Prolactin mediates neuroprotection against excitotoxicity in primary cell cultures of hippocampal neurons via its receptor
Por:
Vergara-Castaneda, E., Grattan, D. R., Pasantes-Morales, H., Perez-Dominguez, M., Cabrera-Reyes, E. A., Morales, T., Cerbon, M.
Publicada:
1 abr 2016
Resumen:
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. (C) 2016 Elsevier B.V. All rights reserved.
Filiaciones:
Vergara-Castaneda, E.:
Univ Nacl Autonoma Mexico, Fac Quim, Inst Nacl Perinatol, Unidad Invest Reprod Humana, Mexico City 04510, DF, Mexico
Grattan, D. R.:
Univ Otago, Ctr Neuroendocrinol, Dunedin, New Zealand
Univ Otago, Dept Anat, Dunedin, New Zealand
Pasantes-Morales, H.:
Inst Fisiol Celular, Mexico City, DF, Mexico
Perez-Dominguez, M.:
Inst Fisiol Celular, Mexico City, DF, Mexico
Cabrera-Reyes, E. A.:
Univ Nacl Autonoma Mexico, Fac Quim, Inst Nacl Perinatol, Unidad Invest Reprod Humana, Mexico City 04510, DF, Mexico
Morales, T.:
Univ Nacl Autonoma Mexico, Inst Neurobiol, Mexico City, DF, Mexico
Cerbon, M.:
Univ Nacl Autonoma Mexico, Fac Quim, Inst Nacl Perinatol, Unidad Invest Reprod Humana, Mexico City 04510, DF, Mexico
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