Vinicius Miessler de Andrade Carvalho

All Publications

• Gene network analysis identifies dysregulated pathways in an autism spectrum disorder caused by mutations in Transcription Factor 4 SCIENTIFIC REPORTS de Carvalho, L. M., Carvalho, V. M. A., Camargo, A. P., Papes, F. 2025; 15 (1): 4993

  Abstract

  Pitt-Hopkins syndrome (PTHS) is a rare neurodevelopmental monogenic disorder in the autistic spectrum caused by mutations in the Transcription Factor 4 gene. Even though the genetic etiology is known, the molecular mechanisms underlying PTHS remain poorly understood. To gain insight into the disease's pathophysiology, we set out to identify genes and pathways putatively involved in the pathology through co-expression and gene hub analyses using transcriptomic data from neural progenitor cells, neurons, and brain organoids derived from PTHS patients. Our results revealed several groups of co-expressed genes that are differentially regulated in PTHS neural cells compared to controls. These groups were enriched for genes involved in neural development and function, including synaptic transmission, membrane excitability, and cell adhesion. We identified several hub genes (highly connected nodes within gene networks that are central in these modules), including some that encode proteins involved in histone modification, synaptic vesicle trafficking, and cell signaling. Furthermore, we found that the differential expression of hub genes in PTHS neural cells was associated with altered cellular processes linked to neurodevelopment, such as cell-cell communication and irregular synaptic networks. Notably, we identified a set of hub genes related to the histone gene family, which is associated with neuronal differentiation and may contribute to PTHS pathogenesis and potentially serve as a biomarker for disease prognosis. Our results support the notion that PTHS involves alterations in neural development and function, particularly in excitatory neurons. The groups of co-expressed genes and hub genes we identified provide new insights into the molecular mechanisms underlying PTHS pathogenesis and could potentially be targeted for therapeutic intervention.

  View details for DOI 10.1038/s41598-025-89334-0

  View details for Web of Science ID 001418722300031

  View details for PubMedID 39929970

  View details for PubMedCentralID PMC11811132

• A neural circuit for male sexual behavior and reward. Cell Bayless, D. W., Davis, C. O., Yang, R., Wei, Y., de Andrade Carvalho, V. M., Knoedler, J. R., Yang, T., Livingston, O., Lomvardas, A., Martins, G. J., Vicente, A. M., Ding, J. B., Luo, L., Shah, N. M. 2023

  Abstract

  Male sexual behavior is innate and rewarding. Despite its centrality to reproduction, a molecularly specified neural circuit governing innate male sexual behavior and reward remains to be characterized. We have discovered a developmentally wired neural circuit necessary and sufficient for male mating. This circuit connects chemosensory input to BNSTprTac1 neurons, which innervate POATacr1 neurons that project to centers regulating motor output and reward. Epistasis studies demonstrate that BNSTprTac1 neurons are upstream of POATacr1 neurons, and BNSTprTac1-released substance P following mate recognition potentiates activation of POATacr1 neurons through Tacr1 to initiate mating. Experimental activation of POATacr1 neurons triggers mating, even in sexually satiated males, and it is rewarding, eliciting dopamine release and self-stimulation of these cells. Together, we have uncovered a neural circuit that governs the key aspects of innate male sexual behavior: motor displays, drive, and reward.

  View details for DOI 10.1016/j.cell.2023.07.021

  View details for PubMedID 37572660

• Transcription Factor 4 loss-of-function is associated with deficits in progenitor proliferation and cortical neuron content NATURE COMMUNICATIONS Papes, F., Camargo, A. P., de Souza, J. S., Carvalho, V. M. A., Szeto, R. A., LaMontagne, E., Teixeira, J. R., Avansini, S. H., Sanchez-Sanchez, S. M., Nakahara, T. S., Santo, C. N., Wu, W., Yao, H., Araujo, B. M. P., Velho, P. F., Haddad, G. G., Muotri, A. R. 2022; 13 (1): 2387

  Abstract

  Transcription Factor 4 (TCF4) has been associated with autism, schizophrenia, and other neuropsychiatric disorders. However, how pathological TCF4 mutations affect the human neural tissue is poorly understood. Here, we derive neural progenitor cells, neurons, and brain organoids from skin fibroblasts obtained from children with Pitt-Hopkins Syndrome carrying clinically relevant mutations in TCF4. We show that neural progenitors bearing these mutations have reduced proliferation and impaired capacity to differentiate into neurons. We identify a mechanism through which TCF4 loss-of-function leads to decreased Wnt signaling and then to diminished expression of SOX genes, culminating in reduced progenitor proliferation in vitro. Moreover, we show reduced cortical neuron content and impaired electrical activity in the patient-derived organoids, phenotypes that were rescued after correction of TCF4 expression or by pharmacological modulation of Wnt signaling. This work delineates pathological mechanisms in neural cells harboring TCF4 mutations and provides a potential target for therapeutic strategies for genetic disorders associated with this gene.

