Cloning and Expression of the Thyrotropin-releasing Hormone Receptor from GH3 Rat Anterior Pituitary Cells

Overview

Journal Biochem J

Specialty Biochemistry

Date 1992 Jun 15

PMID 1377915

Citations 17

Authors

P de la Pena

L M Delgado

D del Camino

F Barros

Affiliations

Soon will be listed here.

Abstract

Functional thyrotropin-releasing hormone (TRH) receptors have been expressed in Xenopus laevis oocytes following the microinjection of total and poly(A)+ RNA from GH3 rat anterior pituitary tumour cells. Under voltage-clamp conditions, application of the peptide induced a biphasic Ca(2+)-dependent chloride current. The amplitude of the initial, fast, component of the response was dependent on the concentration of the hormone and on the amount of mRNA injected. Size fractionation of poly(A)+ RNA on a continuous sucrose gradient and Northern blot analysis indicated that the receptor was encoded by an mRNA of approx. 3.5 kb. A 3.28 kbp cDNA encoding the TRH receptor has been cloned and sequenced. Full functionality of the predicted 412-amino-acid receptor protein was demonstrated by functional expression of cell surface receptors in Xenopus oocytes after both cytoplasmic injection of sense RNA transcribed in vitro from this cDNA and nuclear injection of the cDNA under the control of the Herpes simplex virus thymidine kinase promoter. The predicted protein contains seven putative membrane-spanning domains and shows significant sequence identify with some G-protein-coupled receptors. RNA blot analysis indicates that the mRNA for the TRH receptor is exclusively expressed in the pituitary gland. Expression studies performed with clones in which the 3' region of the mRNA has been successively shortened indicate that the 3' terminal region is not an important determinant for efficient functional expression in oocytes.

Citing Articles

Influence of Kv11.1 (hERG1) K channel expression on DNA damage induced by the genotoxic agent methyl methanesulfonate.

Fernandez-Villabrille S, Alvarez-Gonzalez E, Barros F, de la Pena P, Sierra L Pflugers Arch. 2021; 473(2):197-217.

PMID: 33452554 DOI: 10.1007/s00424-021-02517-2.

Cell type influences the molecular mechanisms involved in hormonal regulation of ERG K+ channels.

Carretero L, Barros F, Miranda P, Fernandez-Trillo J, Machin A, de la Pena P Pflugers Arch. 2012; 463(5):685-702.

PMID: 22415214 DOI: 10.1007/s00424-012-1094-y.

Molecular determinants of interactions between the N-terminal domain and the transmembrane core that modulate hERG K+ channel gating.

Fernandez-Trillo J, Barros F, Machin A, Carretero L, Dominguez P, de la Pena P PLoS One. 2011; 6(9):e24674.

PMID: 21935437 PMC: 3174182. DOI: 10.1371/journal.pone.0024674.

Distribution and axonal projections of neurons coexpressing thyrotropin-releasing hormone and urocortin 3 in the rat brain.

Wittmann G, Fuzesi T, Liposits Z, Lechan R, Fekete C J Comp Neurol. 2009; 517(6):825-40.

PMID: 19844978 PMC: 2849936. DOI: 10.1002/cne.22180.

Participation of HERG channel cytoplasmic structures on regulation by the G protein-coupled TRH receptor.

Alonso-Ron C, Barros F, Manso D, Gomez-Varela D, Miranda P, Carretero L Pflugers Arch. 2008; 457(6):1237-52.

PMID: 19002712 DOI: 10.1007/s00424-008-0599-x.

References

Hinkle P . Pituitary TRH receptors. Ann N Y Acad Sci. 1989; 553:176-87. DOI: 10.1111/j.1749-6632.1989.tb46639.x. View

Gunning P, Ponte P, Okayama H, Engel J, Blau H, Kedes L . Isolation and characterization of full-length cDNA clones for human alpha-, beta-, and gamma-actin mRNAs: skeletal but not cytoplasmic actins have an amino-terminal cysteine that is subsequently removed. Mol Cell Biol. 1983; 3(5):787-95. PMC: 368601. DOI: 10.1128/mcb.3.5.787-795.1983. View

Lechleiter J, Hellmiss R, Duerson K, Ennulat D, David N, Clapham D . Distinct sequence elements control the specificity of G protein activation by muscarinic acetylcholine receptor subtypes. EMBO J. 1990; 9(13):4381-90. PMC: 552228. DOI: 10.1002/j.1460-2075.1990.tb07888.x. View

