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. 1985 May;4(5):1257–1260. doi: 10.1002/j.1460-2075.1985.tb03769.x

Quantitation of proenkephalin A messenger RNA in bovine brain, pituitary and adrenal medulla: correlation between mRNA and peptide levels.

C W Pittius, N Kley, J P Loeffler, V Höllt
PMCID: PMC554333  PMID: 4006917

Abstract

Concentrations of mRNA coding for the opioid peptide precursor proenkephalin A were measured in bovine brain areas, pituitary and adrenal medulla. In all tissues, a single hybridizable species of 1400 bases in size was found by Northern blot analysis using as a probe a single-stranded (ss) cDNA complementary to bovine proenkephalin A mRNA. In solution hybridization experiments the distribution of the mRNA was quantified. Considerable differences were found for the abundance of proenkephalin A mRNA in the various tissues: from 0.023% in the adrenal medulla to 0.00002% in the adenohypophysis. Relative abundance in the various brain areas varied greater than 20-fold, being highest in the caudate nucleus (0.0025%) and lowest in the thalamus and substantia nigra (0.0001%). Comparison with immunoreactive peptide concentrations in these tissues showed a close correlation between the levels of proenkephalin A mRNA and the immunoreactive peptides.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bloch B., Baird A., Ling N., Benoit R., Guillemin R. Immunohistochemical evidence that brain enkephalins arise from a precursor similar to adrenal preproenkephalin. Brain Res. 1983 Mar 21;263(2):251–257. doi: 10.1016/0006-8993(83)90317-7. [DOI] [PubMed] [Google Scholar]
  3. Bloch B., Milner R. J., Baird A., Gubler U., Reymond C., Bohlen P., le Guellec D., Bloom F. E. Detection of the messenger RNA coding for preproenkephalin A in bovine adrenal by in situ hybridization. Regul Pept. 1984 Jul;8(4):345–354. doi: 10.1016/0167-0115(84)90045-4. [DOI] [PubMed] [Google Scholar]
  4. Bloom F., Battenberg E., Rossier J., Ling N., Guillemin R. Neurons containing beta-endorphin in rat brain exist separately from those containing enkephalin: immunocytochemical studies. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1591–1595. doi: 10.1073/pnas.75.3.1591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boarder M. R., Lockfeld A. J., Barchas J. D. Measurement of methionine-enkephalin [Arg6,Phe7] in rat brain by specific radioimmunoassay directed at methionine sulphoxide enkephalin[Arg6,Phe7]. J Neurochem. 1982 Feb;38(2):299–304. doi: 10.1111/j.1471-4159.1982.tb08629.x. [DOI] [PubMed] [Google Scholar]
  6. Civelli O., Birnberg N., Herbert E. Detection and quantitation of pro-opiomelanocortin mRNA in pituitary and brain tissues from different species. J Biol Chem. 1982 Jun 25;257(12):6783–6787. [PubMed] [Google Scholar]
  7. Comb M., Seeburg P. H., Adelman J., Eiden L., Herbert E. Primary structure of the human Met- and Leu-enkephalin precursor and its mRNA. Nature. 1982 Feb 25;295(5851):663–666. doi: 10.1038/295663a0. [DOI] [PubMed] [Google Scholar]
  8. Cuello A. C., Paxinos G. Evidence for a long Leu-enkephalin striopallidal pathway in rat brain. Nature. 1978 Jan 12;271(5641):178–180. doi: 10.1038/271178a0. [DOI] [PubMed] [Google Scholar]
  9. Del Fiacco M., Paxinos G., Cuello A. C. Neostriatal enkephalin-immunoreactive neurones project to the globus pallidus. Brain Res. 1982 Jan 7;231(1):1–17. doi: 10.1016/0006-8993(82)90003-8. [DOI] [PubMed] [Google Scholar]
  10. Durnam D. M., Palmiter R. D. A practical approach for quantitating specific mRNAs by solution hybridization. Anal Biochem. 1983 Jun;131(2):385–393. doi: 10.1016/0003-2697(83)90188-4. [DOI] [PubMed] [Google Scholar]
  11. Gubler U., Seeburg P., Hoffman B. J., Gage L. P., Udenfriend S. Molecular cloning establishes proenkephalin as precursor of enkephalin-containing peptides. Nature. 1982 Jan 21;295(5846):206–208. doi: 10.1038/295206a0. [DOI] [PubMed] [Google Scholar]
  12. Jingami H., Nakanishi S., Imura H., Numa S. Tissue distribution of messenger RNAs coding for opioid peptide precursors and related RNA. Eur J Biochem. 1984 Aug 1;142(3):441–447. doi: 10.1111/j.1432-1033.1984.tb08306.x. [DOI] [PubMed] [Google Scholar]
  13. Kakidani H., Furutani Y., Takahashi H., Noda M., Morimoto Y., Hirose T., Asai M., Inayama S., Nakanishi S., Numa S. Cloning and sequence analysis of cDNA for porcine beta-neo-endorphin/dynorphin precursor. Nature. 1982 Jul 15;298(5871):245–249. doi: 10.1038/298245a0. [DOI] [PubMed] [Google Scholar]
  14. Legon S., Glover D. M., Hughes J., Lowry P. J., Rigby P. W., Watson C. J. The structure and expression of the preproenkephalin gene. Nucleic Acids Res. 1982 Dec 20;10(24):7905–7918. doi: 10.1093/nar/10.24.7905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nakanishi S., Inoue A., Kita T., Nakamura M., Chang A. C., Cohen S. N., Numa S. Nucleotide sequence of cloned cDNA for bovine corticotropin-beta-lipotropin precursor. Nature. 1979 Mar 29;278(5703):423–427. doi: 10.1038/278423a0. [DOI] [PubMed] [Google Scholar]
  16. Noda M., Furutani Y., Takahashi H., Toyosato M., Hirose T., Inayama S., Nakanishi S., Numa S. Cloning and sequence analysis of cDNA for bovine adrenal preproenkephalin. Nature. 1982 Jan 21;295(5846):202–206. doi: 10.1038/295202a0. [DOI] [PubMed] [Google Scholar]
  17. Pittius C. W., Seizinger B. R., Pasi A., Mehraein P., Herz A. Distribution and characterization of opioid peptides derived from proenkephalin A in human and rat central nervous system. Brain Res. 1984 Jun 18;304(1):127–136. doi: 10.1016/0006-8993(84)90868-0. [DOI] [PubMed] [Google Scholar]
  18. Rossier J., Battenberg E., Pittman Q., Bayon A., Koda L., Miller R., Guillemin R., Bloom F. Hypothalamic enkephalin neurones may regulate the neurohypophysis. Nature. 1979 Feb 22;277(5698):653–655. doi: 10.1038/277653a0. [DOI] [PubMed] [Google Scholar]
  19. Tang F., Costa E., Schwartz J. P. Increase of proenkephalin mRNA and enkephalin content of rat striatum after daily injection of haloperidol for 2 to 3 weeks. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3841–3844. doi: 10.1073/pnas.80.12.3841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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