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Topic:Transcriptome

The transcriptome refers to the complete set of RNA transcripts produced by the genome at any given time in a specific cell or tissue type. In horses, transcriptome analysis provides insights into gene expression patterns under various physiological and pathological conditions. This approach allows researchers to identify key regulatory genes and pathways involved in equine development, health, and disease. Transcriptome studies in horses can encompass a range of topics, including developmental biology, immune response, and metabolic processes. These studies utilize technologies such as RNA sequencing to generate comprehensive datasets that can be used to understand the genetic basis of traits and conditions in horses. This page aggregates peer-reviewed research and scholarly articles that explore the methodologies, findings, and implications of transcriptome analysis in equine research.
Comparison of Poly-A+ Selection and rRNA Depletion in Detection of lncRNA in Two Equine Tissues Using RNA-seq.
Non-coding RNA    August 21, 2020   Volume 6, Issue 3 32 doi: 10.3390/ncrna6030032
Dahlgren AR, Scott EY, Mansour T, Hales EN, Ross PJ, Kalbfleisch TS, MacLeod JN, Petersen JL, Bellone RR, Finno CJ.Long non-coding RNAs (lncRNAs) are untranslated regulatory transcripts longer than 200 nucleotides that can play a role in transcriptional, post-translational, and epigenetic regulation. Traditionally, RNA-sequencing (RNA-seq) libraries have been created by isolating transcriptomic RNA via poly-A selection. In the past 10 years, methods to perform ribosomal RNA (rRNA) depletion of total RNA have been developed as an alternative, aiming for better coverage of whole transcriptomic RNA, both polyadenylated and non-polyadenylated transcripts. The purpose of this study was to determine which librar...
Genome-wide transcriptome analysis reveals equine embryonic stem cell-derived tenocytes resemble fetal, not adult tenocytes.
Stem cell research & therapy    May 19, 2020   Volume 11, Issue 1 184 doi: 10.1186/s13287-020-01692-w
Paterson YZ, Cribbs A, Espenel M, Smith EJ, Henson FMD, Guest DJ.Tendon injuries occur frequently in human and equine athletes. Treatment options are limited, and the prognosis is often poor with functionally deficient scar tissue resulting. Fetal tendon injuries in contrast are capable of healing without forming scar tissue. Embryonic stem cells (ESCs) may provide a potential cellular therapeutic to improve adult tendon regeneration; however, whether they can mimic the properties of fetal tenocytes is unknown. To this end, understanding the unique expression profile of normal adult and fetal tenocytes is crucial to allow validation of ESC-derived tenocytes...
Investigating the epithelial barrier and immune signatures in the pathogenesis of equine insect bite hypersensitivity.
PloS one    April 28, 2020   Volume 15, Issue 4 e0232189 doi: 10.1371/journal.pone.0232189
Cvitas I, Oberhänsli S, Leeb T, Dettwiler M, Müller E, Bruggman R, Marti EI.Insect bite hypersensitivity (IBH) is a Th-2, IgE-mediated dermatitis of horses caused by bites of insects of the genus Culicoides that has common features with human atopic dermatitis. Together with Th-2 cells, the epithelial barrier plays an important role in development of type I hypersensitivities. In order to elucidate the role of the epithelial barrier and of the skin immune response in IBH we studied the transcriptome of lesional whole skin of IBH-horses (IBH-LE; n = 9) in comparison to non-lesional skin (IBH-NL; n = 8) as well as to skin of healthy control horses (H; n = 9). To study t...
Effect of oral urea supplementation on the endometrial transcriptome of mares.
Animal reproduction science    April 18, 2020   Volume 216 106464 doi: 10.1016/j.anireprosci.2020.106464
Linhares Boakari Y, El-Sheikh Ali H, Dini P, Loux S, Barbosa Fernandes C, Esteller-Vico A, Scoggin K, Lawrence L, Ball B.An intravenous large dose of protein led to an increased blood urea nitrogen (BUN), resulting in a lesser uterine pH and altered uterine gene expression in mares. The objective of the present study was to evaluate effects of a more physiological methodology to increase BUN on the endometrium of mares. Mares were fed hay and a treatment or control diet (n = 11 mares/treatment) in a crossover design starting at time of ovulation detection (D0) and continuing until D7. Mares of the treated group were fed urea (0.4 g/kg BW) with sweet feed and molasses, and those of the control group were fe...
