Vertebrate Tissues Research Articles

Extraembryonic tissues: exploring concepts, definitions and functions across the animal kingdom.

Extraembryonic tissues: exploring concepts, definitions and functions across the animal kingdom.

New Chemical Dualities Illustrated by Meso and C2 Symmetrical (CTS) Compounds

The most important relationship between chemistry and genetics is nonetheless the corpuscular nature of their objects, molecules (or atoms) and genes, respectively. On the other hand, one states, without a substantial proof, that philosophy should be one step ahead all sciences. Here is a proof that the reverse can also be true. Two internal enantiomeric halves of meso compounds or the two chiral halves of ­C2 symmetrical isomers constitute pairs of entities suitable to work as duality phenomena in science. Four types of isomers have been identified: (A) meso, (B) C2 symmetrical (CTS), (C) irregular chiral (irrechi) and (D) constitutional (constit.). Meso and CTS are characteristic to plants and microorganisms. Almost all natural micromolecular compounds from vertebrate tissues are asymmetric, i.e. they are constit. isomers. An exception to this rule is meso-inositol, an isomer of hexoses, which are themselves, as their congeners asymmetric. By comparing the real (envisaged) meso isomers of these compounds with the asymmetric ones of vertebrate tissues, the reason for nature selected the latter became quite evident: it is the omission of a suite of structural restrictions. Delivering of meso isomers of natural compounds discloses a huge chemical philosophical potential of this issue. An intrinsic property of meso combinations is their character of dimerism, hence their molecule is formed of two entities that are contrary in a spatial, chemical and optical sense, i.e. good candidates for a duality concept. Moreover, a good deal of material is indicated, i.e. CTS isomers, whose sides are chiral and identical, for a new type of duality in philosophy, strongly expressed in nature by a chemical language.

The ecology of modern and fossil vertebrates revisited by lithium isotopes

The vertebrate fossil record documents a plethora of transitions between aquatic and terrestrial environments but their causes are still debated. Quantifying the salinity of living environments is therefore crucial for precising the sequence of ecological transitions. Here, we measured lithium stable isotope composition of mineralized tissues (δ7Limt) of extant and extinct vertebrates from various aquatic environments: seawater, freshwater/terrestrial, and “transitional environments” (i.e. brackish waters, or seasonal access to freshwater and seawater). We report statistically higher δ7Limt values for seawater vertebrates than freshwater ones, taxonomic groups considered separately. Moreover, vertebrates living in transitional environments have intermediate δ7Limt values. Therefore, we show that δ7Limt values of both extant and extinct vertebrates can discriminate their aquatic habitat.

Defining domain-specific orientational order in the desmosomal cadherins

Desmosomes are large, macromolecular protein assemblies that mechanically couple the intermediate filament cytoskeleton to sites of cadherin-mediated cell adhesion, thereby providing structural integrity to tissues that routinely experience large forces. Proper desmosomal adhesion is necessary for the normal development and maintenance of vertebrate tissues, such as epithelia and cardiac muscle, while dysfunction can lead to severe disease of the heart and skin. Therefore, it is important to understand the relationship between desmosomal adhesion and the architecture of the molecules that form the adhesive interface, the desmosomal cadherins (DCs). However, desmosomes are embedded in two plasma membranes and are linked to the cytoskeletal networks of two cells, imposing extreme difficulty on traditional structural studies of DC architecture, which have yielded conflicting results. Consequently, the relationship between DC architecture and adhesive function remains unclear. To overcome these challenges, we utilized excitation-resolved fluorescence polarization microscopy to quantify the orientational order of the extracellular and intracellular domains of three DC isoforms: desmoglein 2, desmocollin 2, and desmoglein 3. We found that DC ectodomains were significantly more ordered than their cytoplasmic counterparts, indicating a drastic difference in DC architecture between opposing sides of the plasma membrane. This difference was conserved among all DCs tested, suggesting that it may be an important feature of desmosomal architecture. Moreover, our findings suggest that the organization of DC ectodomains is predominantly the result of extracellular adhesive interactions. We employed azimuthal orientation mapping to show that DC ectodomains are arranged with rotational symmetry about the membrane normal. Finally, we performed a series of mathematical simulations to test the feasibility of a recently proposed antiparallel arrangement of DC ectodomains, finding that it is supported by our experimental data. Importantly, the strategies employed here have the potential to elucidate molecular mechanisms for diseases that result from defective desmosome architecture.

