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1.
Dev Genes Evol ; 230(1): 39-45, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31960123

RESUMO

The molluscan larval shell formation is a complicated process. There is evidence that the mantle of the primary larva (trochophore) contains functionally different cell populations with distinct gene expression profiles. However, it remains unclear how these cells are specified. In the present study, we identified three cell populations from the shell gland in earlier stages (gastrula) from the bivalve mollusc Crassostrea gigas. These cell populations were determined by analyzing the co-expression relationships among six potential shell formation (pSF) genes using two-color hybridization. The three cell populations, which we designated as SGCPs (shell gland cell populations), formed a concentric-circle pattern from outside to inside of the shell gland. SGCP I was located in the outer edge of the shell gland and the cells expressed pax2/5/8, gata2/3, and bmp2/4. SGCP II was located more internally and the cells expressed two engrailed genes. The last population, SGCP III, was located in the central region of the shell gland and the cells expressed lox4. Determination of the gene expression profiles of SGCPs would help trace their origins and fates and elucidate how these cell populations are specified. Moreover, potential roles of the SGCPs, e.g., development of sensory cells and shell biogenesis, are suggested. Our results reveal the internal organization of the embryonic shell gland at the molecular level and add to the knowledge of larval shell formation.


Assuntos
Crassostrea/citologia , Exoesqueleto/citologia , Exoesqueleto/metabolismo , Animais , Crassostrea/genética , Crassostrea/crescimento & desenvolvimento , Crassostrea/metabolismo , Glândulas Exócrinas/citologia , Glândulas Exócrinas/metabolismo , Feminino , Masculino , Fatores de Transcrição/metabolismo
2.
J Chem Ecol ; 45(11-12): 901-913, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31773376

RESUMO

Trail pheromones deposited by ants lead nestmates to food sources. Based on previous evidence that the trail pheromone of the carpenter ant Camponotus modoc originates from the hindgut, our objective in this study was to identify the key component(s) of the pheromone. We collected C. modoc colonies from conifer forests and maintained them in an outdoor enclosure near our laboratory for chemical analyses and behavioral experiments. In gas chromatographic-electroantennographic detection and gas chromatography-mass spectrometric analyses of worker ant hindgut extracts, we identified five candidate components: 2,4-dimethylhexanoic acid, 2,4-dimethyl-5-hexanolide, pentadecane, dodecanoic acid and 3,4-dihydro-8-hydroxy-3,5,7-trimethylisocoumarin. In a series of trail-following experiments, ants followed trails of synthetic 2,4-dimethyl-5-hexanolide, a blend of the five compounds, and hindgut extract over similar distances, indicating that the hexanolide accounted for the entire behavioral activity of the hindgut extract. The hexanolide not only mediated orientation of C. modoc foragers on trails, it also attracted them over distance, indicating a dual function. Further analyses and bioassays with racemic and stereoselectively synthesized hexanolides revealed that the ants produce, and respond to, the (2S,4R,5S)-stereoisomer. The same stereoisomer is a trail pheromone component in several Camponotus congeners, indicating significant overlap in their respective trail pheromone communication systems.


Assuntos
Misturas Complexas/análise , Feromônios/análise , Alcanos/análise , Animais , Formigas , Comportamento Animal , Técnicas Biossensoriais/métodos , Caproatos/análise , Cumarínicos/análise , Glândulas Exócrinas/metabolismo , Cromatografia Gasosa-Espectrometria de Massas/métodos , Intestinos/química , Ácidos Láuricos/análise , Estereoisomerismo
3.
Endocrinology ; 160(11): 2587-2599, 2019 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-31373638

RESUMO

The prolactin receptor (Prlr) mediates not only the multiple effects of prolactin, but also those of the placental lactogens and, in humans, some actions of growth hormone. Although Prlr expression has been reported to be widespread in the body, specific cellular expression patterns within tissues are undefined for many organs. One persisting problem in investigating Prlr function is that the protein is difficult to detect using conventional methods. To allow investigation of Prlr expression with a single cell resolution, we have recently developed a knock-in mouse strain in which Cre recombinase is expressed together with the long isoform of the Prlr using an internal ribosome entry site. When crossed to a Cre-dependent reporter mouse strain, Cre-mediated recombination will genetically label cells that acutely express the Prlr as well as cells that have transiently expressed the Prlr during development. We report here the anatomical distribution of cells which express the fluorescent reporter τ green fluorescent protein in a total of 38 organs prepared from young adult male and female Prlr reporter mice. Our results establish a resource for dissecting the functional role of Prlr in multiple murine tissues.


