Abstract
We examined the effect of cyclical mechanical stretch on the regulation of cardiac myosin heavy chain genes using an isolated neonatal rat cardiocyte culture system. Cultured cardiocytes grown on a flexible membrane were deformed by vacuum to 20% of maximum elongation, at 60 cycles/min in a serum-free medium. Cyclical stretch did not cause myocyte damage as assessed by supernatant LDH measurement and trypan blue exclusion test. The levels of myosin heavy chain (MyHC) mRNA increased as early as 1 h after stretch, reaching 12-fold over the control in 24 h, as shown by Northern blot analysis. However, the proximal 5'-flanking regions of the α- and β-MyHC gene which were linked to chloramphenicol acetyltransferase (CAT) reporter gene did not exhibit enhanced CAT activity following cyclical stretch. Deletion of the chimeric constructs to shorten the 5'-flanking regions of the MyHC genes generated by polymerase chain reaction amplification did not enhance the CAT activity under cyclical stretch. This finding suggests that the stretch-reponse element of the α- and β-MyHC gene promoter is probably not present in the proximal region of either the α- or β-MyHC genes.
Original language | English |
---|---|
Pages (from-to) | 567-573 |
Number of pages | 7 |
Journal | Biochemical and Biophysical Research Communications |
Volume | 210 |
Issue number | 2 |
DOIs | |
Publication status | Published - 1995 |
Externally published | Yes |
Fingerprint
ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Cell Biology
- Molecular Biology
Cite this
Regulation of human cardiac myosin heavy chain genes by cyclical mechanical stretch in cultured cardiocytes. / Shyu, K. G.; Chen, J. J.; Shih, N. L.; Wang, D. L.; Chang, H.; Lien, W. P.; Liew, C. C.
In: Biochemical and Biophysical Research Communications, Vol. 210, No. 2, 1995, p. 567-573.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Regulation of human cardiac myosin heavy chain genes by cyclical mechanical stretch in cultured cardiocytes
AU - Shyu, K. G.
AU - Chen, J. J.
AU - Shih, N. L.
AU - Wang, D. L.
AU - Chang, H.
AU - Lien, W. P.
AU - Liew, C. C.
PY - 1995
Y1 - 1995
N2 - We examined the effect of cyclical mechanical stretch on the regulation of cardiac myosin heavy chain genes using an isolated neonatal rat cardiocyte culture system. Cultured cardiocytes grown on a flexible membrane were deformed by vacuum to 20% of maximum elongation, at 60 cycles/min in a serum-free medium. Cyclical stretch did not cause myocyte damage as assessed by supernatant LDH measurement and trypan blue exclusion test. The levels of myosin heavy chain (MyHC) mRNA increased as early as 1 h after stretch, reaching 12-fold over the control in 24 h, as shown by Northern blot analysis. However, the proximal 5'-flanking regions of the α- and β-MyHC gene which were linked to chloramphenicol acetyltransferase (CAT) reporter gene did not exhibit enhanced CAT activity following cyclical stretch. Deletion of the chimeric constructs to shorten the 5'-flanking regions of the MyHC genes generated by polymerase chain reaction amplification did not enhance the CAT activity under cyclical stretch. This finding suggests that the stretch-reponse element of the α- and β-MyHC gene promoter is probably not present in the proximal region of either the α- or β-MyHC genes.
AB - We examined the effect of cyclical mechanical stretch on the regulation of cardiac myosin heavy chain genes using an isolated neonatal rat cardiocyte culture system. Cultured cardiocytes grown on a flexible membrane were deformed by vacuum to 20% of maximum elongation, at 60 cycles/min in a serum-free medium. Cyclical stretch did not cause myocyte damage as assessed by supernatant LDH measurement and trypan blue exclusion test. The levels of myosin heavy chain (MyHC) mRNA increased as early as 1 h after stretch, reaching 12-fold over the control in 24 h, as shown by Northern blot analysis. However, the proximal 5'-flanking regions of the α- and β-MyHC gene which were linked to chloramphenicol acetyltransferase (CAT) reporter gene did not exhibit enhanced CAT activity following cyclical stretch. Deletion of the chimeric constructs to shorten the 5'-flanking regions of the MyHC genes generated by polymerase chain reaction amplification did not enhance the CAT activity under cyclical stretch. This finding suggests that the stretch-reponse element of the α- and β-MyHC gene promoter is probably not present in the proximal region of either the α- or β-MyHC genes.
UR - http://www.scopus.com/inward/record.url?scp=0029065392&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0029065392&partnerID=8YFLogxK
U2 - 10.1006/bbrc.1995.1697
DO - 10.1006/bbrc.1995.1697
M3 - Article
C2 - 7755635
AN - SCOPUS:0029065392
VL - 210
SP - 567
EP - 573
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 2
ER -