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To examine if brain neurons involved in the efferent control of the kidneys possess melanocortin-4 receptor (MC4-R) and/or tryptophan hydroxylase (TPH). Retrograde tracing pseudorabies virus (PRV)-614 was injected into the kidneys in adult male MC4R-green fluorescent protein (GFP) transgenic mice. After a survival time of 3-7 days, spinal cord and brain were removed and sectioned, and processed for PRV-614 visualization. The neurochemical phenotype of PRV-614-positive neurons was identified using double or triple immunocytochemical labeling against PRV-614, MC4R, or TPH. Double and triple labeling was quantified using microscopy. The majority of PRV-614 immunopositive neurons which also expressed immunoreactivity for MC4R were located in the ipsilateral intermediolateral cell column (IML) of the thoracic spinal cord, the paraventricular nucleus (PVN) of the hypothalamus, and raphe pallidus (RPa), nucleus raphe magnus (NRM) and ventromedial medulla (VMM) of the brainstem. Triple-labeled MC4R/PRV-614/TPH neurons were concentrated in the PVN, RPa, NRM and VMM. These data strongly suggest that central MC4R and TPH are involved in the efferent neuronal control of the kidneys.

Delineation of the central melanocortin circuitry controlling the kidneys by a virally mediated transsynaptic tracing study in transgenic mouse model