Assuntos
Receptores da Prolactina/metabolismo , Animais , Glândulas Endócrinas/metabolismo , Glândulas Exócrinas/metabolismo , Feminino , Trato Gastrointestinal/metabolismo , Sistema Linfático/metabolismo , Masculino , Camundongos , Sistema Respiratório/metabolismo , Sistema Urogenital/metabolismo
4.
J Chem Ecol ; 45(8): 673-683, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31407198

RESUMO

Animals modulate intraspecific signal shape and intensity, notably during reproductive periods. Signal variability typically follows a seasonal scheme, traceable through the expression of visual, acoustic, chemical and behavioral patterns. The chemical channel is particularly important in lizards, as demonstrated by well-developed epidermal glands in the cloacal region that secrete lipids and proteins recognized by conspecifics. In males, the seasonal pattern of gland activity is underpinned by variation of circulating androgens. Changes in the composition of lipid secretions convey information about the signaler's quality (e.g., size, immunity). Presumably, individual identity is associated with a protein signature present in the femoral secretions, but this has been poorly investigated. For the first time, we assessed the seasonal variability of the protein signal in relation to plasma testosterone level (T), glandular activity and the concentration of provitamin D3 in the lipid fraction. We sampled 174 male common wall lizards (Podarcis muralis) over the entire activity season. An elevation of T was observed one to two months before the secretion peak of lipids during the mating season; such expected delay between hormonal fluctuation and maximal physiological response fits well with the assumption that provitamin D3 indicates individual quality. One-dimensional electrophoretic analysis of proteins showed that gel bands were preserved over the season with an invariant region; a result in agreement with the hypothesis that proteins are stable identity signals. However, the relative intensity of bands varied markedly, synchronously with that of lipid secretion pattern. These variations of protein secretion suggest additional roles of proteins, an issue that requires further studies.


Assuntos
Glândulas Exócrinas/metabolismo , Lipídeos/análise , Lagartos/fisiologia , Proteínas de Répteis/análise , Animais , Desidrocolesteróis/análise , Eletroforese em Gel de Campo Pulsado , Cromatografia Gasosa-Espectrometria de Massas , Metabolismo dos Lipídeos , Lipídeos/química , Masculino , Análise de Componente Principal , Estações do Ano , Comportamento Sexual Animal , Testosterona/sangue
5.
Artigo em Inglês | MEDLINE | ID: mdl-31276814

RESUMO

The aim of the study was to compare cellular effects of xenobiotic cadmium and biogenic copper in ampullate silk glands of false black widow Steatoda grossa spider after long-term exposure via ingestion under laboratory conditions. Both the level of selected detoxification parameters (glutathione S-transferase, catalase, and the level of total antioxidant capacity) and degree of genotoxic changes (comet assay) were determined in the silk glands. Additionally the contents of selected amino acids (L-Ala, L-Pro, L-His, L-Phe, DL-Ile, and DL-Asn) in the hunting webs produced by spiders of this species were assessed. The ability of S. grossa females to accumulate cadmium was higher than that for copper. Long-term exposure of spiders to copper did not change the level of detoxification parameters, and the level of DNA damage in the cells of ampullate silk glands was also low. Cadmium had a stronger prooxidative and genotoxic effect than copper in the cells of the analyzed silk glands. However, regardless of the type of metal used, no significant changes in the level of amino acids in silk were found. The obtained results confirmed the effectiveness of metal neutralization mechanisms in the body of the studied spider species, which results in the protection of the function of ampullate silk glands.


