Human corticotropin-releasing factor type 1 receptor analysis with white blood cells mRNAs and DNA.
The human corticotropin-releasing factor receptor type 1 (hCRF-R1) functional transcript is mainly expressed in the anterior pituitary corticotrophs, a tissue usually not available for clinical investigation. Splice variants translated into defective isoforms of the receptor have been described in few peripheral tissues. The aim of this work was to determine whether peripheral white blood cells from healthy individuals, an accessible tissue for clinical investigation, were suitable for the analysis of the hCRF-R1 transcript and gene. We report that: (i) specific amplification of the hCRF-R1 transcript from peripheral white blood cells mRNAs is feasible; (ii) this transcript is similar to the functional transcript; (iii) the draft sequence of chromosome 17 and unrelated sequences allow direct sequencing of all 14 exons of the gene, adjacent splice sites and related branch points. In conclusion, these approaches would be suitable for studies in patients having isolated secondary glucocorticoids deficiency to implicate the hCRH-R1 in the etiology of the disease.
Ben Simon
Elsa
E
Laboratoire de Biologie et Pathologie des Communications Cellulaires dans les Glandes Endocrines, INSERM U369, Faculté de Médecine, RTH Laënnec, 7 rue Guillaume Paradin, 69372 Lyon Cedex 08, France. ebensimon@yahoo.fr
Besancon
Roger
R
Benetollo
Claire
C
Nicolino
Marc
M
Saez
José-Maria
JM
Li
Jacques-Yuan
JY
D004247:DNA / Q000302:isolation & purification*; D017931:DNA Primers; D006801:Humans; D007962:Leukocytes; D012333:RNA, Messenger / Q000302:isolation & purification*; D018019:Receptors, Corticotropin-Releasing Hormone / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D000099052:CRF Receptor, Type 1
D017931:DNA Primers; D012333:RNA, Messenger; D018019:Receptors, Corticotropin-Releasing Hormone; D000099052:CRF Receptor, Type 1; D004247:DNA