Multivariate analysis of RNA levels from postmortem human brains as measured by three different methods of RT-PCR

Nancy L. Johnston, Juraj Cerevnak, Andrew D. Shore, E. Fuller Torrey, Robert H Yolken

Research output: Contribution to journalArticle

Abstract

The analysis of RNA from postmortem human brain tissue by reverse transcription-polymerase chain reaction (RT-PCR) provides a practical method to measure both normal and abnormal brain gene expression. A major limitation in using human material is that yields can vary dramatically from individual to individual, making comparisons between samples difficult. In this report, we study the association of pH and several pre- and postmortem factors on the RNA yields from 89 postmortem human occipital cortices. Glyceraldehyde phosphate dehydrogenase (GAPdH) mRNA levels were measured by RT-PCR. A major variant in this method is the priming used in the reverse transcription reaction. Three different methods of reverse transcription were performed and the resultant levels of products compared against the pre- and postmortem factors and pH. The levels of GAPdH correlated significantly to pH and pH itself to the rapidity of death (ROD) (agonal state) indicating that premortem factors may play the greatest role in determining postmortem RNA levels. The three methods of priming showed different sensitivities, most notably that oligo dT priming alone is vulnerable to long freezer intervals (FI). We conclude that premortem factors are the major affectors of RNA levels variations and that the polyA tail region of the molecule appears to be adversely affected by extended freezer storage.

Original languageEnglish (US)
Pages (from-to)83-92
Number of pages10
JournalJournal of Neuroscience Methods
Volume77
Issue number1
DOIs
StatePublished - Nov 7 1997

Fingerprint

Reverse Transcription
Multivariate Analysis
RNA
Glyceraldehyde
Polymerase Chain Reaction
Brain
Oxidoreductases
Phosphates
Occipital Lobe
Tail
Gene Expression
Messenger RNA

Keywords

  • Glyceraldehyde phosphate dehydrogenase
  • PH
  • Postmortem human brain
  • Psychiatric disorders
  • RNA
  • RT-PCR

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Multivariate analysis of RNA levels from postmortem human brains as measured by three different methods of RT-PCR. / Johnston, Nancy L.; Cerevnak, Juraj; Shore, Andrew D.; Fuller Torrey, E.; Yolken, Robert H.

In: Journal of Neuroscience Methods, Vol. 77, No. 1, 07.11.1997, p. 83-92.

Research output: Contribution to journalArticle

Johnston, Nancy L. ; Cerevnak, Juraj ; Shore, Andrew D. ; Fuller Torrey, E. ; Yolken, Robert H. / Multivariate analysis of RNA levels from postmortem human brains as measured by three different methods of RT-PCR. In: Journal of Neuroscience Methods. 1997 ; Vol. 77, No. 1. pp. 83-92.
@article{2ac20ee4d07c45e1b003e903dd339b4f,
title = "Multivariate analysis of RNA levels from postmortem human brains as measured by three different methods of RT-PCR",
abstract = "The analysis of RNA from postmortem human brain tissue by reverse transcription-polymerase chain reaction (RT-PCR) provides a practical method to measure both normal and abnormal brain gene expression. A major limitation in using human material is that yields can vary dramatically from individual to individual, making comparisons between samples difficult. In this report, we study the association of pH and several pre- and postmortem factors on the RNA yields from 89 postmortem human occipital cortices. Glyceraldehyde phosphate dehydrogenase (GAPdH) mRNA levels were measured by RT-PCR. A major variant in this method is the priming used in the reverse transcription reaction. Three different methods of reverse transcription were performed and the resultant levels of products compared against the pre- and postmortem factors and pH. The levels of GAPdH correlated significantly to pH and pH itself to the rapidity of death (ROD) (agonal state) indicating that premortem factors may play the greatest role in determining postmortem RNA levels. The three methods of priming showed different sensitivities, most notably that oligo dT priming alone is vulnerable to long freezer intervals (FI). We conclude that premortem factors are the major affectors of RNA levels variations and that the polyA tail region of the molecule appears to be adversely affected by extended freezer storage.",
keywords = "Glyceraldehyde phosphate dehydrogenase, PH, Postmortem human brain, Psychiatric disorders, RNA, RT-PCR",
author = "Johnston, {Nancy L.} and Juraj Cerevnak and Shore, {Andrew D.} and {Fuller Torrey}, E. and Yolken, {Robert H}",
year = "1997",
month = "11",
day = "7",
doi = "10.1016/S0165-0270(97)00115-5",
language = "English (US)",
volume = "77",
pages = "83--92",
journal = "Journal of Neuroscience Methods",
issn = "0165-0270",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Multivariate analysis of RNA levels from postmortem human brains as measured by three different methods of RT-PCR