Shikonin Inhibits Fin Regeneration in Zebrafish Larvae.

Shikonin is a naphthoquinone compound extracted from Chinese comfrey for treating cancer. However, there are few reports on its research on vertebrate tissue regeneration. Zebrafish is an ideal model for studying organ regeneration. In this study, we found that 3-dpf of zebrafish larvae exposed to shikonin at concentrations of 0.2, 0.3, and 0.4 mg/L showed increasingly inhibited regeneration of the tail fin. Immunohistochemical staining showed that shikonin exposure from 6 to 12 hpa increased the number of apoptotic cells in the caudal fin wound of larvae and decreased the number of proliferating cells. Shikonin exposure was found to up-regulate oxidative stress, increase ROS levels, and reduce neutrophil recruitment in the early stage of wound repair. Moreover, shikonin exposure caused disordered expression of fin regeneration blastemal-related genes. The use of astaxanthin to down-regulate oxidative stress was found to significantly reduce the inhibition of caudal fin regeneration. Mixed exposure of AMPK inhibitors or fullerenes (C60) with shikonin also showed the similar rescue effect. Collectively, our study showed that shikonin inhibited fin regeneration in zebrafish larvae by the upregulation of oxidative stress level and AMPK signaling pathway. This research provides valuable information on the mechanism of action of shikonin for its safe application.

In situ cell-type-specific cell-surface proteomic profiling in mice

Cell-surface proteins (CSPs) mediate intercellular communication throughout the lives of multicellular organisms. However, there are no generalizable methods for quantitative CSP profiling in specific cell types in vertebrate tissues. Here, we present in situ cell-surface proteome extraction by extracellular labeling (iPEEL), a proximity labeling method in mice that enables spatiotemporally precise labeling of cell-surface proteomes in a cell-type-specific environment in native tissues for discovery proteomics. Applying iPEEL to developing and mature cerebellar Purkinje cells revealed differential enrichment in CSPs with post-translational protein processing and synaptic functions in the developing and mature cell-surface proteomes, respectively. A proteome-instructed invivo loss-of-function screen identified a critical, multifaceted role for Armh4 in Purkinje cell dendrite morphogenesis. Armh4 overexpression also disrupts dendrite morphogenesis; this effect requires its conserved cytoplasmic domain and is augmented by disrupting its endocytosis. Our results highlight the utility of CSP profiling in native mammalian tissues for identifying regulators of cell-surface signaling.

PSIII-A-4 Analysis of Galectin Gene Variants from Diverse Sheep and Goat Populations Genotyped with the Illumina Beadchip

Abstract Galectins are a highly conserved family of β-galactoside binding lectins encoded by lectin, galactosidebinding soluble genes (LGALS). They are found on a variety of cells and tissues of vertebrates and invertebrates. Understanding the roles of galectins in host–pathogen interaction and activation of host innate, and adaptive immunity has become an important research question. Galectins are known to play a role in parasitic infection in small ruminants. However, little is known about LGALS variants in sheep and goat breeds across the world. In this study, we investigated galectin single nucleotide polymorphism (SNP) variants on five distinct sheep breeds from different geographical regions of Ethiopia. A total of 88 sheep (Arsi-Bale=12, Horro=24, Menz=18, Wolayita=18 and Blackhead Somali=16) were genotyped with Ovine Infinium HD SNP BeadChip (600K). An In Silico approach with Ensembl database (https://useast.ensembl.org/index.html) and SNP BeadChip (perfect matched reference SNP along with their genomic position) identified a total of nineteen SNPs from LGALS-1, -2, -3, -4, -7, -8, -9, -11 and -12 with a consequence of upstream, downstream, and missense gene variant. For all these variants, a highly variable minor allele frequency (MAF) was observed across five sheep populations. We also investigated a total of 308 Chinese goat (Qinggeli=48, Nanjiang=48, Luoping=48, Liaoning=116, and Guangfeng=48) genotyped with Illumina Goat 50K BeadChip. The goat chip only contained a SNP from LGal-4 (rs268252010) which was also found at variable MAF level across the said populations. These results have implications for animal health and production and warrant further investigation using targeted sequencing of galectin genes across different local and international sheep and goat breeds.

