Discrimination of heterogenous mRNAs encoding strychnine-sensitive glycine receptors in Xenopus oocytes by antisense oligonucleotides

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 86; no. 20; pp. 8103 - 8107
• Main Authors: Akagi, H, Patton, D E, Miledi, R
• Format: Journal Article
• Language: English
• Published: United States National Acad Sciences 01.10.1989
• Subjects: Aging; Animals; Base Sequence; beta-Alanine - pharmacology; Female; gamma-Aminobutyric Acid - pharmacology; Glycine - pharmacology; Glycine - physiology; Kainic Acid - pharmacology; Membrane Potentials - drug effects; Molecular Sequence Data; Oligonucleotide Probes; Oocytes - drug effects; Oocytes - physiology; Poly A - genetics; Poly A - isolation & purification; Protein Biosynthesis; Rats; Receptors, Glycine; Receptors, Neurotransmitter - genetics; Receptors, Neurotransmitter - physiology; RNA - genetics; RNA, Antisense; RNA, Messenger - antagonists & inhibitors; RNA, Messenger - genetics; RNA, Messenger - isolation & purification; Serotonin - pharmacology; Spinal Cord - growth & development; Strychnine - pharmacology; Xenopus
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.86.20.8103

Three synthetic oligodeoxynucleotides complementary to different parts of an RNA encoding a glycine receptor subunit were used to discriminate heterogenous mRNAs coding for glycine receptors in adult and neonatal rat spinal cord. Injection of the three antisense oligonucleotides into Xenopus oocytes specifically inhibited the expression of glycine receptors by adult spinal cord mRNA. In contrast, the antisense oligonucleotides were much less potent in inhibiting the expression of glycine receptors encoded by neonatal spinal cord mRNA. Northern blot analysis revealed that the oligonucleotides hybridized mostly to an adult cord transcript of approximately 10 kilobases in size. This band was also present in neonatal spinal cord mRNA but its density was about one-fourth of the adult cord message. There was no intense band in the low molecular weight position (approximately 2 kilobases), the existence of which was expected from electrophysiological studies with size-fractionated mRNA of neonatal spinal cord. Our results suggest that in the rat spinal cord there are at least three different types of mRNAs encoding functional strychnine-sensitive glycine receptors.