Assuntos
Cádmio/toxicidade , Cobre/toxicidade , Glândulas Exócrinas/efeitos dos fármacos , Aranhas/efeitos dos fármacos , Animais , Cádmio/metabolismo , Cobre/metabolismo , Dano ao DNA , Glândulas Exócrinas/metabolismo , Feminino , Inativação Metabólica , Aranhas/genética , Aranhas/metabolismo
6.
J Chem Ecol ; 45(7): 588-597, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31342233

RESUMO

Honey bee (Apis mellifera) queens produce pheromones responsible for mediating both male mating behavior and many critical facets of worker social organization within their colony. These pheromones are dynamic multi-component blends, allowing the communication of detailed information. Indeed, variation in the queen's mating and reproductive state is associated with significant changes in her pheromone profiles, and these different pheromone profiles elicit different behavioral and physiological responses in female workers. Here we evaluate behavioral responses of male drones to the chemical blends produced by two exocrine glands in queens, and determine if the blends and responses are altered by the queen's mating and reproductive state. We find that drone attraction to the chemical blends of mandibular glands produced by mated, laying queens versus virgin queens is reduced, suggesting that the queens produce a reliable signal of their mating receptivity. Interestingly, while the chemical blends of mating, laying queens and virgins queens largely overlap, mated, laying queens produce a greater number of chemicals and greater quantities of certain chemicals than virgin queens, suggesting that these chemicals may serve to inhibit behavioral responses of drones to mated, laying queens. Thus, our results highlight the importance of considering chemical cues and signals that serve to both stimulate and inhibit behavioral responses during social interactions in animals.


Assuntos
Comportamento Animal/efeitos dos fármacos , Feromônios/farmacologia , Comunicação Animal , Animais , Abelhas , Glândulas Exócrinas/química , Glândulas Exócrinas/metabolismo , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Masculino , Feromônios/análise , Reprodução/fisiologia
7.
PLoS One ; 14(7): e0220187, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31329654

RESUMO

The pink bollworm, Pectinophora gossypiella, is a world-wide pest of cotton and in some parts of the cotton growing region is controlled by the mating disruption technique using synthetic sex pheromone. The sex pheromone consists of two compounds, (Z,Z)- and (Z,E)-7,11-hexadecadienyl acetates, in about a 50:50 ratio. However, recently, a population with sex pheromone compound ratios of about 62:38 were found in cotton fields that use mating disruption in Israel. To investigate how the change developed, we compared the pheromone gland transcriptomes between a reference laboratory population and a population obtained from an Israeli cotton field utilizing mating disruption. We analyzed four biological replicates from each population and found transcripts encoding 17 desaturases, 8 reductases, and 17 candidate acetyltransferases in both populations, which could be involved in sex pheromone biosynthesis. The expression abundance of some genes between the two populations was different. Some desaturases and candidate acetyltransferases were found to have mutated in one of the populations. The differentially expressed genes play potential roles in sex pheromone biosynthesis and could be involved in causing altered female sex pheromone ratios in the field population.


Assuntos
Glândulas Exócrinas/metabolismo , Lepidópteros/genética , Atrativos Sexuais/metabolismo , Transcriptoma , Animais , Feminino , Especiação Genética , Masculino
8.
Mol Biol Evol ; 36(9): 1921-1930, 2019 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-31238339

RESUMO

Chemical signaling in animals often plays a central role in eliciting a variety of responses during reproductive interactions between males and females. One of the best-known vertebrate courtship pheromone systems is sodefrin precursor-like factors (SPFs), a family of two-domain three-finger proteins with a female-receptivity enhancing function, currently only known from salamanders. The oldest divergence between active components in a single salamander species dates back to the Late Paleozoic, indicating that these proteins potentially gained a pheromone function earlier in amphibian evolution. Here, we combined whole transcriptome sequencing, proteomics, histology, and molecular phylogenetics in a comparative approach to investigate SPF occurrence in male breeding glands across the evolutionary tree of anurans (frogs and toads). Our study shows that multiple families of both terrestrially and aquatically reproducing frogs have substantially increased expression levels of SPFs in male breeding glands. This suggests that multiple anuran lineages make use of SPFs to complement acoustic and visual sexual signaling during courtship. Comparative analyses show that anurans independently recruited these proteins each time the gland location on the male's body allowed efficient transmission of the secretion to the female's nares.


