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Becchetti, A.

Publications and source records attributed to Becchetti, A..

4 recordsLinked to original sources

Divergent modulation of dopaminergic neurons by hypocretin/orexin receptors-1 and -2 shapes dopaminergic cell activity and socio-emotional behavior

BACKGROUNDMany neuropsychiatric disorders involve dysregulation of the dopaminergic (DA) input to the forebrain. Of particular relevance are DA projections stemming from the midbrain ventral tegmental area (VTA). A key neuromodulatory influence onto DAVTA neurons arises from lateral hypothalamic area hypocretin/orexin (OX) neurons. Despite being a major input, the differential action of orexin peptides A and B (OXA and OXB) on orexin receptors 1 and 2 in DA cells is poorly understood. We recently identified profoundly divergent functions of OX1R vs OX2R in DA cells in regulating sleep/wake architecture, brain oscillations and cognitive behaviors. OX2R, but not OX1R, loss dramatically increased time in EEG theta-rich (alert) wakefulness, reward-driven learning and attentional skills, but impaired inhibitory control. METHODSUsing genetically engineered mice whose DA cells selectively lack OX input via Hcrtr1 (DAOx1R-KO) or Hcrtr2 (DAOx2R-KO), we assessed intrinsic excitability and electrophysiological responses of DAVTA neurons and evaluated behavioral phenotypes across multiple domains. RESULTSWe uncover previously unrecognized effects of OX peptides on DAVTA cell response. In WT and control mice, we show that while OXA enhances, OXB diminishes DAVTA neuronal excitability. OX1R-deficient DA cells lose OXA responding and OX2R-deficient DA cells lose OXB responding. DA Ox1R loss generates anxiety-like behavior and context-dependent hyperactivity. In contrast, OX2R loss decreases sociability and, despite exhibiting enhanced reward-driven learning, mice show highly compromised aversion-driven learning. CONCLUSIONSWe evidence strikingly distinct functions of OX1R vs OX2R signaling in modulating the intrinsic excitability of DAVTA neurons and influencing DA-related behaviors. These data implicate OX[->]DA signaling pathways in neuropsychiatric endophenotypes relevant to obsessive-compulsive, attention-deficit/hyperactivity, and autism spectrum disorders, and raise important considerations for the development of OXR-targeted therapeutics.

neuroscience↗

An integrin centered complex coordinates ion transport and pH to regulate f-actin organization and cell migration in breast cancer.

Reciprocal signaling between the Tumor Microenvironment (TME) and cancer cells regulates abnormal proliferation, migration and pro-metastatic behavior. Major player in such interaction is integrin-mediated cell adhesion to the Extracellular Matrix (ECM). Integrin receptors organize signaling hubs constituted by multiprotein membrane complexes often comprising ion channels and transporters. We studied whether and how integrin-centered multiprotein complexes control cell behavior in Breast Cancer (BCa) cell populations with different molecular characteristics. BCa cells were cultured onto the ECM protein fibronectin (FN), to trigger {beta}1 integrin activation. Through biochemical, immunofluorescence and electrophysiological experiments we provide evidence of a novel signaling pathway that involves a {beta}1 integrin-centered plasma membrane complex formed by different transport proteins: the hERG1 K+ channel, the neonatal form of the Na+ channel NaV1.5 (nNaV1.5) and the Na+/H+ antiporter NHE1. The NHE1/hERG1/{beta}1/nNaV1.5 complex was found on the plasma membrane of BCa cells, and particularly of Triple Negative Breast Cancer (TNBCa). When engaged by cell adhesion to FN, such membrane complex recruited the cytoskeletal actin-binding protein a-actinin1 and stimulated NHE1-mediated cytoplasmic alkalinization. Thus, the multiprotein complex activation affected TNBCa migration and invasiveness by stimulating f-actin organization directly (through -actinin1) and indirectly (by intracellular alkalinization). The contribution of both hERG1 and nNaV1.5 was essential, as the adhesion-dependent signaling pathway and its functional consequences were inhibited by blocking either channel with, respectively, E4031 and TTX, or by applying RNA silencing procedures. The contribution of hERG1 to the structural integrity of the membrane complex appeared to be critical, as the adhesion-dependent signals were hampered by harnessing the hERG1/{beta}1 integrin complex with a single chain bispecific antibody (scDb-hERG1-{beta}1) which disrupts the macromolecular complex without blocking the K+ current, as well as by E4031, which impairs the complex formation by blocking the channel in the open state. In conclusion, we revealed that integrin-centered macromolecular complexes in BCa cells recruit a battery of ion transport proteins that cooperate in modulating different aspects of the downstream signals that lead to malignant behavior. This complex could be targeted to develop novel therapeutic strategies for one of the most difficult-to-treat cancers, i.e. TNBCa.

cancer biology↗

Orexin action on the dopaminergic system modulates theta during REM sleep and wakefulness

Both dopaminergic (DA) and orexinergic (OX) systems establish brain-wide neuromodulatory circuits that profoundly influence brain states and behavioral outputs. To unravel their interactions, we inactivated OX-to-DA neurotransmission by selective disruption of HcrtR1/OxR1, or HcrtR2/OxR2, or both receptors, in DA neurons. Chronic loss of OXR2 in DA neurons (OxR2Dat-CKO mice) dramatically increased electrocorticographic (EcoG) theta rhythms in wakefulness and REM sleep. Episode duration and total times spent in active wakefulness and REMS were prolonged, and theta/fast-gamma wave coupling was enhanced in both states. Increased theta in OxR2DatCKO mice baseline wake was accompanied by diminished infra-theta and increased fast-gamma activities, i.e. the mice exhibited signs of constitutive electrocortical hyperarousal, albeit uncoupled with locomotor activity. These effects were not seen in OxR1-ablated dopaminergic mutants, which tended to show opposite phenotypes, resembling those caused by the loss of both receptors. Our data establish a clear, genetically-defined link between monosynaptic orexin-to-dopaminergic connectivity and the power of theta oscillations, with a differential role of OXR2 in cross-frequency wave coupling and attentional processes.

neuroscience↗

An early Sox2-dependent gene expression program required for hippocampal dentate gyrus development

The hippocampus is a brain area central for cognition. Mutations in the human SOX2 transcription factor cause neurodevelopmental defects, leading to intellectual disability and seizures, together with hippocampal dysplasia. We generated an allelic series of Sox2 conditional mutations in mouse, deleting Sox2 at different developmental stages. Late Sox2 deletion (from E11.5, via Nestin-Cre) affects only postnatal hippocampal development; earlier deletion (from E10.5, Emx1-Cre) significantly reduces the dentate gyrus, and the earliest deletion (from E9.5, FoxG1-Cre) causes drastic abnormalities, with almost complete absence of the dentate gyrus. We identify a set of functionally interconnected genes (Gli3, Wnt3a, Cxcr4, p73 and Tbr2), known to play essential roles in hippocampal embryogenesis, which are downregulated in early Sox2 mutants, and (Gli3 and Cxcr4) directly controlled by SOX2; their downregulation provides plausible molecular mechanisms contributing to the defect. Electrophysiological studies of the Emx1Cre mouse model reveal altered excitatory transmission in CA1 and CA3 regions.

developmental biology↗