scholarly journals The zebrafish as a model system for analyzing mammalian and native α-crystallin promoter function

2017 ◽  
Author(s):  
Mason Posner ◽  
Kelly Murray ◽  
Hayden Eighinger ◽  
Amy Drossman ◽  
Zachary Haley ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Promoter Activity ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Zebrafish Embryos ◽  
Mrna Levels ◽  
Gfp Expression ◽  
Promoter Function ◽  
Lens Crystallin ◽  
Αb Crystallin

Previous studies have used the zebrafish to investigate the biology of lens crystallin proteins and their roles in development and disease. However, little is known about zebrafish α-crystallin promoter function, how it compares to that of mammals, or whether mammalian α-crystallin promoter activity can be assessed using zebrafish embryos. We injected a variety of α-crystallin promoter fragments from each species combined with the coding sequence for green fluorescent protein (GFP) into zebrafish zygotes to determine the resulting spatiotemporal expression patterns in the developing embryo. We also measured mRNA levels and protein abundance for all three zebrafish α-crystallins. Our data showed that mouse and zebrafish αA-crystallin promoters generated similar GFP expression in the lens, but with earlier onset when using mouse promoters. Expression was also found in notochord and skeletal muscle in a small percentage of embryos. Mouse αB-crystallin promoter fragments drove GFP expression primarily in zebrafish skeletal muscle, with less common expression in notochord, lens, heart and in extraocular regions of the eye. A short fragment containing only a lens-specific enhancer region produced no GFP expression, suggesting that these lens responsive elements in the mouse are not used in the zebrafish. The two paralogous zebrafish αB-crystallin promoters produced subtly different expression profiles, with the αBa promoter driving expression equally in notochord and skeletal muscle while the αBb promoter resulted primarily in skeletal muscle expression. Messenger RNA for zebrafish αa, aBa and αBb were all detected by 1 day post fertilization (dpf). Parallel reaction monitoring (PRM) mass spectrometry was used to detect αA, αBa, and αBb peptides in digests of zebrafish embryos. In whole embryos, αA-crystallin was first detected by 2 dpf, peaked in abundance by 4-5 dpf, and was localized to the eye. αBa was also detected in whole embryo at nearly constant levels from 1-6 dpf, was also localized primarily to the eye, and its abundance in extraocular tissues decreased from 4-7 dpf. In contrast, due to its low abundance, no αBb protein could be detected in whole embryo, or dissected eye and extraocular tissues. Our results show that mammalian α-crystallin promoters can be efficiently screened in zebrafish embryos and that their controlling regions are well conserved, although their use in each species may reflect evolutionary changes in developmental roles for α-crystallins. An ontogenetic shift in zebrafish αBa-crystallin promoter activity provides an interesting system for examining the evolution and control of tissue specificity. Future studies that combine these promoter based approaches with the expanding ability to engineer the zebrafish genome via techniques such as CRISPR/Cas9 will allow the manipulation of protein expression to test hypotheses about lens crystallin function and its relation to lens biology and disease.

scholarly journals The zebrafish as a model system for analyzing mammalian and native α-crystallin promoter function

2017 ◽  
Author(s):  
Mason Posner ◽  
Kelly Murray ◽  
Hayden Eighinger ◽  
Amy Drossman ◽  
Zachary Haley ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Promoter Activity ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Zebrafish Embryos ◽  
Mrna Levels ◽  
Gfp Expression ◽  
Promoter Function ◽  
Lens Crystallin ◽  
Αb Crystallin

