- unprecedented transcriptome sequencing of the white-eared opossum. - albiventris can also contribute to the understanding of the molecular mechanisms that govern the different stages of organogenesis. - transcriptome sequences and catalogue of genes for a marsupial species at different neonatal stages, allowing the study of the mechanisms involved in organogenesis.. - 2019 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0. - Full list of author information is available at the end of the article. - Marsupials are one of the three large modern groups of mammals and are considered the closest external group to eutherian mammals (placentals). - Analysis of the D. - albiventris This work generated transcriptomes (RNA-seq) of the white-eared opossum during the first days of its postna- tal development. - We also identified a few transcripts that were exclu- sively expressed in each of the analysed stages: 109 tran- scripts were exclusively expressed at P0, 149 at P5 and 237 at P10 (Additional file 3).. - The BLASTX analysis of the D. - BLASTN analysis of the D. - Although there were a limited number of genes to valid- ate these data, we observed total concordance in the direc- tion of expression but not in the magnitude of the expression change between the RNA-seq and RT-qPCR re- sults (Fig. - At P5, the ventral skeletal muscle of the body wall (Additional file 5 d) and longitudinal and trans- versal sections of limb skeletal muscle (Additional file 5 f) were strongly stained by ISH. - The dia- phragm (Additional file 5a, e and h) showed moder- ate staining of the skeletal muscle. - Homogeneity of the biological triplicates determined by PCA (a) and the colourmap matrix b based on transcript expression values between P0, P5 and P10. - the extrinsic muscles of the eye showed strong stain- ing (Additional file 5 J).. - In this work, we sequenced the transcriptomes of whole neonates of the white-eared opossum (Didelphis albiven- tris) in three postnatal developmental stages to reveal the mechanisms of development in this South American marsupial species. - 3 Validation of the RNA-seq expression profiles by RT-qPCR. - domestica, revealed the importance of the opossum transcriptome for under- standing the biology of this species and the evolution of the marsupial group. - Mucosal immune response PIGR Facilitation of the transcytosis of soluble polymeric isoforms of immunoglobulin A and immune complexes. - SOBP Regulates the cellular fate and standardization of the organ of Corti. - Crystallins are required for the development and maintenance of the transparency of the eye lens. - Midbrain development MSX1 Related to the development of the midline structure of the forebrain, expressed in the spinal cord. - Limbs Embryonic hindlimb morphogenesis FGF4 Expressed in the thoracic and pelvic limbs and the apical ectodermal ridge of the limb bud. - C6 Component of the complement cascade. - Mucosal immune response PLA2G1B Secreted member of the phospholipase A2 (PLA2) class of enzymes produced by the pancreatic acinar cells. - The strong staining for titin hybridization reflected the pres- ence of mature skeletal muscle fibres in the tongue at P0 and P10, in the body wall at P0, in the limbs at P5, and in the extrinsic muscles of the eye at P10. - The moderate hybridization of titin at P10 in the heart is consistent with reports that most ventricular cardiac muscle cells are fully differentiated by the end of the fifteenth postna- tal week [72]. - Thus, our data confirmed the specificity of the synthesized probe. - The elucidation of the. - The main tissues and organs in the development of the white-eared opossum. - Consequently, we could obtain some idea of the most important genic events controlling each stage.. - Gene ontology terms associated with the development of the immune system were found to be enriched in our analysis of the D. - the complement protein network, an essential compo- nent of the innate and humoural immune system [76], participates in several non-inflammatory processes, such as coagulation [77], along with components of the co- agulation and fibrinolysis systems [76]. - Regarding the use of the primary jaw joint, the marsu- pial neonate approximates the embryonic condition of mammalian ancestors [80]. - The differenti- ation of the inner ear hair cells, located in the cochlea, is negatively regulated by HES1 [87], and the stereocilia orientation in these cells is guided by PTK7 [88, 89].. - The SOBP gene is expressed mainly in the cochlear duct, regulating the cellular fate and standardization of the organ of Corti [90, 91]. - The gastrointestinal system of postnatal marsupials has been explored, and the results contribute to telling the story of the evolution of this system in vertebrates [98 – 101]. - and that the milk regulates the microbiota of the neo- nate intestine [103]. - Furthermore, studies indicate that the milk plays a role in accelerating the development and maturation of the stomach [102]. - However, the mo- lecular mechanisms involved in these changes during the development of the marsupial stomach are not yet. - Hence, the enriched genes linked to the di- gestive system identified in this study, such as SLC9A4 and MOGAT3, could help to understand the develop- ment of the mammalian gastrointestinal system. - The development of the mammal kidneys is another topic that is still