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Gonzalez, S.

Publications and source records attributed to Gonzalez, S..

At least 19 recordsLinked to original sources

Dynamic structural equation models for ecosystem-based fisheries management: case studies and a practical guide

Once central challenge in adopting ecosystem-based fisheries management (EBFM) is understanding and predicting the interconnected processes, feedbacks, and temporal dependencies that characterize ecosystem dynamics. Structural equation models and structural causal models (SEM and SCM) quantify interconnected relationships and evaluate causality from observational data but cannot readily accommodate the incomplete and irregularly sampled data typical of complex systems. Dynamic structural equation models (DSEM) estimate SEMs but accommodate lagged effects, missing data, and non-linear and non-stationary effects. However, there is little guidance regarding how DSEM can be used to support EBFM, particularly within governance and decision-making contexts. Here, we present three case studies that highlight distinct inferential goals for understanding ecosystem processes in the context of EBFM. First, we quantify ecosystem-level indicators to support monitoring of climate-driven ecosystem changes. Second, we examine whether hypothesized direct and indirect ecosystem linkages improve predictions of recruitment. Finally, we compare support for hypotheses of causal drivers of fish productivity. To provide practical advice, we synthesize lessons learned into a workflow to for others navigating DSEM. We suggest that the first step is identifying the inferential objective, which establishes the foundation for the remainder of the workflow, guiding the development of graphical models, model specification, and the selection of statistical and scientific validation steps (e.g., testing conditional independencies) needed to evaluate hypothesized causal relationships. By integrating observational data, expert knowledge, and mechanistic hypotheses within a unified framework, DSEM is a powerful tool for advancing EBFM, and more broadly, understanding the processes that govern complex ecosystems.

ecology↗

An adhesion GPCR regulates cell adhesion and mating in the closest living relatives of metazoans

The transition to metazoan multicellularity required the evolution of cell-surface receptors that coordinate adhesion and signaling under changing environmental conditions. We investigated potential regulators of cell interactions in the choanoflagellate Salpingoeca rosetta, one of the closest living relatives of metazoans. Here, we identify Cupidon, an adhesion G protein-coupled receptor that acts as a dual-function regulator of cell adhesion and mating. Under well-fed (i.e., nutrient-replete) conditions, Cupidon suppresses cell aggregation by inhibiting N-acetylglucosamine-dependent collar-mediated adhesion. Starvation of S. rosetta triggers gametogenesis, resulting in anisogametes: female gametes with an elongated collar and male gametes that form a basal protrusion, the "fertilopod." Cupidon undergoes concurrent changes in proteolytic processing and localization, ultimately concentrating at the gamete contact interface, where it promotes gamete fusion. Together, our findings reveal that aGPCR-mediated regulation of cell adhesion predates the origin of metazoans.

evolutionary biology↗

Molecular Basis of Histone H3 Reading and Writing by Legionella pneumophila SET Domain Lysine Methyltransferases

RomA and its highly conserved strain ortholog LegAS4 are SET and ankyrin domain-containing effector proteins of the intracellular bacterial pathogen Legionella pneumophila. These enzymes are secreted into host cells, where they translocate to the nucleus and methylate Lys14 in histone H3 (H3K14), a novel post-translation modification (PTM) that reprograms gene expression and promotes bacterial replication. To elucidate their H3K14 substrate specificity, we determined the crystal structures of LegAS4 and RomA bound to histone H3 peptides and characterized nucleosome binding and methylation by the enzymes. The results reveal a distinctive bipartite engagement of the histone H3 N-terminal tail by the enzymes SET and ankyrin domains. Further, the ankyrin domain distinguishes different PTMs at H3R2 and H3K4, which is crucial for nucleosome engagement and H3K14 methylation. Together, these studies yield new insights into histone H3 reading and writing by the L. pneumophila histone lysine methyltransferases during host cell infection.

biochemistry↗

Multiple-Context Training Enhances Generalization of Cued Fear Extinction and Requires the Dorsal Hippocampus