Ephrin Micropatterns Exogenously Modulate Cell Organization in Organoid‐Derived Intestinal Epithelial Monolayers

AbstractEph‐ephrin signaling acts as spatial cue to define the tissue boundaries, the axonal growth, or the organization of compartmentalized tissues in vertebrates. By the regulation of tension, adhesion, and repulsion, intermingling of cells expressing the membrane‐tethered ligand and cells expressing the membrane‐tethered receptor is prevented. Despite being surface‐bound, most of the studies addressing Eph‐ephrin interactions use soluble ligands, which lack the spatial component needed to study tissue patterning. Here, it is shown that spatial patterns of ephrin ligands can modulate the organization of the different compartments in organoid‐derived intestinal epithelial monolayers. A modified version of the microcontact printing technique is used to create spatial cues of ephrin ligand on basement membrane surrogates. It is shown that both ligand concentration and cellular density can impact the strength of the repulsive effect triggered by Eph‐ephrin signaling. Finally, it is demonstrated that by using micropatterned ephrin spatial cues one can modify the orientation of intestinal crypts and align them in the direction set by the pattern. The approach presented here is believed to be an excellent tool to study exogenous signal with relevant spatial distribution in vivo.

Functions of Membrane Progesterone Receptors (mPRs, PAQRs) in Nonreproductive Tissues.

Gender differences in a wide variety of physiological parameters have implicated the ovarian hormones, estrogens and progesterone, in the regulation of numerous nonreproductive tissue functions. Rapid, nongenomic (nonclassical) progesterone actions mediated by membrane progesterone receptors (mPRs), which belong to the progestin and adipoQ receptor family, have been extensively investigated in reproductive and nonreproductive tissues since their discovery in fish ovaries 20 years ago. The 5 mPR subtypes (α, β, γ, δ, ε) are widely distributed in vertebrate tissues and are often expressed in the same cells as the nuclear progesterone receptor (PR) and progesterone receptor membrane component 1, thereby complicating investigations of mPR-specific functions. Nevertheless, mPR-mediated progesterone actions have been identified in a wide range of reproductive and nonreproductive tissues and distinguished from nuclear PR-mediated ones by knockdown of these receptors with siRNA in combination with a pharmacological approach using mPR- and PR-specific agonists. There are several recent reviews on the roles of the mPRs in vertebrate reproduction and cancer, but there have been no comprehensive assessments of mPR functions in nonreproductive tissues. Therefore, this article briefly reviews mPR functions in a broad range of nonreproductive tissues. The evidence that mPRs mediate progesterone and progestogen effects on neuroprotection, lordosis behavior, respiratory control of apnea, olfactory responses to pheromones, peripheral nerve regeneration, regulation of prolactin secretion in prolactinoma, immune functions, and protective functions in vascular endothelial and smooth muscle cells is critically reviewed. The ubiquitous expression of mPRs in vertebrate tissues suggests mPRs regulate many additional nonreproductive functions that remain to be identified.

Vertebrate extracellular matrix protein hemicentin-1 interacts physically and genetically with basement membrane protein nidogen-2

