Localization of the mRNAs for Two Dopamine D 2 Receptor Isoforms in the Rat Brain

Localization of the mRNAs for Two Dopamine D 2 Receptor Isoforms in the Rat Brain Abstract: Two molecular forms of the dopamine D., receptor were generated by alternative RNA splicing. To investigate the relative distributions of the two mRNAs encoding the D2 receptor isoforms, D2(415) and D2(444), we performed in situ hybridization histochemistry in the rat brain with the two oligonucleotide probes. An insert probe complementary to an additional fragment of the D, receptor mRNA cloned from the rat brain, and a spanning probe complementary to its contiguous sequence were used. These 48 base probes were 3′‐end labeled with (35S)dATP. The brains were dissected from male SD rats and frozen in dry ice and acetone. Cryostat sections (16 um) were collected on gelatin coated slides and stored at – 20oC. In situ hybridization studies were conducted with a probe concentration of 1 × 10e dpm/100 ^1 of buffer per brain slice at 37oC for 18–20 h in a humid chamber. The slides were washed, dried and exposed to tritium sensitive film for one week. The autoradiograph showed that both mRNA were present at high levels in the corpus striatum, accumbens nucleus and substantia nigra (pars compacta). Identical patterns of labeling were obtained in the rat brain using both the insert and spanning probes, although the optical densities detected with the insert probe were higher than those with the spanning probe in the corpus striatum. This suggests that both D2 receptor mRNAs are expressed similarly in each region of the rat brain and D2(14) expressed dominantly in the corpus striatum. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Psychiatry and Clinical Neurosciences Wiley

Localization of the mRNAs for Two Dopamine D 2 Receptor Isoforms in the Rat Brain

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Publisher
Wiley
Copyright
Copyright © 1991 Wiley Subscription Services, Inc., A Wiley Company
ISSN
1323-1316
eISSN
1440-1819
D.O.I.
10.1111/j.1440-1819.1991.tb00530.x
Publisher site
See Article on Publisher Site

Abstract

Abstract: Two molecular forms of the dopamine D., receptor were generated by alternative RNA splicing. To investigate the relative distributions of the two mRNAs encoding the D2 receptor isoforms, D2(415) and D2(444), we performed in situ hybridization histochemistry in the rat brain with the two oligonucleotide probes. An insert probe complementary to an additional fragment of the D, receptor mRNA cloned from the rat brain, and a spanning probe complementary to its contiguous sequence were used. These 48 base probes were 3′‐end labeled with (35S)dATP. The brains were dissected from male SD rats and frozen in dry ice and acetone. Cryostat sections (16 um) were collected on gelatin coated slides and stored at – 20oC. In situ hybridization studies were conducted with a probe concentration of 1 × 10e dpm/100 ^1 of buffer per brain slice at 37oC for 18–20 h in a humid chamber. The slides were washed, dried and exposed to tritium sensitive film for one week. The autoradiograph showed that both mRNA were present at high levels in the corpus striatum, accumbens nucleus and substantia nigra (pars compacta). Identical patterns of labeling were obtained in the rat brain using both the insert and spanning probes, although the optical densities detected with the insert probe were higher than those with the spanning probe in the corpus striatum. This suggests that both D2 receptor mRNAs are expressed similarly in each region of the rat brain and D2(14) expressed dominantly in the corpus striatum.

Journal

Psychiatry and Clinical NeurosciencesWiley

Published: Dec 1, 1991

References

  • Dopamine receptors: A classification
    Creese, Creese; Leff, Leff

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