Cellular localization of messenger RNA for beta-1 and beta-2 adrenergic receptors in rat brain: An in situ hybridization study

doi:10.1016/0306-4522(93)90148-9

Neuroscience

Volume 56, Issue 4, October 1993, Pages 1023-1039

https://doi.org/10.1016/0306-4522(93)90148-9 Get rights and content

Abstract

Selective,³⁵S-labeled, oligonucleotide probes were designed from sequences of the rat beta-1 and beta-2 adrenoceptor messenger RNAs for use in in situ hybridization experiments on sections of unfixed rat brain and spinal cord. After hybridized sections were exposed to film or dipped in autoradiographic emulsion, specific and selective labeling patterns characteristic for each receptor messenger RNA and region of the central nervous system were observed. For example, labeling for beta-1 messenger RNA was found in the anterior olfactory nucleus, cerebral cortex, lateral intermediate septal nucleus, reticular thalamic nucleus, oculomotor complex, vestibular nuclei, deep cerebellar nuclei, trapezoid nucleus, abducens nucleus, ventrolateral pontine and medullary reticular formations, the intermediate gray matter of the spinal cord and in the pineal gland, while beta-2 messenger RNA labeling was strongest in the olfactory bulb, piriform cortex, hippocampal formation, thalamic intralaminar nuclei and cerebellar cortex. In some of these regions the beta-1 labeling seemed mainly confined to the cell nucleus. Whether or not this apparently nuclear labeling is specific, i.e. indicates synthesis of beta-1 receptor, remains to be established. However, all labeling patterns described disappeared when excess unlabeled probes were added to their respective radiolabeled probes or when sense probes were employed.

Since the in situ method labels only cell bodies that produce the messenger RNA for these two beta receptor subtypes, a comparison between these maps and those of past autoradiographic studies mapping the location of central beta receptors using drugs as radioligands may produce further insights regarding the pre- and postsynaptic localization of these receptors in the various parts of the central nervous system circuitry.

References (96)

AokiC. et al.
Ultrastructural localization of β-adrenergic receptor-like immunoreactivity in the cortex and neostriatum of rat brain
Brain Res.
(1987)
AokiC. et al.
Cytoplasmic loop of β-adrenergic receptors: synaptic and intracellular localization and relation to catecholaminergic neurons in the nuclei of the solitary tracts
Brain Res.
(1989)
ArakiT. et al.
Localization of GABA_A-receptor β₂-subunit mRNA-containing neurons in the rat central nervous system
Neuroscience
(1992)
ArangoV. et al.
Demonstration of high- and low-affinity β-adrenergie receptors in slide-mounted section of rat and human brain
Brain Res.
(1990)
BloomF.E. et al.
Studies on the norepinephrine-containing afferents to Purkinje cells of the rat cerebellum.I. Localization of the fibers and their synapses
Brain Res.
(1971)
ChangR.S. et al.
Neurotransmitter receptor localizations: brain lesion induced alterations in benzodiazepine, GABA, beta-adrenergic and histamine H 1-receptor binding
Brain Res.
(1980)
ChidaK. et al.
Lesions of rostral ventrolateral medulla abolish some cardio- and cerebrovascular components of the cerebellar fastigal pressor and depressor responses
Brain Res.
(1990)
ErnsbergerP. et al.
Astrocytes cultured from specific brain regions differ in their expression of adrenergic binding sites
Brain Res.
(1990)
GraybielA.M. et al.
Fiber connections of the basal ganglia
Prog. Brain Res.
(1979)
GrimmL.J. et al.
Chronic reserpine administration selectively up-regulates β₁ and α_1b-adrenergic receptors in rat brain: an autoradiographic study
Neuroscience
(1992)

HerkenhamM.

Mismatches between neurotransmitter and receptor localizations in brain: observations and implications

Neuroscience

(1987)

HokfeltT. et al.

Immunohistochemical evidence for the existence of adrenaline neurons in the rat brain

Brain Res.

