Isolation and Pharmacological Characterization of Two Functional Splice Variants of Corticotropin‐Releasing Factor Type 2 Receptor from Tupaia belangeri

Isolation and Pharmacological Characterization of Two Functional Splice Variants of... From brain, heart and muscle tissue of the tree shrew (Tupaia belangeri), a higher order mammal, cDNA clones were isolated that encoded two functional splice variants of the corticotropin‐releasing factor (CRF) type 2 receptor (CRF‐R2). The first, full‐length splice variant, amplified from brain and heart tissue, encoded a CRF receptor protein that is 410 amino acids in length and ≈96% homologous to human CRF‐R2α. The second, full‐length splice variant, derived from skeletal muscle tissue, encoded a 437‐amino acid CRF receptor protein that is ≈92% homologous to human CRF‐R2β. Semiquantitative reverse transcriptase polymerase chain reaction (RT‐PCR) amplifications and RNase protection analyses, showed that tree shrew CRF‐R2α (tCRF‐R2α) and tree shrew CRF‐R2β (tCRF‐R2β) were coexpressed in brain tissue but not in heart and skeletal muscle tissue. Finally, human embryonic kidney 293 (HEK293) cells stably transfected with tCRF‐R2α and tCRF‐R2β were used to demonstrate that the CRF analogs urocortin and sauvagine bind with significantly greater affinity (21‐ to 140‐fold) to these two CRF‐R2 splice variants than do human/rat and ovine CRF analogs. In keeping with these results of our CRF binding studies, EC50 values were substantially lower for urocortin‐and sauvagine‐stimulated than for h/rCRF‐and oCRF‐stimulated cyclic AMP accumulation in HEK293 cells stably transfected with tCRF‐R2α or tCRF‐R2β cDNAs. The tree shrew therefore constitutes an important animal model in which to investigate the role of CRF receptor subtypes in the stress response. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Neuroendocrinology Wiley

Isolation and Pharmacological Characterization of Two Functional Splice Variants of Corticotropin‐Releasing Factor Type 2 Receptor from Tupaia belangeri

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Publisher
Wiley
Copyright
Copyright © 1999 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0953-8194
eISSN
1365-2826
DOI
10.1046/j.1365-2826.1999.00348.x
Publisher site
See Article on Publisher Site

Abstract

From brain, heart and muscle tissue of the tree shrew (Tupaia belangeri), a higher order mammal, cDNA clones were isolated that encoded two functional splice variants of the corticotropin‐releasing factor (CRF) type 2 receptor (CRF‐R2). The first, full‐length splice variant, amplified from brain and heart tissue, encoded a CRF receptor protein that is 410 amino acids in length and ≈96% homologous to human CRF‐R2α. The second, full‐length splice variant, derived from skeletal muscle tissue, encoded a 437‐amino acid CRF receptor protein that is ≈92% homologous to human CRF‐R2β. Semiquantitative reverse transcriptase polymerase chain reaction (RT‐PCR) amplifications and RNase protection analyses, showed that tree shrew CRF‐R2α (tCRF‐R2α) and tree shrew CRF‐R2β (tCRF‐R2β) were coexpressed in brain tissue but not in heart and skeletal muscle tissue. Finally, human embryonic kidney 293 (HEK293) cells stably transfected with tCRF‐R2α and tCRF‐R2β were used to demonstrate that the CRF analogs urocortin and sauvagine bind with significantly greater affinity (21‐ to 140‐fold) to these two CRF‐R2 splice variants than do human/rat and ovine CRF analogs. In keeping with these results of our CRF binding studies, EC50 values were substantially lower for urocortin‐and sauvagine‐stimulated than for h/rCRF‐and oCRF‐stimulated cyclic AMP accumulation in HEK293 cells stably transfected with tCRF‐R2α or tCRF‐R2β cDNAs. The tree shrew therefore constitutes an important animal model in which to investigate the role of CRF receptor subtypes in the stress response.

Journal

Journal of NeuroendocrinologyWiley

Published: Jun 1, 1999

References

  • Chronic psychosocial stress induces morphological alterations in hippocampal pyramidal neurons of the tree shrew
    Fuchs, Fuchs; Uno, Uno; Flügge, Flügge

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