

Authors:  Sean J O'Sullivan; Evangelia Malahias; James Park; Ankita Srivastava; Beverly A S Reyes; Jon Gorky; Rajanikanth Vadigepalli; Elisabeth J Van Bockstaele; James S Schwaber
Journal:  Front Neurosci       Date:  2019-07-03       Impact factor: 4.677

9.  Single-Cell Transcriptional Analysis Reveals Novel Neuronal Phenotypes and Interaction Networks Involved in the Central Circadian Clock.

Authors:  James Park; Haisun Zhu; Sean O'Sullivan; Babatunde A Ogunnaike; David R Weaver; James S Schwaber; Rajanikanth Vadigepalli
Journal:  Front Neurosci       Date:  2016-10-25       Impact factor: 4.677

  9 in total
  1 in total

Review 1.  Similarities in alcohol and opioid withdrawal syndromes suggest common negative reinforcement mechanisms involving the interoceptive antireward pathway.

Authors:  Sean J O'Sullivan; James S Schwaber
Journal:  Neurosci Biobehav Rev       Date:  2021-02-26       Impact factor: 9.052

  1 in total

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