The cardiac mutant Mexican axolotl is a unique animal model for evaluation of cardiac myofibrillogenesis.
Hearts from cardiac mutant Mexican axolotl, Ambystoma mexicanum, do not form organized myofibrils and fail to beat. Though previous biochemical and immunohistochemical experiments showed a possible reduction of cardiac tropomyosin it was not clear that this caused the lack of organized myofibrils in mutant hearts. We used cationic liposomes to introduce both rabbit and chicken tropomyosin protein into whole hearts of embryonic axolotls in whole heart organ cultures. The mutant hearts had a striking increase in the number of well-organized sarcomeric myofibrils when treated with rabbit or chicken tropomyosin. FITC-labeled rabbit tropomyosin was used to examine the kinetics of incorporation of the exogenous protein into mutant hearts and confirmed the uptake of exogenous protein by the cells of live hearts in culture. By 4 h of transfection, both normal and mutant hearts were found to incorporate FITC-labeled tropomyosin into myofibrils. We also delivered an anti-tropomyosin antibody (CH 1) into normal hearts to disrupt the existing cardiac myofibrils which also resulted in reduced heartbeat rates. CH1 antibody was detected within the hearts and disorganization of the myofibrils was apparent when compared to normal controls. Introduction of a C-protein monoclonal antibody (ALD 66) did not result in a disruption of organized myofibrils. The results show clearly that chicken or rabbit tropomyosin could be incorporated by the mutant hearts and that it was sufficient to overcome the factors causing a lack of myofibril formation in the mutant. This finding also suggests that a lack of organized myofibrils is caused primarily by either inadequate levels of tropomyosin or endogenous tropomyosin in mutant hearts is unsuitable for myofibril formation, which we were able to duplicate with the introduction of tropomyosin antibody. Furthermore, incorporation of a specific exogenous protein or antibody into normal and mutant hearts of the Mexican axolotl in whole heart organ culture offers an unique model to evaluate functionalroles of contractile proteins necessary for cardiac development and differentiation.
Zajdel
R W
RW
Department of Anatomy and Cell Biology, SUNY Health Science Center at Syracuse, Syracuse, New York, 13210, USA.
Dube
D K
DK
Lemanski
L F
LF
D000558:Ambystoma mexicanum / Q000235:genetics*; D000818:Animals; D002645:Chickens; D006330:Heart Defects, Congenital / Q000235:genetics*; D018613:Microscopy, Confocal; D009024:Morphogenesis; D009124:Muscle Proteins / Q000378:metabolism; D009154:Mutation; D009200:Myocardial Contraction / Q000235:genetics*; D009210:Myofibrils / Q000235:genetics*; D010714:Phosphatidylethanolamines; D011817:Rabbits; D014335:Tropomyosin / Q000172:deficiency* / Q000494:pharmacology
D009124:Muscle Proteins; D010714:Phosphatidylethanolamines; D014335:Tropomyosin; C020888:1,2-dielaidoylphosphatidylethanolamine