Assessment of gene expression and peptide secretion from individual cells.
We have developed an assay that allows one to monitor gene expression in and peptide secretion from individual cells. By combining the reverse hemolytic plaque with in situ hybridization, investigators can quantitate simultaneously the level of gene expression and the level of secretion of a peptide. The method can be used in any system in which an appropriate antibody for the reverse hemolytic plaque assay and probes complementary to the mRNA of interest are available. It can be used to monitor the level of mRNA and secretion of the peptide product, or expression of one gene and the secretion of another peptide. In this paper we will describe the major steps of the method. We have used the pituitary lactotroph as a model to demonstrate the power of this technique. However, we believe that this method may be an important approach to answer many questions regarding the cellular and molecular mechanisms that regulate the coupling of peptide secretion and gene expression at the single cell level.
Wise
P M
PM
Department of Physiology, School of Medicine, University of Maryland, Baltimore 21201.
Scarbrough
K
K
Larson
G
G
Chiu
S
S
Weiland
N G
NG
Lloyd
J M
JM
Hinkle
D A
DA
Cai
A
A
D000818:Animals; D002417:Cattle; D015870:Gene Expression; D006462:Hemolytic Plaque Technique; D017403:In Situ Hybridization; D010903:Pituitary Gland, Anterior / Q000378:metabolism*; D011388:Prolactin / Q000235:genetics / Q000378:metabolism*; D012313:RNA; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats
D012333:RNA, Messenger; D012313:RNA; D011388:Prolactin