2-deoxyglucose transiently inhibits yeast AMPK signaling and triggers glucose transporter endocytosis, potentiating the drug toxicity.

2-deoxyglucose is a glucose analog that impacts many aspects of cellular physiology. After its uptake and its phosphorylation into 2-deoxyglucose-6-phosphate (2DG6P), it interferes with several metabolic pathways including glycolysis and protein N-glycosylation. Despite this systemic effect, resistance can arise through strategies that are only partially understood. In yeast, 2DG resistance is often associated with mutations causing increased activity of the yeast 5'-AMP activated protein kinase (AMPK), Snf1. Here we focus on the contribution of a Snf1 substrate in 2DG resistance, namely the alpha-arrestin Rod1 involved in nutrient transporter endocytosis. We report that 2DG triggers the endocytosis of many plasma membrane proteins, mostly in a Rod1-dependent manner. Rod1 participates in 2DG-induced endocytosis because 2DG, following its phosphorylation by hexokinase Hxk2, triggers changes in Rod1 post-translational modifications and promotes its function in endocytosis. Mechanistically, this is explained by a transient, 2DG-induced inactivation of Snf1/AMPK by protein phosphatase 1 (PP1). We show that 2DG-induced endocytosis is detrimental to cells, and the lack of Rod1 counteracts this process by stabilizing glucose transporters at the plasma membrane. This facilitates glucose uptake, which may help override the metabolic blockade caused by 2DG, and 2DG export-thus terminating the process of 2DG detoxification. Altogether, these results shed a new light on the regulation of AMPK signaling in yeast and highlight a remarkable strategy to bypass 2DG toxicity involving glucose transporter regulation.

Character variation in separate body regions of Gerreidae (Osteichthyes: Teleostei) fishes inferred from geometric morphometrics.

Body morphology is a valuable feature for distinguishing teleostean fishes. However, the utility of character variation in separate body regions has yet to be tested. The taxonomy of the Gerreidae family is controversial due to character overlapping among its fish species. This work aims to analyze and compare the body shape variation in three regions, cephalic, trunk, and caudal peduncle, using landmark data and geometric morphometric methods in 17 species and five genera of the family Gerreidae. The pattern of shape variation for the cephalic region consisted of well-defined character states exclusive of each species analyzed. Shape variation in the trunk and caudal peduncle regions does not distinguish all species in this study. This study showed that the dorsal cephalic profile is highly variable among the species, therefore, shape variation in this region is useful for distinguishing Gerreidae species. In contrast, some species within the same genus share similar shape states in the trunk and caudal peduncle regions, with the most shape variation in the dorsal profile and anal fin for the trunk and in the middle of the caudal peduncle.

Alike but genetically divergent: The resurrection of Urotrygon asterias (Jordan & Gilbert, 1883) from its closest relatives, the Munda and the Blotched stingray.

The genus Urotrygon comprises small- to medium-sized endemic round rays on the American continent and has undergone several synonymization processes. Here, we used an integrative taxonomic approach, including meristic, morphometric, and mtDNA analyses, to resolve the particularly intricate relationship among Urotrygon munda Gill, 1863, Urotrygon chilensis (Günther, 1872), and Urotrygon asterias (Jordan & Gilbert, 1883). The latter species is currently a synonym of U. munda but is also considered the U. chilensis "northern morphotype." These taxonomic entities have historically been confounded, mainly due to their phenotypical resemblance along their geographic distribution in the eastern Pacific. We assessed 78 specimens (43 "northern" and 30 "southern morphotypes" of U. chilensis, as well as 5 U. munda) using 19 external variables for taxonomic and morphometric analysis. Distinct meristic patterns, including pectoral and pelvic ceratotrichia, vertebrae number, and thorn distribution along the dorsal midline, were observed in the series-type specimens of the three taxonomic entities. Our multivariate morphometric analyses consistently differentiated the three groups as distinct taxonomic entities, with an overall classification accuracy of 66.7%. The meristic results also provided reliable information distinguishing the three entities. Based on the nicotinamide adenine dinucleotide (NADH2) and cytochrome oxidase subunit I (COI) genes, our phylogenetic analysis were consistent with the morphometric and meristic data, supporting these three entities as distinct species having their own evolutionary lineages. Our comprehensive approach confidently demonstrated that the northern U. chilensis morphotype matched and corresponded to the description of the Starry round ray, U. asterias, confirming its taxonomic resurrection as a valid species distinct from U. chilensis and U. munda. The geographic distribution of U. asterias spans from the tropical west coast of Mexico (including the Gulf of California) to Costa Rica, revealing that microevolutionary processes have well-defined population clades within this range. Furthermore, U. chilensis is unequivocally established as the sole Urotrygon species occurring south of the Guayaquil marine ecoregion. In addition, the public COI and NADH2 sequences available for Urotrygon hosted in the ad hoc online databases were found to be misidentified, emphasizing the need for rigorous taxonomic scrutiny in this group. Finally, our research underscores the significance of an integrative approach that combines morphometric, meristic, and molecular techniques with historical data to disentangle the complexities of closely related taxa.