Modulation of stress and immune response by Amblyomin-X results in tumor cell death in a horse melanoma model.
Scientific reports    April 14, 2020   Volume 10, Issue 1 6388 doi: 10.1038/s41598-020-63275-2
Lichtenstein F, Iqbal A, de Lima Will SEA, Bosch RV, DeOcesano-Pereira C, Goldfeder MB, Chammas R, Trufen CEM, Morais KLP, de Souza JG, Natalino RJM....We have investigated Amblyomin-X-treated horse melanomas to better understand its mode of action through transcriptome analysis and the in vivo model. Amblyomin-X is a Kunitz-type homologous protein that selectively leads to the death of tumor cells via ER stress and apoptosis, currently under investigation as a new drug candidate for cancer treatment. Melanomas are immunogenic tumors, and a better understanding of the immune responses is warranted. Equine melanomas are spontaneous and not so aggressive as human melanomas are, as this study shows that the in vivo treatment of encapsulated hors...
Asynchronous Embryo Transfer Followed by Comparative Transcriptomic Analysis of Conceptus Membranes and Endometrium Identifies Processes Important to the Establishment of Equine Pregnancy.
International journal of molecular sciences    April 7, 2020   Volume 21, Issue 7 2562 doi: 10.3390/ijms21072562
Gibson C, de Ruijter-Villani M, Bauersachs S, Stout TAE.Preimplantation horse conceptuses require nutrients and signals from histotroph, the composition of which is regulated by luteal progesterone and conceptus-secreted factors. To distinguish progesterone and conceptus effects we shortened the period of endometrial progesterone-priming by asynchronous embryo transfer. Day 8 embryos were transferred to synchronous (day 8) or asynchronous (day 3) recipients, and RNA sequencing was performed on endometrium and conceptuses recovered 6 and 11 days later (embryo days 14 and 19). Asynchrony resulted in many more differentially expressed genes (DEGs) in ...
Transcriptomic analysis reveals the key regulators and molecular mechanisms underlying myometrial activation during equine placentitis†.
Biology of reproduction    February 18, 2020   Volume 102, Issue 6 1306-1325 doi: 10.1093/biolre/ioaa020
El-Sheikh Ali H, Boakari YL, Loux SC, Dini P, Scoggin KE, Esteller-Vico A, Kalbfleisch T, Ball BA.The key event in placentitis-induced preterm labor is myometrial activation with the subsequent initiation of labor. However, the molecular mechanisms underlying myometrial activation are not fully understood in the mares. Therefore, the equine myometrial transcriptome was characterized during placentitis (290.0 ± 1.52 days of GA, n = 5) and the prepartum period (330 days of GA, n = 3) in comparison to normal pregnant mares (289.8 ± 2.18 days of GA, n = 4). Transcriptome analysis identified 596 and 290 DEGs in the myometrium during placentitis and the prepartum period, respectively...
Transcriptomic Analysis of Testicular Gene Expression in Normal and Cryptorchid Horses.
Animals : an open access journal from MDPI    January 8, 2020   Volume 10, Issue 1 doi: 10.3390/ani10010102
Han H, Dong H, Chen Q, Gao Y, Li J, Li W, Dang R, Lei C.Testes produce sperm, and investigations into gene expression in the testes will enhance the understanding of the roles of testicular genes in male reproduction. Cryptorchidism, the failure of one or both testes to descend into the scrotal sac, is a common congenital malformation in horses. The major clinical consequence of this abnormality is impaired fertility. The aim of this study was to analyze the expression patterns of testicular genes and to identify the differentially expressed genes (DEGs) in testes between cryptorchid and normal horses. In this study, the gene expression patterns in...
Transcriptomic hallmarks of bone remodelling revealed by RNA-Seq profiling in blood of Arabian horses during racing training regime.
Gene    July 24, 2018   Volume 676 256-262 doi: 10.1016/j.gene.2018.07.040
Stefaniuk-Szmukier M, Ropka-Molik K, Piórkowska K, Żukowski K, Bugno-Poniewierska M.The impact of exercises on young developing organisms is still of interest to researchers. Similarly like Thoroughbreds, Arabian horses competing at the race track. The high percent of lameness and loss of days in training are often the result of weakness in the condition of the musculoskeletal system. The objective of the presented study was to identify by RNA-Seq method, the possible skeletal system originating transcriptomic profile in peripheral blood of Arabian horses undergoing race training. Obtained results showed that one of the most significantly deregulated pathway involved in bone ...
