Christian Pinali, Nadim Malik, J. Bernard Davenport, Laurence J. Allan, Lucy Murfitt, Mohammad M. Iqbal, Mark R. Boyett, Elizabeth J. Wright, Rachel Walker, Yu Zhang, Halina Dobryznski, Cathy M. Holt, Ashraf Kitmitto
Journal of the American Heart Association. 2017;6:e004834, originally published May 4, 2017
Christian Pinali, Nadim Malik, J. Bernard Davenport, Laurence J. Allan, Lucy Murfitt, Mohammad M. Iqbal, Mark R. Boyett, Elizabeth J. Wright, Rachel Walker, Yu Zhang, Halina Dobryznski, Cathy M. Holt, Ashraf Kitmitto
Journal of the American Heart Association May 2017, 6 (5) e004834; DOI: https://doi.org/10.1161/JAHA.116.004834
By ChristianPinali, NadimMalik, J. BernardDavenport, Laurence J.Allan, LucyMurfitt, Mohammad M.Iqbal, Mark R.Boyett, Elizabeth J.Wright, RachelWalker, YuZhang, HalinaDobryznski, Cathy M.Holt and AshrafKitmitto
Pig t‐ts form a radial distribution in control cardiac myocytes A, An exemplar portion of a serial SEM image from a volumetric stack illustrating the...Show More
Pig t‐ts form a radial distribution in control cardiac myocytes A, An exemplar portion of a serial SEM image from a volumetric stack illustrating the details of the cardiac myocyte ultrastructure. M, mitochondria; N, nucleus. Arrows indicate t‐ts. Scale bar=5 μm. B, Segmentation of the sarcolemma (pink) and t‐tubules (green) illustrating a radial organization extending from the exterior to interior of the cell. C, View of the t‐t network orthogonal to the sarcolemmal plane illustrating the regular spacing and organization of the t‐ts. Scale bar=2 μm. SEM indicates scanning electron microscopy.Show Less
Christian Pinali, Nadim Malik, J. Bernard Davenport, Laurence J. Allan, Lucy Murfitt, Mohammad M. Iqbal, Mark R. Boyett, Elizabeth J. Wright, Rachel Walker, Yu Zhang, Halina Dobryznski, Cathy M. Holt, Ashraf Kitmitto
Journal of the American Heart Association May 2017, 6 (5) e004834; DOI: https://doi.org/10.1161/JAHA.116.004834
By ChristianPinali, NadimMalik, J. BernardDavenport, Laurence J.Allan, LucyMurfitt, Mohammad M.Iqbal, Mark R.Boyett, Elizabeth J.Wright, RachelWalker, YuZhang, HalinaDobryznski, Cathy M.Holt and AshrafKitmitto
Tissue‐level characterization of the porcine heart post‐MI. A, Image of an explanted pig heart 4 weeks post‐...
Tissue‐level characterization of the porcine heart post‐MI. A, Image of an explanted pig heart 4 weeks post‐MI showing an area of infarct distinguished by a pale region of ischemic tissue; the peri‐infarct region (also termed border zone) directly adjacent to the infarct and remote region of the myocardium. B, Histological analysis of pig myocardium 4 weeks following infarction. (i) Remote area of myocardium showing normal cardiac myocytes. (ii) Section showing border zone (BZ) adjacent to infarct zone (IZ) with the dashed line illustrating a clear demarcation between the zones. The infarct zone is characterized by pale stained tissue containing inflammatory cells and neovessels. (iii) Infarct zone showing pale stained necrotic tissue with some inflammatory cell infiltration. Scale bar=100 μm. C, Fibroblast recruitment post‐MI in the pig. Fibroblasts immunopositive for vimentin stained red and alpha‐actin (green). Example immunostaining in the infarct zone 4 weeks postinfarct in the infarct zone (i) and corresponding border zone (ii). Scale bar=25 μm. D, Quantification of the myofibroblasts reveals a greater proportion within the infarct region compared to border and remote zones (***P<0.001). MI indicates myocardial infarction.