Morphological abnormalities in seven American round ray specimens: A review of America's batomorph anomalies.

Although morphological abnormalities in several rays and skate species around the American continents have frequently mentioned, their numbers are unknown. The present work record morphological abnormalities in four Urotrygonidae species. Two anophthalmic specimens were detected (Urotrygon microphthalmum and Urobatis halleri). Two individuals lacked caudal fins (Urobatis maculatus and Urotrygon chilensis). Two round rays showed incomplete fusion of the pectoral fin to the head (U. microphthalmum and U. chilensis). Vertebral compression and fusion were found in a 6-year-old female Urotrygon rogersi. In addition, 118 abnormal batomorph specimens were gathered from the available bibliography, spanning the last six decades (1959-2021). Amblyraja doellojuradoi was the species with the highest number of abnormalities (18). The most common anomaly was an incomplete fusion of the pectoral fin with the head. Since 2010, at least 30 anomalous batomorphs have been recorded every 5 years. Sixty-nine abnormal specimens occurred in the Northern Hemisphere (1.00-60.00 N). The Cortezian (Pacific) and Southeastern Brazil (Atlantic) marine ecoregions stood out with the highest number of these specimens. Mexico recorded 58 anomalous specimens, followed by Brazil (n = 36). Biological, abiotic and anthropogenic factors are probably the leading causes. However, additional studies are necessary to elucidate these speculations.

New microsatellite loci for estimating genetic diversity and structure in Octopus hubbsorum from Nayarit, México.

BACKGROUND: Octopus hubbsorum Berry, 1953 is the most important species for commercial fishing in the Mexican Pacific. However, there is a lack of information regarding population structure that could have important management implications. We tested 44 microsatellite loci in O. hubbsorum by cross-amplification from O. bimaculatus. METHODS AND RESULTS: Genetic diversity and structure was tested over 30 octopus sampled from Santa Cruz de Miramar (Nayarit, México). A total of 11 loci were successfully amplified. All loci were polymorphic with the number of effective alleles ranging from 2.13 to 23.14, while three loci significantly deviated from Hardy-Weinberg equilibrium. No significant LD was observed between pairs of loci (P ≥ 0.05). The application of the new markers in a O. hubbsorum population from Santa Cruz de Miramar Nayarit, México, did not showed Wahlund or isolate breaking effects due to the mixing of distinct populations. CONCLUSIONS: The loci were useful to estimate levels of pairwise relatedness and to discard the presence of recent demographic bottlenecks in the population. We consider that eight microsatellites are adequate from the 11 amplified loci.

Cytoplasmic LSM-1 protein regulates stress responses through the insulin/IGF-1 signaling pathway in Caenorhabditis elegans.

Genes coding for members of the Sm-like (LSm) protein family are conserved through evolution from prokaryotes to humans. These proteins have been described as forming homo- or heterocomplexes implicated in a broad range of RNA-related functions. To date, the nuclear LSm2-8 and the cytoplasmic LSm1-7 heteroheptamers are the best characterized complexes in eukaryotes. Through a comprehensive functional study of the LSm family members, we found that lsm-1 and lsm-3 are not essential for C. elegans viability, but their perturbation, by RNAi or mutations, produces defects in development, reproduction, and motility. We further investigated the function of lsm-1, which encodes the distinctive protein of the cytoplasmic complex. RNA-seq analysis of lsm-1 mutants suggests that they have impaired Insulin/IGF-1 signaling (IIS), which is conserved in metazoans and involved in the response to various types of stress through the action of the FOXO transcription factor DAF-16. Further analysis using a DAF-16::GFP reporter indicated that heat stress-induced translocation of DAF-16 to the nuclei is dependent on lsm-1. Consistent with this, we observed that lsm-1 mutants display heightened sensitivity to thermal stress and starvation, while overexpression of lsm-1 has the opposite effect. We also observed that under stress, cytoplasmic LSm proteins aggregate into granules in an LSM-1-dependent manner. Moreover, we found that lsm-1 and lsm-3 are required for other processes regulated by the IIS pathway, such as aging and pathogen resistance.

