Neurokinin 1 receptors and neprilysin modulation of mouse bladder gene regulation

Igor Dozmorov, Marcia R. Saban, Norma P. Gerard, Bao Lu, Ngoc Bich Nguyen, Michael Centola, Ricardo Saban

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Neurokinin 1 (NK1) receptors play a fundamental role in neurogenic inflammation. We sought to determine the mechanisms down-stream from NK1 receptor (NK1R) activation using cDNA arrays and a novel statistical method to analyze gene expression. We used female NK 1R-/- and wild-type (WT) mice that were sensitized actively by intraperitoneal injections of dinitrophenol 4 (DNP 4)-human serum albumin. Cystitis was induced by intravesical instillation of antigen of DNP4-ovalbumin, and control mice were challenged with saline. At 1, 4, and 24 h after instillation, bladders were removed for 1) RNA extraction (n = 3), 2) replicate of RNA extraction (n = 3), and 3) morphological analysis (n = 6). For cDNA array experiments, three bladders from each group were homogenized, and total RNA was obtained. DNase-treated RNA was reverse-transcribed to cDNA, labeled with [α- 32P]dATP and hybridized to Atlas Mouse 1.2 Arrays (Clontech). After calculating the mean and SD for background spots, each experimental value was assigned a normalized score S using the formula S′ = (S - Av)/SD, where S′ is the original pixel value, and Av and SD are the mean and standard deviation of background spots, respectively. Only genes that expressed 3 SD values above background were used. Hypervariable genes were sorted by cluster analysis. Matrices of correlation coefficients were calculated and represented in a connectivity mosaic. As results, we found that in WT mice the most prominent gene cluster had neprilysin in a central position and positively correlated to a group of activator protein-1 (AP-1)-responsive genes, including laminin-α3, tissue plasminogen activator 11, fos-B, and TNF-β. In WT mice, antigen-induced bladder inflammation led to a downregulation in neprilysin expression. In contrast, NK1R-/- mice failed to mount an inflammatory reaction and presented neprilysin negatively correlated with the same genes described in WT. In conclusion, this work indicates an overriding participation of NK1R and neprilysin in bladder inflammation, provides a working model for the involvement of AP-1 transcription factor, and evokes testable hypotheses regarding the role of NK1R and neprilysin in inflammation.

Original languageEnglish (US)
Pages (from-to)239-250
Number of pages12
JournalPhysiological Genomics
Volume12
StatePublished - Apr 2003

Fingerprint

Neurokinin-1 Receptors
Neprilysin
Urinary Bladder
RNA
Genes
Intravesical Administration
Transcription Factor AP-1
Inflammation
Oligonucleotide Array Sequence Analysis
Neurogenic Inflammation
Dinitrophenols
Antigens
Cystitis
Deoxyribonucleases
Atlases
Ovalbumin
Laminin
Tissue Plasminogen Activator
Multigene Family
Intraperitoneal Injections

Keywords

  • cDNA arrays
  • Cystitis animal model
  • Gene regulation
  • Inflammation
  • Neutral endopeptidase
  • Substance P
  • Transgenic/knockout

ASJC Scopus subject areas

  • Physiology
  • Genetics

Cite this

Dozmorov, I., Saban, M. R., Gerard, N. P., Lu, B., Nguyen, N. B., Centola, M., & Saban, R. (2003). Neurokinin 1 receptors and neprilysin modulation of mouse bladder gene regulation. Physiological Genomics, 12, 239-250.

Neurokinin 1 receptors and neprilysin modulation of mouse bladder gene regulation. / Dozmorov, Igor; Saban, Marcia R.; Gerard, Norma P.; Lu, Bao; Nguyen, Ngoc Bich; Centola, Michael; Saban, Ricardo.

In: Physiological Genomics, Vol. 12, 04.2003, p. 239-250.

Research output: Contribution to journalArticle

Dozmorov, I, Saban, MR, Gerard, NP, Lu, B, Nguyen, NB, Centola, M & Saban, R 2003, 'Neurokinin 1 receptors and neprilysin modulation of mouse bladder gene regulation', Physiological Genomics, vol. 12, pp. 239-250.
Dozmorov I, Saban MR, Gerard NP, Lu B, Nguyen NB, Centola M et al. Neurokinin 1 receptors and neprilysin modulation of mouse bladder gene regulation. Physiological Genomics. 2003 Apr;12:239-250.
Dozmorov, Igor ; Saban, Marcia R. ; Gerard, Norma P. ; Lu, Bao ; Nguyen, Ngoc Bich ; Centola, Michael ; Saban, Ricardo. / Neurokinin 1 receptors and neprilysin modulation of mouse bladder gene regulation. In: Physiological Genomics. 2003 ; Vol. 12. pp. 239-250.
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