Messenger RNA phenotyping for semi-quantitative comparison of glucocorticoid receptor transcript levels in the developing embryonic mouse palate.
A specific complex of glucocorticoid and receptor interacts with responsive genes in target tissues of developing palate and controls its morphogenesis. In order to examine developmental changes in glucocorticoid receptor gene expression during embryonic palate formation, variations in transcript levels have been determined using a simple semi-quantitative reverse transcription-polymerase chain reaction (PCR) procedure. Oligo (dT)16 was used as the primer for reverse transcription, and the amount of PCR product for glucocorticoid receptor was normalized against those for glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and beta-actin. We found that either GAPDH or beta-actin were suitable internal standards, and that the mRNA levels of glucocorticoid receptor undergoes a significant decrease of 30% at the developmental stage of embryonic mouse palatal shelf re-orientation. Immunolocalization of the receptor demonstrated a significant change in the distribution pattern of glucocorticoid receptor during the re-orientation stage of secondary palate formation. Our present data suggest that developmental variations in glucocorticoid receptor level control morphogenetic events of secondary palate formation.
Lau
E C
EC
University of Southern California, Center for Craniofacial Molecular Biology, Los Angeles 90033.
Li
Z Q
ZQ
Santos
V
V
Slavkin
H C
HC
D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D017931:DNA Primers; D004622:Embryo, Mammalian; D005314:Embryonic and Fetal Development; D005260:Female; D015870:Gene Expression; D007150:Immunohistochemistry; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D009024:Morphogenesis; D010159:Palate / Q000166:cytology / Q000196:embryology / Q000378:metabolism*; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism*; D011965:Receptors, Glucocorticoid / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D014158:Transcription, Genetic
D017931:DNA Primers; D012333:RNA, Messenger; D011965:Receptors, Glucocorticoid