Genetic Analysis System Research Articles

Peripheral Neuropathic Pain.

This article synthesizes current knowledge on neuropathic pain, with a brief review of mechanisms, diagnostic approaches, and treatment strategies to help neurologists provide effective and individualized care for patients with this complex condition. The most promising developments in peripheral neuropathic pain are related to the molecular biology of the peripheral nervous system. Systematic molecular and genetic analyses of peripheral nerve terminals and dorsal root ganglia have advanced our understanding of the genetics of function and disease of peripheral nerves, as well as their physiology and clinical manifestations. Peripheral neuropathic pain, similar to central neuropathic pain, is primarily influenced by the biology and pathophysiology of the underlying structures, peripheral sensory nerves, and their central pathways. The clinical course is widely variable in sensory symptoms and intensities, natural history, and response to treatments.

Estimation of economic equations as a management tool in feedlot dairy farming

The aim of this study was to evaluate Pearson's linear correlations between productive and economic indicators as well as to estimate economic equations as tools in dairy farming. We used the database of the Research Group, registered with CNPq under the title ‘Animal and Forage Production in Piaui’, which consisted of two experiments with feedlot lactating cows. Economic and productive indicators were employed. Correlations were determined by Pearson’s linear correlation analysis, using the ‘t’ test. Equations were estimated via regression, using Saeg software (Statistical and Genetic Analysis System), considering α = 0.05. Based on concepts related to Pearson's linear correlation and dairy farming management, we were able to find correlations that allowed us to estimate simple, yet very informative equations, which facilitated the interpretation of the management of a dairy farm. Among the studied variables, net present value, internal rate of return, the cost per liter of milk produced and net margin were those which most contributed to the estimation of equations.

Characterization of the cystargolide biosynthetic gene cluster and functional analysis of the methyltransferase CysG

Cystargolides are natural products originally isolated from Kitasatospora cystarginea NRRL B16505 as inhibitors of the proteasome. They are composed of a dipeptide backbone linked to a β-lactone warhead. Recently, we identified the cystargolide biosynthetic gene cluster but systematic genetic analyses had not been carried out because of the lack of a heterologous expression system. Here, we report the discovery of a homologous cystargolide biosynthetic pathway in Streptomyces durhamensis NRRL-B3309 by genome mining. The gene cluster was cloned via transformation-associated recombination and heterologously expressed in Streptomyces coelicolor M512. We demonstrate that it contains all genes necessary for the production of cystargolide A and B. Single gene deletion experiments reveal that only five of the eight genes from the initially proposed gene cluster are essential for cystargolide synthesis. Additional insights into the cystargolide pathway could be obtained from in vitro assays with CysG and chemical complementation of the respective gene knockout. This could be further supported by the in vitro investigation of the CysG homolog BelI from the belactosin biosynthetic gene cluster. Thereby, we confirm that CysG and BelI catalyze a cryptic SAM-dependent transfer of a methyl group that is critical for the construction of the cystargolide and belactosin β-lactone warheads. Introduction

A MinION-based Long-Read Sequencing Application With One-Step PCR for the Genetic Diagnosis of 21-Hydroxylase Deficiency.

Recently developed long-read sequencing (LRS) technology has been considered an option for CYP21A2 analysis. However, the clinical use of LRS for CYP21A2 analysis is limited. This study's objective is to develop an efficient and low-cost LRS system for CYP21A2 screening. A DNA fragment library was prepared in a single polymerase chain reaction (PCR) that covers the entire CYP21A2 gene and all known junctions caused by TNXB gene structural rearrangements, yielding a single 8-kb product of CYP21A2 or CYP21A1P/CYP21A2 chimera. After barcoding, the PCR products were sequenced on a MinION-based platform with Flongle Flow Cell R9.4.1 and R10.4.1. The reference genotypes of 55 patients with 21-hydroxylase deficiency (21OHD) were established using the conventional method with multiplex ligation-dependent probe amplification (MLPA) and nested PCR. LRS using Flongle Flow Cell R9.4.1 yielded consistent results. Additionally, the recently updated LRS "duplex" analysis with Flongle flow cell R10.4.1 was tested to reveal an advantage of accurately sequencing a variant located on the homopolymer region. By introducing a barcode system, the cost was reduced to be comparable to that of conventional analysis. A novel single-nucleotide variation was discovered at the acceptor site of intron 7, c.940-1G > C. We also identified a subtype of the classical chimeric junction CH2, "CH2a," in the region from the latter part of intron 5 to exon 6. We successfully established a novel low-cost and highly accurate LRS system for 21OHD genetic analysis. Our study provides insight into the feasibility of LRS for diagnosing 21OHD and other genetic diseases caused by structural rearrangements.

