Application of the nonradioactive in situ hybridization for the localization of acetylcholinesterase mRNA in the central nervous system of the rat; comparison to the radioactive technique.
In this preliminary report nonradioactive digoxigenine-based and radioactive in situ hybridization procedures for the localization of acetylcholinesterase mRNA were tested and compared in rat brain. General patterns of Ache mRNA localization observed by both techniques did not differ significantly and were practically the same as reported in previous in situ studies on the mammalian brain. Shorter procedure time and avoidance of precautions necessary at work with radioactive materials are major advantages of nonradioactive technique. Under- and over- staining can be prevented by direct examination of coloring reaction. Faint staining in the control experiment with heterologous DNA suggests that proper stringency is essential for the specificity of staining.
Kreft
S
S
Institute of Pathophysiology, School of Medicine, Ljubljana, Slovenia.
Zajc-Kreft
K
K
Zivin
M
M
Sket
D
D
Grubic
Z
Z
D000110:Acetylcholinesterase / Q000096:biosynthesis*; D000818:Animals; D001923:Brain Chemistry / Q000502:physiology; D002490:Central Nervous System / Q000201:enzymology*; D017403:In Situ Hybridization; D015345:Oligonucleotide Probes; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D013462:Sulfur Radioisotopes
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D013462:Sulfur Radioisotopes; D000110:Acetylcholinesterase