Assuntos
Anuros/metabolismo , Atrativos Sexuais/metabolismo , Animais , Anuros/genética , Glândulas Exócrinas/metabolismo , Feminino , Masculino , Oligopeptídeos/metabolismo , Filogenia , Caracteres Sexuais , Sequenciamento Completo do Exoma
9.
Int J Mol Sci ; 20(10)2019 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-31137550

RESUMO

The silk gland synthesizes and secretes a large amount of protein and stores liquid silk protein at an extremely high concentration. Interestingly, silk proteins and serine protease inhibitors are orderly arranged in the silk gland lumen and cocoon shells. Silk fiber formation and the spinning mechanism have not been fully elucidated. Therefore, we conducted a comparative transcriptome analysis of seven segments of the single silk gland to characterize internal changes in the silk gland during the 5th instar of mature larvae. In total, 3121 differentially expressed genes were identified in the seven segments. Genes highly expressed in the middle silk gland (MSG) were mainly involved in unsaturated fatty acid biosynthesis, fatty acid metabolism, apoptosis-fly, and lysosome pathways, whereas genes highly expressed in the posterior silk gland (PSG) were mainly involved in ribosome, proteasome, citrate cycle, and glycolysis/gluconeogenesis pathways. Thus, the MSG and PSG differ greatly in energy source use and function. Further, 773 gradually upregulated genes (from PSG to MSG) were involved in energy metabolism, silk protein synthesis, and secretion, suggesting that these genes play an important role in silk fiber formation. Our findings provide insights into the mechanism of silk protein synthesis and transport and silk fiber formation.


Assuntos
Bombyx/genética , Glândulas Exócrinas/metabolismo , Seda/genética , Transcriptoma , Animais , Seda/biossíntese
10.
J Insect Physiol ; 116: 1-9, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30953617

RESUMO

The hypopharyngeal glands (HGs) of honey bee nurse workers secrete the major protein fraction of jelly, a protein and lipid rich substance fed to developing larvae, other worker bees, and queens. A hallmark of poorly nourished nurses is their small HGs, which actively degrade due to hormone-induced autophagy. To better connect nutritional stress with HG degradation, we looked to honey bees and other insect systems, where nutrient stress is often accompanied by fat body degradation. The fat body contains stored lipids that are likely a substrate for ecdysteroid synthesis, so we tested whether starvation caused increased fat body lipolysis. Ecdysteroid signaling and response pathways and IIS/TOR are tied to nutrient-dependent autophagy in honey bees and other insects, and so we also tested whether and where genes in these pathways were differentially regulated in the head and fat body. Last, we injected nurse-aged bees with the honey bee ecdysteroid makisterone A to determine whether this hormone influenced HG size and autophagy. We find that starved nurse aged bees exhibited increased fat body lipolysis and increased expression of ecdysteroid production and response genes in the head. Genes in the IIS/TOR pathway were not impacted by starvation in either the head or fat body. Additionally, bees injected with makisterone A had smaller HGs and increased expression of autophagy genes. These data support the hypothesis that nutritional stress induces fat body lipolysis, which may liberate the sterols important for ecdysteroid production, and that increased ecdysteroid levels induce autophagic HG degradation.


