Muscle stem cell or myoblast, method for screening substances that participate in metabolic conversion using same, and pharmaceutical composition comprising substance obtained from said screening method
The object of the present invention is to provide a method for screening a substance involved in a metabolic shift of skeletal muscle, and a kit for screening a substance involved in a metabolic shift of skeletal muscle. The object can be solved by a muscle stem cell or myoblast comprising at least one myosin-heavy chain fusion gene selected from the group consisting of a myosin-heavy chain I fusion gene wherein a myosin-heavy chain I gene and a fluorescent protein or photoprotein gene are fused, a myosin-heavy chain IIa fusion gene wherein a myosin-heavy chain IIa gene and a fluorescent protein or photoprotein gene are fused, a myosin-heavy chain IId/x fusion gene wherein a myosin-heavy chain IId/x gene and a fluorescent protein or photoprotein gene are fused, and a myosin-heavy chain IIb fusion gene wherein a myosin-heavy chain IIb gene and a fluorescent protein or photoprotein gene are fused.
1. A muscle stem cell or myoblast comprising at least one myosin-heavy chain fusion gene selected from the group consisting of:
a myosin-heavy chain I gene and a fluorescent protein or photoprotein gene,
a myosin-heavy chain IIa gene and a fluorescent protein or photoprotein gene,
a myosin-heavy chain IIdix gene and a fluorescent protein or photoprotein gene, and
a myosin-heavy chain Hb gene and a fluorescent protein or photoprotein gene.
2. The muscle stem cell or myoblast according to claim 1 , wherein the cell is an immortalized cell.
3. The muscle stern cell or myoblast according to claim 1 , wherein the fluorescent protein or photoprotein gene is linked 5′ to the myosin heavy chain I, IIa, IIdx, or IIb gene.
4. The muscle stem cell or myoblast according to claim 1 , wherein the fluorescent protein or photoprotein gene encodes at least one fluorescent protein or photoprotein selected from the group consisting of Sirius, EBFP, SBP2, ESP2, Azurite, mKalamal, TagBFP, mBlueberry, mTurquoise, ECFP, Cerulean, mCerulean, TagCFP, AmCyan, mTP1, MiCy (Midoriishi Cyan), TurboGFP, CFP, AcGFP, TagGFP, AG (Azami-Green), mAG1, ZsGreen, EmGFP (Emerald), EGFP, GP2, T-Sapphire, HyPer, TagYFP, mAmetrine, EYFP, YFP, Venus, Citrine, PhiYFP, PhiYFP-m, turboYFP, ZsYellow, mBanana, mKO1, KO(Kusabira Orange), mOrange, mOrange2, mKO2, Keima570, TurboRFP, DsRed-Express, DsRed, DsRed2, TagRFP, TagRFP-T, DsRed-Monomer, mApple, AsRed2, mStrawberry, TurboFP602, mRP1, JRed, KillerRed, mCherry, KeimaRed, HcRed, mRasberry, mKate2, TagFP635, mPlum, egFP650, Neptune, mNeptune, egFP670, and luciferase.
5. A method for screening a test substance affecting myosin heavy chain expression, said method comprising the steps of:
a) providing a muscle stem cell or myoblast comprising at least one myosin-heavy chain fusion gene selected from the group consisting of:
a myosin-heavy chain I gene and a fluorescent protein or photoprotein gene,
a myosin-heavy chain IIa gene and a fluorescent protein or photoprotein gene,
a myosin-heavy chain Ild/x gene and a fluorescent protein or photoprotein gene. and
a myosin-heavy chain IIb gene and a fluorescent protein or photoprotein gene;
b) contacting said muscle stem cell or myoblast with a test substance; and
c) analyzing the expression of at least one myosin-heavy chain fusion gene in said muscle stem cell or myoblast;
wherein an increase in myosin-heavy chain fusion gene expression indicates that the test substance increases myosin heavy chain gene expression, or a decrease in myosin-heavy chain fusion gene expression indicates that the test substance decreases myosin heavy chain gene expression.