Brachydanio Rerio (Hamilton) and Danio Dangila (Hamilton) - Promising Species for Ornamental Fisheri
Structure, function, and temporal expression of Fibrillin in Danio rerio.
description
Transcript of Structure, function, and temporal expression of Fibrillin in Danio rerio.
![Page 1: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/1.jpg)
Structure, function, and temporal expression of Fibrillin in Danio rerio.
Christian Francom, Jeffrey Caldwell, Barbara Trask, Ph.D.
![Page 2: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/2.jpg)
Extracellular MatrixFunctions
Structure—tense or elasticCellular AdhesionCellular MotilityMolecular Reservoir
![Page 3: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/3.jpg)
Elastic Extracellular MatricesComponent parts
ElastinMicrofibrils
Fibrillins MAGPs Fibulins Other proteins and proteoglycans
Disorders involving the Elastic ECMElastin SVASMicrofibrils Marfan Syndrome
![Page 4: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/4.jpg)
QuestionHow do the microfibrillar proteins contribute
to the function of the elastic extracellular matrix?When and where is each expressed?What inter-molecular interactions are
maintained?
![Page 5: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/5.jpg)
Model OrganismZebrafish (Danio rerio)
External fertilization and development
Colorless during the first few days of development
Genome sequenced (although not assembled)
**first evolutionary organism to have functional elastic extracellular matrix
Easily maintainedhttp://www.pdn.cam.ac.uk/staff/lewis/zfish3AgesSlide.jpg
![Page 6: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/6.jpg)
Stages of Z-fish Development
![Page 7: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/7.jpg)
Sequences and PrimersFibrillin
atggcagcctgtcctgaattaatcttggaaagagagggaggagaaggagtggatgggggattctttaaggcgaagatgactaaggatttggtggaacgggagattcgcatcacagatgtgcagccggcaaatctgcagcaactgcgtgatgctgtcatctcaaaattggccaaaatctctgaggaatatttccagtaccttgttgaatctttgccacaaagaattgaggcatttgtgaaagcaaaaaggatgtccaacctggagggagccccaggctccaggatattctcagctcaggatgtgaacgagtgcaacgaccccaacacctgcatcagcggtctgtgtctgaacacccctggcagctacatctgcagctgcccccccgactttgagctcaatcccactggagtcggctgcgtcgacactcgctctggaaactgctacctggacgtccagtaccgaggcgatttctctgagagtctgagctgctccaatgagattggagtgggagtgtccaaagcgtcctgctgctgttctaggggtgtggcctggggacagccctgcgaggcctgtccatcactcaactccaagatctgtttcactgcacaacctgaaatctccagtctggtgcagggagagaccagccagtgcttccccaggaggagtatccagtgctgggacacattagcggctcattcccttgggcaagcctggagctcagatggccagctgcacgtttccactcttcatctttgttgtggtgtgaaagaggagaaggagggtttaggaaaacaaaaaagcagtggacagagtgatgttacaaagacaacaggcttcacacagagctctgacagtcgcgtcaaagcgagtgcgctgttattaaaatggtaccgtcccagtgacagctctcacagctttatttttgcagtgtcaggactacggtccaatcatgcaatagtagaggacatccgtctgttcaacagtaagcaatatggagctgagccgctgaaagaactgctcaaactactaccagaagcagatgaggtgaagagattgagagaattcaaaggagacgcaaataagctgacactggttgattccttcatgttccagctcattcaagtaccacgctttgatgtgcgtatcgaggtcatggtacttcaagaagagtttttcccattatgtactgccatgagtcgtgacatcaacattgtgcgccaggcaactgaagctgtgcctgtattagccttctctaagaaaccaatggctcatgaccttttagtcctgacaggtggctatgcaggaaacgctgtaggcttcaaattgtcctctcttctctctctggctgacacaaaggccaacaagcctggcatgaacctgctccattttgtggcattactctcgaggatcttctctgtgttttcacagggtggctatgcaggaaacgctgtaggcttcaaattgtcctctcttctctctctggctgacacaaaggccaacaagcctggcatgaacctgctccattttgtggcattagaagcacagaaaaaggatgaggcacttttaaagttcccagagaagctaacacatattcagagtgcagccagaatttcagtagaaaatatagaaggagagttttcctcactctatgtgaaaacacggtctttggaacagaagattcaagatgacgatgagctgcaaaagcagctagatccatttttgcagagctctgcaaaagccttgcaggaccttaagcaccgcaggctggacttgcgcaaagagggtaatgccctcattgacttcttctgtgaagacaaagacaccttcaaattggatgagtgcttcagaatcttccaagatttctgtctaaagttcaaaaaagctgtgaaggacaactcggacagagaactgaaagaggaagctagacagcgccgcttacgggaactagaggagagacggtgtgcctggggtgcagccaatattgaaacaggtggtaatggaggttttggtcgcagcagcagtgaaaatgatgttgacatcttaaccaaagatggcctcctagacttcctgcttcaatcaagacctcaaagcccccacagtcctttaggacgctcagctagtgctcgccgctaccaccacacatctagtgaccgcagcctccgtggctatatggagctggtctctgggtcttcagcaagtgaatacgccaaattcagcagcctgccacgttcaagcaggaaccatcaaagaaagaccatgccatggcttgtcttgcaggatgataatagagagttgattccacaaagtcaggtttaccaagagaagttagcagtttcacaaagggttgagactgaaccaatcagtcctttggcaagatactcaaatttaggctacaacttgaacagtgatccctatagtaataataataacaattatagttctctttctgaaggtggtcttacacctcgattaaataaaacaaagaatgtctttcaggggacatctggtcaaggcattggacacatgaatgtttctgttgagaaacgcaaactagtgcctgggcttcaaccctttgagataaccagccagatcaataataacagtcatgtttctttggacaatcaacatgaagtgtgtgtgactgatctggaacgagaagaccttcccaaagcttttgttcttgatacaccaccatctagtagaagttctgaggaggggcataagctagaaaaagtatggaatgacaaaatcatctcatcacagagagacgaagcagacacaagtaccttctcttcaacatcttgtgacaccccattgccccttgatccctccagtaaaaaacctgtgttttatgtcatggactgcactgaaacggattgttccattgtgttggactgttctgagattgagagttctcctattataaaagaaggacctggacttatcccaaagttacaggatgtcaactttccaagaatcattcaagacaccaattctctttcatccaattttgagtcttcagactgtcactttgtgtccacatcaagtgatgaaccatttgtaggaagggtcgatcaaacttcagaatcagtcagtccatctgtaacaactgaagaagcagagacagacagctgtgacacagctgaagggcaaagagttgatgagaaaggtgttcagacaggtagccccaaaacgacacttatcaaaaccaagacttcatccaaagggatgactgctcacaccagccgtcctacaagaaccctcacagctactgagactcagaacatgcgaaaagtagtgcccattacaaggcaaaatcgatctagtagcagcatgagaaaggttgataaaccacctgtacatgagagctctgaacccagacgtccatttcgagaccaaagcatgccagccagaagaggcgagagacaaggtagacctacaagacattcaagccttccaccagaggatcctaggacaccaagaggttcgacaccaagcagctctcgttggggaagggactcaattcaacacaggccctccatcaagaaagccaatgccaaacctgttcgtaacattcctaaaccagcaccagaggaaaagatgtgtcgctctacaatgcgtgccttggctcaagctcaggctcaaggaacctcagaaggaggtaccgctgacagccctaacaccaccccaaaaagttcttcacctttgcctagctttgctcgcaacactgtggcatctacttctagatctaaaaaagacatgccatcagcaactgtaaccggaacaccatcaaagagcctatcgagaactagctctcagagattgcctggtggtagaacagagtctgtcagtagcgggcctttaccaaggagtgaggataaactaggaggctcgattaggagggttcaaagtgtaagagcttccagtcgaagcagtgacacaccttcacaatctggtcgggatcactcccttaaaagtagcagtttctcagacagatccacaccaactagagacaccatcatctccagcaggaccagcaaacccacatggaaatag
MAGPatgagactctgttacctgcttctgctgtcactgccgggtctgctggctcaggtgcagaactatgatgaggattatacagacatcttcggtaacagcaacaactacgacaaccccttcttctaccagcagcagatattacagacgcagacgtcagggcctgaacagaacgcgctggatggacagacagagccgactgaacctggaccgctggactgtcgtgaggagcagtatccctgcactcggctctattctgtccatcagccgtgtaaacagtgtctgaacagcatctgcttctacagtctgcgcaggatgtatgtgatcaataaggaggtgtgtgtgcggacggtgtgtgcacatgaggagctcttgagagcggatctgtgtcgtgaccagttcccccgctgtggtgtaatggcgcacattggccagtgtggggcgatggggagcaactgcgccaggagttgtggatcctgctaa
![Page 8: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/8.jpg)
Expression DataFibrillin MAGP
Fibrillin
Actin Control
MAGP
0
50
100
150
200
8 Som 21 Som 24 hpf 35 hpf 52 hpf 75 hpf 96 hpf 120 hpf
Developmental Timepoint
Rela
tive
Inte
nsity
Fibrillin
0
50
100
150
200
8 Som 21Som
24 hpf 35 hpf 52 hpf 75 hpf 96 hpf 120 hpf
Developmental Timepoint
Aver
age
Rela
tive
Inte
nsity
MAGPGAPDH Control
![Page 9: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/9.jpg)
Future PossibilitiesMore replicates should be done
Analysis with Real-Time PCRAlternative primers should be madeIn situ hybridization should localize
expression
![Page 10: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/10.jpg)
QuestionHow do the microfibrillar proteins contribute
to the function of the elastic extracellular matrix?When and where is each expressed?What inter-molecular interactions are
maintained?