Newly Established Genetic System for Functional Analysis of MetSV.

The linear chromosome of the Methanosarcina spherical virus with 10,567 bp exhibits 22 ORFs with mostly unknown functions. Annotation using common tools and databases predicted functions for a few genes like the type B DNA polymerase (MetSVORF07) or the small (MetSVORF15) and major (MetSVORF16) capsid proteins. For verification of assigned functions of additional ORFs, biochemical or genetic approaches were found to be essential. Consequently, we established a genetic system for MetSV by cloning its genome into the E. coli plasmid pCR-XL-2. Comparisons of candidate plasmids with the MetSV reference based on Nanopore sequencing revealed several mutations of yet unknown provenance with an impact on protein-coding sequences. Linear MetSV inserts were generated by BamHI restriction, purified and transformed in Methanosarcina mazei by an optimized liposome-mediated transformation protocol. Analysis of resulting MetSV virions by TEM imaging and infection experiments demonstrated no significant differences between plasmid-born viruses and native MetSV particles regarding their morphology or lytic behavior. The functionality of the genetic system was tested by the generation of a ΔMetSVORF09 mutant that was still infectious. Our genetic system of MetSV, the first functional system for a virus of methanoarchaea, now allows us to obtain deeper insights into MetSV protein functions and virus-host interactions.

The Validation of a Precursor Lesion of Epithelial Ovarian Cancer in Fancd2-KO Mice.

Ovarian cancer (OC) has the highest mortality rate of all gynaecological malignancies. The asymptomatic nature and limited understanding of early disease hamper research into early-stage OC. Therefore, there is an urgent need for models of early-stage OC to be characterised to improve the understanding of early neoplastic transformations. This study sought to validate a unique mouse model for early OC development. The homozygous Fanconi anaemia complementation group D2 knock-out mice (Fancd2-/-) develop multiple ovarian tumour phenotypes in a sequential manner as they age. Using immunohistochemistry, our group previously identified purported initiating precursor cells, termed 'sex cords', that are hypothesised to progress into epithelial OC in this model. To validate this hypothesis, the sex cords, tubulostromal adenomas and equivalent controls were isolated using laser capture microdissection for downstream multiplexed gene expression analyses using the Genome Lab GeXP Genetic Analysis System. Principal component analysis and unbiased hierarchical clustering of the resultant expression data from approximately 90 OC-related genes determined that cells from the sex cords and late-stage tumours clustered together, confirming the identity of the precursor lesion in this model. This study, therefore, provides a novel model for the investigation of initiating neoplastic events that can accelerate progress in understanding early OC.

Hb Q-Thailand heterozygosity unlinked with the (–α4.2/) α+-thalassemia deletion allele identified by long-read SMRT sequencing: hematological and molecular analyses