Hemicentins are large proteins of the extracellular matrix that belong to the fibulin family and play pivotal roles during development and homeostasis of a variety of invertebrate and vertebrate tissues. However, bona fide interaction partners of hemicentins have not been described as yet. Here, applying surface plasmon resonance spectroscopy and co-immunoprecipitation, we identify the basement membrane protein nidogen-2 (NID2) as a binding partner of mouse and zebrafish hemicentin-1 (HMCN1), in line with the formerly described essential role of mouse HMCN1 in basement membrane integrity. We show that HMCN1 binds to the same protein domain of NID2 (G3) as formerly shown for laminins, but with an approximately 3.5-fold lower affinity and in a competitive manner. Furthermore, immunofluorescence and immunogold labeling revealed that HMCN1/Hmcn1 is localized close to basement membranes and in partial overlap with NID2/Nid2a in different tissues of mouse and zebrafish. Genetic knockout and antisense-mediated knockdown studies in zebrafish further show that loss of Nid2a leads to similar defects in fin fold morphogenesis as the loss of Laminin-α5 (Lama5) or Hmcn1. Finally, combined partial loss-of-function studies indicated that nid2a genetically interacts with both hmcn1 and lama5. Together, these findings suggest that despite their mutually exclusive physical binding, hemicentins, nidogens, and laminins tightly cooperate and support each other during formation, maintenance, and function of basement membranes to confer tissue linkage.

Microplastics in Internal Tissues of Companion Animals from Urban Environments.

Simple SummaryMicroplastics are widespread anthropogenic contaminants, imposing a potential threat to organisms. A preliminary study was conducted to assess microplastics in postmortem samples of internal tissues of companion animals. Suspected microplastics were observed in the internal tissues of cats and dogs. Suspected microplastics were found in 35 out of 49 animals and 80 out of 242 samples. Particles sized 1–10 µm comprised 50.3% of the suspected microplastics. The number of particles found was very low and analytical methods must still be developed to improve the characterization and quantification of smaller-sized factions of microplastics. Moreover, this study suggests that microplastics may be internalized and distributed to the internal tissues of terrestrial vertebrates.Companion animals living in urban areas are exposed to environmental contaminants, which may include microplastics. A preliminary study was conducted by collecting postmortem samples from the internal tissue (lungs, ileum, liver, kidney, and blood clots) of 25 dogs (Canis familiaris) and 24 cats (Felis catus) living in an urban environment in Porto metropolitan area, Portugal. Suspected microplastics were found in 80 samples from 35 animals (18 cats and 17 dogs), often occurring in more than one tissue of the same animal (71.4%), primarily under small sizes (50.3% as 1–10 µm). Micro-Raman spectroscopy confirmed a fraction of particles as common polymer types (e.g., polyethylene terephthalate). However, the number of particles was very low. This study highlights the possibilities of the internalization and distribution of microplastics in the internal tissues of terrestrial vertebrates.

Responses of hematopoietic cells after ionizing-irradiation in anemic adult medaka (Oryzias latipes)

Purpose Hematopoietic tissues of vertebrates are highly radiation sensitive and the effects of ionizing radiation on the hematopoiesis have been studied in mammals and teleosts for decades. In this study, radiation responses in the kidney, the main hematopoietic organ in teleosts, were investigated in Japanese medaka (Oryzias latipes), which has been a model animal and a large body of knowledge has been accumulated in radiation biology. Methods Kidney, the main hematopoietic tissue of adult medaka fish, was locally irradiated using proton and carbon ion beams irradiation system of Takasaki Ion Accelerator for Advanced Radiation Application (TIARA), QST, and the effects on peripheral blood cells and histology of the kidney were investigated. Results When only kidneys were locally irradiated with proton or carbon ion beam (15 Gy), the hematopoietic cells in the irradiated kidney and cell density in the peripheral blood decreased 7 days after the irradiation in the same manner as after the whole-body irradiation with γ-rays (15 Gy). These results demonstrate that direct irradiation of the hematopoietic cells in the kidney induced cell death and/or cell cycle arrest and stopped the supply of erythroid cells. Then, the cell density in the peripheral blood recovered to the control level within 4 days and 7 days after the γ-ray and proton beam irradiation (15 Gy), respectively, while the cell density in the peripheral blood did not recover after the carbon ion beam irradiation (15 Gy). The hematopoietic cells in the irradiated kidneys temporarily decreased and recovered to the control level within 21 days after the γ-ray or proton beam irradiation (15 Gy), while it did not recover after the carbon ion beam irradiation (15 Gy). In contrast, the recovery of the cell density in the peripheral blood delayed when anemic medaka were irradiated 1 day after the administration of phenylhydrazine. With and without γ-ray irradiation, a large number of hematopoietic cells was still proliferating in the kidney 7 days after the anemia induction. Conclusions The results obtained strongly suggest that the hematopoietic stem cells in medaka kidney prioritize to proliferate and increase peripheral blood cells to eliminate anemia, even when they are damaged by high-dose irradiation.

