FIG. 6.

RT-PCR coamplification of leptin receptor Ob-Rb and GAPDH genes during the acute stage of infection. Total RNAs were extracted from the hippocampi, cortexes, cerebella, and hypothalami of three sham-inoculated and three infected mice. RT was performed on 0.5 μg of total pooled RNA, which was then diluted 1/10 and subjected to 28 cycles of PCR (95°C for 45 s, 60°C for 45 s, and 72°C for 1 min), using 3 mM MgCl₂. Note the upregulation of Ob-Rb expression in the hypothalami of infected mice compared to that in sham-inoculated mice. The results for each brain structure (ratio of Ob-Rb counts per minute to GAPDH counts per minute in infected structures relative to those in sham-inoculated mouse structures) are given in Table 5.