Differential expression of the α1C adrenergic receptor subtype in rat tissues

Alonso‐Llamazares, Ana; Zamanillo, Daniel; Fernández, A.; Chinchetru, Miguel A.; Calvo, Pedro

doi:10.1097/00001756-199309000-00014

Cited by 12 publications

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“…This conclusion is now supported by high correlative associations between the affinity of ligands for the cloned bovine olc-adrenoceptor and their binding affinity for x1A-adrenoceptors in submaxillary gland of rat as well as other rat tissues (values abstracted from the literature). Radioligand binding studies reported recently by others (Faure et al, 1994;Forray et al, 1994;Laz et al, 1994) also support the equivalency of the MIA-adrenoceptor with the mic-adrenoceptor, and considerable molecular biological evidence now exists for the presence of the mlcadrenoceptor gene in rat (Alonso-Llamazares et al, 1993;Laz et al, 1994;Rokosh et al, 1994;Faure et al, 1994;Price et al, 1994) despite an initial negative report (Schwinn & Lomasney, 1992).…”

Section: Discussionmentioning

confidence: 84%

Functional evidence equating the pharmacologically‐defined α_1A‐ and cloned α_1C‐adrenoceptor: studies in the isolated perfused kidney of rat

Blue¹,

Bonhaus²,

Ford³

et al. 1995

British J Pharmacology

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1 The present study characterizes and classifies ax-adrenoceptor-mediated vasoconstriction in the isolated perfused kidney of rat using quantitative receptor pharmacology and compares the results to radioligand binding studies (made in cloned al-adrenoceptor subtypes, native aIA-adrenoceptors in submaxillary gland of rat, and MIA-adrenoceptors in several other tissues of rat).2 Concentration-effect curves to noradrenaline in the presence of 5-methyl-urapidil were biphasic, indicating al-adrenoceptor heterogeneity. The al-adrenoceptor subtype mediating the first phase (low affinity for 5-methyl-urapidil) could not be 'isolated' for detailed pharmacological characterization but was defined by a sensitivity to inhibition by chloroethylclonidine and an inability of methoxamine to activate the site. Additionally, vasoconstriction mediated by this aI-adrenoceptor subtype or subtypes was abolished by nitrendipine (1 JLM), thereby allowing characterization of the second, high affinity site for 5-methyl-urapidil.3 The following antagonists interacted competitively with noradrenaline at the al-adrenoceptor for which 5-methyl-urapidil exhibits high affinity (pKB value): WB 4101 (10.3)>prazosin (9.5)--HV 723 (9.3) -5-methyl-urapidil (9.2)>phentolamine (8.6)>spiperone (pA2 = 8.1)toxymetazoline (7.9). In contrast, insurmountable antagonism was seen with S(+)-and R(-)-niguldipine, the S(+ )-isomer being approximately 30 fold more potent than the R(-)-isomer. Receptor protection experiments indicated that S(+)-niguldipine interacted directly with a,-adrenoceptors. Dehydroniguldipine acted as a competitive antagonist (pKB =9.0). Thus, the results with antagonists define the aI-adrenoceptor as an cXIA-adrenoceptor. 4 An agonist 'fingerprint' was constructed in the presence of nitrendipine to define further the aIA-adrenoceptor. The following order and relativity of agonist potency was obtained: cirazoline (1)t adrenaline (2)>noradrenaline (5) 6 The present study demonstrates that the MlA-adrenoceptor is the predominant a-adrenoceptor subtype mediating vasoconstrictor responses to exogenously administered noradrenaline in the isolated perfused kidney of rat. More importantly, OIA-adrenoceptors mediating vasoconstrictor responses to noradrenaline exhibited a pharmacological equivalency to the cloned bovine xic-adrenoceptor. Thus, definitive functional pharmacological data are provided for equating the two receptors and support results derived recently from molecular and radioligand binding studies.

