Functional Diversity Research Articles

Nutrient contents and microbial communities as mediators of the effects of land-use in ecosystem functioning in alpine ecosystems from Central Mexico

Abstract Purpose The interplay between microbial taxonomy, functional diversity, and land-use changes across diverse ecosystems must be better understood. This study explores, through a statistical modeling approach, the intricate relationships between microbial communities, soil properties (nutrient contents and physicochemical parameters), and land-use changes, using stable isotopes ratio (δ13C/δ15N) as ecosystem function indicator. Methods Using Structural Equation Modeling (SEM) this study aims to investigate how soil microbial communities, soil properties and ecosystem function are interconnected in three ecosystems undergoing land-use change in Nevado de Toluca, Mexico. Results Multivariate analyses of microbial communities and soil parameters for the different land-uses reveal significant variation in both microbial composition and soil properties (i.e. nutrient contents) mainly explained by the land-use history of the studied ecosystems. Through the SEM approach, it was possible to disentangle the relative contribution of land-use change, microbial community composition and nutrient contents to the ecosystem function defined as the δ13C/δ15N ratio. Conclusions Microbial communities are the main drivers of soil ecosystem functioning. However, through SEM approach it is possible to formally test direct and indirect relationships impacting microbial communities and the resulting functional consequences, contributing to a mechanistic understanding of the ecological implications of land-use change, aiding in sustainable land-use decision-making.

Direct a-Hydroxy Acid Loading onto a Bacterial Thiotemplate Assembly Line via a Multienzyme Gateway.

Various nonribosomal peptide synthetases (NRPS) create structural and functional diversity by incorporating a-hydroxy acids into peptide backbones. Trigonic acid, an unusual cyclopropanol-substituted hydroxy acid, is the source of the molecular warhead of malleicyprol, a critical virulence factor of human and animal pathogens of the Burkholderia pseudomallei (BP) group. The process of selecting and loading this building block remained enigmatic as the NRPS module designated for this task is noncanonical. Using a combination of bioinformatics, mutational analyses, targeted metabolomics, and in vitro biochemical assays, we show that two trans-acting enzymes are required to load this central building block onto the modular assembly line. An adenylation-thiolation didomain enzyme (BurJ) activates trigonic acid, followed by the translocation of the enzyme-bound α-hydroxy acid thioester by an FkbH-like protein with a mutated phosphatase domain (BurH). This specialized gateway is the first reported direct loading of a hydroxy acid onto a bona fide NRPS module in bacteria and expands the synthetic biology toolbox for the site-specific incorporation of non-canonical building blocks. Moreover, insight into the biochemical basis of virulence factor biosynthesis can provide a foundation for developing enzyme inhibitors as anti-virulence therapeutics against BP pathogen infections.

The Heterogeneous Habitat of Taiga Forests Changes the Soil Microbial Functional Diversity

The soil contains abundant and diverse microorganisms, which interrelate closely with the aboveground vegetation and impact the structure and function of the forest ecosystem. To explore the effect of vegetation diversity on soil microbial functional diversity in taiga forests, we selected significantly different important values of Larix gmelinii as experimental grouping treatments based on plant investigation from fixed plots in Da Xing’anling Mountains. Following that, we collected soil samples and applied the Biolog-ECO microplate method to investigate differences in carbon source utilization, features of functional diversity in soil microorganisms, and factors influencing them in taiga forests. The AWCD decreased as the important value of Larix gmelinii grew, and soil microorganisms preferred carboxylic acids, amino acids, and carbohydrates over polymers, phenolic acids, and amines. The Shannon and McIntosh indexes decreased significantly with the increase of the important value of Larix gmelinii (p < 0.05) and were positively correlated with soil SOC, MBC, C/N, and pH, but negatively with TN, AP, and AN. Redundancy analysis revealed significant effects on soil microbial functional diversity from soil C/N, SOC, AP, MBC, TN, pH, AN, and WC. To sum up, heterogeneous habitats of taiga forests with different important values altered soil microbial functional diversity.

