Journal: Frontiers in Endocrinology

Article Title: miR-29a/b 1 Regulates the Luteinizing Hormone Secretion and Affects Mouse Ovulation

doi: 10.3389/fendo.2021.636220

Figure Lengend Snippet: Central mechanism in miR-29a/b 1 KO mice. (A, B) GnRHR immunoreactivity in pituitary of wild-type and miR-29a/b 1 KO females. The receptor was not detectable on the plasma membrane of control. (immunohistochemical: wild-type: 13.43 ± 0.7927, miR-29a/b 1 KO: 25.47 ± 0.534, p=0.0249; immunofluorescence: wild-type: 37.79 ± 1.858, miR-29a/b 1 KO: 56.64 ± 2.767, p=0.0045 , n=3). (C) Serum LH and FSH levels in miR-29a/b 1 KO females and controls following ovariectomy (OVX) and sham-operated controls (Sham). (LH: wild-type: p=0.0401 , miR-29a/b 1 KO: p=0.9249 ; FSH: wild-type: p=0.0016 , miR-29a/b 1 KO: p=0.0185 , n=6). (D) Expression of Kiss1 and Gnrh1 in hypothalamus (Gnrh1: wild-type: 1 ± 0.1912, miR-29a/b 1 KO: 0.7287 ± 0.06234, p = 0.1874 , Kiss1: wild-type: 1 ± 0.1305, miR-29a/b 1 KO: 0.8142 ± 0.0757, p=0.8405 , n=15). (E) Normal distribution of GnRH neurons in miR-29a/b 1 KO mice compared to control littermates. OVLT, organum vasculosum of the lamina terminalis. Scale bars, 200μm. (wild-type: 9.827 ± 1.547, miR-29a/b 1 KO: 8.597 ± 0.8466, p=0.5238 , n=3). * p < 0.05, ** p < 0.01 and **** p < 0.0001.

Article Snippet: After permeabilization, the sections were washed three times in PBST, and blocked with 5% normal donkey serum in PBS for 1h at room temperature, then were incubated with mouse anti-Lutropin beta antibody (1:1000, SANTA CRUZ, sc-373941), rabbit anti-GnRH1 (1:500, Immunostar, PA1-121) or rabbit anti-GnRHR antibody ( ) (1:100, Proteintech, 19950-1-AP) overnight at 4°C, followed by staining with Alexa Fluor 647-conjugated donkey anti-mouse antibody (Invitrogen Molecular Probes) or Alexa Fluor 594-conjugated donkey anti-rabbit (Invitrogen Molecular Probes) antibody and DAPI dye to stain nuclei.

Techniques: Immunohistochemical staining, Immunofluorescence, Expressing

Journal: The Journal of Neuroscience

Article Title: Kisspeptin Signaling Is Required for Peripheral But Not Central Stimulation of Gonadotropin-Releasing Hormone Neurons by NMDA

doi: 10.1523/JNEUROSCI.5486-09.2010

Figure Lengend Snippet: Absence of LH release after peripheral NMDA injection in Kiss1- and Gpr54-null mice. A, Bar graph showing the mean ± SEM of LH levels in wild-type (+/+), Kiss1-null (Kiss−/−), and Gpr54-null (Gpr54−/−) adult male mice 10 min after PBS, NMDA, or Kp10 intraperitoneal injection. B, Bar graph showing the mean ± SEM of plasma LH levels in prepubertal mice of each genotype 10 min after PBS, NMDA, or Kp10 intraperitoneal injection. **p < 0.01 (one-way ANOVA, followed by Student–Newman–Keuls test). Numbers in brackets indicate animal number used in each treatment group. C, Representative coronal sections of dual-labeled immunocytochemistry showing the absence of c-Fos staining (black nuclei) in GnRH neurons (brown) in hypothalamus from wild-type or Kiss1-null mice that showed high plasma LH after peripheral stimulation with NMDA (left) or Kp10 (middle and right). Scale bar, 100 μm. All photographs are the same scale. Inset box is 4× magnification. 3V, Third ventricle.

Article Snippet: Rabbit polyclonal anti-GnRH antibody was a generous gift from Prof. G. Tramu (University of Bordeaux 1, Talence, France) ( Beauvillain and Tramu, 1980 ) provided by Dr V. Prevot (Inserm Unit 837, Lille, France).

Techniques: Injection, Clinical Proteomics, Labeling, Immunocytochemistry, Staining

Journal: The Journal of Neuroscience

Article Title: Kisspeptin Signaling Is Required for Peripheral But Not Central Stimulation of Gonadotropin-Releasing Hormone Neurons by NMDA

doi: 10.1523/JNEUROSCI.5486-09.2010

Figure Lengend Snippet: Induction of c-Fos after peripheral or central injection of NMDA in wild-type mice. Representative low-magnification photomicrographs showing immunocytochemical c-Fos staining (black nuclei) in three different coronal sections. A–C, c-Fos staining 2 h after PBS intraperitoneal injection. D–F, c-Fos staining 2 h after NMDA intraperitoneal injection. Note the intense labeling in Arc and moderate labeling in OVLT. G–I, c-Fos staining 2 h after PBS intracerebroventricular injection. Staining is very similar to intraperitoneal PBS. J–L, c-Fos staining 2 h after NMDA intracerebroventricular injection. Note that c-Fos labeling is intense in every periventricular hypothalamic region. Regions containing GnRH cell bodies, such as MnPO, MPO, and MPA (in light brown in A, D, G, J), show high concentration of black nuclei, indicating activated cells, only after intracerebroventricular NMDA (J). Regions known to regulate GnRH neuron activity have high c-Fos staining after intracerebroventricular NMDA only (J, K). Regions known to contain Kp neurons, such as the Arc, show high c-Fos staining after intraperitoneal and intracerebroventricular NMDA (F, L). The sexually dimorphic AVPV containing a Kp neuron population has high c-Fos staining after intracerebroventricular NMDA exclusively (K). 3V, Third ventricle; OT, optic tract. Scale bar, 600 μm. All photographs are the same scale.

