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Lacoste, B.

Publications and source records attributed to Lacoste, B..

11 recordsLinked to original sources

Control of Male Mouse Copulatory Behavior by Midbrain Dopamine Neurons

Ejaculation in male mice and humans requires intravaginal thrusting movements following penile intromission, yet the underlying neural mechanisms remain poorly understood. In this study, we used fiber photometry to record real-time activity of dopamine (DA) neurons in the ventral tegmental area (VTA) of male mice during copulation across six sexual interactions, in order to investigate how these dynamics change with experience and to examine their functional significance. Contrary to expectations, phasic VTA DA activity remained stable across sessions, showing time-locked responses to mounts, penile intromission, individual thrusts, and ejaculation, indicating experience-independent neural representations of all copulatory events. Notably, thrusting-related activity sharply declined just before ejaculation. Optogenetic inhibition of DA neurons disrupted thrusting behavior, but did not block ejaculation, suggesting DA signaling is critical for sustaining copulatory actions rather than triggering ejaculation itself. While prevailing theories frame VTA DA activity as primarily experience-dependent or centered on reward prediction, we here highlight its role in behavioral persistence during copulation. More broadly, our findings extend understanding of DA function beyond classical reward paradigms to the regulation of innate complex, goal-directed behaviors.

neuroscience↗

Metabolic, epigenetic and transcriptomic alterations in postnatal 16p11.2 deficient murine astrocytes

Autism Spectrum Disorders (ASD) are associated with metabolic dysregulation. While astrocytes are integral to cerebral metabolism, their molecular and functional changes in ASD are poorly known. Using early postnatal primary cortical astrocytes from a mouse model of 16p11.2 deletion ASD syndrome (16p11.2df/+ mice), we observed core molecular alterations with sex-specific profiles, suggesting divergent energetic pathways and epigenetic regulation. Targeted metabolomics revealed opposing phenotypes in male versus female 16p11.2df/+ astrocytes, particularly for alpha-ketoglutaric acid. Functionally, 16p11.2df/+astrocytes exhibited elevated phosphorylation in low glucose culture conditions, and reduced glycolysis in high glucose. Epigenetic profiling of male 16p11.2df/+astrocytes revealed differentially hydroxymethylated and methylated regions, with foci on chromosomes 3 and 13. Finally, bulk RNA sequencing in male and female mutant astrocytes indicated differential gene expression with profound sex differences, mostly affecting pathways related to cellular morphology. By establishing 16p11.2df/+ astroglial molecular signatures, this study refines our understanding of glial changes in ASD.

cell biology↗

Purinergic receptor activation rectifies autism-associated endothelial dysfunction

Early cerebrovascular alterations affect brain maturation by impacting trophic support and energy supply. Recent evidence in a 16p11.2 deletion mouse model of autism spectrum disorder (ASD) revealed brain endothelial abnormalities postnatally. Yet, the endothelial alterations eliciting these changes remain unknown. Isolation of brain endothelial cells (ECs) from 14-day old male 16p11.2-deficient and wild-type mice revealed that 16p11.2 deletion-induced endothelial dysfunction is linked to a bioenergetic failure, with reduced intracellular ATP. Intra- or extra-cellular ATP supplementation rescued the function of 16p11.2-deficient ECs in vitro via P2 purinergic receptor activation, specifically P2Y2 receptors. Activating P2Y2 receptors restored cerebrovascular reactivity in 16p11.2-deficient parenchymal arterioles ex vivo and rescued 16p11.2 deletion-associated mouse behaviors. Taken together, this study demonstrates that metabolic reprogramming of brain ECs via purinergic receptor engagement represents a possible therapeutic avenue for ASD.

cell biology↗

Zebrafish glial-vascular interactions progressively expand over the course of brain development

Glial-vascular interactions are critical for the formation and maintenance of brain blood vessels and the blood-brain barrier (BBB) in mammals, but their role in zebrafish is not well understood. Our previous work has detailed the timeline of BBB functional maturation in zebrafish, revealing a conserved mechanism of BBB induction through the suppression of endothelial transcytosis. Yet, as opposed to extensive research on glial-vascular interactions in rodents, such interactions remain largely overlooked in the zebrafish model system. Here, we focus on glial-vascular development in the zebrafish brain, leveraging three glial gene promoters: gfap (glial fibrillary acidic protein), glast (an astrocyte-specific glutamate transporter), and glastini (a new, shortened, equally effective version of the Glast promoter). Using these glial promoters, sparse labeling revealed fewer glial-vascular interactions during early larval stages, with both glial coverage and contact area increasing as the zebrafish brain matured. We then generated stable transgenic lines for both the Glast and Glastini promoters and observed similar increases in glial coverage during larval development, starting at [~]30% coverage at 3 days post-fertilization (dpf) and peaking at [~]60% at 10 dpf. Ultrastructural assessment of glial-vascular interactions using electron microscopy (EM) confirmed a progressive increase in glial coverage over larval development, with maximal coverage reaching [~]70% in adult zebrafish, significantly lower than the nearly 100% coverage observed in mammals. Finally, immunogold-EM labeling confirmed that cells identified as glia in aforementioned morphological analyses were indeed Glast-positive. Taken together, our results identify the temporal profile of glial-vascular maturation in the zebrafish brain.

