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Article ; Online: Whole Animal Imaging of Drosophila melanogaster using Microcomputed Tomography.

Schoborg, Todd A

Journal of visualized experiments : JoVE

2020 , Issue 163

Abstract: Biomedical imaging tools permit investigation of molecular mechanisms across spatial scales, from genes to organisms. Drosophila melanogaster, a well-characterized model organism, has benefited from the use of light and electron microscopy to understand ... ...

Abstract	Biomedical imaging tools permit investigation of molecular mechanisms across spatial scales, from genes to organisms. Drosophila melanogaster, a well-characterized model organism, has benefited from the use of light and electron microscopy to understand gene function at the level of cells and tissues. The application of imaging platforms that allow for an understanding of gene function at the level of the entire intact organism would further enhance our knowledge of genetic mechanisms. Here a whole animal imaging method is presented that outlines the steps needed to visualize Drosophila at any developmental stage using microcomputed tomography (µ-CT). The advantages of µ-CT include commercially available instrumentation and minimal hands-on time to produce accurate 3D information at micron-level resolution without the need for tissue dissection or clearing methods. Paired with software that accelerate image analysis and 3D rendering, detailed morphometric analysis of any tissue or organ system can be performed to better understand mechanisms of development, physiology, and anatomy for both descriptive and hypothesis testing studies. By utilizing an imaging workflow that incorporates the use of electron microscopy, light microscopy, and µ-CT, a thorough evaluation of gene function can be performed, thus furthering the usefulness of this powerful model organism.
MeSH term(s)	Animals ; Drosophila melanogaster/growth & development ; Imaging, Three-Dimensional ; Software ; Whole Body Imaging/methods ; X-Ray Microtomography/methods
Language	English
Publishing date	2020-09-02
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Video-Audio Media
ZDB-ID	2259946-0
ISSN	1940-087X ; 1940-087X
ISSN (online)	1940-087X
ISSN	1940-087X
DOI	10.3791/61515
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: Whole animal imaging of Drosophila melanogaster using microcomputed tomography

Schoborg, Todd A.

Journal of visualized experiments. 2020 Sept. 02, , no. 163

2020

Abstract	Biomedical imaging tools permit investigation of molecular mechanisms across spatial scales, from genes to organisms. Drosophila melanogaster, a well-characterized model organism, has benefited from the use of light and electron microscopy to understand gene function at the level of cells and tissues. The application of imaging platforms that allow for an understanding of gene function at the level of the entire intact organism would further enhance our knowledge of genetic mechanisms. Here a whole animal imaging method is presented that outlines the steps needed to visualize Drosophila at any developmental stage using microcomputed tomography (µ-CT). The advantages of µ-CT include commercially available instrumentation and minimal hands-on time to produce accurate 3D information at micron-level resolution without the need for tissue dissection or clearing methods. Paired with software that accelerate image analysis and 3D rendering, detailed morphometric analysis of any tissue or organ system can be performed to better understand mechanisms of development, physiology, and anatomy for both descriptive and hypothesis testing studies. By utilizing an imaging workflow that incorporates the use of electron microscopy, light microscopy, and µ-CT, a thorough evaluation of gene function can be performed, thus furthering the usefulness of this powerful model organism.
Keywords	Drosophila melanogaster ; animals ; computer software ; dissection ; electron microscopy ; genes ; instrumentation ; light microscopy ; micro-computed tomography ; morphometry ; physiology
Language	English
Dates of publication	2020-0902
Size	p. e61515.
Publishing place	Journal of Visualized Experiments
Document type	Article
ZDB-ID	2259946-0
ISSN	1940-087X
ISSN	1940-087X
DOI	10.3791/61515
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Article: Assessing Anatomical Changes in Male Reproductive Organs in Response to Larval Crowding Using Micro-computed Tomography Imaging

Morimoto, Juliano / Barcellos, Renan / Schoborg, Todd A. / Nogueira, Liebert Parreiras / Colaço, Marcos Vinicius

Neotropical entomology. 2022 Aug., v. 51, no. 4

2022

Abstract: Ecological conditions shape (adaptive) responses at the molecular, anatomical, and behavioral levels. Understanding these responses is key to predict the outcomes of intra- and inter-specific competitions and the evolutionary trajectory of populations. ... ...

