New preprint: ScreenCube: A 3D Printed System For Rapid and Cost-Effective Chemical Screening in Adult Zebrafish

Adrian’s paper “ScreenCube: A 3D Printed System For Rapid and Cost-Effective Chemical Screening in Adult Zebrafish” is now available on BioRxiv! Click here for the preprint.

Phenotype-based small molecule screens in zebrafish embryos and larvae have been successful in accelerating pathway and therapeutic discovery for diverse biological processes. Yet, the application of chemical screens to adult physiologies has been relatively limited due to additional demands on cost, space, and labor associated with drug administration in adult animals. Here, we present a 3D printed system and methods for intermittent drug dosing that enable rapid and cost-effective chemical administration in adult zebrafish. Using pre-filled screening plates, the system enables dosing of 96 fish in ~3 min, with a tenfold-reduction in drug quantity compared to that used in previous chemical screens in adult zebrafish. We characterize water quality kinetics during immersion in the system, and use these kinetics to rationally design intermittent dosing regimens that result in 100% fish survival. As a demonstration of system fidelity, we show the potential to identify two known chemical inhibitors of adult tail fin regeneration, cyclopamine and dorsomorphin. By developing methods for rapid and cost-effective chemical administration in adult zebrafish, this study expands the potential for small molecule discovery in post-embryonic models of development, disease, and regeneration.

New preprint: microCT-Based Skeletal Phenomics in Zebrafish Reveals Virtues of Deep Phenotyping at the Whole-Organism Scale

We have just submitted a preprint of our manuscript, “microCT-Based Skeletal Phenomics in Zebrafish Reveals Virtues of Deep Phenotyping at the Whole-Organism Scale”, on BioRxiv. Click here for the preprint.

Phenomics — in-depth phenotyping at the whole-organism scale — holds promise to enhance our fundamental understanding of genes and genomic variation, yet methods in vertebrates are limited. Here, we demonstrate rapid whole-body profiling of hundreds of traits in the axial skeleton of adult zebrafish. We show the potential for vertebral patterns to confer heightened sensitivity, with similar specificity, in discriminating mutant populations compared to analyzing individual vertebrae in isolation, even when the latter is performed at higher resolution. We identify phenotypes associated with human brittle bone disease and thyroid stimulating hormone receptor hyperactivity. Finally, we develop allometric models and show their potential to discriminate mutant phenotypes masked by growth alterations in growth. Our studies demonstrate virtues of whole-body phenomic pattern analysis in a single organ system. The high sensitivity may increase productivity in genetic screens, and facilitate the study genetic variants of smaller effect size, such as those that underlie complex diseases.

New publication: Neuron subset-specific Pten deletion induces abnormal skeletal activity in mice

We are proud to announce that a paper derived from our collaborative project with the Lugo lab has just been published in Experimental Neurology! Click here for the paper.

Individuals with a history of epilepsy are at higher risk for bone fractures compared to the general population. Although clinical studies support an association between low bone mineral density (BMD) and anti-seizure medications, little is known on whether a history of seizures is linked to altered bone health. Therefore, in this study we tested the hypothesis that bone mass, morphology, and bone mineralization are altered by seizures in genetically epileptic animals and in animals subjected to an episode of status epilepticus. In this study, we used NS-Pten conditional knockout mice (a well-studied genetic model of epilepsy). We used microCT analysis to measure BMD, morphology, and mineralization in NS-Pten+/+ (wildtype) and NS-Pten-/- (knockout) mice at 4 and 8weeks, as well as adult Kv4.2+/+ and Kv4.2-/- mice. We measured BMD, bone morphology, and mineralization in adult NS-Pten+/+ mice that received status epilepticus through kainic acid (20mg/kg intraperitoneal). Further, we measured locomotion for NS-Pten+/+ and NS-Pten-/- mice at 4 and 6weeks. We found that NS-Pten-/- mice exhibited low BMD in the tibial metaphysis and midshaft compared to non-epileptic mice. Morphologically, NS-Pten-/- mice exhibited decreased trabecular volume fraction, and endocortical expansion in both the metaphyeal and diaphyseal compartments. In the midshaft, NS-Pten-/- mice exhibited reduced tissue mineral density, indicating impaired mineralization in addition to morphological deficits. NS-Pten-/- mice exhibited hyperactivity in open field testing, suggesting low bone mass in NS-Pten-/- mice was not attributable to hypoactivity. Differences in BMD were not observed following kainate-induced seizures or in the Kv4.2-/- model of seizure susceptibility. Our findings suggest that deletion of Pten in the brain results in impaired bone mass and mineralization, which may contribute to weaker bones and thereby a higher fracture risk.

