title: Expression of neurokinin B/NK3 receptor and kisspeptin/KISS1 receptor in human granulosa cells.
abstract: OBJECTIVE
  Are neurokinin B (NKB), NK3 receptor (NK3R), kisspeptin (KISS1) and kisspeptin receptor (KISS1R) expressed in human ovarian granulosa cells?
  
  
  CONCLUSIONS
  The NKB/NK3R and kisspeptin/KISS1R systems are co-expressed and functionally active in ovarian granulosa cells.
  
  
  BACKGROUND
  The NKB/NK3R and KISS1/KISS1R systems are essential for reproduction. In addition to their well-recognized role in hypothalamic neurons, these peptide systems may contribute to the control of fertility by acting directly on the gonads, but such a direct gonadal role remains largely unknown.
  
  
  METHODS
  This study analyzed matched mural granulosa cells (MGCs) and cumulus cells (CCs) collected from preovulatory follicles of oocyte donors at the time of oocyte retrieval.
  
  
  METHODS
  The samples were provided by 56 oocyte donor women undergoing ovarian stimulation treatment. Follicular fluid samples containing MGCs and cumulus-oocyte complexes were collected after transvaginal ultrasound-guided oocyte retrieval. RT-PCR, quantitative real-time PCR, immunocytochemistry and western blot were used to investigate the pattern of expression of the NKB/NK3R and KISS/KISS1R systems in MGCs and CCs. Intracellular free Ca(2+) levels, [Ca(2+)]i, in MGCs after exposure to NKB or KISS1, in the presence or not of tachykinin receptor antagonists, were also measured.
  NKB/NK3R and KISS1/KISS1R systems were expressed, at the mRNA and protein levels, in MGCs and CCs, with significantly higher expression in CCs. Kisspeptin increased the [Ca(2+)]i in the cytosol of human MGCs while exposure to NKB failed to induce any change in [Ca(2+)]i. However, the [Ca(2+)]i response to kisspeptin was reduced in the presence of NKB. The inhibitory effect of NKB was only partially mimicked by the NK3R agonist, senktide and marginally suppressed by the NK3R-selective antagonist SB 222200. Yet, a cocktail of antagonists selective for the NK1, NK2 and NK3 receptors blocked the effect of NKB.
  
  
  CONCLUSIONS
  The granulosa and cumulus cells were obtained from oocyte donors undergoing ovarian stimulation, which in comparison with natural cycles, may have affected gene and protein expression in granulosa cells.
  
  
  CONCLUSIONS
  Our data demonstrate that, in addition to their indispensable effects at the central nervous system, the NKB/NK3R and kisspeptin/KISS1R systems are co-expressed and are functionally active in non-neuronal reproductive cells of the female gonads, the ovarian granulosa cells.
  This work was supported by grants from Ministerio de Economía y Competitividad (CTQ2011-25564 and BFI2011-25021) and Junta de Andalucía (P08-CVI-04185), Spain. J.G.-O., F.M.P., M.F.-S., N.P., A.C.-R., T.A.A., M.H., M.R., M.T.-S. and L.C. have nothing to declare.
authors.affiliation: Instituto Valenciano de Infertilidad, Seville, Spain.
authors.forename: J
authors.initials: J
authors.lastname: García-Ortega
authors.affiliation: Instituto de Investigaciones Químicas, CSIC, Seville, Spain.
authors.forename: F M
authors.initials: FM
authors.lastname: Pinto
authors.affiliation: Instituto Valenciano de Infertilidad, Seville, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Fernández-Sánchez
authors.affiliation: Instituto Valenciano de Infertilidad, Seville, Spain.
authors.forename: N
authors.initials: N
authors.lastname: Prados
authors.affiliation: Instituto de Investigaciones Químicas, CSIC, Seville, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Cejudo-Román
authors.affiliation: Instituto de Enfermedades Tropicales y Salud Publica, Universidad de La Laguna, Tenerife, Canary Islands.
authors.forename: T A
authors.initials: TA
authors.lastname: Almeida
authors.affiliation: Instituto de Enfermedades Tropicales y Salud Publica, Universidad de La Laguna, Tenerife, Canary Islands.
authors.forename: M
authors.initials: M
authors.lastname: Hernández
authors.affiliation: Departamento de Biología Celular, Fisiología e Inmunología, Universidad de Cordoba, Cordoba, Spain CIBER Fisiopatología de la Obesidad y la Nutrición, Cordoba, Spain ISCiii and Instituto Maimónides de Investigación Biomédica de Córdoba/Hospital Universitario Reina Sofia, Cordoba, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Romero
authors.affiliation: Departamento de Biología Celular, Fisiología e Inmunología, Universidad de Cordoba, Cordoba, Spain CIBER Fisiopatología de la Obesidad y la Nutrición, Cordoba, Spain ISCiii and Instituto Maimónides de Investigación Biomédica de Córdoba/Hospital Universitario Reina Sofia, Cordoba, Spain luzcandenas@iiq.csic.es fi1tesem@uco.es.
authors.forename: M
authors.initials: M
authors.lastname: Tena-Sempere
authors.affiliation: Instituto de Investigaciones Químicas, CSIC, Seville, Spain luzcandenas@iiq.csic.es fi1tesem@uco.es.
authors.forename: L
authors.initials: L
authors.lastname: Candenas
mesh_terms: D002478:Cells, Cultured; D005260:Female; D006107:Granulosa Cells / Q000378:metabolism*; D006801:Humans; D059648:Kisspeptins / Q000235:genetics / Q000378:metabolism*; D015287:Neurokinin B / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D018039:Receptors, Tachykinin / Q000235:genetics / Q000378:metabolism*
chemical_list: D059648:Kisspeptins; D012333:RNA, Messenger; D018039:Receptors, Tachykinin; C102505:neurokinin receptor 4; D015287:Neurokinin B
keywords: human granulosa cells; kisspeptin; kisspeptin receptor; neurokinin B; tachykinin NK3 receptor