Direct a‐Hydroxy Acid Loading onto a Bacterial Thiotemplate Assembly Line via a Multienzyme Gateway

Various nonribosomal peptide synthetases (NRPS) create structural and functional diversity by incorporating a‐hydroxy acids into peptide backbones. Trigonic acid, an unusual cyclopropanol‐substituted hydroxy acid, is the source of the molecular warhead of malleicyprol, a critical virulence factor of human and animal pathogens of the Burkholderia pseudomallei (BP) group. The process of selecting and loading this building block remained enigmatic as the NRPS module designated for this task is noncanonical. Using a combination of bioinformatics, mutational analyses, targeted metabolomics, and in vitro biochemical assays, we show that two trans‐acting enzymes are required to load this central building block onto the modular assembly line. An adenylation‐thiolation didomain enzyme (BurJ) activates trigonic acid, followed by the translocation of the enzyme‐bound α‐hydroxy acid thioester by an FkbH‐like protein with a mutated phosphatase domain (BurH). This specialized gateway is the first reported direct loading of a hydroxy acid onto a bona fide NRPS module in bacteria and expands the synthetic biology toolbox for the site‐specific incorporation of non‐canonical building blocks. Moreover, insight into the biochemical basis of virulence factor biosynthesis can provide a foundation for developing enzyme inhibitors as anti‐virulence therapeutics against BP pathogen infections.

Bacteriocin diversity, function, discovery and application as antimicrobials.

Bacteriocins are potent antimicrobial peptides that are produced by bacteria. Since their discovery almost a century ago, diverse peptides have been discovered and described, and some are currently used as commercial food preservatives. Many bacteriocins exhibit extensively post-translationally modified structures encoded on complex gene clusters, whereas others have simple linear structures. The molecular structures, mechanisms of action and resistance have been determined for a number of bacteriocins, but most remain incompletely characterized. These gene-encoded peptides are amenable to bioengineering strategies and heterologous expression, enabling metagenomic miningand modification of novel antimicrobials. The ongoing global antimicrobial resistance crisis demands that novel therapeutics be developed to combat infectious pathogens. New compounds that are target-specific and compatible with the resident microbiota would be valuable alternatives to current antimicrobials. As bacteriocins can be broad or narrow spectrum in nature, they are promising tools for this purpose. However, few bacteriocins have gone beyond preclinical trials and none is currently used therapeutically in humans. In this Review, we explore the broad diversity in bacteriocin structure and function, describe identification and optimization methods and discuss the reasons behind the lack of translation beyond the laboratory of these potentially valuable antimicrobials.

Dental Dynamics: a fast new tool for quantifying tooth and jaw biomechanics in 3D Slicer

Abstract Teeth reveal how organisms interact with their environment. Biologists have long looked at the diverse form and function of teeth to study the evolution of feeding, fighting, and development. The exponential rise in the quantity and accessibility of computed tomography (CT) data has enabled morphologists to study teeth at finer resolutions and larger macroevolutionary scales. Measuring tooth function is no easy task, in fact, much of our mechanical understanding is derived from dental shape. Categorical descriptors of tooth shape such as morphological homodonty and heterodonty, overlook nuances in function by reducing tooth diversity for comparative analysis. The functional homodonty method quantitatively assesses the functional diversity of whole dentitions from tooth shape. This method uses tooth surface area and position to calculate the transmission of stress and estimates a threshold for functionally homodont teeth through bootstrapping and clustering techniques. However, some vertebrates have hundreds or thousands of teeth and measuring the shape and function of every individual tooth can be a painstaking task. Here we present Dental Dynamics, a module for 3D slicer that allows for the fast and precise quantification of dentitions and jaws. The tool automates the calculation of several tooth traits classically used to describe form and function (i.e., aspect ratio, mechanical advantage, force, etc.). To demonstrate the usefulness of our module we used Dental Dynamics to quantify 745 teeth across 20 salamanders that exhibit diverse ecologies. We coupled these data with the functional homodonty method to investigate the hypothesis that arboreal Aneides salamanders have novel tooth functions. Dental Dynamics provides a new and fast way to measure teeth and increases the accessibility of the functional homodonty method. We hope Dental Dynamics will encourage further theoretical and methodological development for quantifying and studying teeth.