Previous studies have used the zebrafish to investigate the biology of lens crystallin proteins and their roles in development and disease. However, little is known about zebrafish α-crystallin promoter function, how it compares to that of mammals, or whether mammalian α-crystallin promoter activity can be assessed using zebrafish embryos. We injected a variety of α-crystallin promoter fragments from each species combined with the coding sequence for green fluorescent protein (GFP) into zebrafish zygotes to determine the resulting spatiotemporal expression patterns in the developing embryo. We also measured mRNA levels and protein abundance for all three zebrafish α-crystallins. Our data showed that mouse and zebrafish αA-crystallin promoters generated similar GFP expression in the lens, but with earlier onset when using mouse promoters. Expression was also found in notochord and skeletal muscle in a small percentage of embryos. Mouse αB-crystallin promoter fragments drove GFP expression primarily in zebrafish skeletal muscle, with less common expression in notochord, lens, heart and in extraocular regions of the eye. A short fragment containing only a lens-specific enhancer region produced no GFP expression, suggesting that these lens responsive elements in the mouse are not used in the zebrafish. The two paralogous zebrafish αB-crystallin promoters produced subtly different expression profiles, with the αBa promoter driving expression equally in notochord and skeletal muscle while the αBb promoter resulted primarily in skeletal muscle expression. Messenger RNA for zebrafish αa, aBa and αBb were all detected by 1 day post fertilization (dpf). Parallel reaction monitoring (PRM) mass spectrometry was used to detect αA, αBa, and αBb peptides in digests of zebrafish embryos. In whole embryos, αA-crystallin was first detected by 2 dpf, peaked in abundance by 4-5 dpf, and was localized to the eye. αBa was also detected in whole embryo at nearly constant levels from 1-6 dpf, was also localized primarily to the eye, and its abundance in extraocular tissues decreased from 4-7 dpf. In contrast, due to its low abundance, no αBb protein could be detected in whole embryo, or dissected eye and extraocular tissues. Our results show that mammalian α-crystallin promoters can be efficiently screened in zebrafish embryos and that their controlling regions are well conserved, although their use in each species may reflect evolutionary changes in developmental roles for α-crystallins. An ontogenetic shift in zebrafish αBa-crystallin promoter activity provides an interesting system for examining the evolution and control of tissue specificity. Future studies that combine these promoter based approaches with the expanding ability to engineer the zebrafish genome via techniques such as CRISPR/Cas9 will allow the manipulation of protein expression to test hypotheses about lens crystallin function and its relation to lens biology and disease.

scholarly journals The zebrafish as a model system for analyzing mammalian and native α-crystallin promoter function

PeerJ ◽  
2017 ◽  
Vol 5 ◽  
pp. e4093 ◽  
Author(s):  
Mason Posner ◽  
Kelly L. Murray ◽  
Matthew S. McDonald ◽  
Hayden Eighinger ◽  
Brandon Andrew ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Promoter Activity ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Zebrafish Embryos ◽  
Mrna Levels ◽  
Gfp Expression ◽  
Promoter Function ◽  
Lens Crystallin ◽  
Αb Crystallin