Article Snippet: Rabbit polyclonal anti-GnRH antibody was a generous gift from Prof. G. Tramu (University of Bordeaux 1, Talence, France) ( Beauvillain and Tramu, 1980 ) provided by Dr V. Prevot (Inserm Unit 837, Lille, France).

Techniques: Injection, Staining, Labeling, Concentration Assay, Activity Assay

Journal: The Journal of Neuroscience

Article Title: Kisspeptin Signaling Is Required for Peripheral But Not Central Stimulation of Gonadotropin-Releasing Hormone Neurons by NMDA

doi: 10.1523/JNEUROSCI.5486-09.2010

Figure Lengend Snippet: Absence of c-Fos induction in GnRH neurons after central injection of NMDA. Representative photomicrographs of dual-labeled immunocytochemistry coronal sections showing GnRH neurons (brown) and c-Fos staining (black nuclei) in the MS/DBB region. A, Absence of c-Fos staining in GnRH neurons of PBS-treated wild-type mice. B, High c-Fos staining density but absence of c-Fos expression in GnRH neurons of NMDA-treated wild-type mice. C, GnRH neurons expressing c-Fos (arrows) in Kp10-treated wild-type mice. D, Absence of c-Fos staining in GnRH neurons of PBS-treated Kiss1-null mice. E, High c-Fos staining density but absence of c-Fos expression in GnRH neurons of NMDA-treated Kiss1-null mice. F, GnRH neurons expressing c-Fos (arrows) in Kp10-treated Kiss1-null mice. G, Absence of c-Fos staining in GnRH neurons of PBS-treated Gpr54-null mice. H, High c-Fos staining density but absence of c-Fos expression in GnRH neurons of NMDA-treated Gpr54-null mice. I, Absence of c-Fos staining in GnRH neurons of Kp10-treated Gpr54-null mice. 3V, Third ventricle. Scale bar, 100 μm. All photographs are the same scale.

Article Snippet: Rabbit polyclonal anti-GnRH antibody was a generous gift from Prof. G. Tramu (University of Bordeaux 1, Talence, France) ( Beauvillain and Tramu, 1980 ) provided by Dr V. Prevot (Inserm Unit 837, Lille, France).

Techniques: Injection, Labeling, Immunocytochemistry, Staining, Expressing

Journal: The Journal of Neuroscience

Article Title: Kisspeptin Signaling Is Required for Peripheral But Not Central Stimulation of Gonadotropin-Releasing Hormone Neurons by NMDA

doi: 10.1523/JNEUROSCI.5486-09.2010

Figure Lengend Snippet: Quantification of GnRH neurons with c-Fos expression after central injections. Bar graphs represent the percentage of GnRH neurons expressing c-Fos (mean ± SEM) in the MS/DBB (left) or in the OVLT/POA (right) regions. Arrows indicate gray regions on the brain map. In each region, bar graphs show percentage of GnRH neurons (mean ± SEM) with c-Fos in wild-type (+/+), Kiss1-null (Kiss−/−), and Gpr54-null (Gpr54−/−) mice before (0) and 10 min after PBS (white bars), NMDA (black bars), or Kp10 (gray bars) intracerebroventricular injection. *p < 0.05, **p < 0.01, ***p < 0.001 when different treatments on same genotype were compared. a vs c, p < 0.001; b vs c, p < 0.01; a vs b, not significant when different genotypes were compared with the same treatment in the same subpopulation (one-way ANOVA, followed with Student–Newman–Keuls test in both comparison tests). Numbers in brackets indicate animal number used in each condition.

Article Snippet: Rabbit polyclonal anti-GnRH antibody was a generous gift from Prof. G. Tramu (University of Bordeaux 1, Talence, France) ( Beauvillain and Tramu, 1980 ) provided by Dr V. Prevot (Inserm Unit 837, Lille, France).

Techniques: Expressing, Injection, Comparison

Journal: The Journal of Neuroscience

Article Title: Kisspeptin Signaling Is Required for Peripheral But Not Central Stimulation of Gonadotropin-Releasing Hormone Neurons by NMDA

doi: 10.1523/JNEUROSCI.5486-09.2010

Figure Lengend Snippet: A schematic representation of the possible effect of peripheral versus central injection of NMDA on the GnRH neuronal network. A, Peripherally injected NMDA does not reach the POA in which the GnRH neuron cell bodies are located. NMDA can penetrate into the Arc through the fenestrated capillaries of the ME. NMDA requires Kp neuron activation, revealed by c-Fos expression, in the Arc, and Kp release then activates GnRH release from the nerve terminals. B, Central injection of NMDA activates nNOS- and TH-containing neurons in the POA, revealed by c-Fos expression. This may result in production of the diffusible gas NO and the release of catecholamines (CAT), which might stimulate GnRH release. Question mark denotes an undetermined mechanism. NMDAR, NMDA receptor.

Article Snippet: Rabbit polyclonal anti-GnRH antibody was a generous gift from Prof. G. Tramu (University of Bordeaux 1, Talence, France) ( Beauvillain and Tramu, 1980 ) provided by Dr V. Prevot (Inserm Unit 837, Lille, France).

Techniques: Injection, Activation Assay, Expressing