BACKGROUNDMultiple-session fear extinction (FE) models exposure therapy, which also occurs over many sessions. In humans, conducting extinction training across multiple contexts improves fear suppression as compared to a single context, but rodent work delineating boundary conditions and neural mechanisms remains sparse. METHODSWe compared single-context FE (SCFE) versus multiple-context FE (MCFE) in 129S1 and C57BL/6J mice, manipulating context familiarity (familiar vs novel FE contexts). Outcomes included FE acquisition and tests for fear relapse such as recent (2 d) and remote (30 d) recall in trained, extinction, and novel contexts, as well as reinstatement tests. We mapped extinction-related neural activity with dual Fos labeling (TRAPxFos immunofluorescence) and graph analyses of regional co-activation, and used chemogenetic inhibition to selectively silence dorsal (dHP) or ventral hippocampus (vHP) during extinction. RESULTSMCFE produced greater across-session reduction in freezing than SCFE. The MCFE advantage did not require contextual novelty. MCFE also enhanced short- (2 d) and long-term (30 d) recall in a novel context, but did not prevent recovery over time or US-induced reinstatement. Dual-labeling revealed that MCFE strengthened hippocampal-prefrontal co-activation (with relatively weaker prefrontal-amygdala coupling than SCFE). Chemogenetic silencing of dHP, but not vHP, selectively impaired between-session extinction under MCFE and abolished the MCFE benefit; hippocampal silencing did not improve SCFE. CONCLUSIONSConducting extinction across multiple contexts enhances acquisition and recall relative to SCFE, without requiring novelty, and engages distinct hippocampal-prefrontal circuit dynamics. dHP activity is necessary for the MCFE-specific improvement, highlighting a circuit mechanism in which distributing extinction across contexts augments fear reduction.

neuroscience↗

Asymmetric increase in episodic and procedural memory interference in older adults

In younger adults, newly formed procedural memories are weakened by the subsequent formation of episodic memories (E[->]P interference) and vice versa (P[->]E interference; "cross-memory interference"). Older adults experience significant decline in episodic memory but maintain relatively intact procedural memory. This asymmetric decline in memory may also cause an asymmetric change in cross-memory interference compared to younger adults. For example, older adults may experience a significant increase in one type of cross-memory interference while leaving the other unchanged. Additionally, decline in episodic memory may cause E[->]P interference to either increase or decrease depending on how the episodic and procedural memory systems interact. However, no study to our knowledge has compared cross-memory interference between younger and older adults. We investigated cross-memory interference in younger and older adults by measuring E[->]P (Exp. 1) and P[->]E (Exp. 2) interference in 40 younger (18-40 years old) and 40 older ([≥] 55 years old) adults. Compared to younger adults, the results show that older adults experience significantly stronger E[->]P interference while P[->]E interference was statistically indistinguishable between groups. These results confirm that older adults experience an asymmetric increase in cross-memory interference and suggest that the increase in E[->]P interference is related to the asymmetric decline in episodic memory relative to procedural memory.

neuroscience↗

Failure to reproduce the effect of procedural memory interference on wakeful consolidation of episodic memory in younger and older adults

Brown and Robertson (2007) revealed that skill learning interferes with the wakeful consolidation of episodic memories in young adults. This finding is commonly used as evidence that episodic and procedural memories should not be learned in close temporal proximity but has not been reproduced by an independent laboratory. Additionally, older adults experience episodic memory deficits, but it is unknown whether this group is also vulnerable to this type of interference. We aimed to reproduce Brown and Robertsons (2007) finding in younger adults, while also comparing the magnitude of interference between younger and older adults. Forty younger (18-40 years; n =20) and older adults ([≥]55 years; n = 20) visited the laboratory in the morning and acquired episodic memories (a list of words) immediately before a procedural finger-tapping (procedural) task. Half of all participants were exposed to a learnable sequential structure. In the afternoon of the same day, participants were asked to recall the episodic memories from the morning session. We found weak evidence of interference for both age groups and no statistical difference in interference between groups. Our results suggest that the interfering effects of these memory types may be negligible or overestimated, and that these memory types can be acquired together without interference.

