Photoperiodic modulation of puberty through melatonin-kisspeptin-GnRH signalling in female Wistar rats.

Pubertal onset results from a complex interaction between neuroendocrine signalling and environmental factors such as photoperiod. This study examined how variations in melatonin and estradiol levels, individually and in combination, affect hypothalamic gene expression to identify neuroendocrine regulators of the hypothalamic-pituitary-gonadal (HPG) axis. Juvenile female Wistar rats were exposed to distinct photoperiodic conditions to evaluate pubertal timing, and primary hypothalamic neurons were treated with graded doses of melatonin, estradiol, and the melatonin receptor antagonist luzindole. Expression of key genes, including Kiss1, GnRH, Mtnr1a, Esr2, and Fshr, was analysed using qRT-PCR, and immunocytochemistry validated the observed patterns. Long-day photoperiods significantly advanced puberty, indicated by earlier vaginal opening, higher gonadosomatic index, and elevated estradiol and luteinizing hormone levels (p < 0.05), whereas short-day conditions delayed these effects. In vitro, estradiol upregulated Kiss1 and GnRH, while melatonin suppressed their expression via MTNR1A-dependent mechanisms, partially reversed by luzindole. These results demonstrate that photoperiod-dependent melatonin signalling and estradiol synergistically modulate hypothalamic gene networks crucial for pubertal regulation. Despite lacking full hypothalamic complexity, the in vitro model enabled targeted analysis of direct cellular responses, enhancing understanding of environmental and hormonal convergence in pubertal control.