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A: Neonatal GnRH-GFP transgene expression within cholinergic neurons of the caudate-putamen (CPU) indicates that newborn transgenic mice may serve as animal model to study GnRH effects with slice electrophysiology. To reveal receptor mechanisms, the selective GnRHR1 antagonist Antide, the membrane-impermeable G Protein-Coupled Receptor inhibitor <t>GDP-β-S</t> and the action potantial inhibitor tetrodotoxin (TTX) were used in whole-cell patch-clamp experiments. B: Postnatal week 1 (PNW1; postnatal day 4-7) GnRH-GFP neurons responded to GnRH with reduced resting membrane potential (V rest ) and decreased rates in current pulse-induced firing activity. C: The same inhibitory responses could also be elicited from cholinergic interneurons of newborn ChAT-Cre/zsGreen. GnRH acted via its specific receptor GnRHR1 because inhibitory responses could be prevented with Antide. GnRHR1 mediating GnRH effects was localized within CPU cholinergic neurons. First, GnRH was unable to inhibit cholinergic neurons if the internal electrode solution contained GDP-β-S. Second, GnRH was still able to hyperpolarize cholinergic neurons in the presence of TTX to eliminate activity-dependent indirect actions (TTX+GnRH). D: In contrast to the newborn mice, adult ChAT-Cre/zsGreen animals did not respond to GnRH with reduced V rest or firing rate. E: Medium spiny projection neurons which receive input from cholinergic interneurons were studied in neonatal GAD65-GFP transgenic mice. GnRH did not change the V rest but decreased the firing rate of these neurons, indicating together that the inhibitory response is indirect. F, G: Scatter dot plots summarize the results of measurements in the different treatment groups. *p<0.05 with ANOVA. Scale bar: 25 µm. See also Figure 3 – Source Data for recordings.
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A: Neonatal GnRH-GFP transgene expression within cholinergic neurons of the caudate-putamen (CPU) indicates that newborn transgenic mice may serve as animal model to study GnRH effects with slice electrophysiology. To reveal receptor mechanisms, the selective GnRHR1 antagonist Antide, the membrane-impermeable G Protein-Coupled Receptor inhibitor <t>GDP-β-S</t> and the action potantial inhibitor tetrodotoxin (TTX) were used in whole-cell patch-clamp experiments. B: Postnatal week 1 (PNW1; postnatal day 4-7) GnRH-GFP neurons responded to GnRH with reduced resting membrane potential (V rest ) and decreased rates in current pulse-induced firing activity. C: The same inhibitory responses could also be elicited from cholinergic interneurons of newborn ChAT-Cre/zsGreen. GnRH acted via its specific receptor GnRHR1 because inhibitory responses could be prevented with Antide. GnRHR1 mediating GnRH effects was localized within CPU cholinergic neurons. First, GnRH was unable to inhibit cholinergic neurons if the internal electrode solution contained GDP-β-S. Second, GnRH was still able to hyperpolarize cholinergic neurons in the presence of TTX to eliminate activity-dependent indirect actions (TTX+GnRH). D: In contrast to the newborn mice, adult ChAT-Cre/zsGreen animals did not respond to GnRH with reduced V rest or firing rate. E: Medium spiny projection neurons which receive input from cholinergic interneurons were studied in neonatal GAD65-GFP transgenic mice. GnRH did not change the V rest but decreased the firing rate of these neurons, indicating together that the inhibitory response is indirect. F, G: Scatter dot plots summarize the results of measurements in the different treatment groups. *p<0.05 with ANOVA. Scale bar: 25 µm. See also Figure 3 – Source Data for recordings.
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A: Neonatal GnRH-GFP transgene expression within cholinergic neurons of the caudate-putamen (CPU) indicates that newborn transgenic mice may serve as animal model to study GnRH effects with slice electrophysiology. To reveal receptor mechanisms, the selective GnRHR1 antagonist Antide, the membrane-impermeable G Protein-Coupled Receptor inhibitor <t>GDP-β-S</t> and the action potantial inhibitor tetrodotoxin (TTX) were used in whole-cell patch-clamp experiments. B: Postnatal week 1 (PNW1; postnatal day 4-7) GnRH-GFP neurons responded to GnRH with reduced resting membrane potential (V rest ) and decreased rates in current pulse-induced firing activity. C: The same inhibitory responses could also be elicited from cholinergic interneurons of newborn ChAT-Cre/zsGreen. GnRH acted via its specific receptor GnRHR1 because inhibitory responses could be prevented with Antide. GnRHR1 mediating GnRH effects was localized within CPU cholinergic neurons. First, GnRH was unable to inhibit cholinergic neurons if the internal electrode solution contained GDP-β-S. Second, GnRH was still able to hyperpolarize cholinergic neurons in the presence of TTX to eliminate activity-dependent indirect actions (TTX+GnRH). D: In contrast to the newborn mice, adult ChAT-Cre/zsGreen animals did not respond to GnRH with reduced V rest or firing rate. E: Medium spiny projection neurons which receive input from cholinergic interneurons were studied in neonatal GAD65-GFP transgenic mice. GnRH did not change the V rest but decreased the firing rate of these neurons, indicating together that the inhibitory response is indirect. F, G: Scatter dot plots summarize the results of measurements in the different treatment groups. *p<0.05 with ANOVA. Scale bar: 25 µm. See also Figure 3 – Source Data for recordings.
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A: Neonatal GnRH-GFP transgene expression within cholinergic neurons of the caudate-putamen (CPU) indicates that newborn transgenic mice may serve as animal model to study GnRH effects with slice electrophysiology. To reveal receptor mechanisms, the selective GnRHR1 antagonist Antide, the membrane-impermeable G Protein-Coupled Receptor inhibitor <t>GDP-β-S</t> and the action potantial inhibitor tetrodotoxin (TTX) were used in whole-cell patch-clamp experiments. B: Postnatal week 1 (PNW1; postnatal day 4-7) GnRH-GFP neurons responded to GnRH with reduced resting membrane potential (V rest ) and decreased rates in current pulse-induced firing activity. C: The same inhibitory responses could also be elicited from cholinergic interneurons of newborn ChAT-Cre/zsGreen. GnRH acted via its specific receptor GnRHR1 because inhibitory responses could be prevented with Antide. GnRHR1 mediating GnRH effects was localized within CPU cholinergic neurons. First, GnRH was unable to inhibit cholinergic neurons if the internal electrode solution contained GDP-β-S. Second, GnRH was still able to hyperpolarize cholinergic neurons in the presence of TTX to eliminate activity-dependent indirect actions (TTX+GnRH). D: In contrast to the newborn mice, adult ChAT-Cre/zsGreen animals did not respond to GnRH with reduced V rest or firing rate. E: Medium spiny projection neurons which receive input from cholinergic interneurons were studied in neonatal GAD65-GFP transgenic mice. GnRH did not change the V rest but decreased the firing rate of these neurons, indicating together that the inhibitory response is indirect. F, G: Scatter dot plots summarize the results of measurements in the different treatment groups. *p<0.05 with ANOVA. Scale bar: 25 µm. See also Figure 3 – Source Data for recordings.
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A: Neonatal GnRH-GFP transgene expression within cholinergic neurons of the caudate-putamen (CPU) indicates that newborn transgenic mice may serve as animal model to study GnRH effects with slice electrophysiology. To reveal receptor mechanisms, the selective GnRHR1 antagonist Antide, the membrane-impermeable G Protein-Coupled Receptor inhibitor <t>GDP-β-S</t> and the action potantial inhibitor tetrodotoxin (TTX) were used in whole-cell patch-clamp experiments. B: Postnatal week 1 (PNW1; postnatal day 4-7) GnRH-GFP neurons responded to GnRH with reduced resting membrane potential (V rest ) and decreased rates in current pulse-induced firing activity. C: The same inhibitory responses could also be elicited from cholinergic interneurons of newborn ChAT-Cre/zsGreen. GnRH acted via its specific receptor GnRHR1 because inhibitory responses could be prevented with Antide. GnRHR1 mediating GnRH effects was localized within CPU cholinergic neurons. First, GnRH was unable to inhibit cholinergic neurons if the internal electrode solution contained GDP-β-S. Second, GnRH was still able to hyperpolarize cholinergic neurons in the presence of TTX to eliminate activity-dependent indirect actions (TTX+GnRH). D: In contrast to the newborn mice, adult ChAT-Cre/zsGreen animals did not respond to GnRH with reduced V rest or firing rate. E: Medium spiny projection neurons which receive input from cholinergic interneurons were studied in neonatal GAD65-GFP transgenic mice. GnRH did not change the V rest but decreased the firing rate of these neurons, indicating together that the inhibitory response is indirect. F, G: Scatter dot plots summarize the results of measurements in the different treatment groups. *p<0.05 with ANOVA. Scale bar: 25 µm. See also Figure 3 – Source Data for recordings.
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A: Neonatal GnRH-GFP transgene expression within cholinergic neurons of the caudate-putamen (CPU) indicates that newborn transgenic mice may serve as animal model to study GnRH effects with slice electrophysiology. To reveal receptor mechanisms, the selective GnRHR1 antagonist Antide, the membrane-impermeable G Protein-Coupled Receptor inhibitor <t>GDP-β-S</t> and the action potantial inhibitor tetrodotoxin (TTX) were used in whole-cell patch-clamp experiments. B: Postnatal week 1 (PNW1; postnatal day 4-7) GnRH-GFP neurons responded to GnRH with reduced resting membrane potential (V rest ) and decreased rates in current pulse-induced firing activity. C: The same inhibitory responses could also be elicited from cholinergic interneurons of newborn ChAT-Cre/zsGreen. GnRH acted via its specific receptor GnRHR1 because inhibitory responses could be prevented with Antide. GnRHR1 mediating GnRH effects was localized within CPU cholinergic neurons. First, GnRH was unable to inhibit cholinergic neurons if the internal electrode solution contained GDP-β-S. Second, GnRH was still able to hyperpolarize cholinergic neurons in the presence of TTX to eliminate activity-dependent indirect actions (TTX+GnRH). D: In contrast to the newborn mice, adult ChAT-Cre/zsGreen animals did not respond to GnRH with reduced V rest or firing rate. E: Medium spiny projection neurons which receive input from cholinergic interneurons were studied in neonatal GAD65-GFP transgenic mice. GnRH did not change the V rest but decreased the firing rate of these neurons, indicating together that the inhibitory response is indirect. F, G: Scatter dot plots summarize the results of measurements in the different treatment groups. *p<0.05 with ANOVA. Scale bar: 25 µm. See also Figure 3 – Source Data for recordings.
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Image Search Results