Assuntos
Abelhas/fisiologia , Ecdisteroides/metabolismo , Corpo Adiposo/fisiopatologia , Hormônios de Inseto/metabolismo , Lipólise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Abelhas/genética , Ecdisteroides/biossíntese , Glândulas Exócrinas/crescimento & desenvolvimento , Glândulas Exócrinas/metabolismo , Feminino , Hipofaringe/crescimento & desenvolvimento , Hipofaringe/metabolismo , Hormônios de Inseto/biossíntese , Transcrição Genética
11.
G3 (Bethesda) ; 9(5): 1339-1353, 2019 05 07.
Artigo em Inglês | MEDLINE | ID: mdl-30885922

RESUMO

Regulation of luminal diameter is critical to the function of small single-celled tubes, of which the seamless tubular excretory canals of Caenorhabditis elegans provide a tractable genetic model. Mutations in several sets of genes exhibit the Exc phenotype, in which canal luminal growth is visibly altered. Here, a focused reverse genomic screen of genes highly expressed in the canals found 18 genes that significantly affect luminal outgrowth or diameter. These genes encode novel proteins as well as highly conserved proteins involved in processes including gene expression, cytoskeletal regulation, and vesicular and transmembrane transport. In addition, two genes act as suppressors on a pathway of conserved genes whose products mediate vesicle movement from early to recycling endosomes. The results provide new tools for understanding the integration of cytoplasmic structure and physiology in forming and maintaining the narrow diameter of single-cell tubules.


Assuntos
Proteínas de Caenorhabditis elegans/genética , Caenorhabditis elegans/genética , Caenorhabditis elegans/metabolismo , Glândulas Exócrinas/metabolismo , Animais , Transporte Biológico , Caenorhabditis elegans/ultraestrutura , Proteínas de Caenorhabditis elegans/metabolismo , Glândulas Exócrinas/ultraestrutura , Técnicas de Silenciamento de Genes , Estudos de Associação Genética , Genótipo , Mutação , Fenótipo , Interferência de RNA
12.
Stem Cell Reports ; 12(2): 366-380, 2019 02 12.
Artigo em Inglês | MEDLINE | ID: mdl-30713042

RESUMO

Understanding how epithelial progenitors within exocrine glands establish specific cell lineages and form complex functional secretory units is vital for organ regeneration. Here we identify the transcription factor Sox10 as essential for both the maintenance and differentiation of epithelial KIT+FGFR2b+ progenitors into secretory units, containing acinar, myoepithelial, and intercalated duct cells. The KIT/FGFR2b-Sox10 axis marks the earliest multi-potent and tissue-specific progenitors of exocrine glands. Genetic deletion of epithelial Sox10 leads to loss of secretory units, which reduces organ size and function, but the ductal tree is retained. Intriguingly, the remaining duct progenitors do not compensate for loss of Sox10 and lack plasticity to properly form secretory units. However, overexpression of Sox10 in these ductal progenitors enhances their plasticity toward KIT+ progenitors and induces differentiation into secretory units. Therefore, Sox10 controls plasticity and multi-potency of epithelial KIT+ cells in secretory organs, such as mammary, lacrimal, and salivary glands.


Assuntos
Plasticidade Celular/fisiologia , Células Epiteliais/metabolismo , Glândulas Exócrinas/metabolismo , Fatores de Transcrição SOXE/metabolismo , Animais , Diferenciação Celular/fisiologia , Linhagem da Célula/fisiologia , Deleção de Genes , Masculino , Camundongos , Organogênese/fisiologia , Receptor Tipo 2 de Fator de Crescimento de Fibroblastos/metabolismo , Glândulas Salivares/metabolismo
13.
Poult Sci ; 98(5): 2260-2271, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30624718

RESUMO

The blue-shelled egg not only plays a key role in helping birds to avoid predation as a result of crypsis and mimetism, but it also provides eggshell strength and filters solar radiation; moreover, it has an important economic trait for poultry. However, the source of biliverdin for blue-shelled egg remains unsolved in ducks. The current study detected the biliverdin content and localization of heme oxygenase 1 (HMOX1) in duck shell gland; moreover, RNA-seq analysis was performed in the shell gland of blue-shelled and white-shelled ducks. Results indicated that biliverdin is a primary pigment for blue-shelled egg in ducks, and the HMOX1 protein showed high expression in ciliated epithelial cells of shell gland between blue-shelled and white-shelled ducks. In the pathway of biliverdin synthesis, only 5-aminolevulinate synthase 1 expression level was significantly upregulated in blue-shelled ducks, and nuclear factor, erythroid 2 like 1 and period circadian clock 2 may be the essential elements in biliverdin synthesis of duck shell gland. Furthermore, some of the transporter genes, such as activator-Like and solute carrier family 13 member 5, may be involved in the formation of blue egg in duck. Results of the current study suggested that the biliverdin is most likely synthesized and secreted from epithelial cells of shell gland. In addition, ALAS1 may play a key role in the formation of blue egg in ducks.