ABSTRACTObjectiveIn the present study, two unrelated cases of Hb Q-Thailand heterozygosity unlinked with the (–α4.2/) α+-thalassemia deletion allele were identified by long-read single molecule real-time (SMRT) sequencing in southern China. The aim of this study was to report the hematological and molecular features as well as diagnostic aspects of the rare manifestation.MethodsHematological parameters and hemoglobin analysis results were recorded. A suspension array system for routine thalassemia genetic analysis and long-read SMRT sequencing were applied in parallel for thalassemia genotyping. Traditional methods, including Sanger sequencing, multiplex gap-polymerase chain reaction (gap-PCR) and multiplex ligation-dependent probe amplification (MLPA), were used together to confirm the thalassemia variants.ResultsLong-read SMRT sequencing was used to diagnose two Hb Q-Thailand heterozygous patients for whom the hemoglobin variant was unlinked to the (–α4.2/) allele for the first time. The hitherto undescribed genotypes were verified by traditional methods. Hematological parameters were compared with those of Hb Q-Thailand heterozygosity linked with the (–α4.2/) deletion allele in our study. For the positive control samples, long-read SMRT sequencing revealed a linkage relationship between the Hb Q-Thailand allele and the (–α4.2/) deletion allele.ConclusionsIdentification of the two patients confirms that the linkage relationship between the Hb Q-Thailand allele and the (−α4.2/) deletion allele is a common possibility but not a certainty. Remarkably, as it is superior to traditional methods, SMRT technology may eventually serve as a more comprehensive and precise method that holds promising prospects in clinical practice, especially for rare variants.

Deciphering the Genetic Basis of Silkworm Cocoon Colors Provides New Insights into Biological Coloration and Phenotypic Diversification.

The genetic basis of phenotypic variation is a long-standing concern of evolutionary biology. Coloration has proven to be a visual, easily quantifiable, and highly tractable system for genetic analysis and is an ever-evolving focus of biological research. Compared with the homogenized brown-yellow cocoons of wild silkworms, the cocoons of domestic silkworms are spectacularly diverse in color, such as white, green, and yellow-red; this provides an outstanding model for exploring the phenotypic diversification and biological coloration. Herein, the molecular mechanism underlying silkworm green cocoon formation was investigated, which was not fully understood. We demonstrated that five of the seven members of a sugar transporter gene cluster were specifically duplicated in the Bombycidae and evolved new spatial expression patterns predominantly expressed in silk glands, accompanying complementary temporal expression; they synergistically facilitate the uptake of flavonoids, thus determining the green cocoon. Subsequently, polymorphic cocoon coloring landscape involving multiple loci and the evolution of cocoon color from wild to domestic silkworms were analyzed based on the pan-genome sequencing data. It was found that cocoon coloration involved epistatic interaction between loci; all the identified cocoon color-related loci existed in wild silkworms; the genetic segregation, recombination, and variation of these loci shaped the multicolored cocoons of domestic silkworms. This study revealed a new mechanism for flavonoids-based biological coloration that highlights the crucial role of gene duplication followed by functional diversification in acquiring new genetic functions; furthermore, the results in this work provide insight into phenotypic innovation during domestication.

Development of a clinical microarray system for genetic analysis screening

ObjectivesResearch on the relationship between diseases and genes and the advancement of genetic analysis technologies have made genetic testing in medical care possible. There are various methods for genetic testing, including PCR-based methods and next-generation sequencing; however, screening tests in clinical laboratories are becoming more diverse; therefore, novel measurement systems and equipment are required to meet the needs of each situation. In this study, we aimed to develop a novel microarray-based genetic analysis system that uses a Peltier element to overcome the issues of conventional microarrays, such as the long measurement time and high cost. MethodsWe constructed a microarray system to detect the UDP-glucuronosyltransferase gene polymorphisms UGT1A1*6 and UGT1A1*28 in patients eligible for irinotecan hydrochloride treatment for use in clinical laboratories. To evaluate the performance of the system, the hybridization temperature and reaction time were determined, and the results were compared with those obtained using a conventional hybridization oven. ResultsThe hybridization temperature reached its target in 1/27th of the time required by the conventional system. We assessed 111 human clinical samples and found that our results agreed with those obtained using existing methods. The total time for the newly developed device was reduced by 85 min compared to that for existing methods, as the automated DNA microarray eliminates the time that existing methods spend on manual operation. ConclusionsThe surface treatment technology used in our system enables high-density and strong DNA fixation, allowing the construction of a measurement system suitable for clinical applications.

An immunogenetic investigation of 30 autoimmune and autoinflammatory diseases and their links to psychiatric disorders in a nationwide sample.