Christian Pinali, Nadim Malik, J. Bernard Davenport, Laurence J. Allan, Lucy Murfitt, Mohammad M. Iqbal, Mark R. Boyett, Elizabeth J. Wright, Rachel Walker, Yu Zhang, Halina Dobryznski, Cathy M. Holt, Ashraf Kitmitto
Journal of the American Heart Association May 2017, 6 (5) e004834; DOI: https://doi.org/10.1161/JAHA.116.004834
By ChristianPinali, NadimMalik, J. BernardDavenport, Laurence J.Allan, LucyMurfitt, Mohammad M.Iqbal, Mark R.Boyett, Elizabeth J.Wright, RachelWalker, YuZhang, HalinaDobryznski, Cathy M.Holt and AshrafKitmitto
Infarct region is devoid of cardiac myocytes. An exemplar portion of a serial SEM image from a volumetric stack illustrating a portion of the infarct...Show More
Infarct region is devoid of cardiac myocytes. An exemplar portion of a serial SEM image from a volumetric stack illustrating a portion of the infarct region and the surrounding peri‐infarct cardiac myocytes. The infarct is composed primarily of collagen and fibroblasts. Scale bar=20 μm. BV indicates blood vessel; Co, collagen; SEM, scanning electron microscopy.Show Less
Christian Pinali, Nadim Malik, J. Bernard Davenport, Laurence J. Allan, Lucy Murfitt, Mohammad M. Iqbal, Mark R. Boyett, Elizabeth J. Wright, Rachel Walker, Yu Zhang, Halina Dobryznski, Cathy M. Holt, Ashraf Kitmitto
Journal of the American Heart Association May 2017, 6 (5) e004834; DOI: https://doi.org/10.1161/JAHA.116.004834
By ChristianPinali, NadimMalik, J. BernardDavenport, Laurence J.Allan, LucyMurfitt, Mohammad M.Iqbal, Mark R.Boyett, Elizabeth J.Wright, RachelWalker, YuZhang, HalinaDobryznski, Cathy M.Holt and AshrafKitmitto
Highly branched enlarged t‐tubules form within the peri‐infarct cardiac myocytes. A, Exemplar serial SEM image showing a cardiac myocyte bordering the...Show More
Highly branched enlarged t‐tubules form within the peri‐infarct cardiac myocytes. A, Exemplar serial SEM image showing a cardiac myocyte bordering the infarct. Arrows highlight t‐ts with dimensions typically found in control, whereas solid arrow heads indicate what appear in 2D to be vacuoles. 3D reconstruction shows that these “vacuoles” are part of enlarged highly branched remodeled t‐ts. M, mitochondria; Z, Z‐line; Co, collagen; Fb, fibroblast process. Scale bar=5 μm. B, 3D reconstruction of the t‐ts within peri‐infarct cardiac myocytes. The t‐ts form “overgrown” structures; each separate “overgrown” t‐t is segmented in a different color; the sarcolemma is segmented in pink. Scale bar=5 μm. The dashed line indicates the position of the cross‐section through the cell shown in (C). C, In this view, the remodeled t‐ts can be seen to be attached to the sarcolemma (gray) as indicated by the arrows. There are regions where there are no t‐ts and the spacing of the remodeled t‐ts is irregular. The t‐ts enclosed by the dashed ellipsoid indicate t‐ts with a morphology comparable to control t‐ts. Scale bar=2 μm. D, (i) Shows a remodeled t‐t (blue) adjacent to a t‐t (green) with dimensions similar to control. (ii) Top view of the enlarged, branched invagination revealing how it is formed from multiple, smaller, t‐ts. Scale bar=500 nm. 2D indicates two‐dimensional; 3D, three‐dimensional; SEM, scanning electron microscopy.Show Less