Genetic and functional diversity of β-N-acetylgalactosamine-targeting glycosidases expanded by deep-sea metagenome analysis

β-N-Acetylgalactosamine-containing glycans play essential roles in several biological processes, including cell adhesion, signal transduction, and immune responses. β-N-Acetylgalactosaminidases hydrolyze β-N-acetylgalactosamine linkages of various glycoconjugates. However, their biological significance remains ambiguous, primarily because only one type of enzyme, exo-β-N-acetylgalactosaminidases that specifically act on β-N-acetylgalactosamine residues, has been documented to date. In this study, we identify four groups distributed among all three domains of life and characterize eight β-N-acetylgalactosaminidases and β-N-acetylhexosaminidase through sequence-based screening of deep-sea metagenomes and subsequent searching of public protein databases. Despite low sequence similarity, the crystal structures of these enzymes demonstrate that all enzymes share a prototype structure and have diversified their substrate specificities (oligosaccharide-releasing, oligosaccharide/monosaccharide-releasing, and monosaccharide-releasing) through the accumulation of mutations and insertional amino acid sequences. The diverse β-N-acetylgalactosaminidases reported in this study could facilitate the comprehension of their structures and functions and present evolutionary pathways for expanding their substrate specificity.

Influence of the leaf-cutting ant Atta sexdens Linnaeus (1758) on the taxonomic and functional diversity of woody plants in Atlantic Forest fragments

Influence of the leaf-cutting ant Atta sexdens Linnaeus (1758) on the taxonomic and functional diversity of woody plants in Atlantic Forest fragments

Does fortune follow function? Exploring how consumer preferences drive the functional trait composition of the global songbird trade

Abstract Defaunation and extinction undermine the resilience and functioning of ecological communities and ecosystems. Relative to other disturbances, overexploitation for the global wildlife trade presents a unique case of trait‐based selection, as demand for specific individuals is often tied to unique morphological or aesthetic traits desired by consumers (‘market traits’). Because evolutionary history leads to species that share both market and functional traits, we posit that non‐random patterns of exploitation will result in non‐random loss of functional diversity. We applied a trait‐based framework to the global songbird trade for 4616 species, 148 of which are plausibly threatened by the trade. We quantified select market traits, such as unique coloration and song quality, and ecological traits related to body size, diet, and foraging strategy to assess whether the trade disproportionately threatens particular functional groups. We additionally looked for patterns of association between market traits and functional traits to assess whether selection on certain market traits could drive selection on associated functional traits. We found that overexploited birds are a distinct functional subset of the global songbird pool, with the trade disproportionately threatening large bodied, frugivorous, and seed eating birds. Across all songbirds, there were multiple non‐random associations between market traits and functional traits, with the strongest associations observed among trade‐threatened birds; this was consistent with our theory that consumer‐driven selection on market traits could theoretically result in selection on functional traits. However, there was mixed evidence for this hypothesis at a global scale, suggesting that disproportionate threat to functional diversity may be more likely in regions where there is heavy demand for associated market traits. Policy implications. Our results highlight the need for increased focus on the mechanistic drivers of trait‐based selection on the consumer side of wildlife trade, and how patterns of overexploitation can systematically affect ecological communities and ecosystem services. Read the free Plain Language Summary for this article on the Journal blog.