Autoimmune and autoinflammatory diseases (AIIDs) involve a deficit in an individual's immune system function, whereby the immune reaction is directed against self-antigens. Many AIIDs have a strong genetic component, but they can also be triggered by environmental factors. AIIDs often have a highly negative impact on the individual's physical and mental wellbeing. Understanding the genetic underpinning of AIIDs is thus crucial both for diagnosis and for identifying individuals at high risk of an AIID and mental illness as a result thereof. The aim of the present study was to perform systematic statistical and genetic analyses to assess the role of human leukocyte antigen (HLA) alleles in 30 AIIDs and to study the links between AIIDs and psychiatric disorders. We leveraged the Danish iPSYCH Consortium sample comprising 65 534 individuals diagnosed with psychiatric disorders or selected as part of a random population sample, for whom we also had genetic data and diagnoses of AIIDs. We employed regression analysis to examine comorbidities between AIIDs and psychiatric disorders and associations between AIIDs and HLA alleles across seven HLA genes. Our comorbidity analyses showed that overall AIID and five specific AIIDs were associated with having a psychiatric diagnosis. Our genetic analyses found 81 significant associations between HLA alleles and AIIDs. Lastly, we show connections across AIIDs, psychiatric disorders and infection susceptibility through network analysis of significant HLA associations in these disease classes. Combined, our results include both novel associations as well as replications of previously reported associations in a large sample, and highlight the genetic and epidemiological links between AIIDs and psychiatric disorders.

Cervical Cancer Diagnosis Using an Integrated System of Principal Component Analysis, Genetic Algorithm, and Multilayer Perceptron.

Cervical cancer is one of the most dangerous diseases that affect women worldwide. The diagnosis of cervical cancer is challenging, costly, and time-consuming. Existing literature has focused on traditional machine learning techniques and deep learning to identify and predict cervical cancer. This research proposes an integrated system of Genetic Algorithm (GA), Multilayer Perceptron (MLP), and Principal Component Analysis (PCA) that accurately predicts cervical cancer. GA is used to optimize the MLP hyperparameters, and the MLPs act as simulators within the GA to provide the prediction accuracy of the solutions. The proposed method uses PCA to transform the available factors; the transformed features are subsequently used as inputs to the MLP for model training. To contrast with the PCA method, different subsets of the original factors are selected. The performance of the integrated system of PCA–GA–MLP is compared with nine different classification algorithms. The results indicate that the proposed method outperforms the studied classification algorithms. The PCA–GA–MLP model achieves the best accuracy in diagnosing Hinselmann, Biopsy, and Cytology when compared to existing approaches in the literature that were implemented on the same dataset. This study introduces a robust tool that allows medical teams to predict cervical cancer in its early stage.

Establishment of a genetic transformation system for Codonopsis pilosula callus.

Codonopsis pilosula, a traditional Chinese medicinal and edible plant, contains several bioactive components. However, the biosynthetic mechanism is unclear because of the difficulties associated with functional gene analysis. Therefore, it is important to establish an efficient genetic transformation system for gene function analysis. In this study, we established a highly efficient Agrobacterium-mediated callus genetic transformation system for C. pilosula using stems as explants. After being pre-cultured for 3 days, the explants were infected with Agrobacterium tumefaciens strain GV3101 harboring pCAMBIA1381-35S::GUS at an OD600 value of 0.3 for 15 min, followed by co-cultivation on MS induction medium for 1 day and delayed cultivation on medium supplemented with 250 mg l-1 cefotaxime sodium for 12 days. The transformed calli were selected on screening medium supplemented with 250 mg l-1 cefotaxime sodium and 2.0 mg l-1 hygromycin and further confirmed by PCR amplification of the GUS gene and histochemical GUS assay. Based on the optimal protocol, the induction and transformation efficiency of calli reached a maximum of 91.07%. The establishment of a genetic transformation system for C. pilosula calli lays the foundation for the functional analysis of genes related to bioactive components through genetic engineering technology.

