Kisspeptin signaling in astrocytes modulates the reproductive axis
Encarnacion Torres, … , Manuel Tena-Sempere, Antonio Romero-Ruiz
Encarnacion Torres, … , Manuel Tena-Sempere, Antonio Romero-Ruiz
Published June 11, 2024
Citation Information: J Clin Invest. 2024;134(15):e172908. https://doi.org/10.1172/JCI172908.
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Research Article Endocrinology Reproductive biology

Kisspeptin signaling in astrocytes modulates the reproductive axis

Abstract

Reproduction is safeguarded by multiple, often cooperative, regulatory networks. Kisspeptin signaling, via KISS1R, plays a fundamental role in reproductive control, primarily by regulation of hypothalamic GnRH neurons. We disclose herein a pathway for direct kisspeptin actions in astrocytes that contributes to central reproductive modulation. Protein-protein interaction and ontology analyses of hypothalamic proteomic profiles after kisspeptin stimulation revealed that glial/astrocyte markers are regulated by kisspeptin in mice. This glial-kisspeptin pathway was validated by the demonstrated expression of Kiss1r in mouse astrocytes in vivo and astrocyte cultures from humans, rats, and mice, where kisspeptin activated canonical intracellular signaling-pathways. Cellular coexpression of Kiss1r with the astrocyte markers GFAP and S100-β occurred in different brain regions, with higher percentage in Kiss1- and GnRH-enriched areas. Conditional ablation of Kiss1r in GFAP-positive cells in the G-KiR-KO mouse altered gene expression of key factors in PGE2 synthesis in astrocytes and perturbed astrocyte-GnRH neuronal appositions, as well as LH responses to kisspeptin and LH pulsatility, as surrogate marker of GnRH secretion. G-KiR-KO mice also displayed changes in reproductive responses to metabolic stress induced by high-fat diet, affecting female pubertal onset, estrous cyclicity, and LH-secretory profiles. Our data unveil a nonneuronal pathway for kisspeptin actions in astrocytes, which cooperates in fine-tuning the reproductive axis and its responses to metabolic stress.

Authors

Encarnacion Torres, Giuliana Pellegrino, Melissa Granados-Rodríguez, Antonio C. Fuentes-Fayos, Inmaculada Velasco, Adrian Coutteau-Robles, Amandine Legrand, Marya Shanabrough, Cecilia Perdices-Lopez, Silvia Leon, Shel H. Yeo, Stephen M. Manchishi, Maria J. Sánchez-Tapia, Victor M. Navarro, Rafael Pineda, Juan Roa, Frederick Naftolin, Jesús Argente, Raúl M. Luque, Julie A. Chowen, Tamas L. Horvath, Vincent Prevot, Ariane Sharif, William H. Colledge, Manuel Tena-Sempere, Antonio Romero-Ruiz

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Figure 2

Evidence for kisspeptin signaling in astrocytes.

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Evidence for kisspeptin signaling in astrocytes. (A) Expression analysis...
(A) Expression analysis of glial markers Gfap/GFAP and Vimentin/Vimentin at mRNA and protein levels in POA of adult Kiss1-KO male mice after i.c.v. Kp-10 stimulation (n = 3–4) versus vehicle (n = 3). Data are the mean ± SEM. Statistical significance was determined by Student’s t test: *P < 0.05 versus KO mice treated with vehicle. (B) Representative gels illustrating the expression of Kiss1r, but not Kiss1 mRNA in 2 pools of primary astrocyte cultures from neonatal rat (upper gel) and mouse (lower gel) hypothalamus are presented. Hypothalamic (HTLA) tissue was used as positive control. MM, molecular markers. Real-time PCR of Kiss1r mRNA in primary mouse and human astrocyte cultures (n = 4 for mouse; n = 6 cortical and 5 hypothalamic human cultures) is also shown; values correspond to Ct data. The blue line represents the mean Ct value of the housekeeping gene. In (C), Western blots of phosphorylated ERK (pERK) and AKT (pAKT) in primary rat hypothalamic astrocytes are shown. Bar graphs show the effect of Kp-10 treatment (10–8 M; n = 3) at 1, 10, and 30 minutes (upper panel); representative blots are shown in the lower panel. Astrocyte cultures treated with vehicle (n = 3) were used as a negative control. Data are the mean ± SEM. Statistical significance was determined by Student’s t test: **P < 0.01 versus astrocytes treated with vehicle. (D) Western blots of pERK, total ERK (totERK), pAKT, total AKT (totAKT), and actin, in primary mouse cerebrocortical and hypothalamic astrocytes treated with Kp-10 (n = 3) or Epidermal Growth Factor (EGF, 50 ng/mL; n = 3), used as a positive control. Vehicle-treated astrocytes (n = 3) were used as a negative controls. Data are the mean ± SEM. Statistical significance was determined by 2-way ANOVA followed by Bonferroni’s post hoc test: ****P < 0.0001, astrocytes treated with Kp-10 versus vehicle; or cortical versus hypothalamic astrocytes.