Voigt M, Kispert J, Chin H . Sequence of a rat brain cDNA encoding an alpha-1B adrenergic receptor. Nucleic Acids Res. 1990; 18(4):1053. PMC: 330370. DOI: 10.1093/nar/18.4.1053. View

Monsma Jr F, Mahan L, McVittie L, Gerfen C, Sibley D . Molecular cloning and expression of a D1 dopamine receptor linked to adenylyl cyclase activation. Proc Natl Acad Sci U S A. 1990; 87(17):6723-7. PMC: 54609. DOI: 10.1073/pnas.87.17.6723. View

Straub R, Frech G, Joho R, Gershengorn M . Expression cloning of a cDNA encoding the mouse pituitary thyrotropin-releasing hormone receptor. Proc Natl Acad Sci U S A. 1990; 87(24):9514-8. PMC: 55202. DOI: 10.1073/pnas.87.24.9514. View

Lamberts S, MacLeod R . Regulation of prolactin secretion at the level of the lactotroph. Physiol Rev. 1990; 70(2):279-318. DOI: 10.1152/physrev.1990.70.2.279. View

Julius D, Huang K, Livelli T, Axel R, Jessell T . The 5HT2 receptor defines a family of structurally distinct but functionally conserved serotonin receptors. Proc Natl Acad Sci U S A. 1990; 87(3):928-32. PMC: 53382. DOI: 10.1073/pnas.87.3.928. View

Ritchie A . Two distinct calcium-activated potassium currents in a rat anterior pituitary cell line. J Physiol. 1987; 385:591-609. PMC: 1192362. DOI: 10.1113/jphysiol.1987.sp016509. View

10.

Meyerhof W, Morley S, Schwarz J, Richter D . Receptors for neuropeptides are induced by exogenous poly(A)+ RNA in oocytes from Xenopus laevis. Proc Natl Acad Sci U S A. 1988; 85(3):714-7. PMC: 279625. DOI: 10.1073/pnas.85.3.714. View

11.

Straub R, Oron Y, Gillo B, Thomson R, Gershengorn M . Receptor number determines latency and amplitude of the thyrotropin-releasing hormone response in Xenopus oocytes injected with pituitary RNA. Mol Endocrinol. 1989; 3(6):907-14. DOI: 10.1210/mend-3-6-907. View

12.

Yokota Y, Sasai Y, Tanaka K, Fujiwara T, Tsuchida K, Shigemoto R . Molecular characterization of a functional cDNA for rat substance P receptor. J Biol Chem. 1989; 264(30):17649-52. View

13.

McFarland K, Sprengel R, PHILLIPS H, Kohler M, Rosemblit N, Nikolics K . Lutropin-choriogonadotropin receptor: an unusual member of the G protein-coupled receptor family. Science. 1989; 245(4917):494-9. DOI: 10.1126/science.2502842. View

14.

ODowd B, Hnatowich M, Caron M, Lefkowitz R, Bouvier M . Palmitoylation of the human beta 2-adrenergic receptor. Mutation of Cys341 in the carboxyl tail leads to an uncoupled nonpalmitoylated form of the receptor. J Biol Chem. 1989; 264(13):7564-9. View

15.

Strader C, Sigal I, Dixon R . Structural basis of beta-adrenergic receptor function. FASEB J. 1989; 3(7):1825-32. DOI: 10.1096/fasebj.3.7.2541037. View

16.

Ben-Jonathan N, Arbogast L, Hyde J . Neuroendocrine [corrected] regulation of prolactin release. Prog Neurobiol. 1989; 33(5-6):399-447. DOI: 10.1016/0301-0082(89)90008-7. View

17.

Oron Y, Straub R, Traktman P, Gershengorn M . Decreased TRH receptor mRNA activity precedes homologous downregulation: assay in oocytes. Science. 1987; 238(4832):1406-8. DOI: 10.1126/science.2825350. View

18.

Baehr W, Falk J, Bugra K, Triantafyllos J, McGinnis J . Isolation and analysis of the mouse opsin gene. FEBS Lett. 1988; 238(2):253-6. DOI: 10.1016/0014-5793(88)80490-3. View

19.

Drummond A . Chlordiazepoxide is a competitive thyrotropin-releasing hormone receptor antagonist in GH3 pituitary tumour cells. Biochem Biophys Res Commun. 1985; 127(1):63-70. DOI: 10.1016/s0006-291x(85)80126-1. View

20.

Bonner T, Buckley N, Young A, Brann M . Identification of a family of muscarinic acetylcholine receptor genes. Science. 1987; 237(4814):527-32. DOI: 10.1126/science.3037705. View