Study on the Expression of Vasoactive intestinal polypeptide, Vasopressin and Oxytocin mRNAs in the Rat Brain using Double in situ Hybridization Technique.
- Author:
Kyeong Han PARK
1
;
Hong Suk PARK
;
Young Bok YOO
;
Wang Jae LEE
;
Douk Ho HWANG
;
Byung Lan LEE
;
Choong Ik CHA
;
Sa Sun CHO
;
Sang Ho BAIK
Author Information
1. Department of Anatomy, Seoul National University Medical College, Korea.
- Publication Type:Original Article
- Keywords:
In situ hybridization;
Double labelling;
Digoxigenin;
Radioisotope;
Vasoactive intestinal polypeptide;
Vasopressin;
Oxytocin;
Aging;
Gender;
Corpus callosum;
Midbrain;
Pons;
Cerebellar vermis;
MRI
- MeSH:
Aging;
Animals;
Autoradiography;
Brain*;
Corpus Callosum;
Digoxigenin;
In Situ Hybridization*;
Magnetic Resonance Imaging;
Mesencephalon;
Neuropeptides;
Neurotransmitter Agents;
Oligonucleotide Probes;
Oligonucleotides;
Oxytocin*;
Pons;
Rats*;
RNA, Messenger*;
Signal Transduction;
Vasoactive Intestinal Peptide*;
Vasopressins*;
X-Ray Film
- From:Korean Journal of Anatomy
1998;31(6):807-815
- CountryRepublic of Korea
- Language:Korean
-
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.