Previous studies have used the zebrafish to investigate the biology of lens crystallin proteins and their roles in development and disease. However, little is known about zebrafish α-crystallin promoter function, how it compares to that of mammals, or whether mammalian α-crystallin promoter activity can be assessed using zebrafish embryos. We injected a variety of α-crystallin promoter fragments from each species combined with the coding sequence for green fluorescent protein (GFP) into zebrafish zygotes to determine the resulting spatiotemporal expression patterns in the developing embryo. We also measured mRNA levels and protein abundance for all three zebrafish α-crystallins. Our data showed that mouse and zebrafish αA-crystallin promoters generated similar GFP expression in the lens, but with earlier onset when using mouse promoters. Expression was also found in notochord and skeletal muscle in a smaller percentage of embryos. Mouse αB-crystallin promoter fragments drove GFP expression primarily in zebrafish skeletal muscle, with less common expression in notochord, lens, heart and in extraocular regions of the eye. A short fragment containing only a lens-specific enhancer region increased lens and notochord GFP expression while decreasing muscle expression, suggesting that the influence of mouse promoter control regions carries over into zebrafish embryos. The two paralogous zebrafish αB-crystallin promoters produced subtly different expression profiles, with the aBa promoter driving expression equally in notochord and skeletal muscle while the αBb promoter resulted primarily in skeletal muscle expression. Messenger RNA for zebrafish αA increased between 1 and 2 days post fertilization (dpf), αBa increased between 4 and 5 dpf, but αBb remained at baseline levels through 5 dpf. Parallel reaction monitoring (PRM) mass spectrometry was used to detect αA, aBa, and αBb peptides in digests of zebrafish embryos. In whole embryos, αA-crystallin was first detected by 2 dpf, peaked in abundance by 4–5 dpf, and was localized to the eye. αBa was detected in whole embryo at nearly constant levels from 1–6 dpf, was also localized primarily to the eye, and its abundance in extraocular tissues decreased from 4–7 dpf. In contrast, due to its low abundance, no αBb protein could be detected in whole embryo, or dissected eye and extraocular tissues. Our results show that mammalian α-crystallin promoters can be efficiently screened in zebrafish embryos and that their controlling regions are well conserved. An ontogenetic shift in zebrafish aBa-crystallin promoter activity provides an interesting system for examining the evolution and control of tissue specificity. Future studies that combine these promoter based approaches with the expanding ability to engineer the zebrafish genome via techniques such as CRISPR/Cas9 will allow the manipulation of protein expression to test hypotheses about lens crystallin function and its relation to lens biology and disease.

22 Characterization of tRNA Expression Profiles in Large Offspring Syndrome

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 13-14
Author(s):  
Anna K Goldkamp ◽  
Yahan Li ◽  
Rocio M Rivera ◽  
Darren Hagen
Keyword(s):  
Skeletal Muscle ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Tumor Development ◽  
Gene Clusters ◽  
Tissue Type ◽  
Trna Genes ◽  
Translational Machinery ◽  
Trna Species

Abstract Differentially methylated regions (DMRs) have been associated with Large Offspring Syndrome (LOS) in cattle. Some DMRs overlap transfer RNA (tRNA) gene clusters, potentially altering tRNA expression patterns uniquely by treatment group or tissue type. tRNAs are classified as adapter molecules, serving a key role in the translational machinery implementing genetic code. Variation in tRNA expression has been identified in several disease pathways suggesting an important role in the regulation of biological processes. tRNAs also serve as a source of small non-coding RNAs. To better understand the role of tRNA expression in LOS, total RNA was extracted from skeletal muscle and liver of 105-day fetuses and the tRNAs sequenced. Although there are nearly three times the number of tRNA genes in cattle as compared to human (1,659 vs 597), there is a shared occurrence of transcriptionally silent tRNA genes in both species. This study detected expression of 474 and 487 bovine tRNA genes in skeletal muscle and liver, respectively, with the remainder being very lowly expressed or transcriptionally silent. Eleven tRNA isodecoders are transcriptionally silent in both skeletal muscle and liver and another isodecoder is silent in the liver (SerGGA). Further, the highest expressed isodecoders differ by treatment or tissue type with roughly half correlated to codon frequency. While the absence of certain isodecoders may be relieved by wobble base pairing, missing tRNA species could likely increase the likelihood of mistranslation or mRNA degradation. Differential expression of tissue- and treatment-specific tRNA genes may modulate translation during protein homeostasis or cellular stress, altering regulatory products targeting genes associated with overgrowth in skeletal muscle and/or tumor development in the liver of LOS individuals.

scholarly journals Enhancer Trapping and Annotation in Zebrafish Mediated with Sleeping Beauty, piggyBac and Tol2 Transposons