Lesch-Nyhan Syndrome with Self-harming and Choreoathetosis after Improvement of Hyperuricemia: a case report

Lesch-Nyhan syndrome (LNS) is an inborn error of metabolism. Choreoathetosis, mental retardation and self-harming are the three symptoms. A genetic test that analyses the gene encoding hypoxanthine-guanine phosphoribosyltransferase (HPRT) and blood tests showing hyperuricaemia support the diagnosis. However, the relevance of these test results to clinical symptoms is unclear. A 25 years-old man consulted our hospital for self-injury and choreoathetosis. He had epilepsy and mental retardation. He was also suffering from hyperuricemia and was prescribed allopurinol. These symptoms appeared with the improvement of hyperuricemia. He was suspected to have LNS. Metabolomic analysis of his urine before and after the allopurinol loading test was performed by gas chromatography-mass spectrometry (GC/MS) using dry filter paper urine samples as samples. The results showed that he was LNS.This case suggests that LNS is a condition that causes epileptic seizures and that rapid fluctuations in uric acid levels can cause choreoathetosis and self-harming. GC/MS using dry filter paper urine samples was a simple and valid test. This report also shows that genetic diagnosis opportunities are available to patients who have difficulty attending hospitals with advanced genetic analysis systems in urban areas.

Genetic relationships between populations of Aegilops tauschii Coss. (Poaceae) using SCoT molecular markers

The genus Aegilops has an important potential utilization in wheat improvement because of its resistance to different biotic and abiotic stresses and close relation with the cultivated wheat. Aegilops tauschii grows in Iran, westward to Turkey and eastward to Afghanistan and China with a distribution center in the south of Caspian Sea. In spite of its very good biochemical characterization, the knowledge about the DNA variability is very limited and no DNA markers were used to analyses the genomic variability of the populations, up to date. In the present study, genetic diversity of 117 Aegilops tauschii, individuals nine populations were studied using 10 Start Codon Targeted (SCoT) markers. High polymorphic bands (96.33%), polymorphic information content (0.48) and allele number (1.024) showed SCoT as a reliable marker system for genetic analysis in Aegilops tauschii. At the species, the percentage of polymorphic loci [P] was 66.30%, Nei’s gene diversity [H] was 0.35, Shannon index [I] was 0.33 and unbiased gene diversity [UHe] was 0.37. Genetic variation within populations (59%) was higher than among populations (41%) based on analysis of molecular variance (AMOVA). We used SCoT molecular marker for our genetic investigation with the following aims: 1— Investigate genetic diversity both among and with date Aegilops tauschii, 2—Identify genetic groups within these nine populations Aegilops tauschii, and 3—produce data on the genetic structure of date Aegilops tauschii populations. The results obtained revealed a high within-population genetic variability.

A Complex of Pyrosequencing-Based Methods for Detection of Somatic Mutations in Codons 600 and 601 of the BRAF gene.

The nucleotide sequence of the BRAF codons 592-602 was identified using the PyroMark Q24 genetic analysis system. The mutations search in codon 600 was conducted using the 600-S primer in line with the following order of adding nucleotides: GCTGTCАTCTGCTAGCTAGAC (corresponding to nucleotides 1799-1786). The K601E mutation was detected using the 601-S primer in line with the following order of nucleotide addition: GCTACTCACTGTAG (corresponding to nucleotides 1801-1793). The analytical characteristics of the proposed methods for somatic mutations' detection were determined using dilutions of plasmid DNA samples containing the BRAF gene region without mutations or with one of the following mutations: V600E, V600R, V600K, V600M, and K601E. Validation was performed on 132 samples of biological material obtained from the thyroid nodules. The developed methods allow to determine 2% of the V600E or V600M mutations, 1% of the V600K and V600R mutations, and 3% of the K601E mutations in samples with high DNA concentration; it is also possible to confidently detect at least 5% of the mutant allele for all mutations in low concentration samples (less than 500 copies/PCR). During biological material testing, 53 samples with the V600E mutation were detected; the proportion of the mutant allele was 4.9-50.0%. A complex of methods for determination of the nucleotide sequence of the BRAF codons 592-601 and the algorithm for testing samples and analyzing mutations in the BRAF codons 600-601 was developed. The method provides sufficient sensitivity to detect frequent mutations in codons 600 and 601 and allows them to be precisely differentiated.

