Insulin-like Growth Factor 1, Growth Hormone, and Anti-Müllerian Hormone Receptors Are Differentially Expressed during GnRH Neuron Development

Author:

Paganoni Alyssa J. J.1ORCID,Cannarella Rossella23ORCID,Oleari Roberto1ORCID,Amoruso Federica1ORCID,Antal Renata1,Ruzza Marco1ORCID,Olivieri Chiara1ORCID,Condorelli Rosita A.2ORCID,La Vignera Sandro2ORCID,Tolaj Fationa1ORCID,Cariboni Anna1ORCID,Calogero Aldo E.2ORCID,Magni Paolo14ORCID

Affiliation:

1. Department of Pharmacological and Biomolecular Sciences, Università degli Studi di Milano, 20133 Milan, Italy

2. Department of Clinical and Experimental Medicine, University of Catania, 95123 Catania, Italy

3. Glickman Urological & Kidney Institute, Cleveland Clinic Foundation, Cleveland, OH 10681, USA

4. IRCCS MultiMedica, Sesto S. Giovanni, 20099 Milan, Italy

Abstract

Gonadotropin-releasing hormone (GnRH) neurons are key neuroendocrine cells in the brain as they control reproduction by regulating hypothalamic-pituitary-gonadal axis function. In this context, anti-Müllerian hormone (AMH), growth hormone (GH), and insulin-like growth factor 1 (IGF1) were shown to improve GnRH neuron migration and function in vitro. Whether AMH, GH, and IGF1 signaling pathways participate in the development and function of GnRH neurons in vivo is, however, currently still unknown. To assess the role of AMH, GH, and IGF1 systems in the development of GnRH neuron, we evaluated the expression of AMH receptors (AMHR2), GH (GHR), and IGF1 (IGF1R) on sections of ex vivo mice at different development stages. The expression of AMHR2, GHR, and IGF1R was assessed by immunofluorescence using established protocols and commercial antibodies. The head sections of mice were analyzed at E12.5, E14.5, and E18.5. In particular, at E12.5, we focused on the neurogenic epithelium of the vomeronasal organ (VNO), where GnRH neurons, migratory mass cells, and the pioneering vomeronasal axon give rise. At E14.5, we focused on the VNO and nasal forebrain junction (NFJ), the two regions where GnRH neurons originate and migrate to the hypothalamus, respectively. At E18.5, the median eminence, which is the hypothalamic area where GnRH is released, was analyzed. At E12.5, double staining for the neuronal marker ß-tubulin III and AMHR2, GHR, or IGF1R revealed a signal in the neurogenic niches of the olfactory and VNO during early embryo development. Furthermore, IGF1R and GHR were expressed by VNO-emerging GnRH neurons. At E14.5, a similar expression pattern was found for the neuronal marker ß-tubulin III, while the expression of IGF1R and GHR began to decline, as also observed at E18.5. Of note, hypothalamic GnRH neurons labeled for PLXND1 tested positive for AMHR2 expression. Ex vivo experiments on mouse sections revealed differential protein expression patterns for AMHR2, GHR, and IGF1R at any time point in development between neurogenic areas and hypothalamic compartments. These findings suggest a differential functional role of related systems in the development of GnRH neurons.

Funder

Italian Ministry of Health

Italian Fondazione Telethon

Fondazione Collegio Ghislieri and Fondazione Telethon

European Union

Publisher

MDPI AG

Subject

Inorganic Chemistry,Organic Chemistry,Physical and Theoretical Chemistry,Computer Science Applications,Spectroscopy,Molecular Biology,General Medicine,Catalysis

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