![Page 11: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/11.jpg)
Intermolecular interactionsFibrillin interacts with: MAGP interacts with:
MAGP CSPG LTGF-β Others?
Fibrillins CSPG Elastin Others?
![Page 12: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/12.jpg)
Mammalian Structure of Fibrillin-1
Fibrillin-1
3 5 6
![Page 13: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/13.jpg)
Isolated Bands of LTGF-β Domains
3 5 6
300bp200bp100bp
![Page 14: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/14.jpg)
LTGF-β Binding Domains….MAACPELILEREGGEGVDGGFFKAKMTKDLVEREIRITDVQPANLQQLRDAVISKLAKISEEYFQYLVESLPQRIEAFVKAKRMSNLEGAPGSRIFSAQDVNECNDPNTCISGLCLNTPGSYICSCPPDFELNPTGVGCVDTRSGNCYLDVQYRGDFSESLSCSNEIGVGVSKASCCCSRGVAWGQPCEACPSLNSKICFTAQPEISSLVQGETSQCFPRRSIQCWDTLAAHSLGQAWSSDGQLHVSTLHLCCGVKEEKEGLGKQKSSGQSDVTKTTGFTQSSDSRVKASALLLKWYRPSDSSHSFIFAVSGLRSNHAIVEDIRLFNSKQYGAEPLKELLKLLPEADEVKRLREFKGDANKLTLVDSFMFQLIQVPRFDVRIEVMVLQEEFFPLCTAMSRDINIVRQATEAVPVLAFSKKPMAHDLLVLTGGYAGNAVGFKLSSLLSLADTKANKPGMNLLHFVALLSRIFSVFSQGGYAGNAVGFKLSSLLSLADTKANKPGMNLLHFVALEAQKKDEALLKFPEKLTHIQSAARISVENIEGEFSSLYVKTRSLEQKIQDDDELQKQLDPFLQSSAKALQDLKHRRLDLRKEGNALIDFFCEDKDTFKLDECFRIFQDFCLKFKKAVKDNSDRELKEEARQRRLRELEERRCAWGAANIETGGNGGFGRSSSENDVDILTKDGLLDFLLQSRPQSPHSPLGRSASARRYHHTSSDRSLRGYMELVSGSSASEYAKFSSLPRSSRNHQRKTMPWLVLQDDNRELIPQSQVYQEKLAVSQRVETEPISPLARYSNLGYNLNSDPYSNNNNNYSSLSEGGLTPRLNKTKNVFQGTSGQGIGHMNVSVEKRKLVPGLQPFEITSQINNNSHVSLDNQHEVCVTDLEREDLPKAFVLDTPPSSRSSEEGHKLEKVWNDKIISSQRDEADTSTFSSTSCDTPLPLDPSSKKPVFYVMDCTETDCSIVLDCSEIESSPIIKEGPGLIPKLQDVNFPRIIQDTNSLSSNFESSDCHFVSTSSDEPFVGRVDQTSESVSPSVTTEEAETDSCDTAEGQRVDEKGVQTGSPKTTLIKTKTSSKGMTAHTSRPTRTLTATETQNMRKVVPITRQNRSSSSMRKVDKPPVHESSEPRRPFRDQSMPARRGERQGRPTRHSSLPPEDPRTPRGSTPSSSRWGRDSIQHRPSIKKANAKPVRNIPKPAPEEKMCRSTMRALAQAQAQGTSEGGTADSPNTTPKSSSPLPSFARNTVASTSRSKKDMPSATVTGTPSKSLSRTSSQRLPGGRTESVSSGPLPRSEDKLGGSIRRVQSVRASSRSSDTPSQSGRDHSLKSSSFSDRSTPTRDTIISSRTSKPTWK
