title: Expression and distribution of cyclic AMP- and cyclic GMP-binding protein kinases in the human vagina- an immunohistochemical study.
abstract: BACKGROUND
  In contrast to research findings describing the localization of nitric oxide synthases (NOS), guanylyl cyclases, and cyclic adenosine monophosphate (cAMP)- and cyclic guanosine monophosphate (cGMP)-degrading phosphodiesterase isoenzymes in the human vagina, the distribution of proteins known as major targets for cyclic nucleotides has not yet been evaluated. cAMP- and cGMP-dependent protein kinases (cAK, cGKI) have been identified as important receptors for cyclic nucleotides downstream the signaling cascades.
  
  
  OBJECTIVE
  To investigate, by means of immunohistochemistry, the expression of cAK and cGKI in relation to endothelial NOS (eNOS), vasoactive intestinal polypeptide (VIP), and protein gene product 9.5 (PGP 9.5) in the human vagina.
  
  
  METHODS
  Expression and distribution of cAK and cGKI(alpha,beta) in relation to eNOS, VIP, and PGP 9.5 in human vaginal tissue.
  
  
  METHODS
  Immunohistochemical techniques were applied to sections of human vaginal full wall specimens in order to evaluate the presence of cAK and cGKI(alpha,beta) in relation to VIP, PGP 9.5, and eNOS, respectively. Western blot analyses were conducted using cytosolic supernatants of homogenized specimens of the vaginal wall and epithelium.
  
  
  RESULTS
  Immunostaining specific for cGKIbeta was observed in vascular and nonvascular smooth muscle of the vagina. In the endothelial layer, cGKIbeta was found colocalized with eNOS. In contrast, no signals indicating cGKIalpha were registered. cAK-positive subepithelial vessels were found to be innervated by a dense meshwork of PGP-containing varicose nerve fibers, some of which presented expression of VIP. The expression of cAK and cGKIbeta was confirmed by Western blotting.
  
  
  CONCLUSIONS
  Our results demonstrate the expression of cAK and cGKIbeta in the human vagina. The colocalization with VIP and eNOS underlines the significance of both the cAMP and GMP pathway in the control of human vaginal vascular and nonvascular smooth muscle.
authors.affiliation: Division of Surgery, Department of Urology and Uro-Oncology, Hannover Medical School, Hannover 30625, Germany. streetgang@gmx.de
authors.forename: Stefan
authors.initials: S
authors.lastname: Uckert
authors.forename: Eginhard S
authors.initials: ES
authors.lastname: Waldkirch
authors.forename: Knut
authors.initials: K
authors.lastname: Albrecht
authors.forename: Julia
authors.initials: J
authors.lastname: Sonnenberg
authors.forename: Kristina
authors.initials: K
authors.lastname: Langnäse
authors.forename: Karin
authors.initials: K
authors.lastname: Richter
authors.forename: Petter
authors.initials: P
authors.lastname: Hedlund
authors.forename: Markus A
authors.initials: MA
authors.lastname: Kuczyk
mesh_terms: D000293:Adolescent; D000328:Adult; D015153:Blotting, Western; D002102:Cadaver; D000242:Cyclic AMP / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000096:biosynthesis* / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism*; D017869:Cyclic GMP-Dependent Protein Kinases / Q000096:biosynthesis* / Q000378:metabolism; D004727:Endothelium / Q000201:enzymology / Q000378:metabolism; D005260:Female; D005629:Frozen Sections; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007150:Immunohistochemistry; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D019529:Sexuality; D015398:Signal Transduction; D043222:Ubiquitin Thiolesterase / Q000378:metabolism; D014621:Vagina / Q000201:enzymology* / Q000502:physiology; D014660:Vasoactive Intestinal Peptide / Q000378:metabolism; D055815:Young Adult
chemical_list: C482860:UCHL1 protein, human; D014660:Vasoactive Intestinal Peptide; D000242:Cyclic AMP; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D017868:Cyclic AMP-Dependent Protein Kinases; D017869:Cyclic GMP-Dependent Protein Kinases; D043222:Ubiquitin Thiolesterase; D006152:Cyclic GMP