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Article ; Online: Rapid Generation of Pigment Free, Immobile Zebrafish Embryos and Larvae in Any Genetic Background Using CRISPR-Cas9 dgRNPs.

Davis, Andrew E / Castranova, Daniel / Weinstein, Brant M

2021 Volume 18, Issue 4, Page(s) 235–242

Abstract: The ability to carry out high-resolution, high-magnification optical imaging of living animals is one of the most attractive features of the zebrafish as a model organism. However, increasing amounts of pigmentation as development proceeds and ... ...

Abstract	The ability to carry out high-resolution, high-magnification optical imaging of living animals is one of the most attractive features of the zebrafish as a model organism. However, increasing amounts of pigmentation as development proceeds and difficulties in maintaining sustained immobilization of healthy, living animals remain challenges for live imaging. Chemical treatments can be used to suppress pigment formation and movement, but these treatments can lead to developmental defects. Genetic mutants can also be used to eliminate pigment formation and immobilize animals, but maintaining these mutants in lines carrying other combinations of transgenes and mutants is difficult and laborious. In this study, we show that CRISPR duplex guide ribonucleoproteins (dgRNPs) targeting the
MeSH term(s)	Animals ; CRISPR-Cas Systems ; Embryo, Nonmammalian ; Genetic Background ; Larva ; Pigmentation/genetics ; Ribonucleoproteins/genetics ; Zebrafish/genetics
Chemical Substances	Ribonucleoproteins
Language	English
Publishing date	2021-06-02
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Intramural
ZDB-ID	2156020-1
ISSN	1557-8542 ; 1545-8547
ISSN (online)	1557-8542
ISSN	1545-8547
DOI	10.1089/zeb.2021.0011
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Article ; Online: Live Imaging of Cutaneous Wound Healing after Rotary Tool Injury in Zebrafish.

Greenspan, Leah J / Ameyaw, Keith K / Castranova, Daniel / Mertus, Caleb A / Weinstein, Brant M

The Journal of investigative dermatology

2023 Volume 144, Issue 4, Page(s) 888–897.e6

Abstract: Cutaneous wounds are common afflictions that follow a stereotypical healing process involving hemostasis, inflammation, proliferation, and remodeling phases. In the elderly and those suffering from vascular or metabolic diseases, poor healing after ... ...

Abstract	Cutaneous wounds are common afflictions that follow a stereotypical healing process involving hemostasis, inflammation, proliferation, and remodeling phases. In the elderly and those suffering from vascular or metabolic diseases, poor healing after cutaneous injuries can lead to open chronic wounds susceptible to infection. The discovery of new therapeutic strategies to improve this defective wound healing requires a better understanding of the cellular behaviors and molecular mechanisms that drive the different phases of wound healing and how these are altered with age or disease. The zebrafish provides an ideal model for visualization and experimental manipulation of the cellular and molecular events during wound healing in the context of an intact, living vertebrate. To facilitate studies of cutaneous wound healing in zebrafish, we have developed an inexpensive, simple, and effective method for generating reproducible cutaneous injuries in adult zebrafish using a rotary tool. We demonstrate that our injury system can be used in combination with high-resolution live imaging to monitor skin re-epithelialization, immune cell recruitment and activation, and vessel regrowth in the same animal over time. This injury system provides a valuable experimental platform to study key cellular and molecular events during wound healing in vivo with unprecedented resolution.
MeSH term(s)	Animals ; Adult ; Humans ; Aged ; Zebrafish ; Skin/diagnostic imaging ; Skin/injuries ; Wound Healing/physiology ; Re-Epithelialization ; Inflammation
Language	English
Publishing date	2023-11-17
Publishing country	United States
Document type	Journal Article
ZDB-ID	80136-7
ISSN	1523-1747 ; 0022-202X
ISSN (online)	1523-1747
ISSN	0022-202X
DOI	10.1016/j.jid.2023.10.015
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Article ; Online: Anatomy and development of the pectoral fin vascular network in the zebrafish.

Paulissen, Scott M / Castranova, Daniel M / Krispin, Shlomo M / Burns, Margaret C / Menéndez, Javier / Torres-Vázquez, Jesús / Weinstein, Brant M

Development (Cambridge, England)

2022 Volume 149, Issue 5

Abstract: The pectoral fins of teleost fish are analogous structures to human forelimbs, and the developmental mechanisms directing their initial growth and patterning are conserved between fish and tetrapods. The forelimb vasculature is crucial for limb function, ...