Genes ◽  
2018 ◽  
Vol 9 (12) ◽  
pp. 630 ◽  
Author(s):  
Dan Shen ◽  
Songlei Xue ◽  
Shuheng Chan ◽  
Yatong Sang ◽  
Saisai Wang ◽  
...  
Keyword(s):  
Expression Profiles ◽  
Expression Patterns ◽  
Genomic Analysis ◽  
Sleeping Beauty ◽  
Comparative Genomic Analysis ◽  
Regulatory Function ◽  
Comparative Genomic ◽  
Gfp Expression ◽  
Enhancer Trapping ◽  
Multiple Expression

Although transposon-mediated enhancer trapping (ET) is successfully applied in diverse models, the efficiency of various transposon systems varies significantly, and little information is available regarding efficiency of enhancer trapping by various transposons in zebrafish. Most potential enhancers (Ens) still lack evidence of actual En activity. Here, we compared the differences in ET efficiency between sleeping beauty (SB), piggyBac (PB) and Tol2 transposons. Tol2 represented the highest germline transfer efficiencies at 55.56% (NF0 = 165), followed by SB (38.36%, NF0 = 151) and PB (32.65%, NF0 = 149). ET lines generated by the Tol2 transposon tended to produce offspring with a single expression pattern per line, while PB and SB tended to generate embryos with multiple expression patterns. In our tests, 10 putative Ens (En1–10) were identified by splinkerette PCR and comparative genomic analysis. Combining the GFP expression profiles and mRNA expression patterns revealed that En1 and En2 may be involved in regulation of the expression of dlx1a and dlx2a, while En6 may be involved in regulation of the expression of line TK4 transgene and rps26, and En7 may be involved in the regulation of the expression of wnt1 and wnt10b. Most identified Ens were found to be transcribed in zebrafish embryos, and their regulatory function may involve eRNAs.

Gene expression analysis of the pro-oestrous-stage rat uterus reveals neuroligin 2 as a novel steroid-regulated gene

2004 ◽  
Vol 16 (8) ◽  
pp. 763 ◽  
Author(s):  
Han-Seung Kang ◽  
Chae-Kwan Lee ◽  
Ju-Ran Kim ◽  
Seong-Jin Yu ◽  
Sung-Goo Kang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Northern Blot ◽  
Blot Analysis ◽  
Northern Blot Analysis ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Oestrous Cycle ◽  
Mrna Levels ◽  
Rat Uterus ◽  
Ovx Rats

In the present study, differential gene expression in the uteri of ovariectomised (OVX) and pro-oestrous rats (OVX v. pro-oestrus pair) was investigated using cDNA expression array analysis. Differential uterine gene expression in OVX rats and progesterone (P4)-injected OVX rats (OVX v. OVX + P4 pair) was also examined. The uterine gene expression profiles of these two sets of animals were also compared for the effects of P4 treatment. RNA samples were extracted from uterine tissues and reverse transcribed in the presence of [α32P]-dATP. Membrane sets of rat arrays were hybridised with cDNA probe sets. Northern blot analysis was used to validate the relative gene expression patterns obtained from the cDNA array. Of the 1176 cDNAs examined, 23 genes showed significant (>two-fold) changes in expression in the OVX v. pro-oestrus pair. Twenty of these genes were upregulated during pro-oestrus compared with their expression in the OVX rat uterus. In the OVX v. OVX + P4 pair, 22 genes showed significant (>two-fold) changes in gene expression. Twenty of these genes were upregulated in the OVX + P4 animals. The genes for nuclear factor I–XI, afadin, neuroligin 2, semaphorin Z, calpain 4, cyclase-associated protein homologue, thymosin β-4X and p8 were significantly upregulated in the uteri of the pro-oestrus and OVX + P4 rats of both experimental pairs compared with the OVX rat uteri. These genes appear to be under the control of P4. One of the most interesting findings of the present study is the unexpected and marked expression of the neuroligin 2 gene in the rat uterus. This gene is expressed at high levels in the central nervous system and acts as a nerve cell adhesion factor. According to Northern blot analysis, neuroligin 2 gene expression was higher during the pro-oestrus and metoestrus stages than during the oestrus and dioestrus stages of the oestrous cycle. In addition, neuroligin 2 mRNA levels were increased by both 17β-oestradiol (E2) and P4, although P4 administration upregulated gene expression to a greater extent than injection of E2. These results indicate that neuroligin 2 gene expression in the rat uterus is under the control of both E2 and P4, which are secreted periodically during the oestrous cycle.

