Ferret Study

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The ferret SA node is located at the junction of the superior vena cava and right atrium near the sulcus terminalis region (Fig. 1, A and B). Cross sections of the right atrium revealed an ovoid shaped SA node positioned epicardially (Fig. 1C). A light microscopic view of the SA node region shows two nodal arteries, centrally located, surrounded by a ring of dark cells (Fig. 1D). Larger, pale cells form a concentric ring around the dark cells (Fig. 1D). Thus, while being smaller, many features of ferret SA node displayed striking similarities to classic descriptions of human SA node (29; 69; 70). To characterize specific anatomical and functional regions of ferret SA node, we next performed fluorescent in situ hybridization (FISH) to detect (i) cardiac and slow twitch forms of troponin I mRNA (TnIC and TnIS, respectively; Fig. 2, A-D), (ii) cytoskeletal middle neurofilament protein (NF; Fig. 2E) and (iii) growth associated protein 43 (GAP43; Fig. 2F). TnIC has been shown to be the dominant troponin I isoform in atrial myocytes, while TnIS is present in the atrioventricular (AV) node (19). Cytoskeletal middle neurofilament protein (NF) is expressed in the SAN (18; 67; 74). GAP-43 is a marker for neurons (1; 13; 25; 73; 77; 78). Signals generated by antisense probes to TnIS (Fig. 2A) and TnIC (Fig. 2B) show that both transcripts are expressed in our SA nodal sections. TnIS expression was prominent in the central region but minimal to absent in the outer peripheral regions. However, TnIC expression was prominent beyond the central region and minimal to absent in the central region. Overlay of these two transcript expression patterns emphasizes the distinct distribution of cells expressing the two isoforms (Fig. 2C). Fig... ... middle of paper ... ...measure of protein expression. As we have previously shown with Kv1.4, the level of mRNA does not necessarily correlate with protein, although this effect does not seem to be common (10; 51). A second potential issue is that mRNAs detected by FISH may not overlap with protein location within a cell, potentially distorting channel distribution in large distended cells. To ascertain whether this might be the case for channels in the SA node, we performed immunolocalization of several K+ channels (Fig. 8). As is true in ferret ventricles (10), Kv1.4 transcript expression is much greater than the protein expression in the ferret SA node (Fig. 8A). In addition, Kv1.5 transcript expression was also greater than protein expression in the SA node (Fig. 8B). For the other transcripts tested, there are close correlations between transcript and protein (Fig. 8, C-H).

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