Abstract	The pectoral fins of teleost fish are analogous structures to human forelimbs, and the developmental mechanisms directing their initial growth and patterning are conserved between fish and tetrapods. The forelimb vasculature is crucial for limb function, and it appears to play important roles during development by promoting development of other limb structures, but the steps leading to its formation are poorly understood. In this study, we use high-resolution imaging to document the stepwise assembly of the zebrafish pectoral fin vasculature. We show that fin vascular network formation is a stereotyped, choreographed process that begins with the growth of an initial vascular loop around the pectoral fin. This loop connects to the dorsal aorta to initiate pectoral vascular circulation. Pectoral fin vascular development continues with concurrent formation of three elaborate vascular plexuses, one in the distal fin that develops into the fin-ray vasculature and two near the base of the fin in association with the developing fin musculature. Our findings detail a complex, yet highly choreographed, series of steps involved in the development of a complete, functional, organ-specific vascular network.
MeSH term(s)	Animal Fins/anatomy & histology ; Animal Fins/growth & development ; Animals ; Zebrafish/anatomy & histology ; Zebrafish/growth & development
Language	English
Publishing date	2022-03-04
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Intramural
ZDB-ID	90607-4
ISSN	1477-9129 ; 0950-1991
ISSN (online)	1477-9129
ISSN	0950-1991
DOI	10.1242/dev.199676
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Article ; Online: Long-term imaging of living adult zebrafish.

Castranova, Daniel / Samasa, Bakary / Venero Galanternik, Marina / Gore, Aniket V / Goldstein, Allison E / Park, Jong S / Weinstein, Brant M

Development (Cambridge, England)

2022 Volume 149, Issue 4

Abstract: The zebrafish has become a widely used animal model due, in large part, to its accessibility to and usefulness for high-resolution optical imaging. Although zebrafish research has historically focused mostly on early development, in recent years the fish ...

Abstract	The zebrafish has become a widely used animal model due, in large part, to its accessibility to and usefulness for high-resolution optical imaging. Although zebrafish research has historically focused mostly on early development, in recent years the fish has increasingly been used to study regeneration, cancer metastasis, behavior and other processes taking place in juvenile and adult animals. However, imaging of live adult zebrafish is extremely challenging, with survival of adult fish limited to a few tens of minutes using standard imaging methods developed for zebrafish embryos and larvae. Here, we describe a new method for imaging intubated adult zebrafish using a specially designed 3D printed chamber for long-term imaging of adult zebrafish on inverted microscope systems. We demonstrate the utility of this new system by nearly day-long observation of neutrophil recruitment to a wound area in living double-transgenic adult casper zebrafish with fluorescently labeled neutrophils and lymphatic vessels, as well as intubating and imaging the same fish repeatedly. We also show that Mexican cavefish can be intubated and imaged in the same way, demonstrating this method can be used for long-term imaging of adult animals from diverse aquatic species.
MeSH term(s)	Animals ; Animals, Genetically Modified/immunology ; Animals, Genetically Modified/metabolism ; Embryo, Nonmammalian/anatomy & histology ; Embryo, Nonmammalian/metabolism ; Larva/anatomy & histology ; Larva/growth & development ; Larva/metabolism ; Luminescent Proteins/genetics ; Luminescent Proteins/metabolism ; Microscopy, Fluorescence/methods ; Neutrophils/cytology ; Neutrophils/immunology ; Neutrophils/pathology ; Printing, Three-Dimensional ; Time-Lapse Imaging ; Zebrafish/anatomy & histology ; Zebrafish/growth & development ; Zebrafish/metabolism
Chemical Substances	Luminescent Proteins
Language	English
Publishing date	2022-02-21
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Intramural
ZDB-ID	90607-4
ISSN	1477-9129 ; 0950-1991
ISSN (online)	1477-9129
ISSN	0950-1991
DOI	10.1242/dev.199667
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Article ; Online: Maternal control of visceral asymmetry evolution in Astyanax cavefish.

Ma, Li / Ng, Mandy / Shi, Janet / Gore, Aniket V / Castranova, Daniel / Weinstein, Brant M / Jeffery, William R

Scientific reports

2021 Volume 11, Issue 1, Page(s) 10312

Abstract: The direction of visceral organ asymmetry is highly conserved during vertebrate evolution with heart development biased to the left and pancreas and liver development restricted to opposing sides of the midline. Here we show that reversals in visceral ... ...