Assuntos
Biliverdina/genética , Patos/genética , Oviductos/metabolismo , Pigmentação/genética , Transcriptoma , Animais , Biliverdina/metabolismo , Cor , Patos/metabolismo , Casca de Ovo/fisiologia , Glândulas Exócrinas/metabolismo , Feminino
14.
Molecules ; 24(2)2019 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-30669405

RESUMO

Amphibian skin is not to be considered a mere tegument; it has a multitude of functions related to respiration, osmoregulation, and thermoregulation, thus allowing the individuals to survive and thrive in the terrestrial environment. Moreover, amphibian skin secretions are enriched with several peptides, which defend the skin from environmental and pathogenic insults and exert many other biological effects. In this work, the beneficial effects of amphibian skin peptides are reviewed, in particular their role in speeding up wound healing and in protection from oxidative stress and UV irradiation. A better understanding of why some species seem to resist several environmental insults can help to limit the ongoing amphibian decline through the development of appropriate strategies, particularly against pathologies such as viral and fungal infections.


Assuntos
Anfíbios/metabolismo , Peptídeos/farmacologia , Substâncias Protetoras/farmacologia , Pele/efeitos dos fármacos , Cicatrização/efeitos dos fármacos , Sequência de Aminoácidos , Animais , Antioxidantes/química , Antioxidantes/farmacologia , Glândulas Exócrinas/metabolismo , Depuradores de Radicais Livres , Humanos , Peptídeos/química , Substâncias Protetoras/química , Pele/metabolismo , Raios Ultravioleta/efeitos adversos
15.
Genomics ; 111(3): 231-241, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30458272

RESUMO

Scorpionfishes (Scorpaenidae) are a relatively common cause of human envenomation. They often enter coastal waters and their stings can be quite hazardous, provoking extreme pain and causing the victims to take days to recover. There are few genomic resources available for the scorpionfishes. In this study, we elucidated the transcriptomic profile of the venom glands from three different scorpionfish species, namely Scorpaenopsis cirrosa, S. neglecta and S. possi. This is the first report of scorpionfish transcriptomes. After functional and pathway annotation, we employed toxin annotation to identify many species-specific (18, 13 and 19 respectively) and overlapping putative toxins among the three species. Our study represents a significant improvement in the genetic information about the venoms from these three species. Moreover, this work also provides an archive for future studies on evolution of fish toxins and can be used for comparative studies of other fishes.


Assuntos
Venenos de Peixe/genética , Peixes/genética , Animais , Glândulas Exócrinas/metabolismo , Venenos de Peixe/química , Peixes/classificação , Filogenia , Alinhamento de Sequência , Transcriptoma
16.
Mar Genomics ; 43: 1-8, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30293672

RESUMO

Tolerance of organisms towards heterogeneous and variable environments is highly related to physiological flexibility. An effective strategy to enhance physiological flexibility is the expression of polymorphic enzymes. This seems to be the case in the brown shrimp Crangon crangon. It shows high reproduction rates, feeds opportunistically on endo- and epibenthic organisms, and is apparently well adapted to variable environmental conditions. Previous electrophoretic studies revealed a high level of polymorphism and no consistent phenotype of digestive enzymes between individuals. In order to understand the underlying biochemical processes, we carried out a transcriptome-based study of digestive enzymes of C. crangon. Detailed sequence analyses of triacylglycerol lipase, phospholipase A2, alpha amylase, chitinase, trypsin and cathepsin L were performed to identify putative isoforms. The number of isoforms, and thus the degree of polymorphism varied among enzymes: lipases and carbohydrases showed higher numbers of isoforms in enzymes that besides their extracellular function also have diverse intracellular functions. Furthermore, cysteine proteinases showed a lower polymorphism than serine proteinases. We suggest that the expression of enzyme isoforms improves the efficiency of C. crangon in gaining energy from different food sources.