Legionella and mitochondria, an intriguing relationship.

Legionella pneumophila is the causative agent of Legionnaires' disease, a severe pneumonia. L. pneumophila injects via a type-IV-secretion-system (T4SS) more than 300 bacterial proteins into macrophages, its main host cell in humans. Certain of these bacterial effectors target organelles in the infected cell and hijack multiple processes to facilitate all steps of the intracellular life cycle of this pathogen. In this review, we discuss the interplay between L. pneumophila, an intracellular bacterium fully armed with virulence tools, and mitochondria, the extraordinary eukaryotic organelles playing prominent roles in cellular bioenergetics, cell-autonomous immunity and cell death. We present and discuss key findings concerning the multiple interactions of L. pneumophila with mitochondria during infection and the mechanisms employed by T4SS effectors that target mitochondrial functions to subvert infected cells.

High-content assay to measure mitochondrial function and bacterial vacuole size in infected human primary macrophages.

Regulation of bioenergetics and cell death are pivotal mitochondrial functions determining the responses of macrophages to infection. Here, we provide a protocol to investigate mitochondrial functions during infection of macrophages by intracellular bacteria. We describe steps for quantifying mitochondrial polarization, cell death, and bacterial infection in infected, living, human primary macrophages at the single-cell level. We also detail the use of the pathogen Legionella pneumophila as model. This protocol can be adapted to investigate mitochondrial functions in other settings. For complete details on the use and execution of this protocol, please refer to Escoll et al. (2021).1.

Geographical variation in the genetic diversity and composition of the endangered queen conch Strombus gigas (Mesogastropoda: Strombidae) from Yucatán, México.

In Mexico and elsewhere in the Caribbean, the queen conch Strombus gigas is an endangered species. Understanding the genetic connectivity of their populations will support management strategies for long-term conservation of the species. Genetic diversity and population differentiation was assessed from samples collected at Banco Chinchorro and Isla Cozumel in the Mexican Caribbean and at Arrecife Alacranes in the Gulf of Mexico. Samples were obtained from the commercial capture at Banco Chinchorro (n = 50) and Isla Cozumel (n = 40) on March 2004. On November 2004, a non-invasive method for the Arrecife Alacranes sampling was applied, taking the hemolymph of live animals (n = 65) and releasing them to the wild. The mitochondrial DNA variation at two genes (COI and Cyt-b) was analyzed. Genetic diversity at the three locations ranged between 0.55-0.65 in COI and 0.87-0.94 in Cyt-b, showing no bottleneck evidences. A non-significant fixation index (F(ST) = 0.019, p = 0.161) and a Maximum Parsimony Network tree that did not show particular clades associated with any of the geographical locations, suggested a lack of statistically significant genetic differentiation among populations. Nevertheless, the cline patterns observed in both genetic diversity and haplotypic frequencies from Banco Chinchorro through Arrecife Alacranes, and the larger genetic distance between these locations from those between Isla Cozumel, Banco Chinchorro and Arrecife Alacranes, suggest the possibility of a pattern of isolation-by distance. The role of the main current systems over the potential genetic differences in S. gigas populations along the Mexican Caribbean, and the conservation management of S. gigas at these locations as discrete units is discussed.

Revision of the diagnostic characters of two morphologically similar snook species, Centropomus viridis and C. nigrescens (Carangiformes: Centropomidae).