(1974)

KuharM.J.

The mismatch problem in receptor mapping studies

Trends Neurosci.

(1985)

LjungdahlA. et al.

Distribution of substance P-like immunoreactivity in the centra nervous system of the rat—I. Cell bodies and nerve terminals

Neuroscience

(1978)

LomasneyJ.W. et al.

Molecular cloning and expression of the cDNA for the α_1A-adrenergic receptor. The gene for which is located on human chromosome 5

J. biol. Chem.

(1991)

MachidaC.A. et al.

Molecular cloning and expression of the rat β 1-adrenergic receptor gene

J. biol. Chem.

(1990)

MinnemanK.P. et al.

Selective changes in the density of β₁-adrenergic receptors in rat striatum following chronic drug treatment and adrenalectomy

Brain Res.

(1982)

NahorskiS.R.

Heterogeneity of cerebral β-adrenoceptor binding sites in various vertebrate species

Eur J. Pharmac.

(1978)

NahorskiS.R. et al.

Loss of beta-adrenoceptor binding sites in rat striatum following kainic acid lesions

Eur. J. Pharmac.

(1979)

NicholasA.P. et al.

Initial observations on the localization of mRNA for α and β adrenergic receptors in brain and peripheral tissues of rat using in situ hybridization

Molec. cell. Neurosci.

(1991)

O'DonohueT.L. et al.

Biochemical mapping of the noradrenergic ventral bundle projection sites: evidence for a noradrenergic-dopaminergic interaction

Brain Res.

(1979)

OkeA. et al.

Dopamine and norepinephrine enhancement in discrete rat brain regions following neonatal 6-hydroxydopamine treatment

Brain Res.

(1978)

PalaciosJ.M. et al.

Beta-adrenergic receptor localization in rat brain by light microscopic autoradiography

Neurochem. Int.

(1982)

PazosA. et al.

β-Adrenoceptor blocking agents recognize a subpopulation ofserotonin receptors in brain

Brain Res.

(1985)

PazosA. et al.

β-Adrenoceptor subtypes in the human brain: autoradiographic localization

Brain Res.

(1985)

PittmanR.N. et al.

Ontogeny of β₁- and β₂-adrenergic receptors in rat cerebellum and cerebral cortex

Brain Res.

(1980)

PompeianoO.

Relationship of noradrenergic locus coeruleus neurons to vestibulospinal reflexes

ReisineT.D. et al.

A role for striatal beta-adrenergic receptors in the regulation ofdopamine release

Brain Res.

(1982)

ReisineT.D. et al.

The localization of receptor binding sites in the substantia nigra and striatum of the rat

Brain Res.

(1979)

RossR.A. et al.

Effects of lesions of locus coeruleus on regional distribution of dopamine-β-hydroxylase activity in rat brain

Brain Res.

(1974)

StoneE.A. et al.

Further evidence for a glial localization of rat cortical β-adrenoceptors: studies of in vivo cyclic AMP responses to catecholamines

Brain Res.

(1991)

StoneE.A. et al.

Glial localization of adenylate-cyclase-coupled beta-adrenoceptors in rat forebrain slices

Brain Res.

(1990)

U'PrichardD.C. et al.

Differential supersensitivity of β-receptor subtypes in rat cortex and cerebellum after central noradrenergic denervation

Life Sci.

(1980)

VersteegD.H.G. et al.

Regional concentrations of noradrenaline and dopamine in rat brain

Brain Res.

(1976)

VoigtM.M. et al.

The rat α₂,-C4 adrenergic receptor gene encodes a novel pharmacological subtype

Fedn. Eur. biochem. Socs Lett.

(1991)

WolfeB.B. et al.

Selective increases in the density of cerebellar β₁-adrenergic receptors

Brain Res.

(1982)

YoungW.S. et al.

Vasopressin and oxytocin mRNAs in adrenalectomized and Brattleboro rats: analysis by quantitative in situ hybridization histochemistry

Molec. Brain Res.