Heat shock stress: Profile of differential expression in Corynebacterium pseudotuberculosis biovar Equi.
Gene    December 13, 2017   Volume 645 124-130 doi: 10.1016/j.gene.2017.12.015
Gomide ACP, de Sá PG, Cavalcante ALQ, de Jesus Sousa T, Gomes LGR, Ramos RTJ, Azevedo V, Silva A, Folador ARC.Transcriptome studies on Corynebacterium pseudotuberculosis have recently contributed to the understanding about this microorganism's survival mechanisms in various hostile conditions. The gene expression profile of the C. pseudotuberculosis strain 1002 (Ovis biovar), has revealed genes that are possible candidates responsible for its maintenance in adverse environments, such as those found in the host. In another strain of this bacterium, 258 (Equi biovar), a high temperature condition was simulated, in order to verify which genes are responsible for promoting the persistence of the bacterium...
The vacuolar H+ ATPase V0 subunit d2 is associated with chondrocyte hypertrophy and supports chondrocyte differentiation.
Bone reports    August 18, 2017   Volume 7 98-107 doi: 10.1016/j.bonr.2017.08.002
Ayodele BA, Mirams M, Pagel CN, Mackie EJ.Chondrocyte hypertrophy makes important contributions to bone development and growth. We have investigated a number of novel cartilage genes identified in a recent transcriptomic study to determine whether they are differentially expressed between different zones of equine foetal growth cartilage. Twelve genes (, , , , , , , , , , and ) were found to be more highly expressed in the zone of hypertrophic chondrocytes than in the reserve or proliferative zones, whereas and were expressed at lower levels in the hypertrophic zone than in the reserve zone. , which encodes vacuolar H ATPase (V-ATP...
Circulating microRNA profiles of Hendra virus infection in horses.
Scientific reports    August 7, 2017   Volume 7, Issue 1 7431 doi: 10.1038/s41598-017-06939-w
Cowled C, Foo CH, Deffrasnes C, Rootes CL, Williams DT, Middleton D, Wang LF, Bean AGD, Stewart CR.Hendra virus (HeV) is an emerging zoonotic pathogen harbored by Australian mainland flying foxes. HeV infection can cause lethal disease in humans and horses, and to date all cases of human HeV disease have resulted from contact with infected horses. Currently, diagnosis of acute HeV infections in horses relies on the productive phase of infection when virus shedding may occur. An assay that identifies infected horses during the preclinical phase of infection would reduce the risk of zoonotic viral transmission during management of HeV outbreaks. Having previously shown that the host microRNA ...
From chromatogram to analyte to metabolite. How to pick horses for courses from the massive web resources for mass spectral plant metabolomics.
GigaScience    May 19, 2017   Volume 6, Issue 7 1-20 doi: 10.1093/gigascience/gix037
Perez de Souza L, Naake T, Tohge T, Fernie AR.The grand challenge currently facing metabolomics is the expansion of the coverage of the metabolome from a minor percentage of the metabolic complement of the cell toward the level of coverage afforded by other post-genomic technologies such as transcriptomics and proteomics. In plants, this problem is exacerbated by the sheer diversity of chemicals that constitute the metabolome, with the number of metabolites in the plant kingdom generally considered to be in excess of 200 000. In this review, we focus on web resources that can be exploited in order to improve analyte and ultimately metabol...
Understanding the response to endurance exercise using a systems biology approach: combining blood metabolomics, transcriptomics and miRNomics in horses.
BMC genomics    February 17, 2017   Volume 18, Issue 1 187 doi: 10.1186/s12864-017-3571-3
Mach N, Ramayo-Caldas Y, Clark A, Moroldo M, Robert C, Barrey E, López JM, Le Moyec L.Endurance exercise in horses requires adaptive processes involving physiological, biochemical, and cognitive-behavioral responses in an attempt to regain homeostasis. We hypothesized that the identification of the relationships between blood metabolome, transcriptome, and miRNome during endurance exercise in horses could provide significant insights into the molecular response to endurance exercise. For this reason, the serum metabolome and whole-blood transcriptome and miRNome data were obtained from ten horses before and after a 160 km endurance competition. We obtained a global regulatory ...