Comparative study and genetic diversity of Salicornia persica (Chenopodiaceae) using SCOT molecular markers

Salicornia is a cosmopolitan genus having a high complicate taxonomy even described as a nightmare. Morphologically members of Salicornia are halophyte, annual, articulated, and succulent herbs with cortical palisade, opposite decussate scale-leaves, thyrsoid cymes, flowers packed in cauline depressions and the diaspore composed of l-seeded utricle. Salicornia is a priceless edible vegetable, considered as a favourable resource for cultivation in arid and semiarid coastal zones. Concerning the molecular data, although various studies were published, the situation is still quite obscure, at least for some taxa. Also, no DNA markers were used to analyses the genomic variability of the populations, up to date. In the present study, the genetic diversity of 102 individuals of S. persica (15 populations) were studied using 10 Start Codon Targeted (SCoT) markers. High polymorphic bands (94.18%), polymorphic information content (0.27), and allele number (1.38) showed SCoT as a reliable marker system for genetic analysis of this species. We used SCoT molecular marker for our genetic investigation with the following aims: 1) investigate genetic diversity both among and with date S. persica, 2) identify genetic groups within the 15 populations studied, and 3) produce data on the genetic structure of S. persica populations. The results obtained revealed a high within-population genetic variability.

A bacterial three-hybrid assay for forward and reverse genetic analysis of RNA-protein interactions.

This protocol describes a bacterial three-hybrid (B3H) assay, an in vivo system that reports on RNA-protein interactions and can be implemented in both forward and reverse genetic experiments. The B3H assay connects the strength of an RNA-protein interaction inside of living Escherichia coli cells to the transcription of a reporter gene (here, lacZ). We present protocols to (1) insert RNA and protein sequences into appropriate vectors for B3H experiments, (2) detect putative RNA-protein interactions with both qualitative and quantitative readouts and (3) carry out forward genetic mutagenesis screens. The B3H assay builds on a well-established bacterial two-hybrid system for genetic analyses. As a result, protein-protein interactions can be assessed in tandem with RNA interactions with a bacterial two-hybrid assay to ensure that protein variants maintain their functionality. The B3H system is a powerful complement to traditional biochemical methods for dissecting RNA-protein interaction mechanisms: RNAs and proteins of interest do not need to be purified, and their interactions can be assessed under native conditions inside of a living bacterial cell. Once cloning has been completed, an assay can be completed in under a week and a screen in 1-2 weeks.

Avian haemosporidian parasites of accipitriform raptors

BackgroundThe order Accipitriformes comprises the largest group of birds of prey with 260 species in four families. So far, 21 haemosporidian parasite species have been described from or reported to occur in accipitriform birds. Only five of these parasite species have been characterized molecular genetically. The first part of this study involved molecular genetic screening of accipitriform raptors from Austria and Bosnia-Herzegovina and the first chromogenic in situ hybridization approach targeting parasites in this host group. The aim of the second part of this study was to summarize the CytB sequence data of haemosporidian parasites from accipitriform raptors and to visualize the geographic and host distribution of the lineages.MethodsBlood and tissue samples of 183 accipitriform raptors from Austria and Bosnia-Herzegovina were screened for Plasmodium, Haemoproteus and Leucocytozoon parasites by nested PCR, and tissue samples of 23 PCR-positive birds were subjected to chromogenic in situ hybridization using genus-specific probes targeting the parasites’ 18S rRNAs. All published CytB sequence data from accipitriform raptors were analysed, phylogenetic trees were calculated, and DNA haplotype network analyses were performed with sequences from clades featuring multiple lineages detected in this host group.ResultsOf the 183 raptors from Austria and Bosnia-Herzegovina screened by PCR and sequencing, 80 individuals (44%) were infected with haemosporidian parasites. Among the 39 CytB lineages detected, 18 were found for the first time in the present study. The chromogenic in situ hybridization revealed exo-erythrocytic tissue stages of Leucocytozoon parasites belonging to the Leucocytozoon toddi species group in the kidneys of 14 infected birds. The total number of CytB lineages recorded in accipitriform birds worldwide was 57 for Leucocytozoon, 25 for Plasmodium, and 21 for Haemoproteus.ConclusionThe analysis of the DNA haplotype networks allowed identifying numerous distinct groups of lineages, which have not yet been linked to morphospecies, and many of them likely belong to yet undescribed parasite species. Tissue stages of Leucocytozoon parasites developing in accipitriform raptors were discovered and described. The majority of Leucocytozoon and Haemoproteus lineages are specific to this host group, but most Plasmodium lineages were found in birds of other orders. This might indicate local transmission from birds kept at the same facilities (raptor rescue centres and zoos), likely resulting in abortive infections. To clarify the taxonomic and systematic problems, combined morphological and molecular genetic analyses on a wider range of accipitriform host species are needed.