Abstract	Ecological conditions shape (adaptive) responses at the molecular, anatomical, and behavioral levels. Understanding these responses is key to predict the outcomes of intra- and inter-specific competitions and the evolutionary trajectory of populations. Recent technological advances have enabled large-scale molecular (e.g., RNAseq) and behavioral (e.g., computer vision) studies, but the study of anatomical responses to ecological conditions has lagged behind. Here, we highlight the role of X-ray micro-computed tomography (micro-CT) in generating in vivo and ex vivo 3D imaging of anatomical structures, which can enable insights into adaptive anatomical responses to ecological environments. To demonstrate the application of this method, we manipulated the larval density of Drosophila melanogaster Meigen flies and applied micro-CT to investigate the anatomical responses of the male reproductive organs to varying intraspecific competition levels during development. Our data is suggestive of two classes of anatomical responses which broadly agree with sexual selection theory: increasing larval density led to testes and ejaculatory duct to be overall larger (in volume), while the volume of accessory glands and, to a lesser extent, ejaculatory duct decreased. These two distinct classes of anatomical responses might reflect shared developmental regulation of the structures of the male reproductive system. Overall, we show that micro-CT can be an important tool to advance the study of anatomical (adaptive) responses to ecological environments.
Keywords	Drosophila melanogaster ; Neotropics ; computer vision ; entomology ; intraspecific competition ; larvae ; males ; micro-computed tomography ; sexual selection
Language	English
Dates of publication	2022-08
Size	p. 526-535.
Publishing place	Springer International Publishing
Document type	Article
ZDB-ID	2105363-7
ISSN	1678-8052 ; 1519-566X
ISSN (online)	1678-8052
ISSN	1519-566X
DOI	10.1007/s13744-022-00976-5
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Article ; Online: Analysis of

Petersen, Courtney E / Tripoli, Benjamin A / Schoborg, Todd A / Smyth, Jeremy T

American journal of physiology. Heart and circulatory physiology

2021 Volume 322, Issue 2, Page(s) H296–H309

Abstract: Heart failure is often preceded by pathological cardiac hypertrophy, a thickening of the heart musculature driven by complex gene regulatory and signaling processes. ... ...

Abstract	Heart failure is often preceded by pathological cardiac hypertrophy, a thickening of the heart musculature driven by complex gene regulatory and signaling processes. The
MeSH term(s)	Animals ; Calcium Signaling ; Cardiomegaly/diagnostic imaging ; Cardiomegaly/genetics ; Drosophila Proteins/metabolism ; Drosophila melanogaster ; Heart/growth & development ; ORAI1 Protein/metabolism ; Stromal Interaction Molecule 1/metabolism ; X-Ray Microtomography/methods ; ras Proteins/metabolism
Chemical Substances	Drosophila Proteins ; ORAI1 Protein ; Stim protein, Drosophila ; Stromal Interaction Molecule 1 ; olf186-F protein, Drosophila ; Ras85D protein, Drosophila (EC 3.6.5.2) ; ras Proteins (EC 3.6.5.2)
Language	English
Publishing date	2021-12-24
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	603838-4
ISSN	1522-1539 ; 0363-6135
ISSN (online)	1522-1539
ISSN	0363-6135
DOI	10.1152/ajpheart.00387.2021
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Article ; Online: Assessing Anatomical Changes in Male Reproductive Organs in Response to Larval Crowding Using Micro-computed Tomography Imaging.