Assuntos
Proteínas de Artrópodes/genética , Crangonidae/genética , Polimorfismo Genético , Animais , Proteínas de Artrópodes/análise , Proteínas de Artrópodes/química , Crangonidae/enzimologia , Glândulas Exócrinas/metabolismo , Trato Gastrointestinal/enzimologia , Perfilação da Expressão Gênica , Análise de Sequência de Proteína
17.
G3 (Bethesda) ; 8(11): 3593-3605, 2018 11 06.
Artigo em Inglês | MEDLINE | ID: mdl-30213867

RESUMO

Targeting gene function with spatial or temporal specificity is a key goal in molecular genetics. CRISPR-Cas9 has greatly facilitated this strategy, but some standard approaches are problematic. For instance, simple tissue-specific or global overexpression of Cas9 can cause significant lethality or developmental delays even in the absence of gRNAs. In particular, we found that Gal4-mediated expression of UAS-Cas9 in the Drosophila prothoracic gland (PG) was not a suitable strategy to disrupt gene expression, since Cas9 alone caused widespread lethality. The PG is widely used for studying endocrine gland function during animal development, but tools validating PG-specific RNAi phenotypes are lacking. Here, we present a collection of modular gateway-compatible CRISPR-Cas9 tools that allow precise modulation of target gene activity with temporal and spatial specificity. We also demonstrate that Cas9 fused to the progesterone ligand-binding domain can be used to activate gene expression via RU486. Using these approaches, we were able to avoid the lethality associated with simple GAL4-mediated overexpression of Cas9 in the PG. Given that the PG is a polytene tissue, we conclude that these tools work effectively in endoreplicating cells where Cas9 has to target multiple copies of the same locus. Our toolkit can be easily adapted for other tissues and can be used both for gain- and loss-of-function studies.


Assuntos
Drosophila/genética , Glândulas Exócrinas/metabolismo , Animais , Sistemas CRISPR-Cas , Sistema Enzimático do Citocromo P-450/genética , Proteínas de Drosophila/genética , Regulação da Expressão Gênica , Oxigenases de Função Mista/genética
18.
Front Immunol ; 9: 1757, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30108587

RESUMO

Collectin liver 1 (CL-L1, alias collectin 10) and collectin kidney 1 (CL-K1, alias collectin 11) are oligomeric pattern recognition molecules associated with the complement system, and mutations in either of their genes may lead to deficiency and developmental defects. The two collectins are reportedly localized and synthesized in the liver, kidneys, and adrenals, and can be found in the circulation as heteromeric complexes (CL-LK), which upon binding to microbial high mannose-like glycoconjugates activates the complement system via the lectin activation pathway. The tissue distribution of homo- vs. heteromeric CL-L1 and -K1 complexes, the mechanism of heteromeric complex formation and in which tissues this occurs, is hitherto incompletely described. We have by immunohistochemistry using monoclonal antibodies addressed the precise cellular localization of the two collectins in the main human tissues. We find that the two collectins have widespread and almost identical tissue distribution with a high expression in epithelial cells in endo-/exocrine secretory tissues and mucosa. There is also accordance between localization of mRNA transcripts and detection of proteins, showing that local synthesis likely is responsible for peripheral localization and eventual formation of the CL-LK complexes. The functional implications of the high expression in endo-/exocrine secretory tissue and mucosa is unknown but might be associated with the activity of MASP-3, which has a similar pattern of expression and is known to potentiate the activity of the alternative complement activation pathway.


