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Exposure to an Antisense Oligonucleotide Decreases Corticotropin‐Releasing Factor Receptor Binding in Rat Pituitary Cultures
Author(s) -
Owens Michael J.,
Mulchahey Jeff J.,
Kasckow John W.,
Plotsky Paul M.,
Nemeroff Charles B.
Publication year - 1995
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1995.64052358.x
Subject(s) - receptor , oligonucleotide , endocrinology , medicine , biology , gene knockdown , anterior pituitary , corticotropin releasing hormone , microbiology and biotechnology , sense (electronics) , hormone , chemistry , biochemistry , gene
Corticotropin‐releasing factor (CRF) appears to integrate the endocrine, autonomic, immunologic, and behavioral responses of mammals to stress. To investigate further the role of CRF in the CNS, we have begun investigating the usefulness of “antisense knockdown” strategies directed against the CRF receptor using rat anterior pituitary gland primary cell cultures. The 15‐mer antisense (5′ CTG‐CGG‐GCG‐CCG‐TCC 3′) and “scrambled” control (5′ CGT‐CCG‐CGC‐GCT‐GCG 3′) oligonucleotides were synthesized based on the rat CRF receptor sequence just downstream of the initiation codon. In each of four separate experiments, exposure to 10 µmol/L of antisense oligonucleotide for 40–67 h resulted in significant (17–36%) decreases in 125 I‐ovine CRF binding to pituitary cells as compared with either control (no oligonucleotide) or 10 µmol/L of “scrambled” oligonucleotide. Moreover, compared with scrambled oligonucleotide, exposure to 10 µmol/L of antisense oligonucleotide, which produced a 22% decrease in CRF receptor binding, also resulted in a significant attenuation of the adrenocorticotrophic hormone response following a 30‐min challenge with 100 pmol/L of CRF. Thus, CRF receptor antisense oligonucleotides apparently reduce functional expression of CRF receptors. This technique may be useful in studying the kinetics of CRF receptor production and the physiological functions of CRF receptors within the CNS.