neuroscience↗

Human striatal progenitor cells that contain inducible safeguards and overexpress BDNF rescue Huntington's disease phenotypes in R6/2 mice

Huntingtons disease (HD) is an autosomal-dominant neurodegenerative disorder characterized by striatal atrophy. Reduced trophic support due to decreased striatal levels of neurotrophins (NTs), mainly brain-derived neurotrophic factor (BDNF), contributes importantly to HD pathogenesis; restoring NTs has significant therapeutic potential. Human pluripotent stem cells (hPSC) offer a scalable platform for NT delivery but has potential safety risks including teratoma formation. We engineered hPSCs to constitutively produce BDNF and contain inducible safeguards to eliminate these cells if safety concerns arise. This study examined the efficacy of intrastriatally transplanted striatal progenitor cells (STRpcs) derived from these hPSCs against HD phenotypes in R6/2 mice. Engrafted STRpcs overexpressing BDNF alleviated motor and cognitive deficits and reduced mutant huntingtin aggregates. Activating the inducible safety switch with rapamycin safely eliminated the engrafted cells. These results demonstrate that BDNF delivery via a novel hPSC-based platform incorporating safety switches could be a safe and effective HD therapeutic.

neuroscience↗

Potentiation of the M1 muscarinic acetylcholine receptor normalizes neuronal activation patterns and improves apnea severity in Mecp2+/- mice

Rett syndrome (RTT) is a neurodevelopmental disorder that is caused by loss-of-function mutations in the methyl-CpG binding protein 2 (MeCP2) gene. RTT patients experience a myriad of debilitating symptoms, which include respiratory phenotypes that are often associated with lethality. Our previous work established that expression of the M1 muscarinic acetylcholine receptor (mAchR) is decreased in RTT autopsy samples, and that potentiation of the M1 receptor improves apneas in a mouse model of RTT; however, the population of neurons driving this rescue is unclear. Loss of Mecp2 correlates with excessive neuronal activity in cardiorespiratory nuclei. Since M1 is found on cholinergic interneurons, we hypothesized that M1-potentiating compounds decrease apnea frequency by tempering brainstem hyperactivity. To test this, Mecp2+/- and Mecp2+/+ mice were screened for apneas before and after administration of the M1 positive allosteric modulator (PAM) VU0453595 (VU595). Brains from the same mice were then imaged for c-Fos, ChAT, and Syto16 using whole-brain light-sheet microscopy to establish genotype and drug-dependent activation patterns that could be correlated with VU595s efficacy on apneas. The vehicle-treated Mecp2+/- brain exhibited broad hyperactivity when coupled with the phenotypic prescreen, which was significantly decreased by administration of VU595, particularly in regions known to modulate the activity of respiratory nuclei (i.e. hippocampus and striatum). Further, the extent of apnea rescue in each mouse showed a significant positive correlation with c-Fos expression in non-cholinergic neurons in the striatum, thalamus, dentate gyrus, and within the cholinergic neurons of the brainstem. These results indicate that Mecp2+/- mice are prone to hyperactivity in brain regions that regulate respiration, which can be normalized through M1 potentiation.