A: Neonatal GnRH-GFP transgene expression within cholinergic neurons of the caudate-putamen (CPU) indicates that newborn transgenic mice may serve as animal model to study GnRH effects with slice electrophysiology. To reveal receptor mechanisms, the selective GnRHR1 antagonist Antide, the membrane-impermeable G Protein-Coupled Receptor inhibitor GDP-β-S and the action potantial inhibitor tetrodotoxin (TTX) were used in whole-cell patch-clamp experiments. B: Postnatal week 1 (PNW1; postnatal day 4-7) GnRH-GFP neurons responded to GnRH with reduced resting membrane potential (V rest ) and decreased rates in current pulse-induced firing activity. C: The same inhibitory responses could also be elicited from cholinergic interneurons of newborn ChAT-Cre/zsGreen. GnRH acted via its specific receptor GnRHR1 because inhibitory responses could be prevented with Antide. GnRHR1 mediating GnRH effects was localized within CPU cholinergic neurons. First, GnRH was unable to inhibit cholinergic neurons if the internal electrode solution contained GDP-β-S. Second, GnRH was still able to hyperpolarize cholinergic neurons in the presence of TTX to eliminate activity-dependent indirect actions (TTX+GnRH). D: In contrast to the newborn mice, adult ChAT-Cre/zsGreen animals did not respond to GnRH with reduced V rest or firing rate. E: Medium spiny projection neurons which receive input from cholinergic interneurons were studied in neonatal GAD65-GFP transgenic mice. GnRH did not change the V rest but decreased the firing rate of these neurons, indicating together that the inhibitory response is indirect. F, G: Scatter dot plots summarize the results of measurements in the different treatment groups. *p<0.05 with ANOVA. Scale bar: 25 µm. See also Figure 3 – Source Data for recordings.