scholarly journals Microfibril-associated Glycoprotein-2 (MAGP-2) Is Specifically Associated with Fibrillin-containing Microfibrils but Exhibits More Restricted Patterns of Tissue Localization and Developmental Expression Than Its Structural Relative MAGP-1

1998 ◽  
Vol 46 (8) ◽  
pp. 871-885 ◽  
Author(s):  
Mark A. Gibson ◽  
Merran L. Finnis ◽  
Jaliya S. Kumaratilake ◽  
Edward G. Cleary
Keyword(s):  
Skeletal Muscle ◽  
Expression Patterns ◽  
Fetal Lung ◽  
Northern Blotting ◽  
Developmental Expression ◽  
Mrna Levels ◽  
Western Blots ◽  
Fibrillin 1 ◽  
Significant Expression ◽  
Nuchal Ligament

SUMMARY We developed an affinity-purified anti-MAGP-2 peptide antibody that specifically identified MAGP-2 on Western blots of purified matrix proteins and extracts of nuchal ligament. Immunolocalization studies on tissues from a 210-day-old fetus and a mature bovine showed that MAGP-2 was located in similar regions to MAGP-1 and fibrillin-1 but that the distribution of MAGP-2 was more restricted. In fetal nuchal ligament, skeletal muscle, and spleen the distribution of MAGP-2 was indistinguishable from that of MAGP-1. In contrast to MAGP-1, MAGP-2 was not detected in the medial layer of fetal thoracic aorta and in much of the peritubular matrix of fetal and mature kidney and in the mature ocular zonule. Some differences in the immunolocalization patterns were also evident in fetal lung, cartilage, skin, and heart. Immunoelectron microscopy confirmed that MAGP-2 was specifically associated with fibrillin-containing microfibrils in nuchal ligament, dermis, adventitia of aorta, glomerular mesangium and perimysium. Northern blotting of RNA from tissues of a 210-day-old fetus indicated that steady-state MAGP-2 mRNA levels were highest in nuchal ligament. Significant expression was also detected in lung, heart, skeletal muscle, skin, and Achilles tendon. The tissue pattern of MAGP-2 expression differed significantly from that of MAGP-1. MAGP-2 expression appeared to be higher in nuchal ligament, heart, and skeletal muscle and lower in aorta and kidney. In nuchal ligament, MAGP-2 mRNA expression appeared to peak around 180 days of fetal development, which correlates with the period of onset of elastinogenesis in this tissue. Overall, the immunolocalization and expression patterns of MAGP-2 appeared to be distinct from those of other microfibrillar components. This is consistent with the view that MAGP-2 plays a unique role in the biology of the microfibrils, perhaps by mediating their interaction with cell surfaces at specific stages of development and differentiation.

Evolutionary aspects of a new MyoD gene in amphioxus (Branchiostoma belcheri) and its promoter specificity in skeletal and cardiac muscles

Biologia ◽  
2014 ◽  
Vol 69 (9) ◽  
Author(s):  
Xungang Tan ◽  
Pei Zhang ◽  
Shao Du
Keyword(s):  
Skeletal Muscle ◽  
Transcription Factors ◽  
Zebrafish Embryos ◽  
Evolutionary Analysis ◽  
Gene Duplications ◽  
Gfp Expression ◽  
Specific Expression ◽  
Branchiostoma Belcheri ◽  
Cardiac Muscles ◽  
Myod Gene

