In situ hybridization of dihydroxyacetone phosphate acyltransferase, the regulating enzyme involved in plasmalogen biosynthesis.
In situ hybridization can be carried out using different methods. The experimenter has to choose various parameters: the type of tissue fixation, the time of incubation, and the duration of the exposure time. All these parameters are determinant for the sensitivity and the resolution of this technique. This publication of technical aspects described different experiments performed for in situ hybridization on liver tissue. We may conclude on the parameters to optimize each step of the hybridization procedure. Moreover, this technique could be transposed to the brain and applied to little structures with a light expression of DHAP-AT.
André
Agnès
A
INRA, Unité de Nutrition Lipidique, 17 rue Sully, BP 86510, 21065 Dijon Cedex, France.
Tessier
Christian
C
Brétillon
Lionel
L
Sébédio
Jean-Louis
JL
Chardigny
Jean-Michel
JM
D000217:Acyltransferases / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000033:anatomy & histology / Q000201:enzymology; D017403:In Situ Hybridization; D066298:In Vitro Techniques; D008099:Liver / Q000201:enzymology; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013347:Subcellular Fractions / Q000201:enzymology; D013997:Time Factors; D016707:Tissue Fixation / Q000379:methods
D012333:RNA, Messenger; D000217:Acyltransferases; C020051:glycerone-phosphate O-acyltransferase