Role of desmin filaments in chicken cardiac myofibrillogenesis.
Desmin filaments are muscle-specific intermediate filaments located at the periphery of the Z-discs, and they have been postulated to play a critical role in the lateral registration of myofibrils. Previous studies suggest that intermediate filaments may be involved in titin assembly during the early stages of myofibrillogenesis. In order to investigate the putative function of desmin filaments in myofibrillogenesis, rabbit anti-desmin antibodies were introduced into cultured cardiomyocytes by electroporation to perturb the normal function of desmin filaments. Changes in the assembly of several sarcomeric proteins were examined by immunofluorescence. In cardiomyocytes incorporated with normal rabbit serum, staining for alpha-actinin and muscle actin displayed the typical Z-line and I-band patterns, respectively, while staining for titin with monoclonal anti-titin A12 antibody, which labels a titin epitope at the A-I junction, showed the periodic doublet staining pattern. Staining for C-protein gave an amorphous pattern in early cultures and identified A-band doublets in older cultures. In contrast, in cardiomyocytes incorporated with anti-desmin antibodies, alpha-actinin was found in disoriented Z-discs and the myofibrils became fragmented, forming mini-sarcomeres. In addition, titin was not organized into the typical A-band doublet, but appeared to be aggregated. Muscle actin staining was especially weak and appeared in tiny clusters. Moreover, in all ages of cardiomyocytes tested, C-protein remained in the disassembled form. The present data suggest the essential role of desmin in myofibril assembly.
Wang
S M
SM
Department of Anatomy, College of Medicine, National Taiwan University, Taipei, Taiwan 100, Republic of China. smwang@ha.mc.ntu.edu.tw
Huang
Y S
YS
Wu
J C
JC
Tseng
Y Z
YZ
D000185:Actinin / Q000378:metabolism; D000199:Actins / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D002352:Carrier Proteins / Q000378:metabolism; D002642:Chick Embryo; D064211:Connectin; D003599:Cytoskeleton / Q000378:metabolism; D003893:Desmin / Q000378:metabolism / Q000502:physiology*; D018274:Electroporation; D000939:Epitopes; D005455:Fluorescent Antibody Technique; D009124:Muscle Proteins / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism*; D009210:Myofibrils / Q000378:metabolism*; D011494:Protein Kinases / Q000378:metabolism; D011817:Rabbits
D000199:Actins; D002352:Carrier Proteins; D064211:Connectin; D003893:Desmin; D000939:Epitopes; D009124:Muscle Proteins; C020723:citrate-binding transport protein; D000185:Actinin; D011494:Protein Kinases