Historically, the taxonomic identification of the two snook species, Centropomus viridis and C. nigrescens, has been challenging due to their morphological similarity and the inconsistency of the characters used for diagnosis. Therefore, this study aimed to evaluate the usefulness of the morphologic, meristic, and morphometric characters currently being used to identify C. viridis and C. nigrescens, based on molecular data. The results showed that the gas-bladder shape (i.e., C. viridis with diverticula and C. nigrescens without diverticula) was the only morphological character univocally related to genetic identification. Likewise, geometric morphometrics separated two groups; each corresponds to only one of two genetically (and gas bladder shape) identified species. Of all the meristic characters examined, only the second dorsal fin ray count (nine for C. viridis and ten for C. nigrescens) was related to the gas bladder shape and genetic identity; therefore, it is the only external character with a diagnostic utility to separate each species.

Genetic homogeneity of the Pacific thread herring (Opisthonema libertate) (Günther, 1867) in the Eastern Pacific, inferred from mtDNA sequences.

In the present study, the population genetic structure of the Pacific thread herring (Opisthonema libertate) was analyzed through mitochondrial DNA (mtDNA) control region sequences. Organisms were collected from June 2015 to July 2015 from four commercial landing sites (Bahia Magdalena, Guaymas and Mazatlan, Mexico, and Puntarenas, Costa Rica) and one artisanal sampling site (Puerto La Libertad, El Salvador). A total of 125 sequences were analyzed. High levels of haplotype (h = 0.990) and nucleotide (π = 0.030) diversity were found. Pairwise Φst comparisons indicated differences attributed mainly to the organisms from El Salvador. However, Bayesian inferences did not support the existence of different populations. The haplotype distribution between locations did not show a clear phylogeographic pattern. Mismatched distribution showed a unimodal pattern for the five sampled areas, indicative of sudden demographic expansion. These results were supported by Bayesian skyline plot. Our results do not support the hypothesis that the Pacific thread herring presents a population genetic structure. Future genetic comparisons should include a larger number of samples as well as more polymorphic molecular markers to further support our results.

Taxonomic assessment of species of the genus Octopus from the northeastern Pacific via morphological, molecular and morphometric analyses.

Species of the genus Octopus from the northeastern Pacific are ecologically and economically important; however, their taxonomy is confusing and has not been comprehensively assessed. In this study, we performed a taxonomic evaluation of these species considering the morphological characteristics of the original descriptions, a molecular analysis of partial COI-gene sequences, and a traditional morphometry analysis of nine body measurements. Several interesting findings were obtained with our results: for instance, we updated the diagnoses of some species by including characters such as the number of lamellae per demibranch and the presence of chromatophores in the visceral sac; we deposited partial COI-gene sequences of species that had not been incorporated into the GenBank repository; and according to the morphometric analysis, we confirmed that the lengths of arms I-IV are relevant to discriminate the species under study. The taxa evaluated were morphologically, molecularly and morphometrically well-delimited; however, features such as funnel organ shape and arm length proportions in regard to dorsal mantle length are either not included in the diagnosis of the genus Octopus or overlap with other genera. Hence, this information, combined with the results obtained from the molecular analysis, supports the generic re-assignation of two of the species evaluated.

Genetic and cellular sensitivity of Caenorhabditis elegans to the chemotherapeutic agent cisplatin.

Cisplatin and derivatives are commonly used as chemotherapeutic agents. Although the cytotoxic action of cisplatin on cancer cells is very efficient, clinical oncologists need to deal with two major difficulties, namely the onset of resistance to the drug and the cytotoxic effect in patients. Here, we used Caenorhabditis elegans to investigate factors influencing the response to cisplatin in multicellular organisms. In this hermaphroditic model organism, we observed that sperm failure is a major cause of cisplatin-induced infertility. RNA sequencing data indicate that cisplatin triggers a systemic stress response, in which DAF-16/FOXO and SKN-1/NRF2, two conserved transcription factors, are key regulators. We determined that inhibition of the DNA damage-induced apoptotic pathway does not confer cisplatin protection to the animal. However, mutants for the pro-apoptotic BH3-only gene ced-13 are sensitive to cisplatin, suggesting a protective role of the intrinsic apoptotic pathway. Finally, we demonstrated that our system can also be used to identify mutations providing resistance to cisplatin and therefore potential biomarkers of innate cisplatin-refractory patients. We show that mutants for the redox regulator trxr-1, ortholog of the mammalian thioredoxin reductase 1 TRXR1, display cisplatin resistance. By CRISPR/Cas9, we determined that such resistance relies on the presence of the single selenocysteine residue in TRXR-1.This article has an associated First Person interview with the first author of the paper.