Exploring the spectral variation hypothesis for α- and β-diversity: a comparison of open vegetation and forests

Airborne hyperspectral imaging holds great promise for estimating plant diversity and composition, given its unprecedented combination of aerial coverage, spatial resolution, and spectral detail. Recently, there has been renewed attention toward the spectral variation hypothesis (SVH), which predicts that higher spectral variation is correlated with greater plant diversity. While several studies have highlighted methodological challenges involved with the SVH, there is little consensus about when it yields strong predictions of taxonomic, functional, and phylogenetic diversity. In part, this may be because prior studies have not explicitly considered how underlying environmental gradients drive changes in spectral and species composition. In this study, we tested the SVH separately in open vegetation (i.e. grasses and shrubs) and in forests at five sites across Canada. Generalized additive models revealed that spectral diversity was a better predictor of functional α-diversity than of taxonomic or phylogenetic α-diversity in both vegetation types. Mantel tests and Procrustes analyses revealed weak to moderate associations between spectral and plant β-diversity and composition in open vegetation, and moderate associations in forests. The better fit in forests appeared to be influenced by the presence of an elevational gradient and associated species turnover (from deciduous to coniferous trees); we observed weaker relationships when examining only a subset of this gradient. We suggest that the high variability in the strength of associations between plant and spectral diversity reported to date might be affected by the presence of environmental gradients. Finally, we found that different wavelength bands contributed to spectral α-diversity in open vegetation vs. forests, suggesting different spectral features are important for different vegetation types. In conclusion, spectral diversity is a potentially powerful tool for biodiversity assessment, but it requires a context-specific approach.

The role of sheltered habitats in biodiversity conservation of species sensitive to drought: a case study using ground beetles (Coleoptera, Carabidae) in the Gorongosa National Park

AbstractIn the aftermath of 2019’ tropical cyclones Idai and Kenneth, we assessed ground beetle communities of the Gorongosa National Park (GNP) in Mozambique. The influence of habitat shelters, namely the buffering role of closed microhabitat conditions on Alpha diversity and community trait values during a long period of drought, was evaluated across the main habitat types of the park: miombo forests, mixed forests, transitional forests, and grasslands (open savannas). These habitat types comprised a distance gradient in relation to lake Urema, in the center of GNP. Miombo forests were farther from the lake while grasslands and transitional forests were in the floodplain area. Ground beetle communities were sampled using pitfall traps set up at 25 sites of each habitat type along an environmental gradient of tree canopy cover during the last twenty days of the dry season. Higher species richness of ground beetles was found in closed habitat shelters along the distance gradient to lake Urema. A higher functional diversity was also found along the canopy gradient, with larger sized and wingless species being more abundant in closed habitats than in open areas. This result highlights the crucial role of habitat closedness in the protection of sensitive ground beetles. In particular, the buffer effect of tree canopy and the protection of the understory plants and the litter layer was critical for the survival of hygrophilous species and will be key in conservation strategies to face climate aridification and habitat fragmentation.

Dryland reservoirs support greater taxonomic and functional beta diversity of zooplankton regardless of hydrological period

Dryland reservoirs support greater taxonomic and functional beta diversity of zooplankton regardless of hydrological period

Neuroprotective amyloid β N-terminal peptides differentially alter human α7- and α7β2-nicotinic acetylcholine (nACh) receptor single-channel properties.

Oligomeric amyloid β 1-42 (oAβ1-42) exhibits agonist-like action at human α7- and α7β2-containing nicotinic receptors. The N-terminal amyloid β1-15 fragment (N-Aβ fragment) modulates presynaptic calcium and enhances hippocampal-based synaptic plasticity via α7-containing nicotinic receptors. Further, the N-Aβ fragment and its core sequence, the N-amyloid-beta core hexapeptide (N-Aβcore), protect against oAβ1-42-associated synapto- and neurotoxicity. Here, we investigated how oAβ1-42, the N-Aβ fragment, and the N-Aβcore regulate the single-channel properties of α7- and α7β2-nicotinic receptors. Single-channel recordings measured the impact of acetylcholine, oAβ1-42, the N-Aβ fragment, and the N-Aβcore on the unitary properties of human α7- and α7β2-containing nicotinic receptors expressed in nicotinic-null SH-EP1 cells. Molecular dynamics simulations identified potential sites of interaction between the N-Aβ fragment and orthosteric α7+/α7- and α7+/β2- nicotinic receptor binding interfaces. The N-Aβ fragment and N-Aβcore induced α7- and α7β2-nicotinic receptor single-channel openings. Relative to acetylcholine, oAβ1-42 preferentially enhanced α7β2-nicotinic receptor single-channel open probability and open-dwell times. Co-application with the N-Aβcore neutralized these effects. Further, administration of the N-Aβ fragment alone, or in combination with acetylcholine or oAβ1-42, selectively enhanced α7-nicotinic receptor open probability and open-dwell times (compared to acetylcholine or oAβ1-42). Amyloid-beta peptides demonstrate functional diversity in regulating α7- and α7β2-nicotinic receptor function, with implications for a wide range of nicotinic receptor-mediated functions in Alzheimer's disease. The effects of these peptides on α7- and/or α7β2-nicotinic receptors revealed complex interactions with these subtypes, providing novel insights into the neuroprotective actions of amyloid β-derived fragments against the toxic effects of oAβ1-42.

