Spectroscopic probing of dynamic changes during stimulation and cell remodeling in the single cardiac myocyte.
Optical microscopy, involving both fluorescence imaging and confocal Raman microspectroscopy, was used to visualize single, isolated, electrically active heart muscle cells. For example, short-term, dynamic changes in Raman bands during the contraction cycle, as well as persistent band changes during structural remodeling (microscopic rearrangements of cellular structures) in culture over longer periods of time, were obtained from the cellular content (sarcoplasm) of the heart cells. The results of the short-term studies, collected during electrical stimulation, showed dynamic changes in the Raman amide I band intensity, which occurred in phase with changes in cell length during cardiomyocyte contraction. The longer term studies of quiescent cardiomyocytes in culture over 3 days revealed a progressive and sustained increase in the intensity of the amide I band. Over the same period of culture, a decrease in the number of t-tubules (invaginations of the cell membrane, sarcolemma, which ensure the spreading of the action potential into the bulk of the sarcoplasm) was observed using confocal z-sections of the fluorescently labeled sarcolemma. The ability to measure both short-term dynamic changes associated with stimulated contraction and longer term persistent remodeling in the structure of intracellular macromolecules is valuable for assessing the physiological state of the cell, in real time.
Inya-Agha
O
O
Department of Electronics, University of Glasgow, Glasgow G12 8LT, Scotland.
Klauke
N
N
Davies
T
T
Smith
G
G
Cooper
J M
JM
D000200:Action Potentials / Q000502:physiology; D000577:Amides / Q000737:chemistry; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002478:Cells, Cultured; D004558:Electric Stimulation; D007424:Intracellular Fluid / Q000737:chemistry / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D009200:Myocardial Contraction / Q000502:physiology*; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000473:pathology / Q000648:ultrastructure; D012508:Sarcolemma / Q000502:physiology; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013997:Time Factors
D000577:Amides