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1.
Nat Methods ; 20(9): 1409-1416, 2023 09.
Article in English | MEDLINE | ID: mdl-37474808

ABSTRACT

Understanding the routing of neuronal information requires the functional characterization of connections. Neuronal projections recruit large postsynaptic ensembles with distinct postsynaptic response types (PRTs). PRT is typically probed by low-throughput whole-cell electrophysiology and is not a selection criterion for single-cell RNA-sequencing (scRNA-seq). To overcome these limitations and target neurons based on specific PRTs for soma harvesting and subsequent scRNA-seq, we created Voltage-Seq. We established all-optical voltage imaging and recorded the PRT of 8,347 neurons in the mouse periaqueductal gray (PAG) evoked by the optogenetic activation of ventromedial hypothalamic (VMH) terminals. PRTs were classified and spatially resolved in the entire VMH-PAG connectome. We built an onsite analysis tool named VoltView to navigate soma harvesting towards target PRTs guided by a classifier that used the VMH-PAG connectome database as a reference. We demonstrated Voltage-seq by locating VMH-driven γ-aminobutyric acid-ergic neurons in the PAG, guided solely by the onsite classification in VoltView.


Subject(s)
Connectome , Mice , Animals , Transcriptome , Neurons/physiology , Periaqueductal Gray/physiology
2.
Eur J Neurosci ; 52(3): 2995-3001, 2020 08.
Article in English | MEDLINE | ID: mdl-32372526

ABSTRACT

GnRH neuron activity is under the influence of multiple stimuli, including those coming from the endocannabinoid and the immune systems. Since it has been previously suggested that some of the main elements controlling the GnRH pulse generator possess the TRPV1 receptor, the aim of the present study was to evaluate the participation of the hypothalamic TRPV1, through its pharmacological blockade, in the activity of the hypothalamic-pituitary-testicular axis in male rats under basal or acute inflammatory conditions. Our hypothesis was based on the idea that the hypothalamic TRPV1 participates in the synthesis of the main neuromodulatory signals controlling GnRH, and therefore the reproductive axis. Our results showed that the hypothalamic TRPV1 blockade induced pro-inflammatory effects by increasing Tnfα and Il-1ß mRNA hypothalamic levels and inhibited the reproductive axis by affecting Gnrh, Kiss1 and Rfrp3 mRNA levels and decreasing plasma levels of luteinizing hormone and testosterone under basal conditions, without significant additive effects in rats exposed to systemic LPS. Altogether, these results suggest that the hypothalamic TRPV1 receptor participates in the regulation of the GnRH system, probably by modulating immune-dependent mechanisms.


Subject(s)
Gonadotropin-Releasing Hormone , Luteinizing Hormone , Animals , Gonadotropin-Releasing Hormone/metabolism , Hypothalamus/metabolism , Male , Neurons/metabolism , Rats , TRPV Cation Channels/genetics , Testosterone
3.
Reprod Toxicol ; 69: 1-12, 2017 04.
Article in English | MEDLINE | ID: mdl-28077272

ABSTRACT

We investigated arsenite exposure on the reproductive axis of dams (during pregnancy and at cyclicity resumption) and their offspring. Pregnant rats were exposed to 5 (A5) or 50ppm (A50) of sodium arsenite in drinking water from gestational day 1 (GD1) until sacrifice at GD18 or two months postpartum. Offspring were exposed to the same treatment as their mothers from weaning to adulthood. A50-pregnant rats gained less weight, showed increased testosterone and estradiol but pregnancy was unaffected. After lactation, arsenic-exposed dams presented compromised cyclicity, decreased estradiol, increased follicle-stimulating hormone (FSH), less preovulatory follicles and presence of ovarian cysts, suggesting impaired reproduction. A50-offspring presented lower body weight; A50-female-offspring showed elevated gonadotropin releasing hormone (GnRH), FSH and testosterone, while A50-males showed diminished GnRH/FSH, but normal testosterone. We conclude that arsenite at the present exposure levels did not compromise pregnancy outcome while it negatively affected reproductive physiology in postpartum dams and their offspring.


Subject(s)
Arsenites/toxicity , Prenatal Exposure Delayed Effects , Sodium Compounds/toxicity , Adrenal Glands/drug effects , Adrenal Glands/metabolism , Animals , Arsenic/metabolism , Female , Hormones/metabolism , Hypothalamus/drug effects , Hypothalamus/metabolism , Lactation , Liver/metabolism , Male , Maternal-Fetal Exchange , Ovary/drug effects , Ovary/metabolism , Pituitary Gland/drug effects , Pituitary Gland/metabolism , Pregnancy , Rats , Rats, Sprague-Dawley , Reproduction/drug effects , Sexual Maturation/drug effects
4.
Food Chem Toxicol ; 100: 207-216, 2017 Feb.
Article in English | MEDLINE | ID: mdl-28017702

ABSTRACT

Drinking water is the main source of arsenic exposure. Chronic exposure has been associated with metabolic disorders. Here we studied the effects of arsenic on glucose metabolism, in pregnant and post-partum of dams and their offspring. We administered 5 (A5) or 50 (A50) mg/L of sodium arsenite in drinking water to rats from gestational day 1 (GD1) until two months postpartum (2MPP), and to their offspring from weaning until 8 weeks old. Liver arsenic dose-dependently increased in arsenite-treated rats to levels similar to exposed population. Pregnant A50 rats gained less weight than controls and recovered normal weight at 2MPP. Arsenite-treated pregnant animals showed glucose intolerance on GD16-17, with impaired insulin secretion but normal insulin sensitivity; they showed dose-dependent increased pancreas insulin on GD18. All alterations reverted at 2MPP. Offspring from A50-treated mothers showed lower body weight at birth, 4 and 8 weeks of age, and glucose intolerance in adult females, probably due to insulin secretion and sensitivity alterations. Arsenic alters glucose homeostasis during pregnancy by altering beta-cell function, increasing risk of developing gestational diabetes. In pups, it induces low body weight from birth to 8 weeks of age, and glucose intolerance in females, demonstrating a sex specific response.


Subject(s)
Arsenites/toxicity , Drinking Water/adverse effects , Glucose Intolerance/etiology , Prenatal Exposure Delayed Effects/etiology , Animals , Body Weight/drug effects , Drinking Water/analysis , Female , Glucose Tolerance Test , Insulin/analysis , Lipid Peroxidation/drug effects , Liver/drug effects , Oxidative Stress/drug effects , Pregnancy , Rats , Rats, Sprague-Dawley
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