under investigation [107]. - This gene is required for the for- mation of intermediate segments of the pronephros [111] and is necessary to maintain the identity of the pronephric territory and define its size [112]. - IRX1 is expressed in the proximal tubular segment and the intermediate tubule segments of the pronephros and exhibits an important function in the differen- tiation of a proximo-medial S-shaped body (SSB) subdo- main [113]. - Because the forebrain of marsupials primarily de- velops postnatally, it is possible to study the brain devel- opment of the young in the pouch in stages that are equivalent to human mid-embryogenesis [115]. - A study in which in-pouch electroporation was performed using newborns of the Australian marsupial fat-tailed dunnart (Sminthopsis crassicaudata) demonstrated that marsupials can be an excellent in vivo model for studying forebrain development and evolution [115].. - While cranial and adult structures have been receiving attention, there is a gap in the knowledge of the evolu- tion of mammalian post-cranial structures, such as the limbs [120]. - albiventris transcripts, enabling detailed analysis of the unique attributes of this native Brazilian marsupial.. - three of the specimens were ran- domly selected for RNA-seq procedures and three for histological procedures. - All animal pro- cedures were conducted following the instructions of the ethical committee (CEUA/UFMG, ethical approval 220/. - Ro- tation and traction movements of the embryos were per- formed to remove the young from the female mammary papillae. - The quality of the isolated total RNA was evaluated in a Bioanalyzer (Agilent). - SuperScript II (Invitrogen) and oligo(dT) were used for reverse transcription of the first cDNA strand.. - The ends of the molecules were treated with T4 DNA Polymerase and Klenow DNA Polymerase (Illumina). - The 3′ end of the synthesized cDNA were phosphorylated with T4 PNK (Illumina) and adenylated with Klenow exo (Illumina).. - RNA-seq sequencing was performed using the HiSeq 2500 platform (Illumina) according to the recommenda- tions of the manufacturer and the paired-end reads protocol. - blast/executables/blast+/LATEST/) with default parame- ters was used to compare the nucleotide sequences of the D. - The probability density graphics of the coverage identity were generated in the gplots and ggplot2 packages of RStudio software.. - cDNA was synthesized using 1 μg of total RNA following the recommendations of the RevertAid™ H Minus First. - Additional file 1: Probability density values for the coverage identity of the D. - According to the analysis of the median and mean values, the coverage. - Sagittal view of the heart (G). - Frontal view of the tongue and mandible skeletal muscle (I). - Frontal view of the eye (J).. - Ultrastructure of the placenta of the tammar wallaby, Macropus eugenii: comparison with the grey short-tailed opossum, Monodelphis domestica. - https://doi.org . - https://doi.org/10.. - https://doi.org/10.1186/s . - On the development of the chondrocranium and the histological anatomy of the head in perinatal stages of marsupial mammals. - https://doi.org/10.1186/. - Taxonomy, affinities, and paleobiology of the tiny metatherian mammal Minusculodelphis, from the early Eocene of South America. - https://doi.org/10.1007/s . - Genome of the marsupial Monodelphis domestica reveals innovation in non- coding sequences. - Genome sequencing and analysis of the Tasmanian devil and its transmissible cancer. - The identification of immune genes in the milk transcriptome of the Tasmanian devil (Sarcophilus harrisii). - https://doi.org/10.1038/s . - Characterisation of the immune compounds in koala milk using a combined transcriptomic and proteomic approach. - Draft De novo Transcriptome of the rat kangaroo Potorous tridactylus as a tool for cell biology. - Transcriptome sequencing of the long-nosed bandicoot (Perameles nasuta) reveals conservation and innovation of immune genes in the marsupial order Peramelemorphia. - https://doi.org/10.1007/. - Transcriptomic changes in the pre-implantation uterus highlight histotrophic nutrition of the developing marsupial embryo. - https://doi.org/10.1038/s z.. - Genome of the Tasmanian tiger provides insights into the evolution and demography of an extinct marsupial carnivore. - https://doi.org/10.1038/s y.. - Morphological aspects of genital female of the opossum (Didelphis sp. - https://doi.org/10.1590/S . - https://doi.org/. - subfunctionalization of the CRX homeobox gene in mammalian evolution.. - The development of the immune tissues in marsupial pouch young. - Ontogenetic and phylogenetic transformations of the ear ossicles in marsupial mammals. - Evolution of the mammalian middle ear and jaw: adaptations and novel structures. - https://doi.org/10.1111/j.. - Retinoic acid signaling is necessary for the development of the organ of Corti. - Molecular insights into evolution of the vertebrate gut: focus on stomach and parietal cells in the marsupial, Macropus eugenii. - https://doi.org/10.1002/. - Cross- fostering of the tammar wallaby (Macropus eugenii) pouch young accelerates fore-stomach maturation. - Impaired gastric acid secretion in mice with a targeted disruption of the NHE4 Na+/H+ exchanger. - Development of the mammalian kidney. - https://doi.org/10.1371/
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