Tissue resolved, gene structure refined equine transcriptome.
BMC genomics    January 20, 2017   Volume 18, Issue 1 103 doi: 10.1186/s12864-016-3451-2
Mansour TA, Scott EY, Finno CJ, Bellone RR, Mienaltowski MJ, Penedo MC, Ross PJ, Valberg SJ, Murray JD, Brown CT.Transcriptome interpretation relies on a good-quality reference transcriptome for accurate quantification of gene expression as well as functional analysis of genetic variants. The current annotation of the horse genome lacks the specificity and sensitivity necessary to assess gene expression especially at the isoform level, and suffers from insufficient annotation of untranslated regions (UTR) usage. We built an annotation pipeline for horse and used it to integrate 1.9 billion reads from multiple RNA-seq data sets into a new refined transcriptome. This equine transcriptome integrates eight d...
LPS-induced modules of co-expressed genes in equine peripheral blood mononuclear cells.
BMC genomics    January 5, 2017   Volume 18, Issue 1 34 doi: 10.1186/s12864-016-3390-y
Pacholewska A, Marti E, Leeb T, Jagannathan V, Gerber V.Lipopolysaccharide (endotoxin, LPS) is a strong inducer of the innate immune response. It is widespread in our environment, e.g. in house dust and contributes to asthma. Compared to humans, horses are even more sensitive to LPS. However, data on LPS effects on the equine transcriptome are very limited. Using RNA-seq we analysed LPS-induced differences in the gene expression in equine peripheral blood mononuclear cells at the gene and gene-network level in two half-sib families and one group of unrelated horses. 24 h-LPS challenge of equine immune cells resulted in substantial changes in the t...
Profiling of exercise-induced transcripts in the peripheral blood cells of Thoroughbred horses.
Journal of equine science    December 15, 2016   Volume 27, Issue 4 157-164 doi: 10.1294/jes.27.157
Tozaki T, Kikuchi M, Kakoi H, Hirota KI, Mukai K, Aida H, Nakamura S, Nagata SI.Transcriptome analyses based on DNA microarray technology have been used to investigate gene expression profiles in horses. In this study, we aimed to identify exercise-induced changes in the expression profiles of genes in the peripheral blood of Thoroughbred horses using DNA microarray technology (15,429 genes on 43,603 probes). Blood samples from the jugular vein were collected from six horses before and 1 min, 4 hr, and 24 hr after all-out running on a treadmill. After the normalization of microarray data, a total of 26,830 probes were clustered into four groups and 11 subgroups showing si...
Equine dendritic cells generated with horse serum have enhanced functionality in comparison to dendritic cells generated with fetal bovine serum.
BMC veterinary research    November 15, 2016   Volume 12, Issue 1 254 doi: 10.1186/s12917-016-0880-8
Ziegler A, Everett H, Hamza E, Garbani M, Gerber V, Marti E, Steinbach F.Dendritic cells are professional antigen-presenting cells that play an essential role in the initiation and modulation of T cell responses. They have been studied widely for their potential clinical applications, but for clinical use to be successful, alternatives to xenogeneic substances like fetal bovine serum (FBS) in cell culture need to be found. Protocols for the generation of dendritic cells ex vivo from monocytes are well established for several species, including horses. Currently, the gold standard protocol for generating dendritic cells from monocytes across various species relies u...
Transcriptome analysis of equine sarcoids.
Veterinary and comparative oncology    October 25, 2016   Volume 15, Issue 4 1370-1381 doi: 10.1111/vco.12279
Semik E, Gurgul A, Ząbek T, Ropka-Molik K, Koch C, Mählmann K, Bugno-Poniewierska M.Equine sarcoids are the most commonly detected skin tumours in Equidae. In the present research, a comparative transcriptomic analysis was performed which aimed at looking inside a tumour biology and identification of the expression profile as a potential source of cancer specific genes useful as biomarkers. We have used Horse Gene Expression Microarray data from matched equine sarcoids and tumour-distant skin samples. In total, 901 significantly differentially expressed genes (DEGs) between lesional and healthy skin samples have been identified (fold change ≥ 2; P < 0.05). The la...