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Section: Discussionmentioning

confidence: 84%

Functional evidence equating the pharmacologically‐defined α_1A‐ and cloned α_1C‐adrenoceptor: studies in the isolated perfused kidney of rat

Blue¹,

Bonhaus²,

Ford³

et al. 1995

British J Pharmacology

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“…In this case, primers were derived from sequences of the fifth and sixth transmembrane domains of the bovine a, a -adrenergic receptor (Schwinn et al, 1990) . The nucleotide sequence of the amplified fragment (310 bp) had 95% identity with the sequence of the homologous rat subtype (Alonso-Llamazares et al ., 1993), and the deduced amino acid sequences were identical except in two positions . RT-PCR assays were carried out to examine the distribution of the a,-adrenergic receptor subtypes in mouse tissues (Fig .…”

Section: Resultsmentioning

confidence: 99%

“…The cDNA fragment was amplified under the following conditions : 3 min at 94°C, followed by 1 min at 94°C, 1 min at 60°C, and 30 s at 72°C, for 40 cycles, using 1 pl of the cDNA solution and 1 .25 units of Taq DNA polymerase (Boehringer Mannheim) . The products of the PCR procedures were resolved on 1 % lowmelting point agarose, digested with EcoRI, cloned into the pUC 119 vector, and sequenced (Alonso-Llamazares et al ., 1993) . The experimental protocols involving laboratory animals were approved by the Institutional Review Committee of this university and meet the guidelines of our responsible governmental agency .…”

Section: Methodsmentioning

confidence: 99%

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Molecular Cloning of α_1d‐Adrenergic Receptor and Tissue Distribution of Three α₁‐Adrenergic Receptor Subtypes in Mouse

Alonso‐Llamazares

Zamanillo

Casanova

et al. 1995

Journal of Neurochemistry

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A partial cDNA encoding most of the third intracellular loop of the mouse a, d -adrenergic receptor subtype was amplified from hippocampus by reverse transcription-polymerase chain reaction (RT-PCR) using degenerate oligodeoxynucleotide primers . This DNA fragment was used as a probe to isolate an a, d -adrenergic receptor cDNA from a mouse brain cDNA library . The deduced amino acid sequence encodes a potential protein of 562 amino acids, and northern hybridization of poly(A)+ RNA isolated from mouse brain detected a single 3 .0-kb transcript . Partial cDNA fragments of the alband a, a -adrenergic receptor subtypes were also amplified from mouse brain and sequenced . Analysis of the mRNA expression by RT-PCR indicated that the a,-adrenergic receptors are widely distributed in mouse tissues . The a,d subtype is expressed in brain areas such as hippocampus, striatum, and brainstem and also in many extracerebral tissues, such as lung, liver, heart, kidney, and spleen . The a,a subtype is also expressed in many tissues, whereas the alb subtype has a more restricted expression, with high levels in striatum, brainstem, and diencephalus . Key Words : a,d -Adrenergic receptor-Mouse-cDNA cloning-Tissue distribution -a,-Adrenergic receptor subtypes . J. Neurochem . 65, 2387Neurochem . 65, -2392Neurochem . 65, (1995 .The adrenergic receptors are members of a large family of membrane proteins that are coupled to guanine nucleotide-binding proteins (G proteins) . They are currently divided into three types (a,, a2 , and ß), each having several receptor members (Lomasney et al., 1991a ;Bylund, 1992) . Radioligand binding data with the a,-adrenergic receptor agonists oxymetazoline and methoxamine and the antagonists WB4101 and phentolamine indicated the existence of two a,-adrenergic receptor subtypes in rat cerebral cortex, which were named a1A and a,B (Battaglia et al., 1983 ; Morrow and Creese, 1986;Han et al., 1987) .Several cDNAs and genes encoding a t-adrenergic receptor subtypes from various sources, and initially named a]A/alD, a,B , and a lc , have been isolated by

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“…Expression of mRNA for each of the three cloned subtypes can be detected in multiple rat (Alonso-Llamazares et al 1993;Faure et al 1994;Perez et al 1994;Price et al 1994a;Rokosh et al 1994) and human tissues (Hirasawa et al 1993;Price et al 1993Price et al , 1994bWeinberg et al 1994) using appropriately sensitive techniques including RNAse protection assays or reverse-transcriptase polymerase chain reaction. Corresponding α 1A -and α 1B -adrenoceptor proteins have been detected in many of those tissues by numerous radioligand binding studies in rats Han and Minneman 1991;Michel et al 1993aMichel et al , 1994a and humans Yamada et al 1994;Garcia-Sainz et al 1995;Michel et al 1996).…”

Section: Introductionmentioning

confidence: 99%

Is α1D-adrenoreptor protein detactable in rat tissues?