Journal: bioRxiv

Article Title: The cryptic gonadotropin-releasing hormone neuronal system of human basal ganglia

doi: 10.1101/2021.03.04.433872

Figure Lengend Snippet: A: Neonatal GnRH-GFP transgene expression within cholinergic neurons of the caudate-putamen (CPU) indicates that newborn transgenic mice may serve as animal model to study GnRH effects with slice electrophysiology. To reveal receptor mechanisms, the selective GnRHR1 antagonist Antide, the membrane-impermeable G Protein-Coupled Receptor inhibitor GDP-β-S and the action potantial inhibitor tetrodotoxin (TTX) were used in whole-cell patch-clamp experiments. B: Postnatal week 1 (PNW1; postnatal day 4-7) GnRH-GFP neurons responded to GnRH with reduced resting membrane potential (V rest ) and decreased rates in current pulse-induced firing activity. C: The same inhibitory responses could also be elicited from cholinergic interneurons of newborn ChAT-Cre/zsGreen. GnRH acted via its specific receptor GnRHR1 because inhibitory responses could be prevented with Antide. GnRHR1 mediating GnRH effects was localized within CPU cholinergic neurons. First, GnRH was unable to inhibit cholinergic neurons if the internal electrode solution contained GDP-β-S. Second, GnRH was still able to hyperpolarize cholinergic neurons in the presence of TTX to eliminate activity-dependent indirect actions (TTX+GnRH). D: In contrast to the newborn mice, adult ChAT-Cre/zsGreen animals did not respond to GnRH with reduced V rest or firing rate. E: Medium spiny projection neurons which receive input from cholinergic interneurons were studied in neonatal GAD65-GFP transgenic mice. GnRH did not change the V rest but decreased the firing rate of these neurons, indicating together that the inhibitory response is indirect. F, G: Scatter dot plots summarize the results of measurements in the different treatment groups. *p<0.05 with ANOVA. Scale bar: 25 µm. See also Figure 3 – Source Data for recordings.

Article Snippet: Intracellularly applied drug: the membrane impermeable G-protein blocker GDP-β-S (2 mM, Merck) ( ; McDermott et al., 2011).

Techniques: Expressing, Transgenic Assay, Animal Model, Patch Clamp, Activity Assay