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Updated: Dec 31, 2025

Indirect Immunofluorescence on Frozen Sections of Mouse Mammary Gland
Published on: December 1, 2015
Prolactin maintains transient melanin-concentrating hormone expression in the medial preoptic area during established
Ilona C Kokay1, David R Grattan1, Joanne F Murray2,3
1Department of Anatomy, School of Biomedical Sciences, Centre for Neuroendocrinology, University of Otago, Dunedin, New Zealand.
A population of neurones in the medial part of the medial preoptic area (mPOA) transiently express melanin-concentrating hormone (MCH) in mid to late lactation in the rat, and this expression disappears on weaning. Prolactin is known to mediate many of the physiological adaptations that occur within the dam associated with lactation and the mPOA is well endowed with prolactin receptors (Prlr); hence, we hypothesised that these transiently MCH-expressing cells may be regulated by prolactin. By in situ hybridisation, we show that approximately 60% of the cells expressing prepro-MCH (Pmch) mRNA in the medial part of the mPOA on day 19 of lactation also express Prlr mRNA. To demonstrate that these transiently MCH-expressing cells can acutely respond to prolactin, dams were treated with bromocriptine on the morning of day 19 of lactation and then given vehicle or prolactin 4 hours later. In the prolactin-treated animals, over 80% of the MCH-immunopositive cells were also immunopositive for phosphorylated signal transducer and activator of transcription 5, an indicator of prolactin receptor activation: double immunopositive cells were rare in vehicle-treated animals. Finally, the effect of manipulating the circulating concentrations of prolactin on days 17, 18 and 19 on the number of MCH-immunopositive cells on day 19 was determined. Reducing circulating concentrations of prolactin over days 17, 18 and 19 of lactation with or without a suckling stimulus resulted in a reduction (P < 0.05) in the number of MCH-immunopositive cells in the medial part of the mPOA on day 19 of lactation. Further research is required to determine the functional role(s) of these prolactin-activated transiently MCH-expressing neurones; however, we suggest the most likely role involves adaptations in maternal metabolism to support the final week of lactation.
A population of neurones in the medial part of the medial preoptic area (mPOA) transiently express melanin-concentrating hormone (MCH) in mid to late lactation in the rat, and this expression disappears on weaning. Prolactin is known to mediate many of the physiological adaptations that occur within the dam associated with lactation and the mPOA is well endowed with prolactin receptors (Prlr); hence, we hypothesised that these transiently MCH-expressing cells may be regulated by prolactin. By in situ hybridisation, we show that approximately 60% of the cells expressing prepro-MCH (Pmch) mRNA in the medial part of the mPOA on day 19 of lactation also express Prlr mRNA. To demonstrate that these transiently MCH-expressing cells can acutely respond to prolactin, dams were treated with bromocriptine on the morning of day 19 of lactation and then given vehicle or prolactin 4 hours later. In the prolactin-treated animals, over 80% of the MCH-immunopositive cells were also immunopositive for phosphorylated signal transducer and activator of transcription 5, an indicator of prolactin receptor activation: double immunopositive cells were rare in vehicle-treated animals. Finally, the effect of manipulating the circulating concentrations of prolactin on days 17, 18 and 19 on the number of MCH-immunopositive cells on day 19 was determined. Reducing circulating concentrations of prolactin over days 17, 18 and 19 of lactation with or without a suckling stimulus resulted in a reduction (P < 0.05) in the number of MCH-immunopositive cells in the medial part of the mPOA on day 19 of lactation. Further research is required to determine the functional role(s) of these prolactin-activated transiently MCH-expressing neurones; however, we suggest the most likely role involves adaptations in maternal metabolism to support the final week of lactation.
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