developmental biology↗

Neuronal lipid droplets play a conserved and sex-biased role in maintaining whole-body energy homeostasis

ABSTRACTLipids are essential for neuron development and physiology. Yet, the central hubs that coordinate lipid supply and demand in neurons remain unclear. Here, we combine invertebrate and vertebrate models to establish the presence and functional significance of neuronal lipid droplets (LD) in vivo. We find that LD are normally present in neurons in a non-uniform distribution across the brain, and demonstrate triglyceride metabolism enzymes and lipid droplet-associated proteins control neuronal LD formation through both canonical and recently-discovered pathways. Appropriate LD regulation in neurons has conserved and male-biased effects on whole-body energy homeostasis across flies and mice, specifically neurons that couple environmental cues with energy homeostasis. Mechanistically, LD-derived lipids support neuron function by providing phospholipids to sustain mitochondrial and endoplasmic reticulum homeostasis. Together, our work identifies a conserved role for LD as the organelle that coordinates lipid management in neurons, with implications for our understanding of mechanisms that preserve neuronal lipid homeostasis and function in health and disease. HIGHLIGHTSO_LILipid droplets (LD) normally form in neurons across species Neuronal LD are regulated by a conserved gene network C_LIO_LINeuronal LD regulation plays a conserved and sex-biased role in maintaining energy homeostasis C_LIO_LILD regulation supports ER and mitochondrial function in hunger-activated neurons C_LI GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=200 SRC="FIGDIR/small/613929v1_ufig1.gif" ALT="Figure 1"> View larger version (54K): org.highwire.dtl.DTLVardef@1936e92org.highwire.dtl.DTLVardef@40478aorg.highwire.dtl.DTLVardef@18d5faorg.highwire.dtl.DTLVardef@882ee9_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗

Ultrastructural analysis of the brain endothelium by electron tomography

Transmission electron microscopy (TEM) is a powerful imaging technique, yielding ultrastructural investigation of organic and non-organic samples. Despite its ability to reach nanoscale resolutions, conventional TEM presents a major disadvantage by only acquiring two-dimensional snapshots, thus hindering our volumetric understanding of samples. Electron tomography (ET) overcomes this limitation by offering detailed views of a thin specimen in 3 dimensions (3D). This technique is widely used in biology and has expanded our understanding of mitochondrial structure or synaptic organization. Proper brain functioning is highly reliable on a constant nutritional support through its microvasculature lined by endothelial cells. These unique cells form a selective and protective barrier, known as the blood-brain barrier (BBB), which limits the entrance of blood-borne molecules into the brain. In pathological conditions, the BBB is disrupted, resulting in neuronal damage. Understanding the fine changes underlying BBB disruption requires advanced imaging tools such as ET, to detect the finest changes in endothelial ultrastructure. This manuscript briefly explains how TEM and ET function, and then provides a detailed, didactic method for sample preparation, tomogram generation and 3D segmentation of brain endothelial cells using ET.

neuroscience↗

Rock2 heterozygosity improves cognitive behavior and endothelial function in a mouse model of 16p11.2 deletion autism syndrome

Rho-associated coiled-coil containing protein kinase-2 (ROCK2) is a critical player in many cellular processes and has been incriminated in cardiovascular and neurological conditions. Recent evidence has shown that non-selective pharmacological ROCK inhibition ameliorates behavioral alterations in an autism mouse model of 16p11.2 haploinsufficiency. We had also revealed that 16p11.2-deficient mice display cerebrovascular abnormalities, including endothelial dysfunction. To investigate whether genetic blockage of ROCK2 also exerts beneficial effects on cognition and angiogenesis, we generated mice with both 16p11.2 and ROCK2 haploinsufficiency (16p11.2df/+;Rock2+/-). We find that Rock2 heterozygosity on a 16p11.2df/+ background rescues recognition memory and slightly reduces repetitive behaviors. Furthermore, brain endothelial cells (ECs) isolated from 16p11.2df/+;Rock2+/- mice display improved angiogenic capacity compared to ECs from 16p11.2df/+ littermates. Overall, this study implicates Rock2 gene as a critical modulator in 16p11.2-associated alterations, highlighting its potential as a target for treatment of autism spectrum disorders.

developmental biology↗

Astroglial Hmgb1 regulates postnatal astrocyte morphogenesis and cerebrovascular maturation.

