Synthetic strands of neonatal mouse cardiac myocytes: structural and electrophysiological properties.
The aim of the present study was to morphologically and electrically characterize synthetic strands of mouse ventricular myocytes. Linear strands of mouse ventricular myocytes with widths of 34.7+/-4.4 microm (W(1)), 57.9+/-2.5 microm (W(2)), and 86.4+/-3. 6 microm (W(3)) and a length of 10 mm were produced on glass coverslips with a photolithographic technique. Action potentials (APs) were measured from individual cells within the strands with cell-attached microelectrodes. Impulse propagation and AP upstrokes were measured with multisite optical mapping (RH237). Immunostaining was performed to assess cell-cell connections and myofibril arrangement with polyclonal antisera against connexin43 and N-cadherins and monoclonal antibodies against cardiac myosin. Light microscopy and myosin staining showed dense growth of well-developed elongated myocytes with lengths of 34.2+/-4.2 microm (W(1)), 36. 9+/-5.8 microm (W(2)), and 43.7+/-6.9 microm (W(3)), and length/width ratios of 3.9+/-0.2. Gap junctions were distributed around the cell borders (3 to 4 junctions/microm(2) cell area). Each cell was connected by gap junctions to 6.5+/-1.1 neighboring cells. AP duration shortened with time in culture (action potential duration at 50% repolarization: day 4, 103+/-34 ms; day 8, 16+/-3 ms; P:<0.01). Minimum diastolic potential and AP amplitude were 71+/-5 and 97.2+/-7.6 mV, respectively. Conduction velocity and the maximum dV/dt of the AP upstroke were 43.9+/-13.6 cm/s and 196+/-67 V/s, respectively. Thus, neonatal ventricular mouse myocytes can be grown in continuous synthetic strands. Gap junction distribution is similar to the neonatal pattern observed in the hearts of larger mammals. Conduction velocity is in the range observed in adult mice and in the higher range for mammalian species probably due to the higher dV/dt(max). This technique will permit the study of propagation, AP, and structure-function relations at cellular resolution in genetically modified mice.
Thomas
S P
SP
Department of Physiology, University of Bern, Switzerland.
Bircher-Lehmann
L
L
Thomas
S A
SA
Zhuang
J
J
Saffitz
J E
JE
Kléber
A G
AG
D000200:Action Potentials; D000818:Animals; D000831:Animals, Newborn; D048429:Cell Size; D002478:Cells, Cultured; D018031:Connexin 43 / Q000378:metabolism; D005456:Fluorescent Dyes; D006352:Heart Ventricles / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D007150:Immunohistochemistry; D007365:Intercellular Junctions / Q000378:metabolism / Q000502:physiology; D051379:Mice; D008839:Microelectrodes; D018613:Microscopy, Confocal; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D055095:Optics and Photonics; D011726:Pyridinium Compounds
D018031:Connexin 43; D005456:Fluorescent Dyes; D011726:Pyridinium Compounds; C048153:4-(6-(4-dibutylaminophenyl)-1,3,5-hexatrienyl)-1-(4'-sulfobutyl)pyridinium