Morimoto, Juliano / Barcellos, Renan / Schoborg, Todd A / Nogueira, Liebert Parreiras / Colaço, Marcos Vinicius

Neotropical entomology

2022 Volume 51, Issue 4, Page(s) 526–535

Abstract	Ecological conditions shape (adaptive) responses at the molecular, anatomical, and behavioral levels. Understanding these responses is key to predict the outcomes of intra- and inter-specific competitions and the evolutionary trajectory of populations. Recent technological advances have enabled large-scale molecular (e.g., RNAseq) and behavioral (e.g., computer vision) studies, but the study of anatomical responses to ecological conditions has lagged behind. Here, we highlight the role of X-ray micro-computed tomography (micro-CT) in generating in vivo and ex vivo 3D imaging of anatomical structures, which can enable insights into adaptive anatomical responses to ecological environments. To demonstrate the application of this method, we manipulated the larval density of Drosophila melanogaster Meigen flies and applied micro-CT to investigate the anatomical responses of the male reproductive organs to varying intraspecific competition levels during development. Our data is suggestive of two classes of anatomical responses which broadly agree with sexual selection theory: increasing larval density led to testes and ejaculatory duct to be overall larger (in volume), while the volume of accessory glands and, to a lesser extent, ejaculatory duct decreased. These two distinct classes of anatomical responses might reflect shared developmental regulation of the structures of the male reproductive system. Overall, we show that micro-CT can be an important tool to advance the study of anatomical (adaptive) responses to ecological environments.
MeSH term(s)	Animals ; Drosophila ; Drosophila melanogaster/physiology ; Genitalia, Male/diagnostic imaging ; Larva ; Male ; X-Ray Microtomography/methods
Language	English
Publishing date	2022-07-05
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	2105363-7
ISSN	1678-8052 ; 1519-566X
ISSN (online)	1678-8052
ISSN	1519-566X
DOI	10.1007/s13744-022-00976-5
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Article ; Online: A Drosophila insulator interacting protein suppresses enhancer-blocking function and modulates replication timing.

Stow, Emily C / Simmons, James R / An, Ran / Schoborg, Todd A / Davenport, Nastasya M / Labrador, Mariano

Gene

2022 Volume 819, Page(s) 146208

Abstract: Insulators play important roles in genome structure and function in eukaryotes. Interactions between a DNA binding insulator protein and its interacting partner proteins define the properties of each insulator site. The different roles of insulator ... ...

Abstract	Insulators play important roles in genome structure and function in eukaryotes. Interactions between a DNA binding insulator protein and its interacting partner proteins define the properties of each insulator site. The different roles of insulator protein partners in the Drosophila genome and how they confer functional specificity remain poorly understood. The Suppressor of Hairy wing [Su(Hw)] insulator is targeted to the nuclear lamina, preferentially localizes at euchromatin/heterochromatin boundaries, and is associated with the gypsy retrotransposon. Insulator activity relies on the ability of the Su(Hw) protein to bind the DNA at specific sites and interact with Mod(mdg4)67.2 and CP190 partner proteins. HP1 and insulator partner protein 1 (HIPP1) is a partner of Su(Hw), but how HIPP1 contributes to the function of Su(Hw) insulator complexes is unclear. Here, we demonstrate that HIPP1 colocalizes with the Su(Hw) insulator complex in polytene chromatin and in stress-induced insulator bodies. We find that the overexpression of either HIPP1 or Su(Hw) or mutation of the HIPP1 crotonase-like domain (CLD) causes defects in cell proliferation by limiting the progression of DNA replication. We also show that HIPP1 overexpression suppresses the Su(Hw) insulator enhancer-blocking function, while mutation of the HIPP1 CLD does not affect Su(Hw) enhancer blocking. These findings demonstrate a functional relationship between HIPP1 and the Su(Hw) insulator complex and suggest that the CLD, while not involved in enhancer blocking, influences cell cycle progression.
MeSH term(s)	Animals ; Carrier Proteins/genetics ; Carrier Proteins/metabolism ; Cell Proliferation ; DNA Replication ; Drosophila/genetics ; Drosophila/metabolism ; Drosophila Proteins/genetics ; Drosophila Proteins/metabolism ; Enhancer Elements, Genetic ; Heterochromatin/metabolism ; Insulator Elements ; Microtubule-Associated Proteins/metabolism ; Mutation ; Nuclear Proteins/metabolism ; Repressor Proteins/genetics ; Repressor Proteins/metabolism
Chemical Substances	CP190 protein, Drosophila ; Carrier Proteins ; Drosophila Proteins ; HIPP1 protein, Drosophila ; Heterochromatin ; Microtubule-Associated Proteins ; Nuclear Proteins ; Repressor Proteins ; su(Hw) protein, Drosophila
Language	English
Publishing date	2022-01-29
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	391792-7
ISSN	1879-0038 ; 0378-1119
ISSN (online)	1879-0038
ISSN	0378-1119
DOI	10.1016/j.gene.2022.146208
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Article ; Online: Use of hare bone for the manufacture of a Clovis bead.