AbstractVertebrate MyoD family of transcription factors contains four members including MyoD, Myf5, Myogenin and MRF4. These myogenic regulatory factors (MRFs) play key roles in regulating skeletal muscle development and growth. Evolutionary analysis suggests that the four vertebrate MRF genes were derived by gene duplications from a single ancestral gene during chordate evolution. Better understanding of the structure and regulation of MyoD expression in amphioxus Branchiostoma belcheri may provide insight into the evolutionary history of myogenic gene duplications because of the unique position of amphioxus in evolution. We report here that isolation and characterization of a new MyoD gene, AmphiMyoD, in B. belcheri. Sequence analysis revealed that the AmphiMyoD is more closely related to myogenic transcription factors in invertebrates and vertebrates compared with the previously identified three MyoD like genes in amphioxus, suggesting that the AmphiMyoD might be the closest relative of the ancestral myogenic gene. To determine if the AmphiMyoD gene promoter controls muscle-specific expression, the AmphiMyoD promoter was linked with the green fluorescence protein (GFP) reporter and the construct was microinjected into zebrafish embryos for transient expression assay. AmphiMyoD promoter directed skeletal muscle-specific GFP expression in zebrafish embryos. In addition, it also drove GFP expression in cardiac muscles of the injected embryos, but not in other non-muscle tissues. These data demonstrated that the AmphiMyoD promoter contained regulatory elements for skeletal and cardiac muscle-specific expression. Moreover, the regulatory element(s) could function across species.

Interactions between ROS and AMP kinase activity in the regulation of PGC-1α transcription in skeletal muscle cells

2009 ◽  
Vol 296 (1) ◽  
pp. C116-C123 ◽  
Author(s):  
Isabella Irrcher ◽  
Vladimir Ljubicic ◽  
David A. Hood
Keyword(s):  
Skeletal Muscle ◽  
Transcriptional Activation ◽  
Promoter Activity ◽  
Muscle Cells ◽  
Peroxisome Proliferator ◽  
Mrna Levels ◽  
Ros Production ◽  
Ampk Activation ◽  
Peroxisome Proliferator Activated Receptor ◽  
Beneficial Effects

Reactive oxygen species (ROS) play an important role in cellular function via the activation of signaling cascades. ROS have been shown to affect mitochondrial biogenesis, morphology, and function. Their beneficial effects are likely mediated via the upregulation of transcriptional regulators such as peroxisome proliferator-activated receptor-γ coactivator-1 protein-α (PGC-1α). However, the ROS signals that regulate PGC-1α transcription in skeletal muscle are not understood. Here we examined the effect of H2O2 on the regulation of PGC-1α expression, and its relationship to AMPK activation. We demonstrate that 24 h of exogenous H2O2 treatment increased PGC-1α promoter activity and mRNA expression. Both effects were blocked with the addition of N-acetylcysteine, a ROS scavenger. These effects were mediated, in part, via upstream stimulatory factor-1/Ebox DNA binding and involved 1) interactions with downstream sequences and 2) the activation of AMPK. Elevated ROS led to the activation of AMPK, likely via a decline in ATP levels. The activation of AMPK using 5-aminoimidazole-4-carboxamide-1-β- d-ribofuranoside increased PGC-1α promoter activity and mRNA levels but reduced ROS production. Thus the net effect of AMPK activation on PGC-1α expression was a result of increased transcriptional activation, counterbalanced by reduced ROS production. The effects of H2O2 on PGC-1α expression differed depending on the level of ROS within the cell. Low levels of ROS result in reduced PGC-1α mRNA in the absence of an effect on PGC-1α promoter activation. In contrast, elevated levels of H2O2 induce PGC-1α transcription indirectly, via AMPK activation. These data identify unique interactions between ROS and AMPK activation on the expression of PGC-1α in muscle cells.

scholarly journals Changes in Gene Expression during Pituitary Morphogenesis and Organogenesis in the Chick Embryo