Characterisation of equine satellite cell transcriptomic profile response to β-hydroxy-β-methylbutyrate (HMB).
The British journal of nutrition    October 3, 2016   Volume 116, Issue 8 1315-1325 doi: 10.1017/S000711451600324X
Szcześniak KA, Ciecierska A, Ostaszewski P, Sadkowski T.β-Hydroxy-β-methylbutyrate (HMB) is a popular ergogenic aid used by human athletes and as a supplement to sport horses, because of its ability to aid muscle recovery, improve performance and body composition. Recent findings suggest that HMB may stimulate satellite cells and affect expressions of genes regulating skeletal muscle cell growth. Despite the scientific data showing benefits of HMB supplementation in horses, no previous study has explained the mechanism of action of HMB in this species. The aim of this study was to reveal the molecular background of HMB action on equine skeletal m...
Characterization of equine CSN1S2 variants considering genetics, transcriptomics, and proteomics.
Journal of dairy science    December 17, 2015   Volume 99, Issue 2 1277-1285 doi: 10.3168/jds.2015-9807
Cieslak J, Pawlak P, Wodas L, Borowska A, Stachowiak A, Puppel K, Kuczynska B, Luczak M, Marczak L, Mackowski M.Currently, research interest is increasing in horse milk composition and its effect on human health. Despite previously published studies describing the presence of intra- and interbreed variability of equine milk components, no investigations have focused on the genetic background of this variation. Among horse caseins and the genes encoding them, least is known about the structure and expression of the α-S2 casein gene, CSN1S2. Herein, based on direct sequencing of the equine CSN1S2 coding sequence, we describe the presence of 51-bp insertion-deletion (in/del) polymorphism, which significan...
Immune gene expression profiling of PBMC isolated from horses vaccinated with attenuated African horsesickness virus serotype 4.
Immunobiology    September 3, 2015   Volume 221, Issue 2 236-244 doi: 10.1016/j.imbio.2015.09.002
Pretorius A, Faber FE, van Kleef M.Development of African horsesickness (AHS) subunit vaccines will have to include a rational approach that uses knowledge of how the virus interacts with the host immune system. The global in vivo immune response induced by attenuated AHSV serotype 4 in horses was characterised using transcriptome sequencing. PBMC were collected with 24h intervals for four days after inoculation and four days after a second boost, 21 days later. Transcriptome data were normalised to the day 0 naïve transcriptome and up- or down-regulated immune genes identified using the CLC workbench. Peak expression was obse...
De Novo Assembly of the Donkey White Blood Cell Transcriptome and a Comparative Analysis of Phenotype-Associated Genes between Donkeys and Horses.
PloS one    July 24, 2015   Volume 10, Issue 7 e0133258 doi: 10.1371/journal.pone.0133258
Xie FY, Feng YL, Wang HH, Ma YF, Yang Y, Wang YC, Shen W, Pan QJ, Yin S, Sun YJ, Ma JY.Prior to the mechanization of agriculture and labor-intensive tasks, humans used donkeys (Equus africanus asinus) for farm work and packing. However, as mechanization increased, donkeys have been increasingly raised for meat, milk, and fur in China. To maintain the development of the donkey industry, breeding programs should focus on traits related to these new uses. Compared to conventional marker-assisted breeding plans, genome- and transcriptome-based selection methods are more efficient and effective. To analyze the coding genes of the donkey genome, we assembled the transcriptome of donke...
Correction: The transcriptome of equine peripheral blood mononuclear cells.
PloS one    April 27, 2015   Volume 10, Issue 4 e0127372 doi: 10.1371/journal.pone.0127372
No abstract available
Behavioral and Transcriptomic Fingerprints of an Enriched Environment in Horses (Equus caballus).
PloS one    December 10, 2014   Volume 9, Issue 12 e114384 doi: 10.1371/journal.pone.0114384
Lansade L, Valenchon M, Foury A, Neveux C, Cole SW, Layé S, Cardinaud B, Lévy F, Moisan MP.The use of environmental enrichment (EE) has grown in popularity over decades, particularly because EE is known to promote cognitive functions and well-being. Nonetheless, little is known about how EE may affect personality and gene expression. To address this question in a domestic animal, 10-month-old horses were maintained in a controlled environment or EE for 12 weeks. The control horses (n = 9) lived in individual stalls on wood shaving bedding. They were turned out to individual paddocks three times a week and were fed three times a day with pellets or hay. EE-treated horses (n =...