(1986)

ZahniserN.R. et al.

Persistence of β-adrenergic receptors in rat striatum following kainic acid administration

Brain Res.

(1979)

AhlquistR.P.

A study of the adrenotropic receptors

Am. J. Physiol.

(1948)

AlexanderR.W. et al.

Direct identification and characterisation of β-adrenergic receptors in rat brain

Nature

(1975)

AokiC.

β-Adrenergic receptors: astrocytic localization in the adult visual cortex and their relation to catecholamine axon terminals as revealed by electron microscopic immunocytochemistry

J. Neurosci.

(1992)

AokiC. et al.

Ultrastructural immunocytochemical evidence for presynaptic localization of β-adrenergic receptors in the striatum and cerebral cortex of rat brain

Ann. N.Y. Acad. Sci.

(1990)

ArangoV. et al.

Autoradiographic demonstration of increased serotonin 5-HT₂ and β-adrenergic receptor binding sites in the brains of suicide victims

Arch. gen. Psychol.

(1990)

BriceA. et al.

Complete sequence of a cDNA encoding an active rat choline acetyltransferase: a tool to investigate the plasticity of cholinergic phenotype expression

J. Neurosci. Res.

(1989)

BucklandP.R. et al.

Primary structure of the rat beta-2 adrenergic receptor gene

Nucleic Acids Res.

(1990)

BylundD.B. et al.

Beta adrenergic receptor binding in membrane preparations from mammalian brain

Molec. Pharmac.

(1976)

CoonsA.H.

Fluorescent antibody methods

DagerlindÅ. et al.

Sensitive mRNA detection using unfixed tissue: combined radioactive and non-radioactive in situ hybridization histochemistry

Histochemistry

(1992)

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Cellular localization of messenger RNA for beta-1 and beta-2 adrenergic receptors in rat brain: An in situ hybridization study

Abstract

Brain Res.

Brain Res.

Neuroscience

Brain Res.

Brain Res.

Brain Res.

Brain Res.

Brain Res.

Prog. Brain Res.

Neuroscience

Neuroscience

Brain Res.

Trends Neurosci.

Neuroscience

J. biol. Chem.

J. biol. Chem.

Brain Res.

Eur J. Pharmac.

Eur. J. Pharmac.

Molec. cell. Neurosci.

Brain Res.

Brain Res.

Neurochem. Int.

Brain Res.

Brain Res.

Brain Res.

Brain Res.

Brain Res.

Brain Res.

Brain Res.

Brain Res.

Life Sci.

Brain Res.

Fedn. Eur. biochem. Socs Lett.

Brain Res.

Molec. Brain Res.

Brain Res.

A study of the adrenotropic receptors

Am. J. Physiol.

Direct identification and characterisation of β-adrenergic receptors in rat brain

Nature

β-Adrenergic receptors: astrocytic localization in the adult visual cortex and their relation to catecholamine axon terminals as revealed by electron microscopic immunocytochemistry

J. Neurosci.

Ultrastructural immunocytochemical evidence for presynaptic localization of β-adrenergic receptors in the striatum and cerebral cortex of rat brain

Ann. N.Y. Acad. Sci.

Autoradiographic demonstration of increased serotonin 5-HT2 and β-adrenergic receptor binding sites in the brains of suicide victims

Arch. gen. Psychol.

Complete sequence of a cDNA encoding an active rat choline acetyltransferase: a tool to investigate the plasticity of cholinergic phenotype expression

J. Neurosci. Res.

Primary structure of the rat beta-2 adrenergic receptor gene

Nucleic Acids Res.

Beta adrenergic receptor binding in membrane preparations from mammalian brain

Molec. Pharmac.

Fluorescent antibody methods

Sensitive mRNA detection using unfixed tissue: combined radioactive and non-radioactive in situ hybridization histochemistry

Histochemistry

Autoradiographic demonstration of increased serotonin 5-HT₂ and β-adrenergic receptor binding sites in the brains of suicide victims