Abstract	The direction of visceral organ asymmetry is highly conserved during vertebrate evolution with heart development biased to the left and pancreas and liver development restricted to opposing sides of the midline. Here we show that reversals in visceral organ asymmetry have evolved in Astyanax mexicanus, a teleost species with interfertile surface-dwelling (surface fish) and cave-dwelling (cavefish) forms. Visceral organ asymmetry is conventional in surface fish but some cavefish have evolved reversals in heart, liver, and pancreas development. Corresponding changes in the normally left-sided expression of the Nodal-Pitx2/Lefty signaling system are also present in the cavefish lateral plate mesoderm (LPM). The Nodal antagonists lefty1 (lft1) and lefty2 (lft2), which confine Nodal signaling to the left LPM, are expressed in most surface fish, however, lft2, but not lft1, expression is absent during somitogenesis of most cavefish. Despite this difference, multiple lines of evidence suggested that evolutionary changes in L-R patterning are controlled upstream of Nodal-Pitx2/Lefty signaling. Accordingly, reciprocal hybridization of cavefish and surface fish showed that modifications of heart asymmetry are present in hybrids derived from cavefish mothers but not from surface fish mothers. The results indicate that changes in visceral asymmetry during cavefish evolution are influenced by maternal genetic effects.
MeSH term(s)	Animals ; Biological Evolution ; Body Patterning ; Characidae/embryology ; Female
Language	English
Publishing date	2021-05-13
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-021-89702-6
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Article: Comparison of Juvenile Feed Protocols on Growth and Spawning in Zebrafish.

Frederickson, Stephen C / Steinmiller, Mark D / Blaylock, Tiffany Rae / Wisnieski, Mike E / Malley, James D / Pandolfo, Lauren M / Castranova, Daniel

Journal of the American Association for Laboratory Animal Science : JAALAS

2021 Volume 60, Issue 3, Page(s) 298–305

Abstract: Over the past 2 decades, zebrafish, ...

Abstract	Over the past 2 decades, zebrafish,
MeSH term(s)	Animal Feed ; Animals ; Artemia ; Diet/veterinary ; Fertility ; Reproduction ; Zebrafish
Language	English
Publishing date	2021-03-02
Publishing country	United States
Document type	Journal Article
ISSN	1559-6109
ISSN	1559-6109
DOI	10.30802/AALAS-JAALAS-20-000105
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Article ; Online: Publisher Correction: Maternal control of visceral asymmetry evolution in Astyanax cavefish.

Ma, Li / Ng, Mandy / Shi, Janet / Gore, Aniket V / Castranova, Daniel / Weinstein, Brant M / Jeffery, William R

Scientific reports

2021 Volume 11, Issue 1, Page(s) 12934

Language	English
Publishing date	2021-06-15
Publishing country	England
Document type	Published Erratum
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-021-92148-5
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Article ; Online: Live Imaging of Intracranial Lymphatics in the Zebrafish.

Castranova, Daniel / Samasa, Bakary / Venero Galanternik, Marina / Jung, Hyun Min / Pham, Van N / Weinstein, Brant M

Circulation research

2020 Volume 128, Issue 1, Page(s) 42–58

Abstract: Rationale: The recent discovery of meningeal lymphatics in mammals is reshaping our understanding of fluid homeostasis and cellular waste management in the brain, but visualization and experimental analysis of these vessels is challenging in mammals. ... ...

Abstract	Rationale: The recent discovery of meningeal lymphatics in mammals is reshaping our understanding of fluid homeostasis and cellular waste management in the brain, but visualization and experimental analysis of these vessels is challenging in mammals. Although the optical clarity and experimental advantages of zebrafish have made this an essential model organism for studying lymphatic development, the existence of meningeal lymphatics has not yet been reported in this species. Objective: Examine the intracranial space of larval, juvenile, and adult zebrafish to determine whether and where intracranial lymphatic vessels are present. Methods and results: Using high-resolution optical imaging of the meninges in living animals, we show that zebrafish possess a meningeal lymphatic network comparable to that found in mammals. We confirm that this network is separate from the blood vascular network and that it drains interstitial fluid from the brain. We document the developmental origins and growth of these vessels into a distinct network separated from the external lymphatics. Finally, we show that these vessels contain immune cells and perform live imaging of immune cell trafficking and transmigration in meningeal lymphatics. Conclusions: This discovery establishes the zebrafish as a important new model for experimental analysis of meningeal lymphatic development and opens up new avenues for probing meningeal lymphatic function in health and disease.
MeSH term(s)	Animals ; Animals, Genetically Modified ; Lymphangiogenesis/drug effects ; Lymphatic Vessels/drug effects ; Lymphatic Vessels/immunology ; Lymphatic Vessels/physiology ; Meninges/immunology ; Meninges/physiology ; Microscopy, Confocal ; Neutrophil Infiltration ; Neutrophils/immunology ; Optical Imaging ; Vascular Endothelial Growth Factor C/pharmacology ; Zebrafish/genetics
Chemical Substances	VEGFC protein, human ; Vascular Endothelial Growth Factor C
Language	English
Publishing date	2020-11-02
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Intramural ; Video-Audio Media
ZDB-ID	80100-8
ISSN	1524-4571 ; 0009-7330 ; 0931-6876
ISSN (online)	1524-4571
ISSN	0009-7330 ; 0931-6876
DOI	10.1161/CIRCRESAHA.120.317372
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Article ; Online: Clearing for Deep Tissue Imaging.