Christian Pinali, Nadim Malik, J. Bernard Davenport, Laurence J. Allan, Lucy Murfitt, Mohammad M. Iqbal, Mark R. Boyett, Elizabeth J. Wright, Rachel Walker, Yu Zhang, Halina Dobryznski, Cathy M. Holt, Ashraf Kitmitto
Journal of the American Heart Association May 2017, 6 (5) e004834; DOI: https://doi.org/10.1161/JAHA.116.004834
By ChristianPinali, NadimMalik, J. BernardDavenport, Laurence J.Allan, LucyMurfitt, Mohammad M.Iqbal, Mark R.Boyett, Elizabeth J.Wright, RachelWalker, YuZhang, HalinaDobryznski, Cathy M.Holt and AshrafKitmitto
3D reconstruction of the t‐t morphology and arrangement in the remote region. A, The t‐ts (green) adopt a radial configuration extending from the sarc...Show More
3D reconstruction of the t‐t morphology and arrangement in the remote region. A, The t‐ts (green) adopt a radial configuration extending from the sarcolemma (pink) toward the center of the cell, stopping where there is a nucleus. B, View of the t‐t organization orthogonal to the sarcolemma illustrating the regular spacing of the invaginations. Scale bar=2 μm. C through E, Morphometric analysis of Z‐line spacing. C, Portion of a serial SEM image of the remote region illustrating how the sarcomere features are well defined with the Z‐lines identifiable as a dark band with a lighter I‐band flanking on either side. The central M‐band is also resolved. ICD; intercalated disc: M; mitochondria. D and E, Shows 2 views of how the sarcomere spacing was measured using IMOD, regions of 5 separate cells (indicated by the different color) were analyzed. Z‐lines were measured at 1‐μm intervals in the Z‐direction. Scale bar=2 μm. 3D indicates three‐dimensional; SEM, scanning electron microscopy.Show Less
Christian Pinali, Nadim Malik, J. Bernard Davenport, Laurence J. Allan, Lucy Murfitt, Mohammad M. Iqbal, Mark R. Boyett, Elizabeth J. Wright, Rachel Walker, Yu Zhang, Halina Dobryznski, Cathy M. Holt, Ashraf Kitmitto
Journal of the American Heart Association May 2017, 6 (5) e004834; DOI: https://doi.org/10.1161/JAHA.116.004834
By ChristianPinali, NadimMalik, J. BernardDavenport, Laurence J.Allan, LucyMurfitt, Mohammad M.Iqbal, Mark R.Boyett, Elizabeth J.Wright, RachelWalker, YuZhang, HalinaDobryznski, Cathy M.Holt and AshrafKitmitto
T‐Tubule Properties Determined From SBF‐SEM and TEM Analyses...
Christian Pinali, Nadim Malik, J. Bernard Davenport, Laurence J. Allan, Lucy Murfitt, Mohammad M. Iqbal, Mark R. Boyett, Elizabeth J. Wright, Rachel Walker, Yu Zhang, Halina Dobryznski, Cathy M. Holt, Ashraf Kitmitto
Journal of the American Heart Association May 2017, 6 (5) e004834; DOI: https://doi.org/10.1161/JAHA.116.004834
By ChristianPinali, NadimMalik, J. BernardDavenport, Laurence J.Allan, LucyMurfitt, Mohammad M.Iqbal, Mark R.Boyett, Elizabeth J.Wright, RachelWalker, YuZhang, HalinaDobryznski, Cathy M.Holt and AshrafKitmitto
Protein profiling of the tissue lysate from control pigs compared to the peri‐infarct and remote regions pos...
