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Published on: January 17, 2016
Heart-selective expression of the chicken FK506-binding protein (FKBP) 12.6 gene during embryonic development
Shigenobu Yazawa1, Koji Obata, Akio Iio
1United Graduate School of Agricultural Science, Gifu University, Gifu, Japan.
Insights
The FKBP12.6 gene in chickens is expressed early in heart development, playing a role in excitation-contraction coupling. Its expression becomes concentrated in the atria as the heart develops.
Area of Science:
- Molecular Biology
- Developmental Biology
- Cardiology
Background:
- FKBP12.6 (FK506-binding protein 12.6) stabilizes the cardiac ryanodine receptor, a critical Ca(2+) release channel in cardiomyocytes.
- Understanding the developmental expression of FKBP12.6 is crucial for elucidating its role in cardiac function.
Purpose of the Study:
- To clone and characterize the chicken FKBP12.6 (chFKBP12.6) cDNA.
- To investigate the temporal and spatial expression patterns of chFKBP12.6 during embryonic development.
Main Methods:
- cDNA cloning and sequencing.
- Northern blot analysis to determine mRNA abundance and distribution.
- In situ hybridization to localize mRNA expression within embryonic tissues.
- Reverse transcription and polymerase chain reaction (RT-PCR) to detect mRNA presence.
Main Results:
- chFKBP12.6 encodes a 108-amino acid protein highly homologous to mammalian FKBP12.6.
- chFKBP12.6 mRNA is initially detected in the precardiac mesoderm and expressed throughout the developing heart.
- mRNA abundance decreases, and expression becomes restricted to the atria before hatching.
- Expression is observed from early gastrulation, peaking post-heartbeat onset.
Conclusions:
- The chFKBP12.6 gene is expressed early in development, prior to heart morphogenesis, suggesting a role in excitation-contraction coupling.
- The functional role of chFKBP12.6 becomes increasingly specific to the atrium during embryonic development.
Abstract:
FKBP12.6, a member of the family of FK506-binding proteins, selectively associates with the cardiac isoform of the ryanodine receptor and thereby stabilizes this Ca(2+) release channel. A chicken FKBP12.6 (chFKBP12.6) cDNA was cloned and shown to encode a protein of 108 amino acids. The deduced amino acid sequence of chFKBP12.6 is 91-92% identical to those of mammalian FKBP12.6 proteins. Northern blot analysis revealed that chFKBP12.6 mRNA is largely restricted to the heart during embryonic development and that the abundance of this mRNA in the heart decreases, and it becomes restricted to the atrium during cardiogenesis. In situ hybridization revealed that chFKBP12.6 mRNA is localized to the precardiac mesoderm before formation of the primitive heart tube. Expression of the chFKBP12.6 gene was initially apparent throughout the developing multichambered heart but became restricted to the atria before hatching. Reverse transcription and polymerase chain reaction analysis demonstrated that chFKBP12.6 mRNA is present in the embryo from early gastrulation and is most abundant immediately after the onset of the heartbeat. These observations suggest that the chFKBP12.6 gene is expressed before heart morphogenesis to play a role in excitation-contraction coupling in cardiomyocytes and that the function of the encoded protein becomes increasingly restricted to the atrium during embryonic development.
