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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

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  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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