Coupled reverse transcription-polymerase chain reaction (RT-PCR) technique is comparative, quantitative, and rapid

Uses in alcohol research involving low abundance mRNA species such as hypothalamic LHRH and GRF

Mark Kelley, J. K. Jurgens, J. Tentler, N. V. Emanuele, S. E. Blutt, M. A. Emanuele

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

The measurement of alterations in low abundance mRNAs such as the hypothalamic hormones luteinizing hormone-releasing hormone (LHRH) and growth hormone-releasing hormone (GHRH or GRF) from individual hypothalamic tissues in rats has previously been difficult and usually required either isolation of poly(A) mRNA or the pooling of numerous animals to obtain a reasonable signal on Northern blots. Although more sensitive detection methods exist, such as the use of RNA probes or solution hybridization (RNase protection), we have found the most reliable, sensitive, rapid, and accurate method is the reverse transcription-polymerase chain reaction (RT-PCR) using histone H3.3 as an internal control for both steps of this procedure. H3.3 is a cell-cycle independent and constitutively expressed gene in all tissues. We have developed an RT-PCR assay for LHRH and GRF mRNA quantitation and comparative analysis for hypothalamic and extrahypothalamic brain tissues and present the use of RT-PCR for LHRH quantitation in ethanol (EtOH) studies.

Original languageEnglish (US)
Pages (from-to)185-189
Number of pages5
JournalAlcohol
Volume10
Issue number3
DOIs
StatePublished - 1993
Externally publishedYes

Fingerprint

Hypothalamic Hormones
Polymerase chain reaction
Transcription
Gonadotropin-Releasing Hormone
Reverse Transcription
alcohol
Alcohols
Tissue
Polymerase Chain Reaction
Messenger RNA
Research
RNA Probes
Growth Hormone-Releasing Hormone
Ribonucleases
Northern Blotting
Histones
Rats
social isolation
Assays
Brain

Keywords

  • (GRF)
  • (LHRH)
  • (RT-PCR) technique
  • Coupled reverse transcription-polymerase chain reaction
  • Growth hormone-releasing hormone
  • Luteinizing hormone-releasing hormone

ASJC Scopus subject areas

  • Biochemistry
  • Medicine(all)
  • Neuroscience(all)
  • Behavioral Neuroscience
  • Toxicology
  • Health(social science)

Cite this

Coupled reverse transcription-polymerase chain reaction (RT-PCR) technique is comparative, quantitative, and rapid : Uses in alcohol research involving low abundance mRNA species such as hypothalamic LHRH and GRF. / Kelley, Mark; Jurgens, J. K.; Tentler, J.; Emanuele, N. V.; Blutt, S. E.; Emanuele, M. A.

In: Alcohol, Vol. 10, No. 3, 1993, p. 185-189.

Research output: Contribution to journalArticle

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