Astrocytes are intimately linked with brain vessels, a relationship that is critical for neuronal health and function. However, astroglial factors driving these physical and functional associations during postnatal brain development have yet to be identified. We characterized structural and transcriptional changes in mouse cortical astrocytes and microvessels during the first two postnatal weeks and found that high-mobility group box 1 (Hmgb1), normally upregulated with injury and involved in adult cerebrovascular repair, was highly expressed in astrocytes at birth and then decreased rapidly. Astrocyte-selective ablation of Hmgb1 at birth affected astrocyte morphology and endfoot placement, altered distribution of endfoot proteins connexin43 and aquaporin-4, induced transcriptional changes in astrocytes related to cytoskeleton remodeling, and profoundly disrupted endothelial ultrastructure. While lack of astroglial Hmgb1 did not affect the blood-brain barrier or angiogenesis postnatally, it impaired neurovascular coupling and behavior in adult mice. These findings identify astroglial Hmgb1 as a key player in postnatal gliovascular maturation.

developmental biology↗

Cyclical control of female rejection behavior by progesterone receptor expressing neurons of the anterior ventromedial hypothalamus

Internal state-dependent behavioral responses, such as the switch between rejection and acceptance of sexual advances depending on a females reproductive capacity, are fundamental to maintain social interactions and wellbeing. Here we characterize a dedicated circuit for the cyclical control of rejection behavior in the mouse, located in the anterior ventrolateral ventromedial hypothalamus (aVMHvl). In vivo recordings reveal that progesterone receptor expressing neurons of the aVMHvl (aVMHvlPr+) are active during rejection but silent when females accept the male. Moreover, we show that aVMHvlPr+ neurons receive reduced excitatory to inhibitory synaptic input balance during the receptive phase of the reproductive cycle. Finally, optogenetic activation of aVMHvlPr+ neurons in receptive females is sufficient to increase rejection behavior, disrupting the probability of copulation, without affecting other male-directed interactions. This population of aVMHvlPr+ neurons is thus a key neural substrate controlling female sexual behavior, providing an additional barrier to mating when fertilization is not possible.

neuroscience↗

Sex-specific acute cerebrovascular response to photothrombotic stroke in mice requires rho-kinase

With high energy consumption and low energy storage, the brain is highly reliant on continuous cerebral blood flow (CBF) that delivers substrates to maintain proper function, which is compromised after a stroke. The current study explores the overlapping roles played by two important modulators of cerebrovascular tone, rho-kinase (ROCK) and endogenous sex hormones, in the acute CBF responses to a photothrombotic (PT) model of ischemic stroke in ROCK2+/- mice and wild-type (WT) littermates. To remove endogenous hormones, male mice were gonadectomized and female mice were ovariectomized, whereas control ("intact") animals received a sham surgery prior to stroke induction. Intact WT males showed a delayed drop in CBF compared to intact WT females, where maximal CBF drop was observed 48 hours following stroke. Gonadectomy in males did not alter this response, however ovariectomy in females produced a "male-like" response. Intact ROCK2+/- males also showed such phenotypic response, which was not altered by gonadectomy. Alternatively, intact ROCK2+/- females showed a striking difference in CBF values compared to intact WT females, where they displayed higher CBF values immediately post-stroke and also showed a maximal CBF drop 48 hours post-stroke, which was not altered by ovariectomy. Overall, there is a marked sex difference in acute CBF responses to PT stroke, which appears to be mediated by endogenous female sex hormones and ROCK2. This study reveals important sex-differences and the involvement of ROCK2 in acute CBF responses to PT stroke in mice. Significance StatementThere are very few mechanistic investigations on disparities between sexes in post-stroke CBF outcome. Rho-kinase, an important regulator of vascular tone, has only been explored in males in terms of its modulation of CBF following stroke. Both rho-kinase and endogenous female sex hormones have a converging role on the regulation of endothelial nitric oxide synthase (eNOS), an important modulator of vascular tone. Rho-kinase is thought to elicit its neuroprotective effects against ischemic stroke through eNOS, however this has never been investigated in both sexes. Elucidating the cellular and molecular bases of sex differences in cerebrovascular pathophysiology is vital for understanding the origins of stroke outcomes, and for designing novel therapeutic strategies to promote functional recovery in both women and men.

physiology↗

A galanin-positive population of lumbar spinal cord neurons modulates sexual behavior and arousal

During sex, male arousal increases to the ejaculatory threshold, allowing genital sensory input to trigger ejaculation. While copulation and sexual arousal are thought to be centrally regulated by the brain, ejaculation is considered a reflex controlled by a spinal circuit. In this framework, the spinal cord is assumed to be strongly inhibited by descending input from the brain until the ejaculatory threshold, playing no role in the regulation of copulatory behavior. However, this remains untested. Here, we mapped the spinal circuit controlling the bulbospongiosus muscle, which is involved in sperm expulsion in mice. Our findings reveal that bulbospongiosus muscle-motor neurons receive input from galanin-expressing interneurons. This galanin-positive population receives genital input and, while its stimulation leads to bulbospongiosus muscle activity, the evoked muscle-potentials are dependent on spinalization, the males internal state and decrease with repeated stimulation. Moreover, ablation of galanin neurons affected the latency to ejaculate and altered the copulatory pattern. These results suggest an unexpected role of spinal circuits in the control of copulation and arousal, in addition to its established role in ejaculation.

neuroscience↗