Isolation and immortalization of macrophages derived from fetal porcine small intestine and their susceptibility to porcine viral pathogen infections.

Macrophages are a heterogeneous population of cells that are present in all vertebrate tissues. They play a key role in the innate immune system, and thus, in vitro cultures of macrophages provide a valuable model for exploring their tissue-specific functions and interactions with pathogens. Porcine macrophage cultures are often used for the identification and characterization of porcine viral pathogens. Recently, we have developed a simple and efficient method for isolating primary macrophages from the kidneys and livers of swine. Here, we applied this protocol to fetal porcine intestinal tissues and demonstrated that porcine intestinal macrophages (PIM) can be isolated from mixed primary cultures of porcine small intestine-derived cells. Since the proliferative capacity of primary PIM is limited, we attempted to immortalize them by transferring the SV40 large T antigen and porcine telomerase reverse transcriptase genes using lentiviral vectors. Consequently, immortalized PIM (IPIM) were successfully generated and confirmed to retain various features of primary PIM. We further revealed that IPIM are susceptible to infection by the African swine fever virus and the porcine reproductive and respiratory syndrome virus and support their replication. These findings suggest that the IPIM cell line is a useful tool for developing in vitro models that mimic the intestinal mucosal microenvironments of swine, and for studying the interactions between porcine pathogens and host immune cells.

Microvascular anatomy suggests varying aerobic activity levels in the adipose tissues of diving tetrapods.

Adipose tissue has many important functions including metabolic energy storage, endocrine functions, thermoregulation and structural support. Given these varied functions, the microvascular characteristics within the tissue will have important roles in determining rates/limits of exchange of nutrients, waste, gases and molecular signaling molecules between adipose tissue and blood. Studies on skeletal muscle have suggested that tissues with higher aerobic capacity contain higher microvascular density (MVD) with lower diffusion distances (DD) than less aerobically active tissues. However, little is known about MVD in adipose tissue of most vertebrates; therefore, we measured microvascular characteristics (MVD, DD, diameter and branching) and cell size to explore the comparative aerobic activity in the adipose tissue across diving tetrapods, a group of animals facing additional physiological and metabolic stresses associated with diving. Adipose tissues of 33 animals were examined, including seabirds, sea turtles, pinnipeds, baleen whales and toothed whales. MVD and DD varied significantly (P < 0.001) among the groups, with seabirds generally having high MVD, low DD and small adipocytes. These characteristics suggest that microvessel arrangement in short duration divers (seabirds) reflects rapid lipid turnover, compared to longer duration divers (beaked whales) which have relatively lower MVD and greater DD, perhaps reflecting the requirement for tissue with lower metabolic activity, minimizing energetic costs during diving. Across all groups, predictable scaling patterns in MVD and DD such as those observed in skeletal muscle did not emerge, likely reflecting the fact that unlike skeletal muscle, adipose tissue performs many different functions in marine organisms, often within the same tissue compartment.

Anthocyanins, Carotenoids and Chlorophylls in Edible Plant Leaves Unveiled by Tandem Mass Spectrometry.

Natural pigments are a quite relevant group of molecules that are widely distributed in nature, possessing a significant role in our daily lives. Besides their colors, natural pigments are currently recognized as having relevant biological properties associated with health benefits, such as anti-tumor, anti-atherogenicity, anti-aging and anti-inflammatory activities, among others. Some of these compounds are easily associated with specific fruits (such as blueberries with anthocyanins, red pitaya with betalain or tomato with lycopene), vegetables (carrots with carotenoids), plant leaves (chlorophylls in green leaves or carotenoids in yellow and red autumn leaves) and even the muscle tissue of vertebrates (such as myoglobin). Despite being less popular as natural pigment sources, edible plant leaves possess a high variety of chlorophylls, as well as a high variety of carotenoids and anthocyanins. The purpose of this review is to critically analyze the whole workflow employed to identify and quantify the most common natural pigments (anthocyanin, carotenoids and chlorophylls) in edible plant leaves using tandem mass spectrometry. Across the literature there, is a lack of consistency in the methods used to extract and analyze these compounds, and this review aims to surpass this issue. Additionally, mass spectrometry has stood out in the context of metabolomics, currently being a widely employed technique in this field. For the three pigments classes, the following steps will be scrutinized: (i) sample pre-preparation, including the solvents and extraction conditions; (ii) details of the chromatographic separation and mass spectrometry experiments (iii) pigment identification and quantification.

