Study on the Expression of Vasoactive intestinal polypeptide, Vasopressin and Oxytocin mRNAs in the Rat Brain using Double in situ Hybridization Technique / 대한해부학회지
Korean Journal of Anatomy
;
: 807-815, 1998.
Article
in Korean
| WPRIM
| ID: wpr-655798
ABSTRACT
The technique of in situ hybridization using synthetic oligonucleotides labelled by non-radioactive method was developed to localize vasoactive intestinal polypeptide, arginine-vasopressin and oxytocin mRNAs in the rat brain. Also double in situ hybridization technique where combination of non-radioactive and radioactive probes were applied was developed to localize 2 neuropeptide mRNAs in single tissue section. The results were as follows; In non-radioactive in situ hybridization methods using digoxigenin-labelled oligonucleotide probe, alkaline-phosphates method using NBT and BCIP as substrates gave the best result that specific hybridization signals were observed. In radioactive in situ hybridization methods using 35S-labelled oligonucleotide probe, specific hybridization signals were observed in both nuclear track emulsion and X-ray film autoradiography. In double in situ hybridization methods using combination of 35S-labelled and digoxigenin-labelled oligonucleotide probes, specific hybridization signals were observed in the group where K5 emulsion was applied as nuclear track emulsion. The technique of in situ hybridization using digoxigenin-labelled oligonucleotide applied in this study will be useful as alternative for radioactive in situ hybridization technique. Moreover, combination of non-radioactive and radioactive labelled probes in double in situ hybridization technique will be a useful tool for the simultaneous localization of various mRNAs in single section for the study of various neurotransmitters, neuropeptides, receptors and signal transduction molecules.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Oligonucleotides
/
Autoradiography
/
X-Ray Film
/
Neuropeptides
/
Brain
/
Aging
/
RNA, Messenger
/
Mesencephalon
/
Oxytocin
/
Pons
Limits:
Animals
Language:
Korean
Journal:
Korean Journal of Anatomy
Year:
1998
Type:
Article
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