  View details for DOI 10.1038/s41467-022-29942-w

  View details for Web of Science ID 000789924700022

  View details for PubMedID 35501322

  View details for PubMedCentralID PMC9061776

• Investigation of Activated Mouse Olfactory Sensory Neurons via Combined Immunostaining and <i>in situ</i> Hybridization JOVE-JOURNAL OF VISUALIZED EXPERIMENTS de Andrade Carvalho, V., Nakahara, T., Papes, F. 2021

  Abstract

  Animals rely on chemical communication to convey and perceive relevant environmental information, ranging from assessment of food quality to detection of available mating partners or threats. In mice, this task is executed primarily by the olfactory system and its underlying subsystems, including the main and accessory olfactory systems. Both have peripheral organs populated by sensory neurons expressing G-protein coupled receptors able to bind chemical cues that reach the nasal cavity. Even though the molecular characteristics of these receptors is well understood, little is known about their cognate specific ligands. The method described here combines in situ hybridization detection of olfactory or vomeronasal receptors with immunodetection of phosphorylated ribosomal protein S6 (pS6) - a marker of neuronal activation. This protocol was devised to identify neurons activated after a single event of exposure to purified or complex chemical stimuli detected by the olfactory organs. Importantly, this technique allows the investigation of neurons triggered in biologically relevant contexts. Ideally, this method should be used to probe the molecular biology of the olfactory system and to study olfactory behaviors.

  View details for DOI 10.3791/62150

  View details for Web of Science ID 000646217000049

  View details for PubMedID 33900298

• Transcription factor 4 and its association with psychiatric disorders TRANSLATIONAL PSYCHIATRY Teixeira, J. R., Szeto, R. A., Carvalho, V. M. A., Muotri, A. R., Papes, F. 2021; 11 (1): 19

  Abstract

  The human transcription factor 4 gene (TCF4) encodes a helix-loop-helix transcription factor widely expressed throughout the body and during neural development. Mutations in TCF4 cause a devastating autism spectrum disorder known as Pitt-Hopkins syndrome, characterized by a range of aberrant phenotypes including severe intellectual disability, absence of speech, delayed cognitive and motor development, and dysmorphic features. Moreover, polymorphisms in TCF4 have been associated with schizophrenia and other psychiatric and neurological conditions. Details about how TCF4 genetic variants are linked to these diseases and the role of TCF4 during neural development are only now beginning to emerge. Here, we provide a comprehensive review of the functions of TCF4 and its protein products at both the cellular and organismic levels, as well as a description of pathophysiological mechanisms associated with this gene.

  View details for DOI 10.1038/s41398-020-01138-0

  View details for Web of Science ID 000610159000002

  View details for PubMedID 33414364

  View details for PubMedCentralID PMC7791034

• Detection of Activated Mouse Neurons with Temporal Resolution via Dual c-Fos Staining STAR PROTOCOLS Nakahara, T., Carvalho, V., Souza, M., Trintinalia, G., Papes, F. 2020; 1 (3): 100153

  Abstract

  This protocol combines fluorescent in situ hybridization and immunostaining to simultaneously detect, in histological sections from the same animal, subpopulations of neurons activated after two episodes of sensory stimulation. It allows the identification of groups of cells singly activated by either stimulus or co-activated by both stimuli. Our method results in nuclear staining for c-Fos mRNA and c-Fos protein, allowing better spatial and temporal resolution than previously published protocols, although it requires quick brain fixation. For complete details on the use and execution of this protocol, please refer to Carvalho et al. (2015, 2020).

  View details for DOI 10.1016/j.xpro.2020.100153

  View details for Web of Science ID 001050058800007

  View details for PubMedID 33377047

  View details for PubMedCentralID PMC7757322

• Peripheral oxytocin injection modulates vomeronasal sensory activity and reduces pup-directed aggression in male mice SCIENTIFIC REPORTS Nakahara, T. S., Camargo, A. P., Magalhaes, P. H. M., Souza, M. A. A., Ribeiro, P. G., Martins-Netto, P. H., Carvalho, V. M. A., Jose, J., Papes, F. 2020; 10 (1): 19943

  Abstract

  Behaviors are shaped by hormones, which may act either by changing brain circuits or by modifying sensory detection of relevant cues. Pup-directed behaviors have been previously shown to change via action of hormones at the brain level. Here, we investigated hormonal control of pup-induced activity in the vomeronasal organ, an olfactory sensory structure involved in the detection of non-volatile chemosignals. Vomeronasal activity decreases as males switch from a pup-aggressive state to a non-aggressive parenting state, after they socially contact a female. RNA sequencing, qPCR, and in situ hybridization were used to identify expression, in the vomeronasal sensory epithelium, of candidate GPCR hormone receptors chosen by in silico analyses and educated guesses. After identifying that oxytocin and vasopressin receptors are expressed in the vomeronasal organ, we injected the corresponding hormones in mice and showed that oxytocin administration reduced both pup-induced vomeronasal activity and aggressive behavior. Conversely, injection of an oxytocin receptor antagonist in female-primed male animals, which normally exhibit reduced vomeronasal activity, significantly increased the number of active vomeronasal neurons. These data link oxytocin to the modulation of olfactory sensory activity, providing a possible mechanism for changes in male behavior after social experience with females.

