Simultaneous visualization of two cellular mRNA species in individual neurons by use of a new double in Situ hybridization method

Daniel L. Marks; Jeffrey N. Wiemann; Kimberly A. Burton; Karin L. Lent; Donald K. Clifton; Robert A. Steiner

doi:10.1016/1044-7431(92)90051-3

Simultaneous visualization of two cellular mRNA species in individual neurons by use of a new double in Situ hybridization method

Daniel L. Marks, Jeffrey N. Wiemann, Kimberly A. Burton, Karin L. Lent, Donald K. Clifton, Robert A. Steiner

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68 Scopus citations

Abstract

We present a new method for the simultaneous detection of two mRNA species within individual neurons. The technique involves the use of radio-labeled and digoxigenin-labeled cRNA probes, the application of which confers a high specificity and sensitivity to the in situ hybridization analysis. We demonstrate the use of this method by illustrating the coexpression of preprogonadotropin-releasing hormone (GnRH) mRNA and preprogalanin mRNA in neurons in the rat forebrain and report a distinct sexual dimorphism in galanin gene expression by GnRH neurons. Coupling this technology with semi-quantitative analysis of the mRNA species hybridized with the isotopically labeled mRNA would permit studies of gene regulation in individual cells among the heterogeneous populations of the brain.

Original language	English (US)
Pages (from-to)	395-405
Number of pages	11
Journal	Molecular and Cellular Neuroscience
Volume	3
Issue number	5
DOIs	https://doi.org/10.1016/1044-7431(92)90051-3
State	Published - Oct 1992
Externally published	Yes

ASJC Scopus subject areas

Molecular Biology
Cellular and Molecular Neuroscience
Cell Biology

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10.1016/1044-7431(92)90051-3

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@article{f938fc7a2f8d4b1aaf2dcce14bdc7af1,

title = "Simultaneous visualization of two cellular mRNA species in individual neurons by use of a new double in Situ hybridization method",

abstract = "We present a new method for the simultaneous detection of two mRNA species within individual neurons. The technique involves the use of radio-labeled and digoxigenin-labeled cRNA probes, the application of which confers a high specificity and sensitivity to the in situ hybridization analysis. We demonstrate the use of this method by illustrating the coexpression of preprogonadotropin-releasing hormone (GnRH) mRNA and preprogalanin mRNA in neurons in the rat forebrain and report a distinct sexual dimorphism in galanin gene expression by GnRH neurons. Coupling this technology with semi-quantitative analysis of the mRNA species hybridized with the isotopically labeled mRNA would permit studies of gene regulation in individual cells among the heterogeneous populations of the brain.",

author = "Marks, {Daniel L.} and Wiemann, {Jeffrey N.} and Burton, {Kimberly A.} and Lent, {Karin L.} and Clifton, {Donald K.} and Steiner, {Robert A.}",

note = "Funding Information: This work was supported by USPHS (NIH) Grants HD-12625 and HD-12629. The authors thank Emilia Kabigting for her excellent technical support and Dr. Maria E. Vrontakis for her assistance with the galanin cDNA. We are also indebted to Pamela Kolb, Dr. Margaret Miller, and Terre Poppe for their advice and assistance in the production of this manuscript.",

year = "1992",

month = oct,

doi = "10.1016/1044-7431(92)90051-3",

language = "English (US)",

volume = "3",

pages = "395--405",

journal = "Molecular and Cellular Neuroscience",

issn = "1044-7431",

publisher = "Academic Press Inc.",

number = "5",

}

TY - JOUR

T1 - Simultaneous visualization of two cellular mRNA species in individual neurons by use of a new double in Situ hybridization method

AU - Marks, Daniel L.

AU - Wiemann, Jeffrey N.

AU - Burton, Kimberly A.

AU - Lent, Karin L.

AU - Clifton, Donald K.

AU - Steiner, Robert A.

N1 - Funding Information: This work was supported by USPHS (NIH) Grants HD-12625 and HD-12629. The authors thank Emilia Kabigting for her excellent technical support and Dr. Maria E. Vrontakis for her assistance with the galanin cDNA. We are also indebted to Pamela Kolb, Dr. Margaret Miller, and Terre Poppe for their advice and assistance in the production of this manuscript.

PY - 1992/10

Y1 - 1992/10

N2 - We present a new method for the simultaneous detection of two mRNA species within individual neurons. The technique involves the use of radio-labeled and digoxigenin-labeled cRNA probes, the application of which confers a high specificity and sensitivity to the in situ hybridization analysis. We demonstrate the use of this method by illustrating the coexpression of preprogonadotropin-releasing hormone (GnRH) mRNA and preprogalanin mRNA in neurons in the rat forebrain and report a distinct sexual dimorphism in galanin gene expression by GnRH neurons. Coupling this technology with semi-quantitative analysis of the mRNA species hybridized with the isotopically labeled mRNA would permit studies of gene regulation in individual cells among the heterogeneous populations of the brain.

AB - We present a new method for the simultaneous detection of two mRNA species within individual neurons. The technique involves the use of radio-labeled and digoxigenin-labeled cRNA probes, the application of which confers a high specificity and sensitivity to the in situ hybridization analysis. We demonstrate the use of this method by illustrating the coexpression of preprogonadotropin-releasing hormone (GnRH) mRNA and preprogalanin mRNA in neurons in the rat forebrain and report a distinct sexual dimorphism in galanin gene expression by GnRH neurons. Coupling this technology with semi-quantitative analysis of the mRNA species hybridized with the isotopically labeled mRNA would permit studies of gene regulation in individual cells among the heterogeneous populations of the brain.

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AN - SCOPUS:0026743005

SN - 1044-7431

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EP - 405

JO - Molecular and Cellular Neuroscience

JF - Molecular and Cellular Neuroscience

IS - 5

ER -

Simultaneous visualization of two cellular mRNA species in individual neurons by use of a new double in Situ hybridization method

Abstract

ASJC Scopus subject areas

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