FishBone 2017

We are proud to help co-organize FishBone 2017!

FishBone 2017 will be held the day prior to the ECTS Congress at the Salzburg Congress Center on May 12th, 2017 as part of the ECTS Pre-Congress Program. The workshop will be a day-long event open to clinicians, biologists and geneticists dedicated to bone research, with a focus on fish models of human skeletal disease. The workshop seeks to encour­age inter­ac­tion between estab­lished and poten­tial fish users and bone researchers from different disciplines. All presenters are encouraged to provide background to bring participants to a basic level of understanding, as well as to present new data.

Attendance of the ECTS Congress is not required for attendance of the FishBone Workshop. However, those attending the full ECTS Congress will have their FishBone abstracts automatically included into the ECTS program, and be able to present them in a ECTS poster cluster dedicated to fish skeletal models. This exciting step represents the first “official” inclusion of fish models as an independent component at a major bone society meeting.

Abstract submission is now open, and will end on February 22, 2017. Online registration, including options for hotel accomodations, will open on February 15, 2017. Please see the fishbone website (http://blogs.uw.edu/fishbone/) for more details.

We look forward to seeing you Austria!

Welcome to Charlotte Gistelinck

IMG_5764We are thrilled to welcome to Charlotte Gistelinck! Charlotte is a visiting PhD student from Ghent University in Belgium. Her work is focused on generating and characterizing zebrafish models of Osteogenesis Imperfecta. Charlotte will be here for the next six months as part of the UW Visit Program.

Claire Watson wins ASBMR Young Investigator Travel Grant

Headshot Option for RonCongrats to Claire Watson, who won an ASBMR Young Investigator Travel Grant for her abstract entitled “Multi-Modal High-Content Imag­ing Reveals Rela­tion­ships Between Cell Sig­nal­ing and Min­er­al­iza­tion in Zebrafish“. The ASBMR awards these grants to the top Young Investigators who are selected to present an oral or plenary poster presentation at the ASBMR Annual Meeting. Congrats Claire!

 

FishBone 2015

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Come join us for FishBone 2015!

FishBone is an international workshop dedicated to advancing zebrafish, medaka, and other laboratory fish models for skeletal research. The inaugural FishBone Workshop will be held on October 8th, 2015 in Seattle the day prior to the ASBMR Annual Meeting. FishBone will bring together estab­lished and poten­tial fish users and ‎musculoskeletal researchers from dif­fer­ent dis­ci­plines to present up-to-date find­ings, fos­ter col­lab­o­ra­tions, ‎and broaden awareness.‎ We have organized an outstanding program on the broad topics of anatomy, genet­ics, tools/methodologies, and trans­la­tional approaches. Keynote speakers include P. Eckhard Witten (Ghent University), Thomas Schilling (UC Irvine), Shan­non Fisher (Uni­ver­sity of Penn­syl­va­nia), and Matthew Harris (Har­vard Med­ical School).

Please visit the FishBone website (http://blogs.uw.edu/fishbone) for more information.