Characterization of Chicken α2A-Adrenoceptor: Molecular Cloning, Functional Analysis, and Its Involvement in Ovarian Follicular Development.

Adrenoceptors are suggested to mediate the functions of norepinephrine (NE) and epinephrine (EPI) in the central nervous system (CNS) and peripheral tissues in vertebrates. Compared to mammals, the functionality and expression of adrenoceptors have not been well characterized in birds. Here, we reported the structure, expression, and functionality of chicken functional α2A-adrenoceptor, named ADRA2A. The cloned chicken ADRA2A cDNA is 1335 bp in length, encoding the receptor with 444 amino acids (a.a.), which shows high amino acid sequence identity (63.4%) with its corresponding ortholog in humans. Using cell-based luciferase reporter assays and Western blot, we demonstrated that the ADRA2A could be activated by both NE and EPI through multiple signaling pathways, including MAPK/ERK signaling cascade. In addition, the mRNA expression of ADRA2A is found to be expressed abundantly in adult chicken tissues including thyroid, lung, ovary and adipose from the reported RNA-Seq data sets. Moreover, the mRNA expression of ADRA2A is also found to be highly expressed in the granulosa cells of 6–8 mm and F5 chicken ovarian follicles, which thus supports that ADRA2A signaling may play a role in ovarian follicular growth and differentiation. Taken together, our data provide the first proof that the α2A-adrenoceptor is functional in birds involving avian ovarian follicular development.

Analysis of the Expression of Neurotrophins and Their Receptors in Adult Zebrafish Kidney.

Neurotrophins and their receptors are involved in the development and maintenance of neuronal populations. Different reports have shown that all neurotrophin/receptor pathways can also play a role in several non-neuronal tissues in vertebrates, including the kidney. These signaling pathways are involved in different events to ensure the correct functioning of the kidney, such as growth, differentiation, and regulation of renal tubule transport. Previous studies in some fish species have identified the neurotrophins and receptors in the kidney. In this study, for the first time, we compare the expression profiles (mRNA and protein) of all neurotrophin/receptor pathways in the kidney of the adult zebrafish. We quantify the levels of mRNA by using qPCR and identify the expression pattern of each neurotrophin/receptor pathway by in situ hybridization. Next, we detect the proteins using Western blotting and immunohistochemistry. Our results show that among all neurotrophins analyzed, NT-3/TrkC is the most expressed in the glomerule and tubule and in the hematopoietic cells, similar to what has been reported in the mammalian kidney.

Constraining Morphologies of Soft Tissues in Extinct Vertebrates Using Multibody Dynamic Simulations: A Case Study on Articular Cartilage of the Sauropod Dreadnoughtus