Yang

Verfürth

Büscher³

et al. 1997

Naunyn-Schmiedeberg's Arch Pharmacol

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We have used the alpha1D-adrenoceptor selective antagonist, BMY 7378, the alpha1D-selective agonists, adrenaline and phenylephrine, the alpha1A-selective antagonists, (+)-niguldipine, SB 216469 and WB4101, and the non-subtype-selective alpha1-adrenoceptor antagonist, nemonapride, to investigate the presence of alpha1D-adrenoceptors in rat tissues at the protein level. Radioligand binding studies using [3H]prazosin as the radioligand were performed in three tissues containing alpha1D-adrenoceptor mRNA, spleen, cerebral cortex and kidney, and in comparison in one tissue not containing alpha1D-adrenoceptor mRNA, liver. Cerebral cortex and kidney were also studied upon alpha1B-adrenoceptor inactivation by chloroethylclonidine treatment (10 microM, 30 min, 37 degrees C). Experiments with cloned rat alpha1-adrenoceptor subtypes transiently expressed in COS cells confirmed the known selectivity of the investigated drugs for alpha1-adrenoceptor subtypes or the lack thereof of nemonapride. Accordingly nemonapride had steep and monophasic competition curves in all native and chloroethylclonidine-treated tissues. BMY 7378 also had steep and monophasic competition curves and low affinity in all native tissues. In contrast, adrenaline and phenylephrine (in the presence of 100 microM GTP) had monophasic competition curves of low affinity in liver and spleen but biphasic competition curves in cerebral cortex and kidney. Following chloroethylclonidine treatment competition curves for adrenaline, phenylephrine, (+)-niguldipine, SB 216469 and WB 4101 remained biphasic in cerebral cortex and kidney while those for nemonapride remained monophasic. We conclude that alpha1D-adrenoceptors are not readily detectable at the protein level in a variety of rat tissues where their mRNA is expressed. The biphasic competition curves of some agonists and antagonists in chloroethylclonidine-treated rat tissues do not represent alpha1D-adrenoceptors and are not readily explained by the present alpha1A/alpha1B/alpha1D-adrenoceptor classification.

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Differential expression of the α1C adrenergic receptor subtype in rat tissues

Cited by 12 publications

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Functional evidence equating the pharmacologically‐defined α_1A‐ and cloned α_1C‐adrenoceptor: studies in the isolated perfused kidney of rat

Functional evidence equating the pharmacologically‐defined α_1A‐ and cloned α_1C‐adrenoceptor: studies in the isolated perfused kidney of rat

Molecular Cloning of α_1d‐Adrenergic Receptor and Tissue Distribution of Three α₁‐Adrenergic Receptor Subtypes in Mouse

Is α1D-adrenoreptor protein detactable in rat tissues?

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Differential expression of the α1C adrenergic receptor subtype in rat tissues

Cited by 12 publications

References 0 publications

Functional evidence equating the pharmacologically‐defined α1A‐ and cloned α1C‐adrenoceptor: studies in the isolated perfused kidney of rat

Functional evidence equating the pharmacologically‐defined α1A‐ and cloned α1C‐adrenoceptor: studies in the isolated perfused kidney of rat

Molecular Cloning of α1d‐Adrenergic Receptor and Tissue Distribution of Three α1‐Adrenergic Receptor Subtypes in Mouse

Is α1D-adrenoreptor protein detactable in rat tissues?

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Functional evidence equating the pharmacologically‐defined α_1A‐ and cloned α_1C‐adrenoceptor: studies in the isolated perfused kidney of rat

Functional evidence equating the pharmacologically‐defined α_1A‐ and cloned α_1C‐adrenoceptor: studies in the isolated perfused kidney of rat

Molecular Cloning of α_1d‐Adrenergic Receptor and Tissue Distribution of Three α₁‐Adrenergic Receptor Subtypes in Mouse