High throughput biological heart rate monitor that is molecularly determined
This invention provides for a chamber and system designed for use in assaying drug effects on heart rate. The chamber consists of a series of wells. Cardiac myocytes disaggregated from neonatal animals are plated onto the bottom of each well and grown under standard tissue culture conditions. When drugs are to be assayed, the cells in each well are loaded with a calcium sensitive dye and the beating rate in each is monitored with a photodiode. A drug is added in graded concentrations to each well, and equilibrated and effects on rate are observed. This construct permits use of a cell based bioassay for the study of drugs or agents that may alter cardiac rate. This invention can be used in high throughput screening of drugs to evaluate/predict their effects on cardiac rate and rhythm.
1. A method of assaying whether an agent affects the beating rate of a cardiac myocyte which comprises:
(a) contacting a cardiac myocyte in vitro with an amount of a composition comprising a nucleic acid which encodes a HCN channel and MiRP1, effective to cause a sustainable beating rate;
(b) measuring the beating rate after step (a);
(c) contacting the cardiac myocyte with an agent to be assayed for its effects on the beating rate;
(d) measuring the beating rate after step (c); and
(e) comparing the difference between step (b) and step (d) thereby determining whether the agent affects the beating rate.
2. The method of claim 1 , wherein the cardiac myocyte is mammalian.
3. The method of claim 1 , wherein the composition is introduced into the cell by an adenovirus infection, viral-mediated infection, liposome-mediated transfer, microinjection, electroporation, or by coculturing the cell with the composition.
4. The method of claim 3 , wherein the HCN is HCN1.
5. The method of claim 3 , wherein the HCN is HCN2.
6. The method of claim 3 , wherein the HCN is HCN4.
7. The method of claim 1 , wherein the HCN channel is HCN2.
8. The method of claim 1 , wherein the HCN channel is HCN1.
9. The method of claim 1 , wherein the HCN channel is HCN4.