The complete mitochondrial genome of Octopus bimaculatus Verrill, 1883 from the Gulf of California.

The complete mitochondrial genome of Octopus bimaculatus is 16 085 bp in length and includes 13 protein-codes genes, 2 ribosomal RNA genes, 22 transfers RNA genes, and a control region. The composition of genome is A (40.9%), T (34.7%), C (16.9%), and G (7.5%). The control region of O. bimaculatus contains a VNTR locus not present in the genomes from other octopus species. A phylogenetic analysis shows a closer relationship between the mitogenomes from O. bimaculatus and O. vulgaris.

Geographical variation in the genetic diversity and composition of the endangered Queen Conch Strombus gigas (Mesogastropoda: Strombidae) from Yucatán, México

In Mexico and elsewhere in the Caribbean, the queen conch Strombus gigas is an endangered species. Understanding the genetic connectivity of their populations will support management strategies for long term conservation of the species. Genetic diversity and population differentiation was assessed from samples collected at Banco Chinchorro and Isla Cozumel in the Mexican Caribbean and at Arrecife Alacranes in the Gulf of Mexico. Samples were obtained from the commercial capture at Banco Chinchorro (n=50) and Isla Cozumel (n=40) on March 2004. On November 2004, a non-invasive method for the Arrecife Alacranes sampling was applied, taking the hemolymph of live animals (n=65) and releasing them to the wild. The mitochondrial DNA variation at two genes (COI and Cyt-b) was analyzed. Genetic diversity at the three locations ranged between 0.55-0.65 in COI and 0.87-0.94 in Cyt-b, showing no bottleneck evidences. A non-significant fixation index (F ST=0.019, p=0.161) and a Maximum Parsimony Network tree that did not show particular clades associated with any of the geographical locations, suggested a lack of statistically significant genetic differentiation among populations. Nevertheless, the cline patterns observed in both genetic diversity and haplotypic frequencies from Banco Chinchorro through Arrecife Alacranes, and the larger genetic distance between these locations from those between Isla Cozumel, Banco Chinchorro and Arrecife Alacranes, suggest the possibility of a pattern of isolation-by distance. The role of the main current systems over the potential genetic differences in S. gigas populations along the Mexican Caribbean, and the conservation management of S. gigas at these locations as discrete units is discussed. Rev. Biol. Trop. 59 (3): 1115-1126. Epub 2011 September 01.

El caracol rosado Strombus gigas es una especie amenzada en México y otros sitios del Caribe. Su conservación a largo plazo requiere la comprensión de la conectividad entre sus poblaciones. En este estudio se evaluó la diversidad y diferenciación genética de muestras recolectadas en tres sitios del Caribe y Golfo de México adyacentes a la Península de Yucatán. Las muestras se obtuvieron de la captura comercial en Banco Chinchorro (n=50) e Isla Cozumel (n=40) en marzo de 2004. En noviembre de 2004 se obtuvieron muestras de Arrecife Alacranes (n=65) de animales vivos, mediante un método no invasivo diseñado para la obtención de hemolinfa; los organismos muestreados se liberaron de vuelta al medio natural. Se analizó la diversidad genética de dos genes del ADN mitocondrial (COI y Cyt-b). La diversidad genética en las tres localidades varió entre 0.55 - 0.65 en COI y 0.87 - 0.94 en Cyt-b no indicando reducción por cuello de botella. Un índice de fijación no significativamente diferente de cero (F ST=0.019, p=0.161) y un árbol en Red de Máxima Parsimonia que no mostró clados particulares asociados con localidades específicas, sugiere que no hay diferencias genéticas significativas entre sitios. Sin embargo, los patrones clinales observados en la diversidad genética y en las frecuencias haplotípicas, así como la mayor distancia genética registrada entre las localidades más alejadas (Banco Chinchorro y Arrecife Alacranes) sugiere un posible un patrón de aislamiento por distancia. Se discute el papel de los sistemas de corrientes principales del Caribe mexicano sobre la posible diferenciación genética de S. gigas. Asimismo, se discute el manejo de las localidades estudiadas como unidades discretas.