In situ hybridization histochemistry: localisation of vasopressin mRNA in rat brain using a biotinylated oligonucleotide probe.
A biotinylated antisense oligonucleotide probe specific for the glycopeptide sequence of arginine vasopressin mRNA has been used with amplified detection for visualisation of arginine vasopressin mRNA in the rat hypothalamus. RNAase pretreatment to destroy arginine vasopressin mRNA and use of excess complementary oligonucleotide (sense) to absorb the biotinylated antisense oligonucleotide demonstrated the reaction is specific for arginine vasopressin mRNA. Further, dehydration of rats using 2% saline resulted in an increase in specific staining. The staining is localized to those neurones in the hypothalamus known to contain arginine vasopressin.
Arai
H
H
Department of Neuroendocrinology, AFRC Institute of Animal Physiology and Genetics Research, Babraham, Cambridge, U.K.
Emson
P C
PC
Agrawal
S
S
Christodoulou
C
C
Gait
M J
MJ
D000818:Animals; D001127:Arginine Vasopressin / Q000378:metabolism*; D001710:Biotin; D004247:DNA; D007031:Hypothalamus / Q000378:metabolism*; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains; D012260:Ribonucleases / Q000494:pharmacology
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D001127:Arginine Vasopressin; D001710:Biotin; D004247:DNA; D012260:Ribonucleases