Fig. 1
The full-length NK-1R isoform is expressed at robust levels in uninfected ex vivo rhesus macaque frontal cortical tissue, and SP levels are elevated in this tissue following B. burgdorferi infection. Cultured NHP brain tissue was uninfected (−) or infected (+) with B. burgdorferi (Bb, 1 × 107 bacteria; n = 4). Panel a At 2 h following infection, tissue expression of mRNA encoding the combined isoforms of NK-1R (NK-1R), the full-length isoform of NK-1R (fNK-1R), and pre-pro-tachykinin (PPT), was determined by RT-PCR and relative expression normalized to GAPDH levels was determined by densitometric analysis. Panel b At 2 h, protein expression of fNK-1R, the truncated NK-1R isoform (tNK-1R), and the housekeeping gene product β-actin, was determined by immunoblot analysis for each of the four brain tissue samples (1 through 4) either constitutively or following ex vivo infection. Expression in HeLa human epithelial (H) and CATH.a mouse neuronal (C) cell lines is included as positive controls. With an extended imaging exposure time, low-level tNK-1R expression could be discerned in the representative blot shown (middle bands). Panel c: At 4 h, fNK-1R protein expression and SP levels were determined in infected and uninfected tissue samples (n = 4) by immunoblot analysis and normalized to β-actin expression and specific capture ELISA, respectively (Panel c). Data is expressed as the mean ± SEM and asterisk indicates a statistically significant difference from uninfected brain tissue (p < 0.05)