Atrial t-tubules adopt a distinct developmental state as Ca2+ handling matures postnatally

Transverse (t)-tubules ensure a uniform rise in cellular Ca 2+ facilitating cardiac contraction. They play a key role in the large mammalian atria (including human) and their loss in heart failure is associated with impaired Ca 2+ release. While t-tubule restoration is therefore an ideal therapeutic target, atrial t-tubule development is not well understood. Here we sought to determine how atrial t-tubules develop and the impact on Ca 2+ handling. Atrial postnatal development was examined in sheep from newborn through to adulthood. T-tubule development was assessed using confocal microscopy and serial block face Scanning Electron Microscopy. Voltage clamp coupled with Ca 2+ epifluorescence was used to assess concomitant functional changes to Ca 2+ handling. Atrial t-tubule density increased until 3 months of age when the t-tubule network was disordered. As development continued t-tubules became more ordered but surprisingly the distance of the cell interior to t-tubule membrane increased due to a lack of additional t-tubules coupled with increased cell width. As t-tubules developed, L-type Ca 2+ current density ( I Ca-L ) and sarcoplasmic reticulum (SR) Ca 2+ content decreased. Although these changes would be expected to decrease Ca 2+ transient amplitude, Ca 2+ buffering was simultaneously reduced which our data suggests maintains Ca 2+ transient amplitude during neonatal development. By understanding how the Ca 2+ transient is preserved despite drastic changes in t-tubule density and structure during development, this study may provide insights into adaptive mechanisms in Ca 2+ cycling that mitigate the impact of reduced t-tubule density.