title: Autocrine/paracrine proliferative effect of ovarian GH and IGF-I in chicken granulosa cell cultures.
abstract: It is known that growth hormone (GH) and its receptor (GHR) are expressed in granulosa cells (GC) and thecal cells during the follicular development in the hen ovary, which suggests GH is involved in autocrine/paracrine actions in the female reproductive system. In this work, we show that the knockdown of local ovarian GH with a specific cGH siRNA in GC cultures significantly decreased both cGH mRNA expression and GH secretion to the media, and also reduced their proliferative rate. Thus, we analyzed the effect of ovarian GH and recombinant chicken GH (rcGH) on the proliferation of pre-hierarchical GCs in primary cultures. Incubation of GCs with either rcGH or conditioned media, containing predominantly a 15-kDa GH isoform, showed that both significantly increased proliferation as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, proliferating cell nuclear antigen (PCNA) quantification and ((3)H)-thymidine incorporation ((3)H-T) assays in a dose response fashion. Both, locally produced GH and rcGH also induced the phosphorylation of Erk1/2 in GC cultures. Furthermore, GH increased IGF-I synthesis and its release into the GC culture incubation media. These results suggest that GH may act through local IGF-I to induce GC proliferation, since IGF-I immunoneutralization completely abolished the GH-induced proliferative effect. These data suggest that GH and IGF-I may play a role as autocrine/paracrine regulators during the follicular development in the hen ovary at the pre-hierarchical stage.
authors.affiliation: Departamento de Neurobiología Celular y Molecular, Instituto de Neurobiología, Campus Juriquilla, Universidad Nacional Autónoma de México, Querétaro 76230, Mexico.
authors.forename: S Marisela
authors.initials: SM
authors.lastname: Ahumada-Solórzano
authors.affiliation: Departamento de Neurobiología Celular y Molecular, Instituto de Neurobiología, Campus Juriquilla, Universidad Nacional Autónoma de México, Querétaro 76230, Mexico.
authors.forename: Carlos G
authors.initials: CG
authors.lastname: Martínez-Moreno
authors.affiliation: Departamento de Neurobiología Celular y Molecular, Instituto de Neurobiología, Campus Juriquilla, Universidad Nacional Autónoma de México, Querétaro 76230, Mexico.
authors.forename: Martha
authors.initials: M
authors.lastname: Carranza
authors.affiliation: Departamento de Neurobiología Celular y Molecular, Instituto de Neurobiología, Campus Juriquilla, Universidad Nacional Autónoma de México, Querétaro 76230, Mexico.
authors.forename: José
authors.initials: J
authors.lastname: Ávila-Mendoza
authors.affiliation: Departamento de Neurobiología Celular y Molecular, Instituto de Neurobiología, Campus Juriquilla, Universidad Nacional Autónoma de México, Querétaro 76230, Mexico.
authors.forename: José Luis
authors.initials: JL
authors.lastname: Luna-Acosta
authors.affiliation: Dept. Physiology, University of Alberta, Edmonton T6G 2H7, Canada.
authors.forename: Steve
authors.initials: S
authors.lastname: Harvey
authors.affiliation: Departamento de Neurobiología Celular y Molecular, Instituto de Neurobiología, Campus Juriquilla, Universidad Nacional Autónoma de México, Querétaro 76230, Mexico.
authors.forename: Maricela
authors.initials: M
authors.lastname: Luna
authors.affiliation: Departamento de Neurobiología Celular y Molecular, Instituto de Neurobiología, Campus Juriquilla, Universidad Nacional Autónoma de México, Querétaro 76230, Mexico. Electronic address: aramburo@unam.mx.
authors.forename: Carlos
authors.initials: C
authors.lastname: Arámburo
mesh_terms: D000818:Animals; D019898:Autocrine Communication; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002645:Chickens; D005260:Female; D042341:Gonadal Hormones / Q000378:metabolism*; D006107:Granulosa Cells / Q000378:metabolism*; D013006:Growth Hormone / Q000378:metabolism*; D007334:Insulin-Like Growth Factor I / Q000378:metabolism*; D010053:Ovary / Q000378:metabolism*; D019899:Paracrine Communication
chemical_list: D042341:Gonadal Hormones; D007334:Insulin-Like Growth Factor I; D013006:Growth Hormone
keywords: Autocrine/paracrine; Extra-pituitary GH; Granulosa cells; Growth hormone; IGF-I; Proliferation