MSBL @ 2015 ASBMR

Congratulations to Claire Watson, Adrian Monstad-Rios, and Brenen Wynd, who had their abstracts accepted for presentation as the 2015 ASBMR Annual Meeting in Seattle! Collectively, the MSBL will present five abstracts at the meeting. Information for each is abstract is given below.

Abstract Title: Multi-Modal High-Content Imaging Reveals Relationships Between Cell Signaling and Mineralization in Zebrafish
Authors: Claire Watson, Edith Gardiner, Werner Kaminsky, Ronald Kwon
Presentation Number: FR0190 / SA0190
Presentation Type: Plenary Sessions / Poster Sessions
Session: Welcome Reception & Plenary Poster Session / Poster Session I & Poster Tours
Session Date/Time: 10/09/15 5:30 PM – 7:00 PM / 10/10/15 12:30 PM – 2:30 PM

Abstract Title: An Optimized In Vivo Chemical Screening Regimen For Osteoactive Compound Discovery in the Regenerating Zebrafish Tail Fin
Authors: Adrian Monstad-Rios, Ronald Kwon
Presentation Number: MO0105
Presentation Type: Poster Sessions
Session: Poster Session III & Poster Tours
Session Date/Time: 10/12/15 12:30 PM – 2:30 PM

Abstract Title: A Dynamic Anesthesia System For Long-Term Imaging in the Adult Zebrafish Skeleton
Authors: Brenen Wynd, Karuna Patil, Claire Watson, George Sanders, Ronald Kwon
Presentation Number:
Presentation Type: Poster Sessions
Session: Poster Session & Poster Tours
Session Date/Time:

Abstract Title: Conserved Dynamics in Genes Associated with Human BMD and Bone Disorders During Zebrafish and Rat Bone Formation
Authors: Leah Worton, Arden Chew, Claire Watson, Edith Gardiner, Dobrawa Naperiala, Amarjit Virdi, D. Rick Sumner, Cole Trapnell, Yi-Hsiang Hsu, Ronald Kwon
Presentation Number: SU0103
Presentation Type: Poster Sessions
Session: Poster Session II & Poster Tours
Session Date/Time: 10/11/15 12:30 PM – 2:30 PM
Presentation Time: 12:30 PM – 2:30 PM

Abstract Title: MicroCT-Based Barcoding Reveals Novel High Bone Mass Mutations in Zebrafish
Authors: Philippe Huber, Jane Lee, Claire Watson, Marjorie Thompson, Sarah McMenamin, David Parichy, Ronald Kwon
Presentation Number: MO0109
Presentation Type: Poster Sessions
Session: Poster Session III & Poster Tours
Session Date/Time: 10/12/15 12:30 PM – 2:30 PM
Presentation Time: 12:30 PM – 2:30 PM

Welcome to Billy Crutcher and Brandon Douglass

MSBL welcomes Billy Crutcher and Brandon Douglass. Billy and Brandon are Medical Students working in the lab this Summer. Billy is working with Claire Watson on a project examining the utility of fin regeneration as a model of human genetic disorders. Brandon is a UW MSRTP Student working on a collaborative project with Dr. Christopher Allan. Welcome aboard guys!

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Bill Crutcher
Medical Student, Class of 2018
Sidney Kimmel Medical College at Thomas Jefferson University (Philadelphia)
B.A., Mathematics, Boston College (2013)
Email: wxc023@jefferson.edu

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Brandon Douglass
Medical Student
B.S., Physiology, University of Wyoming (2013)
Email: bdoug@uw.edu

 

Career Development Award

Ron was awarded a five year NIH K01 Career Development Award from NIAMS ($614,250) for the project entitled “Neuroskeletal Systems Biology in Zebrafish”. The goal of this project is to integrate zebrafish knockdown and screening strategies with quantitative bone imaging to identify novel neural regulators of bone growth and mineralization. Many thanks to Drs. Ted Gross, Chuck Kimmel, Randall Moon, Bill Cresko, Edith Gardiner, and Steve Bain, as well as MSBL members for their efforts on this project.

