title: Prolactin mediates neuroprotection against excitotoxicity in primary cell cultures of hippocampal neurons via its receptor.
abstract: Recently it has been reported that prolactin (PRL) exerts a neuroprotective effect against excitotoxicity in hippocampus in the rat in vivo models. However, the exact mechanism by which PRL mediates this effect is not completely understood. The aim of our study was to assess whether prolactin exerts neuroprotection against excitotoxicity in an in vitro model using primary cell cultures of hippocampal neurons, and to determine whether this effect is mediated via the prolactin receptor (PRLR). Primary cell cultures of rat hippocampal neurons were used in all experiments, gene expression was evaluated by RT-qPCR, and protein expression was assessed by Western blot analysis and immunocytochemistry. Cell viability was assessed by using the MTT method. The results demonstrated that PRL treatment of neurons from primary cultures did not modify cell viability, but that it exerted a neuroprotective effect, with cells treated with PRL showing a significant increase of viability after glutamate (Glu)--induced excitotoxicity as compared with neurons treated with Glu alone. Cultured neurons expressed mRNA for both PRL and its receptor (PRLR), and both PRL and PRLR expression levels changed after the excitotoxic insult. Interestingly, the PRLR protein was detected as two main isoforms of 100 and 40 kDa as compared with that expressed in hypothalamic cells, which was present only as a 30 kDa variant. On the other hand, PRL was not detected in neuron cultures, either by western blot or by immunohistochemistry. Neuroprotection induced by PRL was significantly blocked by specific oligonucleotides against PRLR, thus suggesting that the PRL role is mediated by its receptor expressed in these neurons. The overall results indicated that PRL induces neuroprotection in neurons from primary cell cultures.
authors.affiliation: Unidad de Investigación en Reproducción Humana, Instituto Nacional de Perinatología-Facultad de Química, Universidad Nacional Autónoma de México (UNAM), Mexico.
authors.forename: E
authors.initials: E
authors.lastname: Vergara-Castañeda
authors.affiliation: Center for Neuroendocrinology and Department of Anatomy, University of Otago, New Zealand.
authors.forename: D R
authors.initials: DR
authors.lastname: Grattan
authors.affiliation: Instituto de Fisiología Celular, Mexico.
authors.forename: H
authors.initials: H
authors.lastname: Pasantes-Morales
authors.affiliation: Instituto de Fisiología Celular, Mexico.
authors.forename: M
authors.initials: M
authors.lastname: Pérez-Domínguez
authors.affiliation: Unidad de Investigación en Reproducción Humana, Instituto Nacional de Perinatología-Facultad de Química, Universidad Nacional Autónoma de México (UNAM), Mexico.
authors.forename: E A
authors.initials: EA
authors.lastname: Cabrera-Reyes
authors.affiliation: Instituto de Neurobiología, UNAM, Mexico.
authors.forename: T
authors.initials: T
authors.lastname: Morales
authors.affiliation: Unidad de Investigación en Reproducción Humana, Instituto Nacional de Perinatología-Facultad de Química, Universidad Nacional Autónoma de México (UNAM), Mexico. Electronic address: mcerbon85@yahoo.com.mx.
authors.forename: M
authors.initials: M
authors.lastname: Cerbón
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian; D018690:Excitatory Amino Acid Agonists / Q000633:toxicity; D005786:Gene Expression Regulation / Q000187:drug effects; D018698:Glutamic Acid / Q000633:toxicity; D006624:Hippocampus / Q000166:cytology*; D009474:Neurons / Q000187:drug effects*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D009841:Oligonucleotides / Q000494:pharmacology; D011388:Prolactin / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011981:Receptors, Prolactin / Q000235:genetics / Q000378:metabolism*
chemical_list: D018690:Excitatory Amino Acid Agonists; D018696:Neuroprotective Agents; D009841:Oligonucleotides; D012333:RNA, Messenger; D011981:Receptors, Prolactin; D018698:Glutamic Acid; D011388:Prolactin
keywords: Excitotoxicity; Hippocampus; Neuroprotection; Prolactin