Rarity of soft tissue preservation, including of articular cartilage, in the fossil record hinders creation of biologically-realistic mechanical models. Previous studies of articular cartilage in extant taxa have documented important aspects of cartilage shapes and thicknesses, but these insights remain generalized and have yet to see systematic implementation in biomechanical modeling. Herein, we present a new method for modeling joints that allows for testing of hypotheses about articular cartilage morphology in extinct taxa. Our case study examines the left elbow joint of the sauropod dinosaurDreadnoughtus schraniusing articular cartilage reconstructions constrained by extant phylogenetic bracketing (EPB). EPB investigations of alligator and chicken articular cartilage revealed the presence of a spherical anterior projection of cartilage on the distal humerus which articulates with the radius during flexion. Importantly, this shape does not directly mirror the underlying bone. Using multibody dynamic models created in Adams™ withouta priorirestrictions on joint degrees of freedom, we simulated the effects of three alternative cartilage reconstructions based on these EPB findings which differ in mediolateral placement of a cartilage sphere and its anteroposterior thickness, encompassing a range of possibilities for the condition inDreadnoughtus. Bone kinematics and contact area (calculated in Geomagic®) were tracked. Additionally, we modeled the elbow of an alligator and turkey using the same methodology and compared the results to XROMM (X-ray Reconstruction of Moving Morphology) analysis of the same limbs. Each model produced distinct results but were generally similar supporting our modeling methodology. Based on these findings, we predict thatDreadnoughtus, and presumably other extinct archosaurs, had a spherical projection of cartilage on the anterior face of the distal end of the humerus for articulation with the radius. Though many valuable insights have been gained by existing modeling methodologies, we chose a different approach that focused on joint contact surfaces. Moreover, applying our methods within a quantitative hypothesis-testing framework can advance the field of paleobiology by testing hypotheses relating shape and kinematics that are not possible with prescribed joint motions.

ENVIRONMENTAL RISKS OF MICRO-CONTAMINATION OF THE ENVIRONMENT WITH TETRACYCLINE

Review of environmental risks associated with the use of tetracycline. Micro-contamination with tetracycline has both medical risks associated with the manifestation of antibiotic resistance in bacterial communities, and environmental risks associated with changes in the transformation of substances in ecosystems and the impact on biodiversity by inhibiting certain species of plants, animals and microorganisms.Micro-contamination with tetracycline antibiotics entails a number of environmental risks. Due to the widespread use, the spread of tetracycline antibiotics in the environment has a global scale. The effect of tetracycline is observed in many objects. The spread of tetracycline is promoted by veterinary use, both directly and as a result of the use of animal waste as fertilizers for agricultural crops and pastures. As a result of the study, the following possible environmental consequences of micro-contamination of the environment with tetracycline were identified:1. Disruption of the work of bacteria included in the bacterial complex of activated sludge at sewage treatment plants using this technology, as well as during composting and affecting the processes of denitrification;2. Impacts on soil microbial communities by suppressing a number of processes of substance biodegradation and changing the biodiversity of microorganisms;3. The risks of micro-contamination of plant communities with tetracycline coming from organic fertilizers are associated with the inhibition of a number of root nutrition processes, species-specific for different crops;4. Microcontamination of marine communities with tetracycline is primarily associated with the impact on producers - algae and cyanobacteria, inhibiting their growth. And secondly, to the subsequent food chains by bioaccumulation in the tissues of vertebrates;5. The toxic effect of small doses of tetracycline from the soil on certain invertebrate species has been shown.

Single-cell transcriptome reveals insights into the development and function of the zebrafish ovary.

Zebrafish are an established research organism that has made many contributions to our understanding of vertebrate tissue and organ development, yet there are still significant gaps in our understanding of the genes that regulate gonad development, sex, and reproduction. Unlike the development of many organs, such as the brain and heart that form during the first few days of development, zebrafish gonads do not begin to form until the larval stage (≥5 days post-fertilization). Thus, forward genetic screens have identified very few genes required for gonad development. In addition, bulk RNA-sequencing studies that identify genes expressed in the gonads do not have the resolution necessary to define minor cell populations that may play significant roles in the development and function of these organs. To overcome these limitations, we have used single-cell RNA sequencing to determine the transcriptomes of cells isolated from juvenile zebrafish ovaries. This resulted in the profiles of 10,658 germ cells and 14,431 somatic cells. Our germ cell data represents all developmental stages from germline stem cells to early meiotic oocytes. Our somatic cell data represents all known somatic cell types, including follicle cells, theca cells, and ovarian stromal cells. Further analysis revealed an unexpected number of cell subpopulations within these broadly defined cell types. To further define their functional significance, we determined the location of these cell subpopulations within the ovary. Finally, we used gene knockout experiments to determine the roles of foxl2l and wnt9b for oocyte development and sex determination and/or differentiation, respectively. Our results reveal novel insights into zebrafish ovarian development and function, and the transcriptome profiles will provide a valuable resource for future studies.