Molecular Fingerprint of Endocannabinoid Signaling in the Developing Paraventricular Nucleus of the Hypothalamus as Revealed by Single-Cell RNA-Seq and In Situ Hybridization
The paraventricular nucleus of the hypothalamus (PVN) regulates, among others, the stress response, sexual behavior, and energy metabolism through its magnocellular and parvocellular neurosecretory cells. Within the PVN, ensemble coordination occurs through the many long-range synaptic afferents, wh...
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2025-05-01
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| author | Evgenii O. Tretiakov Zsófia Hevesi Csenge Böröczky Alán Alpár Tibor Harkany Erik Keimpema |
| author_facet | Evgenii O. Tretiakov Zsófia Hevesi Csenge Böröczky Alán Alpár Tibor Harkany Erik Keimpema |
| author_sort | Evgenii O. Tretiakov |
| collection | DOAJ |
| description | The paraventricular nucleus of the hypothalamus (PVN) regulates, among others, the stress response, sexual behavior, and energy metabolism through its magnocellular and parvocellular neurosecretory cells. Within the PVN, ensemble coordination occurs through the many long-range synaptic afferents, whose activity in time relies on retrograde neuromodulation by, e.g., endocannabinoids. However, the nanoarchitecture of endocannabinoid signaling in the PVN, especially during neuronal development, remains undescribed. By using single-cell RNA sequencing, in situ hybridization, and immunohistochemistry during fetal and postnatal development in mice, we present a spatiotemporal map of both the 2-arachidonoylglycerol (2-AG) and anandamide (AEA) signaling cassettes, with a focus on receptors and metabolic enzymes, in both molecularly defined neurons and astrocytes. We find type 1 cannabinoid receptors (<i>Cnr1</i>), but neither <i>Cnr2</i> nor <i>Gpr55</i>, expressed in neurons of the PVN. <i>Dagla</i> and <i>Daglb</i>, which encode the enzymes synthesizing 2-AG, were found in all neuronal subtypes of the PVN, with a developmental switch from <i>Daglb</i> to <i>Dagla</i>. <i>Mgll</i>, which encodes an enzyme degrading 2-AG, was only found sporadically. <i>Napepld</i> and <i>Faah</i>, encoding enzymes that synthesize and degrade AEA, respectively, were sparsely expressed in neurons throughout development. Notably, astrocytes expressed <i>Mgll</i> and both <i>Dagl</i> isoforms. In contrast, mRNA for any of the three major cannabinoid-receptor subtypes could not be detected. Immunohistochemistry validated mRNA expression and suggested that endocannabinoid signaling is configured to modulate the activity of afferent inputs, rather than local neurocircuits, in the PVN. |
| format | Article |
| id | doaj-art-34e8a1ebb29147278bf95077ebd9f983 |
| institution | DOAJ |
| issn | 2073-4409 |
| language | English |
| publishDate | 2025-05-01 |
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| spelling | doaj-art-34e8a1ebb29147278bf95077ebd9f9832025-08-20T03:11:30ZengMDPI AGCells2073-44092025-05-01141178810.3390/cells14110788Molecular Fingerprint of Endocannabinoid Signaling in the Developing Paraventricular Nucleus of the Hypothalamus as Revealed by Single-Cell RNA-Seq and In Situ HybridizationEvgenii O. Tretiakov0Zsófia Hevesi1Csenge Böröczky2Alán Alpár3Tibor Harkany4Erik Keimpema5Department of Molecular Neurosciences, Center for Brain Research, Medical University of Vienna, 1090 Vienna, AustriaDepartment of Molecular Neurosciences, Center for Brain Research, Medical University of Vienna, 1090 Vienna, AustriaDepartment of Molecular Neurosciences, Center for Brain Research, Medical University of Vienna, 1090 Vienna, AustriaDepartment of Anatomy, Histology and Embryology, Semmelweis University, 1085 Budapest, HungaryDepartment of Molecular Neurosciences, Center for Brain Research, Medical University of Vienna, 1090 Vienna, AustriaDepartment of Molecular