![Page 15: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/15.jpg)
LTGF-β Binding Domains….MAACPELILEREGGEGVDGGFFKAKMTKDLVEREIRITDVQPANLQQLRDAVISKLAKISEEYFQYLVESLPQRIEAFVKAKRMSNLEGAPGSRIFSAQDVNECNDPNTCISGLCLNTPGSYICSCPPDFELNPTGVGCVDTRSGNCYLDVQYRGDFSESLSCSNEIGVGVSKASCCCSRGVAWGQPCEACPSLNSKICFTAQPEISSLVQGETSQCFPRRSIQCWDTLAAHSLGQAWSSDGQLHVSTLHLCCGVKEEKEGLGKQKSSGQSDVTKTTGFTQSSDSRVKASALLLKWYRPSDSSHSFIFAVSGLRSNHAIVEDIRLFNSKQYGAEPLKELLKLLPEADEVKRLREFKGDANKLTLVDSFMFQLIQVPRFDVRIEVMVLQEEFFPLCTAMSRDINIVRQATEAVPVLAFSKKPMAHDLLVLTGGYAGNAVGFKLSSLLSLADTKANKPGMNLLHFVALLSRIFSVFSQGGYAGNAVGFKLSSLLSLADTKANKPGMNLLHFVALEAQKKDEALLKFPEKLTHIQSAARISVENIEGEFSSLYVKTRSLEQKIQDDDELQKQLDPFLQSSAKALQDLKHRRLDLRKEGNALIDFFCEDKDTFKLDECFRIFQDFCLKFKKAVKDNSDRELKEEARQRRLRELEERRCAWGAANIETGGNGGFGRSSSENDVDILTKDGLLDFLLQSRPQSPHSPLGRSASARRYHHTSSDRSLRGYMELVSGSSASEYAKFSSLPRSSRNHQRKTMPWLVLQDDNRELIPQSQVYQEKLAVSQRVETEPISPLARYSNLGYNLNSDPYSNNNNNYSSLSEGGLTPRLNKTKNVFQGTSGQGIGHMNVSVEKRKLVPGLQPFEITSQINNNSHVSLDNQHEVCVTDLEREDLPKAFVLDTPPSSRSSEEGHKLEKVWNDKIISSQRDEADTSTFSSTSCDTPLPLDPSSKKPVFYVMDCTETDCSIVLDCSEIESSPIIKEGPGLIPKLQDVNFPRIIQDTNSLSSNFESSDCHFVSTSSDEPFVGRVDQTSESVSPSVTTEEAETDSCDTAEGQRVDEKGVQTGSPKTTLIKTKTSSKGMTAHTSRPTRTLTATETQNMRKVVPITRQNRSSSSMRKVDKPPVHESSEPRRPFRDQSMPARRGERQGRPTRHSSLPPEDPRTPRGSTPSSSRWGRDSIQHRPSIKKANAKPVRNIPKPAPEEKMCRSTMRALAQAQAQGTSEGGTADSPNTTPKSSSPLPSFARNTVASTSRSKKDMPSATVTGTPSKSLSRTSSQRLPGGRTESVSSGPLPRSEDKLGGSIRRVQSVRASSRSSDTPSQSGRDHSLKSSSFSDRSTPTRDTIISSRTSKPTWK
![Page 16: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/16.jpg)
Future DirectionsClone for expressionProtein binding studies
![Page 17: Structure, function, and temporal expression of Fibrillin in Danio rerio.](https://reader036.fdocuments.net/reader036/viewer/2022062501/56816332550346895dd3b880/html5/thumbnails/17.jpg)
AcknowledgementsJosh McBrideAric ElmerOUR