Figure 5

Effect of transfection of TM isoform-specific sense- and anti-sense oligonucleotides on myofibril organization in hearts from normal axolotl hearts. a. Stereo anaglyph of a 24 section confocal laser scanning microscope z-series of normal axolotl heart stained with CH1 anti-tropomyosin antibody. This type of image is shown because it demonstrates branching myofibrils in whole hearts. b. Stereo anaglyph of normal hearts transfected with TPM4α exon 2- specific anti-sense, 5′-T*A*C*T*AGCTCGTCCTCAAGC*T*G*C*-3′, where N* represents the phophothioate blocked oligonucleotide. Myofibirl organization was disrupted in a majority of the cardiomyocytes. Most of the tropomyosin appears to be in amorphous areas when expressed. Some of the cells do not appear to have a detectable level of tropomyosin. Gross morphology was normal and the cells appeared to be intact although myofibril structure was largely disrupted. The contractility was diminished significantly [36]. c. Normal heart transfected with TPM4α exon 2-specific sense chimeric olighonucleotide, 5′-fG*C*A*GCTTGAAGGCGAGCTA*G*T*A*-3, where *N represents phosphothioate blocked nucleotide, and fG represents G tagged with Fluorescein at the 5′end. This image is a compressed z-series of 2 sections that was not stereo offset but used to demonstrate double staining. Isolated pieces of myofibrils that were contained within these sections of the cardiac cells were stained with tropomyosin antibody (re, arrowheads). This image is primarily useful for demonstrating the presence green fluorescence (GFP) within the nuclear area of the cariomyocytes by five days (asterik). Hoescht staining of the nuclei in the same heart coincided with the FITC staining that were localized in a majority of nuclei (figure not shown). The figure was adapted from Spinner et al. (2004) [36].