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To assess the mechanisms controlling the developmental
regulation of tropoelastin expression in vivo, we developed a
reverse-transcription-polymerase chain reaction (RT-PCR) assay to
detect tropoelastin pre-mRNA as an indicator of ongoing transcription
in intact tissue. RNA was isolated from mid-fetal (early-elastogenic),
neonatal (peak tropoelastin expression), and adult (very low
tropoelastin expression) rat lungs and reverse transcribed, and the
cDNA was amplified with intron specific primers. A weak hybridization
signal for tropoelastin pre-mRNA was seen in mid-fetal samples, and
paralleling the increase in steady-state mRNA levels, a strong signal
for pre-mRNA was detected in neonatal samples, indicating
transcriptional regulation. Stimulation of fetal lung tropoelastin
expression by maternal administration of dexamethasone also led to an
increase in pre-mRNA levels. However, signal for tropoelastin pre-mRNA
in adult samples was equal to that detected in neonatal samples, even
though mRNA levels had dropped about 80-fold. Persistence of
tropoelastin transcription in adult tissue was also seen in cell
culture models and was verified by nuclear runoff assay. In addition,
an RT-PCR assay for
Volume 270,
Number 25,
Issue of June 23, pp. 14899-14906, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
EXPRESSION OF TROPOELASTIN PRE-mRNA PERSISTS AFTER DOWN-REGULATION
OF STEADY-STATE mRNA LEVELS
1 (I) procollagen pre-mRNA accurately revealed
the known transcriptional regulation of this gene. Our results
demonstrate that the induction and maintenance of elastogenesis is
controlled by a transcriptional mechanism, whereas, the cessation of
tropoelastin expression is controlled by a post-transcriptional
mechanism.
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