Deer grazing drove an assemblage‐level evolution of plant dwarfism in an insular system

Abstract Plant dwarfism, a syndrome characterised by a significant reduction in plant height and organ size, is a widely observed pattern of stress‐tolerant life‐form evolution that results from local adaptation to harsh environmental conditions. The drivers of assemblage‐level dwarfism have primarily been attributed to abiotic factors, such as low temperature, aridity, poor soil fertility or frequent fires. While biotic factors such as grazing pressure from herbivores can contribute to the establishment of plant dwarfism, these factors have rarely been tested at assemblage levels. Focusing on a dwarf plant assemblage comprising over 80 taxa on a small continental island in Japan with a high deer density, we hypothesised that historical deer grazing could also be a factor contributing to the large‐scale convergent evolution of dwarfism. To test this hypothesis, we measured the size of 1908 individual plants of 40 taxa pairs, comprising both palatable and unpalatable pairs from the island and their counterpart taxa from neighbouring regions, and sought to assess which factors (i.e. low solar radiation, estimated divergence time, low nutrient conditions and grazing pressure from deer) may have contributed to the formation of the dwarf plant assemblage on the island. We also performed genetic analysis to infer the time frames for the establishment of dwarf taxa. Statistical analyses revealed that plant size was significantly reduced mainly among the palatable taxa growing on the island, with preferential grazing by deer being identified as the most significant factor influencing plant size. Furthermore, genetic analyses revealed that dwarf ecotypes may have evolved over tens of thousands of years. Synthesis: To the best of our knowledge, this study is the first to demonstrate that interactions with herbivores can shape the assemblage‐level convergence of plant dwarfism. These findings enhance our current understanding of the formation of plant functional diversity.

Combined effects of border irrigation and super-absorbent polymers on enzyme activity and microbial diversity of poplar rhizosphere soil.

Fast-growing poplar plantations are considered a great benefit to timber production, but water availability is a key factor limiting their growth and development, especially in arid and semi-arid ecosystems. Super-absorbent polymers facilitate more water retention in soil after rain or irrigation, and they are able to release water gradually during plant growth. This study aimed to examine the effects of reduced irrigation (60% and 30% of conventional border irrigation) co-applied with super-absorbent polymers (0, 40 kg/ha) on root exudates, enzyme activities, microbial functional diversity in rhizosphere soil, and volume increments in poplar (Populus euramericana cv. 'Neva'). The results showed that 60% border irrigation co-applied with super-absorbent polymers significantly increased the content of organic acids, amino acids and total sugars in the root exudates, and the activities of invertase, urease, dehydrogenase, and catalase in the rhizosphere soil in comparison to conventional border irrigation without super-absorbent polymers. Meanwhile, this treatment also enhanced the average well-color development, Shannon index, and McIntosh index, but decreased the Simpson index. Additionally, the average volume growth rate and relative water content of leaves reached their maximum using 60% irrigation with super-absorbent polymers, which was significantly higher than other treatments. However, using 30% irrigation with super-absorbent polymers, had a smaller effect on rhizosphere soil and volume growth than 60% irrigation with super-absorbent polymers. Therefore, using an appropriate water-saving irrigation measure (60% conventional border irrigation with super-absorbent polymers) can help to improve enzyme activities and microbial diversity in the rhizosphere soil while promoting the growth of poplar trees.

