Differential Alterations of the Mitochondrial Morphology and Respiratory Chain Complexes during Postnatal Development of the Mouse Lung

Figure 7

Representative immunofluorescence analysis of mitochondrial complex IV subunit I protein in AECII from lung tissue sections of newborn (NB), P15, and adult (AL) animals. Lung tissue samples from the three postnatal stages were embedded into paraffin. Thereafter, 3 μm paraffin sections were cut with a rotation microtome and processed further for indirect double immunofluorescence. The lung sections were incubated overnight for double labelling with primary antibodies against mitochondrial complex IV subunit I and pro-SP-C, a marker for AECII (Table 1). The following morning, the sections were washed and incubated with the secondary antibodies (Table 1) for 2 h at room temperature. Double fluorescence samples were analyzed by confocal laser scanning microscopy (CLSM) with a Leica TCS SP5. (a, d, and g) Double IF stainings of AECII with their marker protein pro-SP-C. (b, e, and h) IF preparations for the mitochondrial complex IV subunit I. (c, f, and i) Double IF overlay for complex IV subunit I combined with pro-SP-C. NB, newborn; P15, postnatal day 15; AL, adult. Bars represent 20 μm.

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)