Muntifering, Michael / Castranova, Daniel / Gibson, Gregory A / Meyer, Evan / Kofron, Matthew / Watson, Alan M

Current protocols in cytometry

2018 Volume 86, Issue 1, Page(s) e38

Abstract: Biologic tissues are generally opaque due to optical properties that result in scattering and absorption of light. Preparation of tissues for optical microscopy often involves sectioning to a thickness of 50-100 µm, the practical limits of light ... ...

Abstract	Biologic tissues are generally opaque due to optical properties that result in scattering and absorption of light. Preparation of tissues for optical microscopy often involves sectioning to a thickness of 50-100 µm, the practical limits of light penetration and recovery. A researcher who wishes to image a whole tissue must acquire potentially hundreds of individual sections before rendering them into a three-dimensional volume. Clearing removes strongly light-scattering and light-absorbing components of a tissue and equalizes the refractive index of the imaging medium to that of the tissue. After clearing, the maximum depth of imaging is often defined by the microscope optics rather than the tissue. Such visibility enables the interrogation of whole tissues and even animals without the need to section. Researchers can study a biological process in the context of its three-dimensional environment, identify rare events in large volumes of tissues, and trace cells and cell-cell interactions over large distances. This article describes four popular clearing protocols that are relevant to a wide variety of scenarios across biologic disciplines: CUBIC, CLARITY, 3DISCO, and SeeDB. © 2018 by John Wiley & Sons, Inc.
MeSH term(s)	Animals ; Decision Trees ; Fluorescence ; Imaging, Three-Dimensional/methods ; Mice ; Solvents ; Staining and Labeling
Chemical Substances	Solvents
Language	English
Publishing date	2018-07-13
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, N.I.H., Intramural
ISSN	1934-9300
ISSN (online)	1934-9300
DOI	10.1002/cpcy.38
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Article ; Online: MicroRNA-mediated control of developmental lymphangiogenesis.

Jung, Hyun Min / Hu, Ciara T / Fister, Alexandra M / Davis, Andrew E / Castranova, Daniel / Pham, Van N / Price, Lisa M / Weinstein, Brant M

eLife

2019 Volume 8

Abstract: The post-transcriptional mechanisms contributing to molecular regulation of developmental lymphangiogenesis and lymphatic network assembly are not well understood. MicroRNAs are important post-transcriptional regulators during development. Here, we use ... ...

Abstract	The post-transcriptional mechanisms contributing to molecular regulation of developmental lymphangiogenesis and lymphatic network assembly are not well understood. MicroRNAs are important post-transcriptional regulators during development. Here, we use high throughput small RNA sequencing to identify miR-204, a highly conserved microRNA dramatically enriched in lymphatic vs. blood endothelial cells in human and zebrafish. Suppressing miR-204 leads to loss of lymphatic vessels while endothelial overproduction of miR-204 accelerates lymphatic vessel formation, suggesting a critical positive role for this microRNA during developmental lymphangiogenesis. We also identify the NFATC1 transcription factor as a key miR-204 target in human and zebrafish, and show that NFATC1 suppression leads to lymphatic hyperplasia. The loss of lymphatics caused by miR-204 deficiency can be largely rescued by either endothelial autonomous expression of miR-204 or by suppression of NFATC1. Together, our results highlight a miR-204/NFATC1 molecular regulatory axis required for proper lymphatic development.
MeSH term(s)	Animals ; Endothelial Cells/physiology ; Gene Expression Regulation, Developmental ; Humans ; Lymphangiogenesis ; MicroRNAs/metabolism ; NFATC Transcription Factors/metabolism ; Zebrafish
Chemical Substances	MIRN204 microRNA, human ; MicroRNAs ; NFATC Transcription Factors
Language	English
Publishing date	2019-09-03
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Intramural
ZDB-ID	2687154-3
ISSN	2050-084X ; 2050-084X
ISSN (online)	2050-084X
ISSN	2050-084X
DOI	10.7554/eLife.46007
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