Surovell, Todd A / Litynski, McKenna L / Allaun, Sarah A / Buckley, Michael / Schoborg, Todd A / Govaerts, Jack A / O'Brien, Matthew J / Pelton, Spencer R / Sanders, Paul H / Mackie, Madeline E / Kelly, Robert L

Scientific reports

2024 Volume 14, Issue 1, Page(s) 2937

Abstract: A tubular bone bead dating to ~ 12,940 BP was recovered from a hearth-centered activity area at the La Prele Mammoth site in Converse County, Wyoming, USA. This is the oldest known bead from the Western Hemisphere. To determine the taxonomic origin of ... ...

Abstract	A tubular bone bead dating to ~ 12,940 BP was recovered from a hearth-centered activity area at the La Prele Mammoth site in Converse County, Wyoming, USA. This is the oldest known bead from the Western Hemisphere. To determine the taxonomic origin of the bead, we extracted collagen for zooarchaeology by mass spectrometry (ZooMS). We also used micro-CT scanning for morphological analysis to determine likely skeletal elements used for its production. We conclude that the bead was made from a metapodial or proximal phalanx of a hare (Lepus sp.). This find represents the first secure evidence for the use of hares during the Clovis period. While the use of hare bone for the manufacture of beads was a common practice in western North America during the Holocene, its origins can now be traced back to at least the terminal Pleistocene.
MeSH term(s)	Animals ; Phylogeny ; Hares ; Mass Spectrometry ; North America ; Lagomorpha
Language	English
Publishing date	2024-02-05
Publishing country	England
Document type	Journal Article
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-024-53390-9
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Article: Differential genetic expression within reward-specific ensembles in mice.

Litif, Carl G / Flom, Levi T / Sandum, Kathryn L / Hodgins, Skylar L / Vaccaro, Lucio / Stitzel, Jerry A / Blouin, Nicolas A / Mannino, Maria Constanza / Gigley, Jason P / Schoborg, Todd A / Bobadilla, Ana-Clara

bioRxiv : the preprint server for biology

2024

Abstract: Maladaptive reward seeking is a hallmark of cocaine use disorder. To develop therapeutic targets, it is critical to understand the neurobiological changes specific to cocaine-seeking without altering the seeking of natural rewards, e.g., sucrose. The ... ...

Abstract	Maladaptive reward seeking is a hallmark of cocaine use disorder. To develop therapeutic targets, it is critical to understand the neurobiological changes specific to cocaine-seeking without altering the seeking of natural rewards, e.g., sucrose. The prefrontal cortex (PFC) and the nucleus accumbens core (NAcore) are known regions associated with cocaine- and sucrose-seeking ensembles, i.e., a sparse population of co-activated neurons. Within ensembles, transcriptomic alterations in the PFC and NAcore underlie the learning and persistence of cocaine- and sucrose-seeking behavior. However, transcriptomes exclusively driving cocaine seeking independent from sucrose seeking have not yet been defined using a within-subject approach. Using Ai14:cFos-TRAP2 transgenic mice in a dual cocaine and sucrose self-administration model, we fluorescently sorted (FACS) and characterized (RNAseq) the transcriptomes defining cocaine- and sucrose-seeking ensembles. We found reward- and region-specific transcriptomic changes that will help develop clinically relevant genetic approaches to decrease cocaine-seeking behavior without altering non-drug reward-based positive reinforcement.
Language	English
Publishing date	2024-02-08
Publishing country	United States
Document type	Preprint
DOI	10.1101/2023.11.02.565378
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Article: A Drosophila insulator interacting protein suppresses enhancer-blocking function and modulates replication timing