Endocrinology ◽  
2011 ◽  
Vol 152 (3) ◽  
pp. 989-1000 ◽  
Author(s):  
Monika Proszkowiec-Weglarz ◽  
Stacy E. Higgins ◽  
Tom E. Porter
Keyword(s):  
Gene Expression ◽  
Mrna Expression ◽  
Anterior Pituitary ◽  
Molecular Mechanisms ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Mrna Levels ◽  
Pituitary Development ◽  
Excellent Model ◽  
Functional Development

The anterior pituitary gland plays an important role in the regulation of many physiological processes. Formation of Rathke's pouch (RP), the precursor of the anterior pituitary, involves evagination of the oral ectoderm in a multi-step process regulated by cell interactions, signaling pathways, and transcription factors. Chickens are an excellent model to study development because of the availability of large sample sizes, accurate timing of development, and embryo accessibility. The aim of this study was to quantify mRNA expression patterns in the developing chicken anterior pituitary to evaluate the chicken embryo as a model for mammalian pituitary development. The expression profiles of 16 genes differentially expressed in RP and neuroectoderm were determined in this study. Among these, Pitx1, Pitx2, and Hesx1 mRNA levels were high on embryonic days (e) 2.5 to e3 in RP and decreased during development. Expression of Pit1 and Tbx19 mRNA in RP reached the highest levels by e7 and e6.5, respectively. Levels of glycoprotein subunit α mRNA increased beginning at e4. FGF8 mRNA showed the highest expression at e3 to e3.5 in neuroectoderm. BMP2 showed slight decreases in mRNA expression in both tissues during development, while Isl1 and Noggin mRNA expression increased in later development. Taken together, we present the first quantitative transcriptional profile of pituitary organogenesis. Our results will help further understanding of the functional development of this gland. Moreover, because of the high similarity in gene expression patterns observed between chicken and mouse, chickens could serve as an excellent model to study genetic and molecular mechanisms underlying pituitary development.

scholarly journals Potential role of miR-155-5p in fat deposition and skeletal muscle development of chicken

2020 ◽  
Vol 40 (6) ◽  
Author(s):  
Sifan Xu ◽  
Yang Chang ◽  
Guanxian Wu ◽  
Wanting Zhang ◽  
Chaolai Man
Keyword(s):  
Skeletal Muscle ◽  
Muscle Development ◽  
Target Genes ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Fat Deposition ◽  
Skeletal Muscle Development ◽  
Stem Loop ◽  
Related Target ◽  
Kegg Pathways

Abstract miR-155 has multiple functions in many physiological and pathological processes. However, little is known about the expression characteristics of avian miR-155. In the present study, partial pri-miR-155 sequences were cloned from AA+ broiler, Sanhuang broiler and Hy-Line Brown layer, respectively. Stem–loop qRT-PCR was performed to detect the miR-155-5p spatiotemporal expression profiles of each chicken breed, and the target genes of miR-155-5p were predicted in Gene Oncology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The results showed that the partial pri-miR-155 sequences of different breeds of chicken were high conserved. The expression patterns of miR-155-5p between broiler and layer were basically similar, and miR-155-5p was expressed highly in immune related tissues (spleen, thymus and bursa). In the same old chicken (14 days old), miR-155-5p expression activity of fat tissue all had higher level in the three chicken breeds, but the expression activities in skeletal muscle of broilers were significantly lower than that of layer (P<0.05). In different development stages of Hy-Line Brown layer, miR-155-5p expression activities in skeletal muscle of 14-day-old and 10-month-old layers were significantly lower than that of 24-month-old layer (P<0.05). Fat related target genes (ACOX1, ACOT7, FADS1, SCD and HSD17B12) and skeletal muscle related target genes (CCNT2, DMD, CFL2, MAPK14, FLNB, ZBTB18 and CDK5) of miR-155-5p were predicted, respectively. The results indicate that miR-155-5p may be an important factor inhibiting the fat deposition and skeletal muscle development in chicken.

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