  View details for DOI 10.1038/s41598-020-77061-7

  View details for Web of Science ID 000604506600010

  View details for PubMedID 33203885

  View details for PubMedCentralID PMC7673031

• Representation of Olfactory Information in Organized Active Neural Ensembles in the Hypothalamus CELL REPORTS de Andrade Carvalho, V., Nakahara, T., de Andrade Souza, M., Cardozo, L., Trintinalia, G., Pissinato, L., Venancio, J., Stowers, L., Papes, F. 2020; 32 (8): 108061

  Abstract

  The internal representation of sensory information via coherent activation of specific pathways in the nervous system is key to appropriate behavioral responses. Little is known about how chemical stimuli that elicit instinctive behaviors lead to organized patterns of activity in the hypothalamus. Here, we study how a wide range of chemosignals form a discernible map of olfactory information in the ventromedial nucleus of the hypothalamus (VMH) and show that different stimuli entail distinct active neural ensembles. Importantly, we demonstrate that this map depends on functional inputs from the vomeronasal organ. We present evidence that the spatial locations of active VMH ensembles are correlated with activation of distinct vomeronasal receptors and that disjunct VMH ensembles exhibit differential projection patterns. Moreover, active ensembles with distinct spatial locations are not necessarily associated with different behavior categories, such as defensive or social, calling for a revision of the currently accepted model of VMH organization.

  View details for DOI 10.1016/j.celrep.2020.108061

  View details for Web of Science ID 000562818000004

  View details for PubMedID 32846119

• From Synapse to Supper: A Food Preference Recipe with Olfactory Synaptic Ingredients NEURON Nakahara, T. S., Carvalho, V. M. A., Papes, F. 2020; 107 (1): 8-11

  Abstract

  C1ql3 protein and its receptor Bai3 are involved in synaptic organization and function. In this issue of Neuron, Wang et al. (2020) report that both are essential for synaptic function between the anterior olfactory nucleus and the olfactory bulb and for the generation, but not recall, of associative olfactory memories determining food preference in mice.

  View details for DOI 10.1016/j.neuron.2020.06.016

  View details for Web of Science ID 000548279900003

  View details for PubMedID 32645309

• The Strange Case of Aggression and the Brain NEURON Carvalho, V. M. A., Nakahara, T. S., Papes, F. 2017; 95 (4): 734-737

  Abstract

  Territorial male mice are aggressive toward intruding males, but socially bonded males are not. Through manipulation of activity in a subset of neurons in the ventromedial hypothalamus, Yang et al. (2017) report that social and physiological factors non-linearly interact to control male aggression.

  View details for DOI 10.1016/j.neuron.2017.07.041

  View details for Web of Science ID 000407859400005

  View details for PubMedID 28817796

• Detection of pup odors by non-canonical adult vomeronasal neurons expressing an odorant receptor gene is influenced by sex and parenting status BMC BIOLOGY Nakahara, T. S., Cardozo, L. M., Ibarra-Soria, X., Bard, A. D., Carvalho, V. M. A., Trintinalia, G. Z., Logan, D. W., Papes, F. 2016; 14: 12

  Abstract

  Olfaction is a fundamental sense through which most animals perceive the external world. The olfactory system detects odors via specialized sensory organs such as the main olfactory epithelium and the vomeronasal organ. Sensory neurons in these organs use G-protein coupled receptors to detect chemosensory stimuli. The odorant receptor (OR) family is expressed in sensory neurons of the main olfactory epithelium, while the adult vomeronasal organ is thought to express other types of receptors.Here, we describe Olfr692, a member of the OR gene family identified by next-generation RNA sequencing, which is highly upregulated and non-canonically expressed in the vomeronasal organ. We show that neurons expressing this gene are activated by odors emanating from pups. Surprisingly, activity in Olfr692-positive cells is sexually dimorphic, being very low in females. Our results also show that juvenile odors activate a large number of Olfr692 vomeronasal neurons in virgin males, which is correlated with the display of infanticide behavior. . In contrast, activity substantially decreases in parenting males (fathers), where infanticidal aggressive behavior is not frequently observed.Our results describe, for the first time, a sensory neural population with a specific molecular identity involved in the detection of pup odors. Moreover, it is one of the first reports of a group of sensory neurons the activity of which is sexually dimorphic and depends on social status. Our data suggest that the Olfr692 population is involved in mediating pup-oriented behaviors in mice.

  View details for DOI 10.1186/s12915-016-0234-9

  View details for Web of Science ID 000370007400001

  View details for PubMedID 26878847

  View details for PubMedCentralID PMC4753656