Adrian Monstad-Rios awarded Undergraduate Travel Award

IMG_0216Congrats to Adrian, who was awarded a UW Undergraduate Travel Award to present his research at the 2015 Annual Meeting of the Orthopaedic Research Society (ORS). His abstract, entitled “Development and Validation of a 3D Printed Chemical Screening System for Osteoactive Compound Discovery in the Regenerating Zebrafish Tail Fin”, was one of only four to be selected for the Bone Biology Walking Tour on 3/29/15.

Welcome to Claire Watson

Headshot Option for RonWe are happy to announce a new lab mem­ber, Claire Wat­son. Claire comes to us from the lab of Bruce Tem­pel, where she received her Ph.D. in 2012 from the UW Depart­ment of Phar­ma­col­ogy. She is an expert in audi­tory genet­ics, and brings a unique per­spec­tive with her expe­ri­ence in mol­e­c­u­lar biol­ogy, chem­istry, genet­ics, and imag­ing. Wel­come aboard!

Write up in IBMS BoneKEy

Two of our zebrafish abstracts from the 2014 ASBMR Annual Meeting got written up in a BoneKEy report on skeletal genomics! An excerpt from the article is below:

“Also, at this year’s meeting, a new animal model was introduced into the musculoskeletal (MSK) genetics field—a zebrafish (Danio rerio). Regenerating zebrafish caudal fin is a well-known model of post-amputation’s de novo membranous bone formation (however, it was not widely studied by the bone experts). Ronald Kwon of University of Washington used a transcriptomic analysis to reveal conserved osteogenic signatures during tail fin regeneration. Both he and a group from Harvard presented a technique of quantitative micro-computed tomography (μCT) analysis of zebrafish bone.”

 

 

Adrian and Ron to present at 2015 ORS Annual Meeting

Great news, the MSBL will be presenting two abstracts at the 2015 ORS Annual Meeting in Las Vegas, including a Spotlight Session talk. See below for more info.

POSTER SESSION #: PS1-029
POSTER #: 0537
POSTER TITLE: Development and Validation of a 3D Printed Chemical Screening System for Osteoactive Compound Discovery in the Regenerating Zebrafish Tail Fin
AUTHORS: Adrian T. Monstad-Rios, Ronald Y. Kwon.
University of Washington, Seattle, WA, USA.

SPOTLIGHT SESSION DATE & TIME: Monday Mar 30 2015 8:00 AM – 9:00 AM
PAPER #: 0147
PAPER TITLE: Cross-Species Transcriptomic Analysis Reveals Homologous Transcriptional Stages and Conserved Gene Co-Expression Dynamics During Zebrafish and Rat Bone Regeneration
AUTHORS: Ronald Y. Kwon1, Amarjit Virdi2, D. Rick Sumner2.
1University of Washington, Seattle, WA, USA
2Rush University Medical Center, Chicago, IL, USA.

 

Research featured in Medscape Medical News

Our work investigating the connection between epilepsy and bone (presented at the 2014 ASBMR Meeting) was recently featured in Medscape Medical News! The article highlights a study by the Misra group demonstrating an association between autism (which has high co-morbidity with epilepsy), low bone mass, and increased fracture risk. Click here to download a pdf of the article.

Zebrafish study featured as “Paper to Watch” in IBMS BoneKEy

Great news! Our recent JBMR paper (currently in press) just got written up in the “Watch” section of IBMS BoneKEy. In the article, the editor states that:

“The zebrafish model of bone regeneration is emerging as an important contributor to studies of new bone formation in mammalian species and humans, since the pathways involved appear to be highly conserved.”

We couldn’t agree more, and are thrilled to be part of efforts to help advance this system for our field. Click here to read the article.

