Methods
Through in-house services and close external collaborators, TCDM provides expertise and research facilities to implement-advanced, at-the-edge studies in reproductive and metabolic neuroendocrinology in suitable mouse (and rat) models, with special attention to characterization of hormonal responses to neuropeptide and metabolic regulators, quantitative and in situ analysis of gene and protein expression changes in brain areas of interest, and experimental manipulation of key central regulatory pathways. The major focus of the Unit is in reproductive (puberty & adult) and metabolic physiology. Hence, it closely collaborates with the Reproductive Biology Unit and the Nutrition and Metabolism Unit. In addition, the Unit maintains close connections with the laboratory of Experimental Neuroendocrinology at the Instituto Maimonides de Investigación Biomédica de Cordoba (IMIBIC) and the University of Cordoba (UCO), where Prof. Tena-Sempere serves also as PI.
Neuroendocrine phenotyping of rodent models
- Monitoring of pubertal onset and maturation by external (vaginal opening, preputial separation) and internal (gonadal histology, follicular dynamics, hormonal levels) indices.
- Analysis of reproductive and metabolic hormones, including gonadotropins, sex steroids, adipokines, pancreatic and gut hormones, in suitable preclinical models.
- Quantitative and in situ expression analyses of neuropeptides and receptors at the gene and protein levels in specific brain, mainly hypothalamic, areas in rodent models (in collaboration with IMIBIC-UCO).
- Analysis of hormonal and gene/protein expression changes after intracerebroventricular and stereotactic administration of compounds and virogenetic manipulation of specific neuronal populations, including DREADDs technology (in collaboration with IMIBIC-UCO).
- Analysis of reproductive and metabolic hormonal responses to pharmacological and nutritional (e.g., obesogenic) interventions (in collaboration with IMIBIC-UCO).
- Metabolic phenotyping (in collaboration with the Nutrition and Metabolic Unit)
- Reproductive phenotyping (in collaboration with the Reproductive Biology Unit)
Major neuroendocrine techniques
- Radioimmunoassay
- Stereotactic surgery (in collaboration with IMIBIC-UCO)
- Brain in situ hybridization (in collaboration with IMIBIC-UCO)
- Brain immunohistochemistry (in collaboration with IMIBIC-UCO)
- Virogenetic manipulation of neuronal populations, including DREADDs (in collaboration with IMIBIC-UCO)
- Electrophysiology and Optogenetics (expected from Jan 2016 on; in collaboration with IMIBIC-UCO)
Detection of GnRH mRNA expression by in situ hybridization
(GnRH neurons: grain clusters) in the preotic area (POA).
Images kindly provided by Juan Roa (IMIBIC-UCO)
Detection of Kiss1 mRNA expression by in situ hybridization
(Kiss1 neurons: grain clusters) in the arcuate nucleus (ARC).
Images kindly provided by Juan Roa (IMIBIC-UCO)