Stow, Emily C. / Simmons, James R. / An, Ran / Schoborg, Todd A. / Davenport, Nastasya M. / Labrador, Mariano

Gene. 2022 Apr. 20, v. 819

2022

Abstract	Insulators play important roles in genome structure and function in eukaryotes. Interactions between a DNA binding insulator protein and its interacting partner proteins define the properties of each insulator site. The different roles of insulator protein partners in the Drosophila genome and how they confer functional specificity remain poorly understood. The Suppressor of Hairy wing [Su(Hw)] insulator is targeted to the nuclear lamina, preferentially localizes at euchromatin/heterochromatin boundaries, and is associated with the gypsy retrotransposon. Insulator activity relies on the ability of the Su(Hw) protein to bind the DNA at specific sites and interact with Mod(mdg4)67.2 and CP190 partner proteins. HP1 and insulator partner protein 1 (HIPP1) is a partner of Su(Hw), but how HIPP1 contributes to the function of Su(Hw) insulator complexes is unclear. Here, we demonstrate that HIPP1 colocalizes with the Su(Hw) insulator complex in polytene chromatin and in stress-induced insulator bodies. We find that the overexpression of either HIPP1 or Su(Hw) or mutation of the HIPP1 crotonase-like domain (CLD) causes defects in cell proliferation by limiting the progression of DNA replication. We also show that HIPP1 overexpression suppresses the Su(Hw) insulator enhancer-blocking function, while mutation of the HIPP1 CLD does not affect Su(Hw) enhancer blocking. These findings demonstrate a functional relationship between HIPP1 and the Su(Hw) insulator complex and suggest that the CLD, while not involved in enhancer blocking, influences cell cycle progression.
Keywords	DNA ; DNA replication ; Drosophila ; cell cycle ; cell proliferation ; eukaryotic cells ; genes ; heterochromatin ; mutation ; nuclear lamina ; retrotransposons
Language	English
Dates of publication	2022-0420
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	391792-7
ISSN	1879-0038 ; 0378-1119
ISSN (online)	1879-0038
ISSN	0378-1119
DOI	10.1016/j.gene.2022.146208
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Article ; Online: Taking Centrioles to the Elimination Round.

Schoborg, Todd A / Rusan, Nasser M

Developmental cell

2016 Volume 38, Issue 1, Page(s) 10–12

Abstract: Two recent papers published in The Journal of Cell Biology (Borrego-Pinto et al., 2016) and Science (Pimenta-Marques et al., 2016) have begun to shed light on the mechanism of centriole elimination during female oogenesis, highlighting a protective role ... ...

Abstract	Two recent papers published in The Journal of Cell Biology (Borrego-Pinto et al., 2016) and Science (Pimenta-Marques et al., 2016) have begun to shed light on the mechanism of centriole elimination during female oogenesis, highlighting a protective role for Polo kinase and the pericentriolar material.
MeSH term(s)	Animals ; Centrioles/physiology ; Female ; Humans ; Oogenesis/physiology ; Protein-Serine-Threonine Kinases/metabolism
Chemical Substances	Protein-Serine-Threonine Kinases (EC 2.7.11.1)
Language	English
Publishing date	2016-06-19
Publishing country	United States
Document type	Journal Article ; Comment
ZDB-ID	2054967-2
ISSN	1878-1551 ; 1534-5807
ISSN (online)	1878-1551
ISSN	1534-5807
DOI	10.1016/j.devcel.2016.06.027
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