The Lamellipodin homologue MIG-10 is not essential for dorsal intercalation in the embryonic epidermis of the C. elegans embryo.

Dorsal intercalation of the embryonic epidermis in the Caenorhabditis elegans embryo is a promising system for genetic analysis of convergent extension, a conserved process in animal embryos. We sought to identify functionally important actin regulators in dorsal epidermal cells. A promising candidate is MIG-10, the single MIG-10/RIAM/Lamellipodin (MRL) family member in C. elegans. We endogenously tagged all mig-10 isoforms with mNeonGreen and analyzed mig-10 mutants using 4-dimensional microscopy. MIG-10::mNG is expressed prominently in muscle progenitors but is not detectable in the dorsal epidermis. mig-10(ct41) homozygotes complete dorsal intercalation in a manner indistinguishable from wildtype, indicating MIG-10 is not essential during dorsal intercalation.

Acoustic and Genetic Data Can Reduce Uncertainty Regarding Populations of Migratory Tree-Roosting Bats Impacted by Wind Energy.

Simple SummaryAlthough wind energy provides valuable environmental and economic benefits, it has unintended negative consequences for wildlife, particularly birds and bats that collide with the rotating blades. In North America, for example, there are increasing concerns that wind turbines threaten the persistence of populations of migratory tree-roosting bats, such as the hoary bat (Lasiurus cinereus). Because it is not possible to monitor population sizes for these solitary and cryptic species using traditional techniques, we must rely on other methods, such as acoustic and genetic monitoring, to provide information regarding population status and trends. Using these approaches, we can build a weight of evidence to assess whether mortality associated with wind turbines is sustainable and determine if and when mitigation measures should be implemented to reduce wind turbine mortality. To this end, we recommend that acoustic data be collected using the North American Bat Monitoring Program (NABat) protocols, and that genetic diversity be monitored at repeated time intervals to document species trends. There are no short-term measures to resolve population-level questions for migratory tree-roosting bats. Nonetheless, we discuss opportunities for relatively short-term investments that will lead to long-term success in reducing uncertainty for these species.Wind turbine-related mortality may pose a population-level threat for migratory tree-roosting bats, such as the hoary bat (Lasiurus cinereus) in North America. These species are dispersed within their range, making it impractical to estimate census populations size using traditional survey methods. Nonetheless, understanding population size and trends is essential for evaluating and mitigating risk from wind turbine mortality. Using various sampling techniques, including systematic acoustic sampling and genetic analyses, we argue that building a weight of evidence regarding bat population status and trends is possible to (1) assess the sustainability of mortality associated with wind turbines; (2) determine the level of mitigation required; and (3) evaluate the effectiveness of mitigation measures to ensure population viability for these species. Long-term, systematic data collection remains the most viable option for reducing uncertainty regarding population trends for migratory tree-roosting bats. We recommend collecting acoustic data using the statistically robust North American Bat Monitoring Program (NABat) protocols and that genetic diversity is monitored at repeated time intervals to show species trends. There are no short-term actions to resolve these population-level questions; however, we discuss opportunities for relatively short-term investments that will lead to long-term success in reducing uncertainty.