Probe-independent and direct quantification of insulin mRNA and growth hormone mRNA in enriched cell preparations.
Task division in multicellular organisms ensures that differentiated cell types produce cell-specific proteins that fulfill tasks for the whole organism. In some cases, the encoded mRNA species is so abundant that it represents a sizeable fraction of total mRNA in the cell. In this study, we have used a probe- and primer-free technique to quantify such abundant mRNA species in order to assess regulatory effects of in vitro and in vivo conditions. As a first example, we were able to quantify the regulation of proinsulin mRNA abundance in beta-cells by food intake or by the glucose concentration in tissue culture. The second example of application of this technique is the effect of corticosteroids on growth hormone mRNA in enriched somatrotrophs. It is anticipated that other examples exist in which measurement of very abundant mRNAs in dedicated cells will help to understand biological processes, monitor disease states, or assist biotechnological manufacturing procedures.
Van Lommel
Leentje
L
Herestraat 49, P.O. Box 901, B-3000 Leuven, Belgium.
Janssens
Kristel
K
Quintens
Roel
R
Tsukamoto
Katsura
K
Vander Mierde
Dirk
D
Lemaire
Katleen
K
Denef
Carl
C
Jonas
Jean-Christophe
JC
Martens
Geert
G
Pipeleers
Daniel
D
Schuit
Frans C
FC
D000199:Actins / Q000235:genetics; D000818:Animals; D001709:Biotechnology / Q000379:methods; D002149:Energy Intake; D007328:Insulin / Q000235:genetics*; D007515:Islets of Langerhans / Q000502:physiology; D007700:Kinetics; D011384:Proinsulin / Q000235:genetics; D018075:RNA, Complementary / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction
D000199:Actins; D007328:Insulin; D018075:RNA, Complementary; D012333:RNA, Messenger; D011384:Proinsulin