Simultaneous monitoring of pituitary hormone secretion and gene expression within individual cells.
We have recently developed a method to simultaneously quantitate the level of gene expression and the level of secretion of a peptide from individual cells. Our approach has been to combine the reverse hemolytic plaque assay sequentially with in situ hybridization. We present data to show how we have used the pituitary lactotroph as a model to demonstrate the power of this technique. However, we are particularly excited about the potential application of this strategy to approach a broad spectrum of questions regarding the cellular and molecular mechanisms that regulate the coupling of peptide secretion and gene expression at the single cell level. 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.
Wise
P M
PM
Department of Physiology, School of Medicine, University of Maryland, Baltimore 21201.
Larson
G H
GH
Scarbrough
K
K
Chiu
S F
SF
Weiland
N G
NG
Lloyd
J M
JM
Hinkle
D A
DA
Cai
A H
AH
D000818:Animals; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D066298:In Vitro Techniques; D010902:Pituitary Gland / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D011388:Prolactin / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D013973:Thyrotropin-Releasing Hormone / Q000494:pharmacology
D012333:RNA, Messenger; D004958:Estradiol; D013973:Thyrotropin-Releasing Hormone; D011388:Prolactin