neuroscience↗

A high-throughput method for quantifying Drosophila fecundity

Measurements of Drosophila fecundity are used in a wide variety of studies, such as investigations of stem cell biology, nutrition, behavior, and toxicology. In addition, because fecundity assays are performed on live flies, they are suitable for longitudinal studies such as investigations of aging or prolonged chemical exposure. However, standard Drosophila fecundity assays have been difficult to perform in a high-throughput manner because experimental factors such as the physiological state of the flies and environmental cues must be carefully controlled to achieve consistent results. In addition, exposing flies to a large number of different experimental conditions (such as chemical additives in the diet) and manually counting the number of eggs laid to determine the impact on fecundity is time-consuming. We have overcome these challenges by combining a new multiwell fly culture strategy with a novel 3D-printed fly transfer device to rapidly and accurately transfer flies from one plate to another; the RoboCam, a low-cost, custom built robotic camera to capture images of the wells automatically; and an image segmentation pipeline to automatically identify and quantify eggs. We show that this method is compatible with robust and consistent egg laying throughout the assay period; and demonstrate that the automated pipeline for quantifying fecundity is very accurate (r2 = 0.98 for the correlation between the automated egg counts and the ground truth) In addition, we show that this method can be used to efficiently detect the effects on fecundity induced by dietary exposure to chemicals. Taken together, this strategy substantially increases the efficiency and reproducibility of high throughput egg laying assays that require exposing flies to multiple different media conditions.

genetics↗

A foundational atlas of autism protein interactions reveals molecular convergence

Translating high-confidence (hc) autism spectrum disorder (ASD) genes into viable treatment targets remains elusive. We constructed a foundational protein-protein interaction (PPI) network in HEK293T cells involving 100 hcASD risk genes, revealing over 1,800 PPIs (87% novel). Interactors, expressed in the human brain and enriched for ASD but not schizophrenia genetic risk, converged on protein complexes involved in neurogenesis, tubulin biology, transcriptional regulation, and chromatin modification. A PPI map of 54 patient-derived missense variants identified differential physical interactions, and we leveraged AlphaFold-Multimer predictions to prioritize direct PPIs and specific variants for interrogation in Xenopus tropicalis and human forebrain organoids. A mutation in the transcription factor FOXP1 led to reconfiguration of DNA binding sites and altered development of deep cortical layer neurons in forebrain organoids. This work offers new insights into molecular mechanisms underlying ASD and describes a powerful platform to develop and test therapeutic strategies for many genetically-defined conditions.

systems biology↗

Nanobody CDR3 mimetics enhance SOS1-catalyzed nucleotide exchange on RAS

RAS proteins control various intracellular signaling networks. Mutations at specific locations were shown to stabilize their active GTP-bound state, which is associated with the development of multiple cancers. An attractive approach to modulate RAS signaling is through its regulatory guanine nucleotide exchange factor (GEF) son of sevenless 1 (SOS1). With the recent discovery of Nanobody14, which potently enhances SOS1-catalyzed nucleotide exchange on RAS, we explored the feasibility of developing peptide mimetics by structurally mimicking the complementarity-determining region 3 (CDR3). Guided by a biochemical GEF assay and X-ray co-crystal structures, successive rounds of optimization and gradual conformational rigidification led to CDR3 mimetics showing half of the maximal activation potential of the native nanobody. Altogether, this study provides the first proof-of-concept that peptides able to functionally modulate a protein-protein interaction can be obtained by structural mimicry of a nanobody paratope.

bioengineering↗

ChILL & DisCO to discover competitive, connective and allosteric Nanobodies that modulate the SOS1-RAS protein-protein interactions and tune the nucleotide exchange rate

Protein-protein interactions (PPIs) are central in cell metabolism but research tools for the structural and functional characterization of these PPIs are often missing. Here we introduce novel and broadly applicable immunization (Cross-link PPIs and immunize llamas, ChILL) and selection strategies (Display and co-selection, DisCO) for the discovery of diverse Nanobodies that either stabilize or disrupt PPIs in a single experiment. We applied ChILL and DisCO to identify competitive, connective or fully allosteric Nanobodies that inhibit or facilitate the formation of the SOS1*RAS complex and modulate the nucleotide exchange rate on this pivotal GTPase in vitro and RAS signalling in cellulo. One of these connective Nanobodies fills a cavity that was previously identified as the binding pocket for a series of therapeutic lead compounds. The long complementarity-determining region (CDR3) that penetrates this binding pocket serves as an innovative pharmacophore for extending the repertoire of potential leads.