Assuntos
Colectinas/genética , Epitélio/metabolismo , Membrana Mucosa/metabolismo , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/metabolismo , Anticorpos Monoclonais/farmacologia , Colectinas/imunologia , Colectinas/metabolismo , Ativação do Complemento/efeitos dos fármacos , Ativação do Complemento/imunologia , Glândulas Endócrinas/metabolismo , Células Epiteliais/metabolismo , Glândulas Exócrinas/metabolismo , Perfilação da Expressão Gênica/métodos , Humanos , Rim/metabolismo , Fígado/metabolismo , Ligação Proteica
19.
Biol Bull ; 235(1): 1-11, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-30160995

RESUMO

High fecundity often contributes to successful invasives. In molluscs, this may be facilitated by the albumen gland-capsule gland complex, which in gastropods secretes the egg perivitelline fluid that nourishes and protects embryos. The biochemistry of the albumen gland-capsule gland complex and its relationship with fecundity remain largely unknown. We addressed these issues in Pomacea canaliculata (Lamarck, 1822), a highly invasive gastropod whose fecundity and reproductive effort exceed those of ecologically similar gastropods. We evaluated the dynamics of its major secretion compounds (calcium, polysaccharides, and total proteins) as well as the gene expression and stored levels of perivitellins during key moments of the reproductive cycle, that is, before and after first copulation and at low, medium, and high reproductive output. Copulation and first oviposition do not trigger the onset of albumen gland-capsule gland complex biosynthesis. On the contrary, soon after an intermediate reproductive effort, genes encoding perivitellins overexpressed. A high reproductive effort caused a decrease in all albumen gland-capsule gland complex secretion components. Right after a high reproductive output, the albumen gland-capsule gland complex restored the main secretion components, and calcium recovered baseline reserves; but proteins and polysaccharides did not. These metabolic changes in the albumen gland-capsule gland complex after multiple ovipositions were reflected in a reduction in egg mass but did not compromise egg quality. At the end of the cycle, egg dry weight almost doubled the initial albumen gland-capsule gland complex weight. Results indicate that albumen gland-capsule gland complex biosynthesis limits a constantly high reproductive output. Therefore, lowering fecundity by targeting biosynthesis could effectively reduce the rate of this species' spread.


Assuntos
Espécies Introduzidas , Caramujos/fisiologia , Animais , Glândulas Exócrinas/metabolismo , Reprodução/fisiologia
20.
J Exp Biol ; 221(Pt 14)2018 07 30.
Artigo em Inglês | MEDLINE | ID: mdl-29880633

RESUMO

Amphibian skin is unique among vertebrate classes, containing a large number of multicellular exocrine glands that vary among species and have diverse functions. The secretions of skin glands contain a rich array of bioactive compounds including antimicrobial peptides (AMPs). Such compounds are important for amphibian innate immune responses and may protect some species from chytridiomycosis, a lethal skin disease caused by the fungal pathogens Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans (Bsal). While the bioactivity of skin secretions against Bd has been assessed for many amphibian taxa, similar studies are lacking for Bsal, a chytrid fungus that is especially pathogenic for salamanders. We studied the skin glands and their potential functions in an aquatic salamander, the three-toed amphiuma (Amphiuma tridactylum). Skin secretions of captive adult salamanders were analyzed by RP-HPLC and tested against the growth of Bd and Bsal using in vitro assays. We found that compounds within collected skin secretions were similar between male and female salamanders and inhibited the growth of Bd and Bsal. Thus, skin secretions that protect against Bd may also provide protection against Bsal. Histological examination of the skin glands of preserved salamanders revealed the presence of enlarged granular glands concentrated within caudal body regions. A site of potential gland specialization was identified at the tail base and may indicate specialized granular glands related to courtship and communication.


Assuntos
Anti-Infecciosos/metabolismo , Quitridiomicetos/efeitos dos fármacos , Glândulas Exócrinas/metabolismo , Micoses/veterinária , Pele , Urodelos , Animais , Anti-Infecciosos/química , Anti-Infecciosos/farmacologia , Quitridiomicetos/fisiologia , Feminino , Masculino , Micoses/microbiologia , Urodelos/fisiologia
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