Linking aerial hyperspectral data to canopy tree biodiversity: An examination of the spectral variation hypothesis

AbstractImaging spectroscopy is emerging as a leading remote sensing method for quantifying plant biodiversity. The spectral variation hypothesis predicts that variation in plant hyperspectral reflectance is related to variation in taxonomic and functional identity. While most studies report some correlation between spectral and field‐based (i.e., taxonomic and functional) expressions of biodiversity, the observed strength of association is highly variable, and the utility in applying spectral community properties to examine environmental drivers of communities remains unknown. We linked hyperspectral data acquired by airborne imaging spectrometers with precisely geolocated field plots to examine the spectral variation hypothesis along a temperate‐to‐boreal forest gradient in southern Québec, Canada. First, we examine the degree of association between spectral and field‐based dimensions of canopy tree composition and diversity. Second, we ask whether the relationships between field‐based community properties and the environment are reproduced when using spectral community properties. We found support for the spectral variation hypothesis with the strength of association generally greater for the functional than taxonomic dimension, but the strength of relationships was highly variable and dependent on the choice of method or metric used to quantify spectral and field‐based community properties. Using a multivariate approach (comparisons of separate ordinations), spectral composition was moderately well correlated with field‐based composition; however, the degree of association increased when univariately relating the main axes of compositional variation. Spectral diversity was most tightly associated with functional diversity metrics that quantify functional richness and divergence. For predicting canopy tree composition and diversity using environmental variables, the same qualitative conclusions emerge when hyperspectral or field‐based data are used. Spatial patterns of canopy tree community properties were strongly related to the turnover from temperate‐to‐boreal communities, with most variation explained by elevation. Spectral composition and diversity provide a straightforward way to quantify plant biodiversity across large spatial extents without the need for a priori field observations. While commonly framed as a potential tool for biodiversity monitoring, we show that spectral community properties can be applied more widely to assess the environmental drivers of biodiversity, thereby helping to advance our understanding of the drivers of biogeographical patterns of plant communities.

Species richness and ecological connectivity of the mammal communities in urban and peri-urban areas at Mexico City

AbstractUrban expansion has emerged as a pervasive driver of biodiversity loss in Mexican cities due to shifts in landscape composition and configuration. Hence, the preservation of forest cover, green spaces and isolated trees assume a critical role in conserving the biodiversity within urban areas. We compared mammal assemblages across 520 sites in Mexico City examining the impact of local vegetation and site characteristics (e.g., patch size, isolation distance) on diversity patterns. Then, we used a generalized linear model to evaluate the relationship between mammal assemblages (including both alpha and beta diversities) and the level of structural and functional connectivity across the landscape. We collected 5,063 records of 38 species from the sampled sites. Mammalian richness ranged from 1 to 9 spp./site, and the mean of taxonomic and functional (Functional Distance [MFD]) alpha diversities were 1.9 ± 1.3 spp./site and 0.41 ± 0.60 MFD/site, respectively. The average size of habitat fragments was 0.41 hectares, and the percent forest cover per site ranged from 0.5 to 100%. Both species and functional alpha diversities were different among the land-use and vegetation types (including protected vs. non-protected areas). There was a clear distinction (β value > 0.40) in species composition across the landscape, which was positively related to geographical distance and negatively related to connectivity between sites. We identified four main patches especially important to connectivity across the city and argue that conserving them is critical for restoring key components of biodiversity and ecosystem services in urban and peri-urban areas of Mexico City.

Combining mesocosms with models reveals effects of global warming and ocean acidification on a temperate marine ecosystem.