Marjorie, Leah, and Ron to present at the 2014 ASMBR Annual Meeting

Congrats to Leah1 and Marjorie, who will be presenting their abstracts at the upcoming 2014 ASBMR Annual Meeting in Houston!

Leah1’s abstract is entitled “Sensory Neuron Differentiation Enhances Osteoblast Differentiation Through Soluble Factors”, and describes the development of a novel cell culture model of sensory neuron/bone cell interactions:

[SA0239] Sensory Neuron Differentiation Enhances Osteoblast Differentiation Through Soluble Factors
Leah Worton, Brandon Park, Anthony Redidoro, Edith Gardiner, Ronald Kwon
September 13, 12:30 PM – 02:30 PM
Session: Poster Session I & Poster Tours

Marjorie’s abstract describes a collaborative project with Joaquin Lugo’s lab at Baylor University is entitled “Brain-Specific PTEN Deletion Induces Abnormal Skeletal Activity in Mice”, and explores the role of cortical brain neurons in regulating skeletal activity.

[SA0181] Brain-Specific PTEN Deletion Induces Abnormal Skeletal Activity in Mice
Marjorie Thompson, Philippe Huber, Gregory Smith, Andrew Holley, Steven Bain, Edith Gardiner, Joaquin Lugo, Ronald Kwon
September 13, 12:30 PM – 02:30 PM
Session: Poster Session I & Poster Tours

Ron will also be presenting two abstracts. The first, entitled “Cross-Species Transcriptomic Analysis Reveals Conserved Osteogenic Signatures During Zebrafish and Rat Bone Regeneration” describes a collaborative project with Rick Sumner and Amarjit Viridi at Rush University in which we are exploring the potential utility of the regenerating zebrafish fin as rapid genetic platform for identifying conserved gene regulatory networks underlying mammalian osteogenesis. The second, entitled “Rotopol and MicroCT Imaging in the Regenerating Zebrafish Fin for BMD Therapeutic Discovery”, describes our ongoing collaboration with Werner Kaminsky in the Department of Chemistry in which we are developing Rotopol imaging-based modalities for high-throughput bone mineralization imaging in the regenerating zebrafish fin.

[SA0475] Rotopol and MicroCT Imaging in the Regenerating Zebrafish Fin for BMD Therapeutic Discovery
Ronald Kwon, Anthony Recidoro, Werner Kaminsky
September 13, 12:30 PM – 02:30 PM
Session: Poster Session I & Poster Tours

[SU0234] Cross-Species Transcriptomic Analysis Reveals Conserved Osteogenic Signatures During Zebrafish and Rat Bone Regeneration
Ronald Kwon, Amarjit Virdi, D. Rick Sumner
September 14, 12:30 PM – 02:30 PM
Session: Poster Session II & Poster Tours

Abstract to be presented at 2014 American Epilepsy Society Meeting

Congrats to the Lugo lab, whose abstract entitled “Neuron subset-specific-PTEN deletion induces abnormal skeletal activity in mice” received a podium presentation at the upcoming 2014 American Epilepsy Society Meeting in Seattle. The abstract reflects a collaborative project with Joaquin Lugo’s lab at Baylor University in which we are working understanding a potential link between cortical brain dysfunction (such that which arises during epilepsy) and impaired skeletal activity.

Marjorie Thompson and Anthony Recidoro to present at 2014 Undergraduate Research Symposium

Congrats to Marjorie and Anthony, who will be presenting their research at the 2014 UW Undergraduate Research Symposium. Marjorie will be giving a talk for her abstract “Brain-specific PTEN deletion induces epilepsy and abnormal bone formation in mice” in Session 2F: Brain Development, Injury, Regeneration and Restoration of Function (242 MGH, 3:30 PM to 5:00 PM). Anthony will be presenting his poster “Neuronal regulation of regenerative bone growth in the zebrafish fin” in Poster Session 4 (Commons East, Easel #48, 4:00 PM to 6:00 PM).