bioengineering↗

Pleiotropy of autism-associated chromatin regulators

Gene ontology analyses of high confidence autism spectrum disorder (hcASD) risk genes have historically highlighted chromatin regulation and synaptic function as major contributors to pathobiology. Our recent functional work in vivo has additionally implicated microtubule biology and identified disrupted cellular proliferation as a convergent ASD phenotype. As many chromatin regulators, including ASD risk genes ADNP and CHD3, are known to directly regulate both tubulins and histones, we studied the five chromatin regulators most strongly associated with ASD (ADNP, CHD8, CHD2, POGZ, and SUV420H1/KMT5B) specifically with respect to microtubule biology. We observe that all five localize to microtubules of the mitotic spindle in vitro and in vivo. Further in-depth investigation of CHD2 provides evidence that patient-derived mutations lead to a range of microtubule-related phenotypes, including disrupted localization of the protein at the mitotic spindle, spindle defects, cell cycle stalling, DNA damage, and cell death. Lastly, we observe that ASD genetic risk is significantly enriched among microtubule-associated proteins, suggesting broader relevance. Together, these results provide further evidence that the role of tubulin biology and cellular proliferation in ASD warrant further investigation and highlight the pitfalls of relying solely on annotated gene functions in the search for pathological mechanisms.

developmental biology↗

Mimicking genuine drought responses using a high throughput plate assay

Soil-free assays that induce water stress are routinely used to investigate drought responses in the plant Arabidopsis thaliana. Due to their ease of use, the research community often relies on polyethylene glycol (PEG), mannitol and salt (NaCl) treatments to reduce the water potential of agar media, and thus induce drought conditions in the laboratory. However, while these types of stress can create phenotypes that resemble those of water deficit experienced by soil-grown plants, it remains unclear how these treatments compare at the transcriptional level. Here, we demonstrate that these different methods of lowering water potential elicit both shared and distinct transcriptional responses in Arabidopsis shoot and root tissue. When we compared these transcriptional responses to those found in Arabidopsis roots subject to vermiculite drying, we discovered many genes induced by vermiculite drying were repressed by low water potential treatments on agar plates (and vice versa). Additionally, we also tested another method for lowering water potential of agar media. By increasing the nutrient content and tensile strength of agar, we show the hard agar (HA) treatment can be leveraged as a high-throughput assay to investigate natural variation in Arabidopsis growth responses to low water potential.

plant biology↗

Copy number footprints of platinum-based anticancer therapies

Recently, distinct mutational footprints observed in metastatic tumors, secondary malignancies and normal human tissues have been demonstrated to be caused by the exposure to several chemotherapeutic drugs. These characteristic mutations originate from specific lesions caused by these chemicals to the DNA of exposed cells. However, it is unknown whether the exposure to these chemotherapies leads to a specific footprint of larger chromosomal aberrations. Here, we address this question exploiting whole genome sequencing data of metastatic tumors obtained from patients exposed to different chemotherapeutic drugs. As a result, we discovered a specific copy number footprint across tumors from patients previously exposed to platinum-based therapies. This footprint is characterized by a significant increase in the number of chromosomal fragments of copy number 1-4 and size smaller than 10 Mb in exposed tumors with respect to their unexposed counterparts (median 14-387% greater across tumor types). The number of chromosomal fragments characteristic of the platinum-associated CN footprint increases significantly with the activity of the well known platinum-related footprint of single nucleotide variants across exposed tumors.

genomics↗

Deploying photons for communication within neuronal networks

Deficiencies in neurotransmission lead to neurological disorders or misinterpretation of perceived threats. To restore defects in cellular communication, we developed a synthetic, photon-assisted synaptic transmission (PhAST) system. PhAST is based on luciferases and channelrhodopsins that enable the transmission of a neuronal state across space, using photons as neurotransmitters. We demonstrate the ability to overcome synaptic barriers and rescue the behavioral deficit of a genetically engineered glutamate mutant with conditional, Ca2+-triggered photon emission between two cognate neurons of the Caenorhabditis elegans nociceptive avoidance circuit. We also deploy these ingredients for asynaptic transmission between two unrelated cells in a sexually dimorphic neuronal network. Functional PhAST could sensitize otherwise poorly responsive males to touch and hence expand the behavioral repertoire. Our study, thus, establishes a powerful framework for complex photon-based communication between neurons in a living animal, that can readily be expanded to synthetic neuronal networks, organoids or non-invasive brain-machine interfaces.