Ocean warming and species exploitation have already caused large-scale reorganization of biological communities across the world. Accurate projections of future biodiversity change require a comprehensive understanding of how entire communities respond to global change. We combined a time-dynamic integrated food web modeling approach (Ecosim) with previous data from community-level mesocosm experiments to determine the independent and combined effects of ocean warming, ocean acidification and fisheries exploitation on a well-managed temperate coastal ecosystem. The mesocosm parameters enabled important physiological and behavioral responses to climate stressors to be projected for trophic levels ranging from primary producers to top predators, including sharks. Through model simulations, we show that under sustainable rates of fisheries exploitation, near-future warming or ocean acidification in isolation could benefit species biomass at higher trophic levels (e.g., mammals, birds, and demersal finfish) in their current climate ranges, with the exception of small pelagic fishes. However, under warming and acidification combined, biomass increases at higher trophic levels will be lower or absent, while in the longer term reduced productivity of prey species is unlikely to support the increased biomass at the top of the food web. We also show that increases in exploitation will suppress any positive effects of human-driven climate change, causing individual species biomass to decrease at higher trophic levels. Nevertheless, total future potential biomass of some fisheries species in temperate areas might remain high, particularly under acidification, because unharvested opportunistic species will likely benefit from decreased competition and show an increase in biomass. Ecological indicators of species composition such as the Shannon diversity index decline under all climate change scenarios, suggesting a trade-off between biomass gain and functional diversity. By coupling parameters from multilevel mesocosm food web experiments with dynamic food web models, we were able to simulate the generative mechanisms that drive complex responses of temperate marine ecosystems to global change. This approach, which blends theory with experimental data, provides new prospects for forecasting climate-driven biodiversity change and its effects on ecosystem processes.

A knock-in allele of Hand2 expressing Dre recombinase.

HAND2 is a basic helix-loop-helix transcription factor with diverse functions during development. To facilitate the investigation of genetic and functional diversity among Hand2-expressing cells in the mouse, we have generated Hand2Dre, a knock-in allele expressing Dre recombinase. To avoid disrupting Hand2 function, the Dre cDNA is inserted at the 3' end of the Hand2 coding sequence following a viral 2A peptide. Hand2Dre homozygotes can therefore be used in complex crosses to increase the proportion of useful genotypes among offspring. Dre expression in mid-gestation Hand2Dre embryos is indistinguishable from wild-type Hand2 expression, and HandDre efficiently recombines rox target sites in vivo. In combination with existing Cre and Flp mouse lines, Hand2Dre will therefore extend the ability to perform genetic intersectional labeling, fate mapping, and functional manipulation of subpopulations of cells characterized by developmental expression of Hand2.

Autism patient-derived SHANK2BY29X mutation affects the development of ALDH1A1 negative dopamine neuron.

Autism spectrum disorder (ASD) encompasses a range of neurodevelopmental conditions. Different mutations on a single ASD gene contribute to heterogeneity of disease phenotypes, possibly due to functional diversity of generated isoforms. SHANK2, a causative gene in ASD, demonstrates this phenomenon, but there is a scarcity of tools for studying endogenous SHANK2 proteins in an isoform-specific manner. Here, we report a point mutation on SHANK2, which is found in a patient with autism, located on exon of the SHANK2B transcript variant (NM_133266.5), hereby SHANK2BY29X. This mutation results in an early stop codon and an aberrant splicing event that impacts SHANK2 transcript variants distinctly. Induced pluripotent stem cells (iPSCs) carrying this mutation, from the patient or isogenic editing, fail to differentiate into functional dopamine (DA) neurons, which can be rescued by genetic correction. Available SMART-Seq single-cell data from human midbrain reveals the abundance of SHANK2B transcript in the ALDH1A1 negative DA neurons. We then show that SHANK2BY29X mutation primarily affects SHANK2B expression and ALDH1A1 negative DA neurons in vitro during early neuronal developmental stage. Mice knocked in with the identical mutation exhibit autistic-like behavior, decreased occupancy of ALDH1A1 negative DA neurons and decreased dopamine release in ventral tegmental area (VTA). Our study provides novel insights on a SHANK2 mutation derived from autism patient and highlights SHANK2B significance in ALDH1A1 negative DA neuron.