TY - JOUR
T1 - Validation of antibodies for neuroanatomical localization of the P2Y receptor in macaque brain
AU - Dreisig, Karin
AU - Degn, Matilda
AU - Sund, Louise
AU - Hadaczek, Piotr
AU - Samaranch, Lluis
AU - San Sebastian, Waldy
AU - Bankiewicz, Krystof
AU - Rahbek Kornum, Birgitte
N1 - Copyright © 2016 Elsevier B.V. All rights reserved.
PY - 2016/12/1
Y1 - 2016/12/1
N2 - Focus on the purinergic receptor P2Y11 has increased following the finding of an association between the sleep disorder narcolepsy and a genetic variant in P2RY11 causing decreased gene expression. Narcolepsy is believed to arise from an autoimmune destruction of the hypothalamic neurons that produce the neuropeptide hypocretin/orexin. It is unknown how a decrease in expression of P2Y11 might contribute to an autoimmune reaction towards the hypocretin neurons and the development of narcolepsy. To advance narcolepsy research it is therefore extremely important to determine the neuroanatomical localization of P2Y11 in the brain with particular emphasis on the hypocretin neurons. In this article we used western blot, staining of blood smears, and flow cytometry to select two antibodies for immunohistochemical staining of macaque monkey brain. Staining was seen in neuron-like structures in cortical and hypothalamic regions. Rats do not have a gene orthologue to the P2Y11 receptor and therefore rat brain was used as negative control tissue. The chromogenic signal observed in macaque monkey brain in neurons was not considered reliable, because the antibodies stained rat brain in a similar distribution pattern. Hence, the neuroanatomical localization of the P2Y11 receptor remains undetermined due to the lack of specific P2Y11 antibodies for brain immunohistochemistry.
AB - Focus on the purinergic receptor P2Y11 has increased following the finding of an association between the sleep disorder narcolepsy and a genetic variant in P2RY11 causing decreased gene expression. Narcolepsy is believed to arise from an autoimmune destruction of the hypothalamic neurons that produce the neuropeptide hypocretin/orexin. It is unknown how a decrease in expression of P2Y11 might contribute to an autoimmune reaction towards the hypocretin neurons and the development of narcolepsy. To advance narcolepsy research it is therefore extremely important to determine the neuroanatomical localization of P2Y11 in the brain with particular emphasis on the hypocretin neurons. In this article we used western blot, staining of blood smears, and flow cytometry to select two antibodies for immunohistochemical staining of macaque monkey brain. Staining was seen in neuron-like structures in cortical and hypothalamic regions. Rats do not have a gene orthologue to the P2Y11 receptor and therefore rat brain was used as negative control tissue. The chromogenic signal observed in macaque monkey brain in neurons was not considered reliable, because the antibodies stained rat brain in a similar distribution pattern. Hence, the neuroanatomical localization of the P2Y11 receptor remains undetermined due to the lack of specific P2Y11 antibodies for brain immunohistochemistry.
KW - Animals
KW - Cerebellum/metabolism
KW - Hypothalamus/metabolism
KW - Immunohistochemistry/methods
KW - Macaca
KW - Neurons/metabolism
KW - Prefrontal Cortex/metabolism
KW - Rats
KW - Receptors, Purinergic P2/metabolism
U2 - 10.1016/j.jchemneu.2016.08.002
DO - 10.1016/j.jchemneu.2016.08.002
M3 - Journal article
C2 - 27515691
SN - 0891-0618
VL - 78
SP - 25
EP - 33
JO - Journal of Chemical Neuroanatomy
JF - Journal of Chemical Neuroanatomy
ER -