Protein profiling of the tissue lysate from control pigs compared to the peri‐infarct and remote regions post‐MI using western blotting techniques. The expression levels of the following proteins were analyzed: A, CaV1.2; ion channel subunit of the L‐type voltage‐gated calcium channel B, Tropomyosin C, Tcap; telethonin D, BIN‐1; Bridging integrator 1 E, JP2; junctophilin‐2 F, Exemplar western blots for each of the proteins analyzed. There is an imbalance between JP2 and BIN‐1 expression levels within the peri‐infarct and remote regions. Separate analysis of the BIN‐1 55‐ and 65‐kDa bands showed the same change as when combined. B; border zone (peri‐infarct region), R; remote region, C; control. (*P<0.05, **P<0.01)
Christian Pinali, Nadim Malik, J. Bernard Davenport, Laurence J. Allan, Lucy Murfitt, Mohammad M. Iqbal, Mark R. Boyett, Elizabeth J. Wright, Rachel Walker, Yu Zhang, Halina Dobryznski, Cathy M. Holt, Ashraf Kitmitto
Journal of the American Heart Association May 2017, 6 (5) e004834; DOI: https://doi.org/10.1161/JAHA.116.004834
By ChristianPinali, NadimMalik, J. BernardDavenport, Laurence J.Allan, LucyMurfitt, Mohammad M.Iqbal, Mark R.Boyett, Elizabeth J.Wright, RachelWalker, YuZhang, HalinaDobryznski, Cathy M.Holt and AshrafKitmitto
Immunolabeling of JP2, EHD2, and RyR2 in tissue sections from control and MI LV (peri‐infarct region). A, Representative confocal image of a tissue se...Show More
Immunolabeling of JP2, EHD2, and RyR2 in tissue sections from control and MI LV (peri‐infarct region). A, Representative confocal image of a tissue section with immunolabeling of JP2. JP2 is distributed in the control LV along each t‐t trajectory, the t‐t network has a radial organization in the transverse orientation in agreement with the higher‐resolution SBF‐SEM analyses. The right panel (peri‐infarct tissue) in agreement with the EM data now shows JP2 labeling along enlarged, branched irregular structures. B, EHD2 is accumulated between cells (highlighted with an asterisk [“*”]) with only weak staining within cardiac myocytes. The staining pattern in places is consistent with putative t‐ts (indicated by arrows). In the peri‐infarct region, the EHD2 labeling shows a complex organization. Dashed ellipsoid highlights putative labeling of remodeled t‐ts. C, Dual labeling of JP2 and RyR2 shows colocalization along the structures typical of t‐ts. In the peri‐infarct region, colocalization is maintained along the remodeled t‐ts. EHD2 indicates Eps 15 homology domain protein, isoform 2; EM, electron microscopy; JP2, junctophilin‐2; LV, left ventricle; MI, myocardial infarction; RyR2, ryanodine receptor 2; SBF‐SEM, serial block face scanning electron microscopy.Show Less
Christian Pinali, Nadim Malik, J. Bernard Davenport, Laurence J. Allan, Lucy Murfitt, Mohammad M. Iqbal, Mark R. Boyett, Elizabeth J. Wright, Rachel Walker, Yu Zhang, Halina Dobryznski, Cathy M. Holt, Ashraf Kitmitto
Journal of the American Heart Association May 2017, 6 (5) e004834; DOI: https://doi.org/10.1161/JAHA.116.004834
By ChristianPinali, NadimMalik, J. BernardDavenport, Laurence J.Allan, LucyMurfitt, Mohammad M.Iqbal, Mark R.Boyett, Elizabeth J.Wright, RachelWalker, YuZhang, HalinaDobryznski, Cathy M.Holt and AshrafKitmitto
T‐ts have an intricate folded inner membrane. A, Exemplar serial SEM image of remodeled t‐ts within the peri‐infarct region. Arrows highlight the fold...Show More
T‐ts have an intricate folded inner membrane. A, Exemplar serial SEM image of remodeled t‐ts within the peri‐infarct region. Arrows highlight the folds within the t‐ts. B, Exemplar serial SEM image of a “twin t‐t” within the remote region post‐MI. Arrows highlight the folds within both t‐ts. C, TEM image of control tissue showing several t‐ts in cross‐section. The inner membrane folds are clear in some t‐ts (indicated by arrows). D, TEM image of a control t‐t (partial transverse view) showing that, in some regions, the basal lamina is well defined, but in other parts, although not empty, the interior appears “fuzzy.” E, 3D reconstruction of a portion of a remodeled t‐t within the peri‐infarct region. The outer membrane envelope is shown in purple contours, and the inner folds are shown as green isosurface. The t‐t as viewed from both ends are shown in the panel below. F, (i–iv) correspond to cross‐sections through the t‐t as indicated by the dashed lines in (E). The inner membrane is formed by a high density of folds. Scale bars=200 nm. 3D indicates three‐dimensional; MI, myocardial infarction; SEM, scanning electron microscopy; TEM, transmission electron microscopy; t‐ts, transverse‐tubules.Show Less
