Synaptic plasticity in the hippocampal CA1 area of AR^fl/Y and AR^NesCre male mice.

<p>(<b>a-b</b>) Comparison of the time course of mean long-term potentiation (LTP) induced by high-frequency stimulation (HFS) (<b>a</b>; 9 slices from 7 animals per genotype) or by theta-burst stimulation (TBS) (<b>b</b>; 12 slices from 7–8 males per genotype). *<i>p</i> < 0.05 <i>versus</i> control. (<b>c</b>) Comparison of the time course of mean long-term depression (LTD) induced by low-frequency stimulation (LFS) in males (10–11 slices from 5–6 animals per genotype).</p

Anxiety state level and corticosterone levels in AR^fl/Y and AR^NesCre male mice.

<p><b>(a-b)</b> Time spent and latency to enter in the open arms of the EPM (<b>a</b>) or the O-maze (<b>b</b>) for controls and mutants (n = 8–11 males per genotype). (<b>f</b>) Corticosterone secretion during the circadian cycle (n = 8–11 males per genotype).</p

Temporal order memory and object recognition in AR^fl/Y and AR^NesCre male mice.

<p>(<b>a-b</b>) Familiarization and testing phases of the temporal order (<b>a</b>) and novelty detection (<b>b</b>) tasks. (<b>c-d</b>) Discrimination Indexes in the temporal order (<b>c</b>) and novelty detection tasks (<b>d</b>) for males (n = 8–9 males per genotype). ***<i>p</i> < 0.001 <i>versus</i> object A; ^#<i>p</i> < 0.05 <i>versus</i> controls.</p

Basal synaptic transmission in the hippocampal CA1 area of AR^fl/Y and AR^NesCre male mice.

<p>(<b>a-b</b>) Input/output (I/O) curves of presynaptic fiber volleys (PFVs) (<b>a</b>) and field excitatory postsynaptic potentials (fEPSPs) (<b>b</b>) induced in control medium by the electrical stimulation of glutamatergic afferents (44 slices from 8 males per genotype). PFV magnitude was significantly smaller in AR^NesCre males only at the highest stimulus intensity (*<i>p</i> < 0.05), whereas fEPSP values were already lower at lower intensities (**<i>p</i> < 0.01). (<b>c</b>) Upper panel, representative paired-pulse facilitation (PPF) of fEPSPs induced in a AR^NesCre male by two successive stimuli (arrows) delivered with a 30 ms inter-stimulus interval. Lower panel, mean PPF magnitude (28–29 slices per genotype).</p

Effect of URP on basal or GnRH-dependant gonadotrophin release in cultured pituitary cells.

<p>URP does not influence basal or GnRH-induced LH release. Rat anterior pituitary cells were cultured for 18 h with increasing concentrations (1 to 1000 nM) of URP in combination or not with 0.1 nM of the GnRH agonist Triptorelin (GnRHa). Control cells were treated or not with 0.1 nM GnRHa for the same period of time. LH release into the incubation medium was determined by radioimmunoassay as indicated in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0026611#s2" target="_blank"><i>Materials and Methods</i></a>. Results are expressed in fold increase over basal LH release in unstimulated cells and are the mean ± SEM of three independent experiments.</p

Co-localization of URP mRNA and GnRH peptide.

<p>Stain separation by color deconvulation method. Neurones are stained for GnRH by immunohistochemistry with DAB precipitate, URP mRNA is detected simultaneously by in situ hybridization with NBT/BCIP precipitate (A). Stain separation for DAB (B) and NBT/BCIP (C) respectively. Bars represent 10 µm.</p

Subcellular co-localization of URP and GnRH.

<p>Two ultrathin sections of a vibratome slice of rat median eminence after treatment with antibodies against URP revealed by <u>immunoperoxidase</u> (A and B). The DAB-labelling is observed on fibres (arrows) of the ME. Co-localisation of URP and GnRH peptides in the same processes (C): colloidal gold particles intensified by silver exhibit GnRH peptide (double arrow) whereas DAB precipitate signs the presence of URP (asterisk). Bar represents 0.5 µm (A, B) and 0.25 µm (C). Arrow head: fenestrated capillary. T: tanycyte process.</p

URP expression in the rat hypothalamus.

<p>Frontal sections of rat brain after treatment with anti-URP antibodies revealed by immunoperoxidase (A) and after <i>in situ</i> hybridization of rat URP-mRNA with Dig-labelled probes (B). Intensely reactive neuronal cell bodies with short labelled process are observed in medial septal nucleus (A) and in the vertical limb of the diagonal band of Broca. Bars represent 100 µm.</p

URP distribution in the rat hypothalamus.

<p>Frontal (A–E) and horizontal (F) sections of rat hypothalamus, including respectively the median eminence (A–C) up to the infundibular stalk of the pituitary gland (D–E) and the <i>organum vasculosum laminae terminalis</i> (F), after treatment with anti-URP antibodies revealed by immunofluorescence. Labelled fibres of variable intensity are detected at all levels of the ME, especially in the outer palissade layer. In OVLT, immunofluorescent fibres are in close contact with the vessels (F). Bars represent 500 µm (A–E) and 100 µm (F). Asterisk: third ventricle.</p