Neurosciences, Center for Brain Research, Medical University of Vienna, 1090 Vienna, AustriaThe paraventricular nucleus of the hypothalamus (PVN) regulates, among others, the stress response, sexual behavior, and energy metabolism through its magnocellular and parvocellular neurosecretory cells. Within the PVN, ensemble coordination occurs through the many long-range synaptic afferents, whose activity in time relies on retrograde neuromodulation by, e.g., endocannabinoids. However, the nanoarchitecture of endocannabinoid signaling in the PVN, especially during neuronal development, remains undescribed. By using single-cell RNA sequencing, in situ hybridization, and immunohistochemistry during fetal and postnatal development in mice, we present a spatiotemporal map of both the 2-arachidonoylglycerol (2-AG) and anandamide (AEA) signaling cassettes, with a focus on receptors and metabolic enzymes, in both molecularly defined neurons and astrocytes. We find type 1 cannabinoid receptors (<i>Cnr1</i>), but neither <i>Cnr2</i> nor <i>Gpr55</i>, expressed in neurons of the PVN. <i>Dagla</i> and <i>Daglb</i>, which encode the enzymes synthesizing 2-AG, were found in all neuronal subtypes of the PVN, with a developmental switch from <i>Daglb</i> to <i>Dagla</i>. <i>Mgll</i>, which encodes an enzyme degrading 2-AG, was only found sporadically. <i>Napepld</i> and <i>Faah</i>, encoding enzymes that synthesize and degrade AEA, respectively, were sparsely expressed in neurons throughout development. Notably, astrocytes expressed <i>Mgll</i> and both <i>Dagl</i> isoforms. In contrast, mRNA for any of the three major cannabinoid-receptor subtypes could not be detected. Immunohistochemistry validated mRNA expression and suggested that endocannabinoid signaling is configured to modulate the activity of afferent inputs, rather than local neurocircuits, in the PVN.https://www.mdpi.com/2073-4409/14/11/788cannabinoid receptorendocannabinoidhypothalamusmonoacylglycerol lipasepostnatal development<i>sn</i>-1-diacylglycerol lipase |
| spellingShingle | Evgenii O. Tretiakov Zsófia Hevesi Csenge Böröczky Alán Alpár Tibor Harkany Erik Keimpema Molecular Fingerprint of Endocannabinoid Signaling in the Developing Paraventricular Nucleus of the Hypothalamus as Revealed by Single-Cell RNA-Seq and In Situ Hybridization Cells cannabinoid receptor endocannabinoid hypothalamus monoacylglycerol lipase postnatal development <i>sn</i>-1-diacylglycerol lipase |
| title | Molecular Fingerprint of Endocannabinoid Signaling in the Developing Paraventricular Nucleus of the Hypothalamus as Revealed by Single-Cell RNA-Seq and In Situ Hybridization |
| title_full | Molecular Fingerprint of Endocannabinoid Signaling in the Developing Paraventricular Nucleus of the Hypothalamus as Revealed by Single-Cell RNA-Seq and In Situ Hybridization |
| title_fullStr | Molecular Fingerprint of Endocannabinoid Signaling in the Developing Paraventricular Nucleus of the Hypothalamus as Revealed by Single-Cell RNA-Seq and In Situ Hybridization |
| title_full_unstemmed | Molecular Fingerprint of Endocannabinoid Signaling in the Developing Paraventricular Nucleus of the Hypothalamus as Revealed by Single-Cell RNA-Seq and In Situ Hybridization |
| title_short | Molecular Fingerprint of Endocannabinoid Signaling in the Developing Paraventricular Nucleus of the Hypothalamus as Revealed by Single-Cell RNA-Seq and In Situ Hybridization |
| title_sort | molecular fingerprint of endocannabinoid signaling in the developing paraventricular nucleus of the hypothalamus as revealed by single cell rna seq and in situ hybridization |
| topic | cannabinoid receptor endocannabinoid hypothalamus monoacylglycerol lipase postnatal development <i>sn</i>-1-diacylglycerol lipase |
| url | https://www.mdpi.com/2073-4409/14/11/788 |
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