AU - Johnston, Nancy L.

AU - Cerevnak, Juraj

AU - Shore, Andrew D.

AU - Fuller Torrey, E.

AU - Yolken, Robert H

PY - 1997/11/7

Y1 - 1997/11/7

N2 - The analysis of RNA from postmortem human brain tissue by reverse transcription-polymerase chain reaction (RT-PCR) provides a practical method to measure both normal and abnormal brain gene expression. A major limitation in using human material is that yields can vary dramatically from individual to individual, making comparisons between samples difficult. In this report, we study the association of pH and several pre- and postmortem factors on the RNA yields from 89 postmortem human occipital cortices. Glyceraldehyde phosphate dehydrogenase (GAPdH) mRNA levels were measured by RT-PCR. A major variant in this method is the priming used in the reverse transcription reaction. Three different methods of reverse transcription were performed and the resultant levels of products compared against the pre- and postmortem factors and pH. The levels of GAPdH correlated significantly to pH and pH itself to the rapidity of death (ROD) (agonal state) indicating that premortem factors may play the greatest role in determining postmortem RNA levels. The three methods of priming showed different sensitivities, most notably that oligo dT priming alone is vulnerable to long freezer intervals (FI). We conclude that premortem factors are the major affectors of RNA levels variations and that the polyA tail region of the molecule appears to be adversely affected by extended freezer storage.

AB - The analysis of RNA from postmortem human brain tissue by reverse transcription-polymerase chain reaction (RT-PCR) provides a practical method to measure both normal and abnormal brain gene expression. A major limitation in using human material is that yields can vary dramatically from individual to individual, making comparisons between samples difficult. In this report, we study the association of pH and several pre- and postmortem factors on the RNA yields from 89 postmortem human occipital cortices. Glyceraldehyde phosphate dehydrogenase (GAPdH) mRNA levels were measured by RT-PCR. A major variant in this method is the priming used in the reverse transcription reaction. Three different methods of reverse transcription were performed and the resultant levels of products compared against the pre- and postmortem factors and pH. The levels of GAPdH correlated significantly to pH and pH itself to the rapidity of death (ROD) (agonal state) indicating that premortem factors may play the greatest role in determining postmortem RNA levels. The three methods of priming showed different sensitivities, most notably that oligo dT priming alone is vulnerable to long freezer intervals (FI). We conclude that premortem factors are the major affectors of RNA levels variations and that the polyA tail region of the molecule appears to be adversely affected by extended freezer storage.

KW - Glyceraldehyde phosphate dehydrogenase

KW - PH

KW - Postmortem human brain

KW - Psychiatric disorders

KW - RNA

KW - RT-PCR

UR - http://www.scopus.com/inward/record.url?scp=0030668729&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030668729&partnerID=8YFLogxK

U2 - 10.1016/S0165-0270(97)00115-5

DO - 10.1016/S0165-0270(97)00115-5

M3 - Article

C2 - 9402561

AN - SCOPUS:0030668729

VL - 77

SP - 83

EP - 92

JO - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

SN - 0165-0270

IS - 1

ER -