Table 1.
Sequence of primers used for PCR analysis of PIV3.
Table 2.
Sequence of primers used for analysis of guinea pig ganglia receptor transcripts.
Table 3.
Sequence of primers used for qPCR analysis of guinea pig neurotrophin transcripts.
Fig 1.
PIV3 infection of epithelium in guinea pig trachea 4 d after inoculation with virus or vehicle (control).
A) PIV3 staining of tracheal epithelium from a vehicle-treated control animal. B) PIV3 staining of tracheal epithelium following viral infection in vivo. Bar = 100 μm. C) Each lane represents RT-PCR amplification with primers for PIV3 genes: nucleocapsid protein (NP), fusion protein (FP), hemagglutinin-neuraminidase (HN), and a positive control- guinea pig β-actin (β).
Fig 2.
Guinea pig cough responses to inhaled capsaicin (0.1, 1, 3, 10 μM), citiric acid (CA; 0.01, 0.1, 0.3 M) and bradykinin (BK; 0.1 mg/ml) 4 d after viral inoculation.
Cough was confirmed in conscious guinea pigs based on changes in pressure and visual confirmation. Comparison of the average number of coughs between infected animals and controls were compared. A) Capsaicin: Control, n = 4; PIV3: n = 4. B) CA: Control, n = 8; PIV3, n = 8. D) Bradykinin: Control, n = 10; PIV3, n = 10. *Significantly different from control (p<0.05).
Fig 3.
TRPV1 expression in the jugular ganglia in control animals compared to those inoculated with PIV3 (day 4).
Increased expression of TRPV1 (qPCR) following PIV3 infection for 4 d. Control: n = 7, 14 ganglia; PIV3: n = 7, 14 ganglia. *Significantly different from control (p<0.05).
Fig 4.
Percentage of guinea pig tracheal-specific nodose neurons that express TRPV1 in controls compared to those inoculated with PIV3 (day 4).
Top, representative mRNA expression of TRPV1 from neurons. Bottom, total TRPV1 mRNA expression in infected animals (49.48 ±5.1%) compared to controls (16.28 ±4.0%). Control: n = 11, 86 neurons; PIV3: n = 10, 97 neurons. *Significantly different from control (p<0.05).
Fig 5.
Vehicle or viral infections of the trachea occurred ex vivo for qPCR analyses of neurotrophin levels.
Upper left, a schematic of the isolated, perfused guinea pig airway apparatus used to virally infect intact epithelium. Lower right graph demonstrates an increased expression of BNDF and artemin (ARTN) in trachea following infection with 1/100 diluted PIV3 virus (stock = 2x107 pfu/ml) for 8 h. The data are the average of 4 separate experiments. *Significantly different from control (p<0.05).