Transcriptome-wide analysis of messenger RNA decay in normal and osteoarthritic human articular chondrocytes.
Arthritis & rheumatology (Hoboken, N.J.)    August 27, 2014   Volume 66, Issue 11 3052-3061 doi: 10.1002/art.38849
Tew SR, McDermott BT, Fentem RB, Peffers MJ, Clegg PD.Messenger RNA (mRNA) decay rates control not only gene expression levels, but also responsiveness to altered transcriptional input. We undertook this study to examine transcriptome-wide posttranscriptional regulation in both normal and osteoarthritic (OA) human articular chondrocytes. Methods: Human articular chondrocytes were isolated from normal or OA tissue. Equine articular chondrocytes were isolated from young or old horses at a commercial abattoir. RNA decay was measured across the transcriptome in human cells by microarray analysis following an actinomycin D chase. Messenger RNA levels ...
Dexamethasone acutely down-regulates genes involved in steroidogenesis in stallion testes.
The Journal of steroid biochemistry and molecular biology    July 7, 2014   Volume 143 451-459 doi: 10.1016/j.jsbmb.2014.07.003
Ing NH, Forrest DW, Riggs PK, Loux S, Love CC, Brinsko SP, Varner DD, Welsh TH.In rodents, livestock and primate species, a single dose of the synthetic glucocorticoid dexamethasone acutely lowers testosterone biosynthesis. To determine the mechanism of decreased testosterone biosynthesis, stallions were treated with 0.1mg/kg dexamethasone 12h prior to castration. Dexamethasone decreased serum concentrations of testosterone by 60% compared to saline-treated control stallions. Transcriptome analyses (microarrays, northern blots and quantitative PCR) of testes discovered that dexamethasone treatment decreased concentrations of glucocorticoid receptor alpha (NR3C1), alpha a...
RNA-seq transcriptome profiling of equine inner cell mass and trophectoderm.
Biology of reproduction    March 20, 2014   Volume 90, Issue 3 61 doi: 10.1095/biolreprod.113.113928
Iqbal K, Chitwood JL, Meyers-Brown GA, Roser JF, Ross PJ.Formation of the inner cell mass (ICM) and trophectoderm (TE) marks the first differentiation event in mammalian development. These two cell types have completely divergent fates for the remainder of the developmental process. The molecular mechanisms that regulate ICM and TE formation are poorly characterized in horses. The objective of this study was to establish the transcriptome profiles of ICM and TE cells from horse blastocysts using RNA sequencing (RNA-seq). A total of 12 270 genes were found to be expressed in either lineage. Global analysis of the transcriptome profiles by unsupervi...
Transcriptomic signatures in cartilage ageing.
Arthritis research & therapy    August 23, 2013   Volume 15, Issue 4 R98 doi: 10.1186/ar4278
Peffers M, Liu X, Clegg P.Age is an important factor in the development of osteoarthritis. Microarray studies provide insight into cartilage aging but do not reveal the full transcriptomic phenotype of chondrocytes such as small noncoding RNAs, pseudogenes, and microRNAs. RNA-Seq is a powerful technique for the interrogation of large numbers of transcripts including nonprotein coding RNAs. The aim of the study was to characterise molecular mechanisms associated with age-related changes in gene signatures. Methods: RNA for gene expression analysis using RNA-Seq and real-time PCR analysis was isolated from macroscopicall...
Equine oviduct explant culture: a basic model to decipher embryo-maternal communication.
Reproduction, fertility, and development    August 2, 2013   Volume 26, Issue 7 954-966 doi: 10.1071/RD13089
Nelis H, D'Herde K, Goossens K, Vandenberghe L, Leemans B, Forier K, Smits K, Braeckmans K, Peelman L, Van Soom A.Equine embryos remain for 6 days in the oviduct and thus there is a need for an in vitro model to study embryo-oviductal interactions in the horse, since this subtle way of communication is very difficult to analyse in vivo. Until now, no equine oviduct explant culture model has been characterised both morphologically and functionally. Therefore, we established a culture system for equine oviduct explants that maintained epithelial morphology during 6 days of culture, as revealed by light microscopy and transmission electron microscopy. We demonstrated the presence of highly differentiated, ta...