Nutritionally responsive PMv DAT neurons are dynamically regulated during pubertal transition
Authors: Sáenz de Miera Cristina , Bellefontaine Nicole , Silveira Marina A , Fortin Chelsea N , Zampieri Thais T , Donato Jose , Williams Kevin W , Mendes-da-Silva Cristiano , Heikkinen Laura , Broberger Christian , Frazao Renata , Elias Carol F 
Published: February 2025

Pubertal development is tightly regulated by energy balance. The crosstalk between metabolism and reproduction is orchestrated by complex neural networks and leptin action in the hypothalamus plays a critical role. The ventral premammillary nucleus (PMv) leptin receptor (LepRb) neurons act as an essential relay for leptin action on reproduction. Here, we show that mouse PMv cells expressing the dopamine transporter (DAT) gene,Slc6a3(PMvDAT) form a novel subpopulation of LepRb neurons. Virtually all PMvDATneurons expressedLeprmRNA and responded to acute leptin treatment. Electrophysiological recordings from DATCRE;tdTomato mice showed that PMvDATcells in prepubertal females have a hyperpolarized resting membrane potential compared to diestrous females.Slc6a3mRNA expression in the PMv was higher in prepubertal than in adult females. In prepubertal femalesSlc6a3mRNA expression was higher in overnourished females from small size litters than in controls. PrepubertalLepobfemales showed decreased PMvSlc6a3mRNA expression, that recovered to control levels after 3 days of leptin injections. Using a tracer adenoassociated virus in the PMv of adult DATCre;Kiss1hrGFPfemales, we observed PMvDATprojections in the anteroventral periventricular and periventricular nucleus (AVPV/PeN), surrounding Kiss1hrGFPneurons, a population critical for sexual maturation and positive estrogen feedback in females. The DATCRE;tdTomato projections to the AVPV were denser in adult than in prepubertal females. In adults, they surrounded tyrosine hydroxylase neurons. Overall, these findings suggest that the DAT expressing PMvLepRbsubpopulation play a role in leptin regulation of sexual maturation via actions on AVPV kisspeptin/tyrosine hydroxylase neurons.

Significance Statement

Women with excess or low energy stores (e.g.,obesity or anorexia) have reproductive deficits, including altered puberty onset, disruption of reproductive cycles and decreased fertility. If able to conceive, they show higher risks of miscarriages and preterm birth. The hypothalamic circuitry controlling the interplay between metabolism and reproduction is poorly defined. Neurons in the ventral premammillary nucleus express the leptin receptor and play a key role in the metabolic control of reproduction. Those neurons are functionally and phenotypically heterogeneous. Here we show that a subset of leptin-sensitive neurons co-expresses the dopamine transporter (DAT), is dynamically regulated during pubertal transition and with nutrition and projects to brain sites relevant for sexual maturation.

Graphical abstract

The ventral premammillary nucleus of the hypothalamus plays an essential role in the metabolic control of reproduction. Puberty brings large changes to a subpopulation of PMvLepRbcells expressing the dopamine transporter (PMvDAT). DAT gene expression is higher in prepubertal than in adults and is regulated by leptin in prepubertal females. Dynamic projections from PMvDATcells contact the kisspeptin and tyrosine hydroxylase (TH) populations in the AVPV/PeN during puberty, a critical time for the appearance of these cells in the AVPV/PeN.