Differential splicing of fibronectin pre-messenger ribonucleic acid during cardiac ontogeny and development of hypertrophy in the rat.

BACKGROUND

Fibronectin, an extracellular matrix protein, exists as multiple isoforms expressed in a time- and cell-dependent manner. Since the developmental pattern of fibronectin expression has not been determined in the heart, the first issue of this study was to investigate the expression of total fibronectin mRNA as well as its isoforms during cardiac ontogeny. In adults, pressure overload induces a shift towards the fetal form of proteins expressed by either muscle or nonmuscle cardiac cells. Fetal forms of fibronectin mRNA being found in smooth and nonmuscle cardiac cells soon after imposition of pressure overload, the pattern of fibronectin expression during the development of pathological growth was analyzed to determine whether the two conditions of cardiac growth resulted in an identical pattern of fibronectin expression.

EXPERIMENTAL DESIGN

Total RNA were isolated from rat heart (a) during in utero and postnatal life and (b) at varying periods of time after imposition of a pressure overload induced by coarctation of the thoracic aorta in 25-day-old rats. Fibronectin-EIIIA+ or -EIIIB+ and total fibronectin mRNAs were quantitated by reverse transcription-polymerase chain reactions and dot-blot analysis, respectively.

RESULTS

Fibronectin mRNA, abundant in the 14-day-old fetal heart, rapidly decreased during cardiac physiologic growth (> 5-fold); no changes in the fibronectin mRNA level was observed during the development of pressure-induced cardiac hypertrophy. The percentages of fibronectin transcripts containing EIIIA or EIIIB exons, very high in the early fetal heart (> 45%), harmoniously decreased during cardiac maturation (< 12%). Aortic coarctation resulted in an early, transient (12 to 48 hours) and preferential expression of fibronectin-EIIIA+ mRNA (approximately 40%).

CONCLUSIONS

In rat heart, neither physiologic nor pressure-induced growth requires increased amounts of fibronectin mRNA but the growth conditions specifically modulated the fibronectin pre-mRNA splicing.