Indomethacin inhibits kisspeptin-induced vasoconstriction.

<p>Female CD1 mice were anaesthetised, injected intradermally via the dorsal skin with 5% NaHCO₃/saline (Veh), 10nmol KP-10 (KP) or 30pmol Endothelin-1 (ET-1) in the presence or absence of 3nmol indomethacin (INDO), each containing an equal amount of ^99mTc. Clearance after ten minutes was assessed and compared to total radioactivity in an uninjected sample. Saline vehicle was adjusted to 100% and results displayed as a % change to controls. Results are shown as mean ± SEM, <i>N</i> = 6. * = p≤0.05 as assessed by one way ANOVA followed by a Bonferroni's post test.</p

Kisspeptin and Kiss1R expression does not alter after Angiotensin II treatment.

<p>C57BL/6 mice were treated with saline (white bars) or angiotensin II (Ang II, black bars) for 14 days. They were then killed and their hearts, kidneys and aortas removed before <i>Kiss1</i> (A) and <i>Kiss1R</i> (B) mRNA expression analysis. Results are displayed as mean copy number ± SEM normalised against HPRT-1, SDHA and PLA2 reference genes. <i>N</i> = 3 per group. No significant difference was observed between groups.</p

Pallor inhibition ring at centre of kisspeptin mediated oedema increases in a dose dependent manner.

<p>Female CD1 mice were anaesthetised, injected with Evans Blue i.v. and treated intradermally in the dorsal skin with 0.3–10nmol Kp-10 (KP), 300pmol substance P (SP) or saline control (vehicle) in the dorsal skin. An uninjected site was also included (Un). Plasma extravasation was allowed to continue for 15 minutes. The skin was removed and the region in which oedema was inhibited (pallor area) measured. <i>N</i> = 6. Results are shown as mean area (mm²) ± SEM. * = p≤0.05, as assessed by one-way ANOVA compared to vehicle control.</p

Kisspeptin-10 causes a dose-dependent increase in oedema formation in the cutaneous vasculature.

<p>Female CD1 mice were anaesthetised, injected with Evans Blue dye i.v. and treated intradermally in the dorsal skin with 0.3–10nmol kisspeptin-10 (KP), 300pmol substance P (SP) or saline control (Veh) for 30 minutes. An uninjected site was also included (Un). Plasma exudation was allowed to continue for 30 minutes post-injection at which point mice were killed, dorsal skin removed and the area of oedema measured (A). The lesion was also scored for colour intensity (B). Results are shown as mean area (mm²) ± SEM, <i>N</i> = 4. * = p≤0.05, ** = P≤0.01, *** = p≤0.001 assessed by one-way ANOVA compared to vehicle control.</p

Kisspeptin induced oedema presents with a pallor ring at injection site in the dorsal skin.

<p>Female CD1 mice were anaesthetised, injected with Evans Blue i.v. and treated intradermally in the dorsal skin with 10pmol CGRP, 300pmol substance P (SP), 10nmol Kp-10 (K10), 10nmol Kp-10 +300pmol substance P (K + SP), 300pmol substance P +10pmol CGRP (SP + CGRP) or 10nmol Kp-10 +10pmol CGRP (K + CGRP). Plasma extravasation was allowed to continue for 30 minutes. Mice were killed, the dorsal skin removed and photographed. Pallor ring present in Kp-10 mediated oedema, but not under other conditions, is indicated.</p

Kisspeptin-10 mediated oedema formation is visible 7.5 minutes post-injection.

<p>Female CD1 mice were anaesthetised, injected with Evans Blue i.v. and treated intradermally with 0.3–10nmol Kp-10 (KP) in the dorsal skin for 7.5, 15 or 30 minutes. Mice were killed, the dorsal skin removed and the area of oedema measured. Results are shown as mean area (mm²) ± SEM (3 s.f.). 7.5, 15 and 30 minute treatment groups consisted of <i>N</i> = 5, <i>N</i> = 6 and <i>N</i> = 4 respectively.</p>*<p> = p≤0.05,</p>**<p> = p≤0.01,</p>***<p> = p≤0.001 compared to vehicle controls as assessed by one way ANOVA followed by a Dunnet's post test.</p

Kisspeptin-10 decreases blood flow in the peripheral vasculature.

<p>Male and female CD1 mice were anaesthetised, injected intradermally via the dorsal skin with 3 or 10nmol KP-10 (KP) or 30pmol Endothelin-1 (ET-1), each containing an equal amount of ^99mTc. Clearance after ten minutes was assessed and compared to total radioactivity in an uninjected sample. Saline vehicle was adjusted to 100% and results displayed as a % change to controls. Results are shown as mean ± SEM, <i>N</i> = 5. * = p≤0.05, ** = p≤0.01 as assessed by two-tailed Student's t-test.</p

Mepyramine inhibits Kp-10 induced oedema formation.

<p>Female CD1 mice were anaesthetised, injected with Evans Blue i.v., and then exposed intradermally to saline (Veh), 20nmol Histamine (His), 300pmol substance P (SP) and 10nmol Kp-10 (KP) +/−5nmol Mepyramine (Mepy) for 15 minutes. Mice were then killed, dorsal skin removed and the oedema area measured (A). Lesion colour intensity was also assessed (B). Co-treatment with saline is indicated by white bars and co-treatment with mepyramine is indicated by black bars. Results shown as mean area (mm²) ± SEM, <i>N</i> = 4. n.s. = no significant difference between treatments, *** = p≤0.001 between mepyramine untreated and treated groups as assessed by two way ANOVA.</p

Kp-10 induced oedema is potentiated by CGRP.

<p>Female CD1 mice were anaesthetised, injected with Evans Blue dye i.v. and treated intradermally in the dorsal skin with 10pmol CGRP, 300pmol substance P (SP), 3nmol Kp-10 (KP), 10pmol CGRP +3pmol SP (S + C), 3nmol Kp-10 +10pmol CGRP (K + C) or saline control (Veh) for 30 minutes. An uninjected site was also included (Un). Plasma exudation was allowed to continue for 30 minutes post-injection, mice were killed, dorsal skin removed and the area of oedema measured (A). Lesion colour intensity was also assessed (B). Results are shown as mean area (mm²) ± SEM. <i>N</i> = 8. * = p≤0.05, ** = p≤0.01, *** = p≤0.001 assessed by one-way ANOVA compared to vehicle control.</p

Effect of intra-arcuate nucleus (ARC) and intra-medial preoptic area (mPOA) administration of kisspeptin-10 (KP) on LH secretion.

<p>Representative examples illustrating the effects of intra-ARC infusion of (A) 400 nl aCSF or (B) 100 pmol KP in ovariectomized 17βestradiol-replaced rats. C, Summary showing the effect of KP on LH secretion, calculated by comparing the mean area of under LH profile 2 h before with 1 h after its administration. Representative examples illustrating the effects of intra-mPOA infusion of (D) 400 nl aCSF or (E) 100 pmol KP in ovariectomized 17βestradiol-replaced rats. F, Summary showing the effect of intra-mPOA KP on LH secretion. LH secretion was dramatically increased immediately after KP treatment in both nuclei, which lasted about 1 h in most experimental animals. *P<0.05 versus aCSF control group at the same time point. ^#P<0.05 versus 10 pmol KP treatment group at the same time point; N = 5–7 per group.</p