synthetic biology↗

Phenotyping the tumor-immune microenvironment (TiME) in vivo by cellular-level optical imaging reveals unique combination phenotypes with variable inflammation and endothelial anergy

Immunotherapies have shown unprecedented clinical benefits in several malignancies1-3. However, clinical responses remain variable and unpredictable, indicating the need to develop predictive platforms that can improve patient stratification4. Phenotyping of tumors into hot, altered, or cold5 based on assessment of only T-cell infiltration in static tumor biopsies provides suboptimal prediction of immunotherapy response6,7. In vivo dynamic mechanisms within the tumor microenvironment such as tumor angiogenesis and leukocyte trafficking5,8,9 also play a central role in modulating anti-tumor immunity and therefore immunotherapy response. Here, we report novel tumor immune microenvironment (TiME) phenotyping in vivo in patients with non-invasive spatially-resolved cellular-level imaging based on endogenous contrast. Investigating skin cancers as a model, with reflectance confocal microscopy (RCM) imaging10, we determined four major phenotypes with variable prevalence of vasculature (Vasc) and inflammation (Inf) features: VaschiInfhi, VaschiInflo, VascloInfhi and Vascmed/hiInflo. The VaschiInfhi phenotype correlates with high immune activation, exhaustion, and vascular signatures while VaschiInflo with endothelial anergy and immune exclusion. Automated quantification of TiME features demonstrates moderate-high accuracy and correlation with corresponding gene expression. Prospectively analyzed response to topical immunotherapy show highest response in VascloInfhi, and reveals the added value of vascular features in predicting treatment response. Our novel in vivo cellular-level imaging and phenotyping approach can potentially advance our fundamental understanding of TiME, develop robust predictors for immunotherapy outcomes and identify novel targetable pathways in future.

cancer biology↗

Post-stroke administration of the p75 neurotrophin receptor modulator, LM11A-31, attenuates chronic changes in brain metabolism, increases neurotransmitter levels, and improves recovery

The aim of this study was to test whether post-stroke oral administration of a small molecule p75 neurotrophin receptor (p75NTR) modulator (LM11A-31) can augment neuronal survival and improve recovery in a mouse model of stroke. Mice were administered LM11A-31 for up to 12 weeks, beginning 1 week after stroke. Metabolomic analysis revealed that after 2 weeks of daily treatment, mice that received LM11A-31 were distinct from vehicle treated mice by principal component analysis and had higher levels of serotonin, acetylcholine, and dopamine in their ipsilateral hemisphere. LM11A-31 treatment also improved redox homeostasis by restoring reduced glutathione. It also offset a stroke induced reduction in glycolysis by increasing acetyl-CoA. There was no effect on cytokine levels in the infarct. At 13 weeks following stroke, adaptive immune cell infiltration in the infarct was unchanged in LM11A-31 treated mice, indicating that LM11A-31 does not alter the chronic inflammatory response to stroke at the site of the infarct. However, LM11A-31 treated mice had less brain atrophy, neurodegeneration, tau pathology, and microglial activation in other regions of the ipsilateral hemisphere. These findings correlated with improved recovery of motor function on a ladder test, improved sensorimotor and cognitive abilities on a nest construction test, and less impulsivity in an open field test. These data support small molecule modulation of the p75 neurotrophin receptor for preserving neuronal health and function during stroke recovery. SIGNIFICANCE STATEMENTThe findings from this study introduce the p75 neurotrophin receptor as a novel small molecule target for promotion of stroke recovery. Given that LM11A-31 is in clinical trials as a potential therapy for Alzheimers disease, it could be considered as a candidate for assessment in stroke or vascular dementia studies.

neuroscience↗