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  • 1
    Online Resource
    Online Resource
    Micro-CT Analysis of Fat in the Killifish Nothobranchius furzeri
    Cold Spring Harbor Laboratory ; 2023
    In:  Cold Spring Harbor Protocols Vol. 2023, No. 8 ( 2023-08), p. pdb.prot107884-
    Details
    In: Cold Spring Harbor Protocols, Cold Spring Harbor Laboratory, Vol. 2023, No. 8 ( 2023-08), p. pdb.prot107884-
    Abstract: Aging is associated with an increase in body fat mass and a concomitant decrease in lean mass and bone density in mammals. Body adiposity can also be redistributed with age, resulting in abdominal fat accumulation and subcutaneous fat reduction. In addition, specific variation in fat distribution is considered to be a risk factor for a number of age-related metabolic disorders. Micro computed tomography (micro-CT) is a nondestructive high-resolution imaging method that uses planar X-ray images captured at various angles around a sample of interest to yield a three-dimensional array of radiodensity values, which can then be used to computationally extract the adipose volume in situ using its innate contrast properties. This method was successfully used to study adipose tissue dynamics in rodents and more recently in zebrafish. The naturally short-lived African turquoise killifish is an emerging model organism to study the biology of aging. Like mammals, killifish also have different fat deposits (visceral and subcutaneous), making them a suitable model to study age-related changes in fat mass and distribution. However, procedures allowing precise quantification of fat content and distribution are missing in this species. Here, we provide an optimized protocol to measure and quantify fat distribution in turquoise killifish by micro-CT scan analysis and show the applicability of the method in young and old animals of both sexes.
    Type of Medium: Online Resource
    ISSN: 1940-3402 , 1559-6095
    URL: Article
    DOI: 10.1101/pdb.prot107884
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2023
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  • 2
    Online Resource
    Online Resource
    A novel nuclear receptor/coregulator complex controls C. elegans lipid metabolism, larval development, and aging
    Cold Spring Harbor Laboratory ; 2004
    In:  Genes & Development Vol. 18, No. 17 ( 2004-09-01), p. 2120-2133
    Details
    In: Genes & Development, Cold Spring Harbor Laboratory, Vol. 18, No. 17 ( 2004-09-01), p. 2120-2133
    Abstract: Environmental cues transduced by an endocrine network converge on Caenorhabditis elegans nuclear receptor DAF-12 to mediate arrest at dauer diapause or continuous larval development. In adults, DAF-12 selects long-lived or short-lived modes. How these organismal choices are molecularly specified is unknown. Here we show that coregulator DIN-1 and DAF-12 physically and genetically interact to instruct organismal fates. Homologous to human corepressor SHARP, DIN-1 comes in long (L) and short (S) isoforms, which are nuclear localized but have distinct functions. DIN-1L has embryonic and larval developmental roles. DIN-1S, along with DAF-12, regulates lipid metabolism, larval stage-specific programs, diapause, and longevity. Epistasis experiments reveal that din-1S acts in the dauer pathways downstream of lipophilic hormone, insulin/IGF, and TGFβ signaling, the same point as daf-12 . We propose that the DIN-1S/DAF-12 complex serves as a molecular switch that implements slow life history alternatives in response to diminished hormonal signals.
    Type of Medium: Online Resource
    ISSN: 0890-9369 , 1549-5477
    URL: Article
    DOI: 10.1101/gad.312604
    RVK:
    WA 15000
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2004
    detail.hit.zdb_id: 1467414-2
    SSG: 12
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  • 3
    Online Resource
    Online Resource
    daf-12 encodes a nuclear receptor that regulates the dauer diapause and developmental age in C. elegans
    Cold Spring Harbor Laboratory ; 2000
    In:  Genes & Development Vol. 14, No. 12 ( 2000-06-15), p. 1512-1527
    Details
    In: Genes & Development, Cold Spring Harbor Laboratory, Vol. 14, No. 12 ( 2000-06-15), p. 1512-1527
    Abstract: The daf-12 gene acts at the convergence of pathways regulating larval diapause, developmental age, and adult longevity in Caenorhabditis elegans . It encodes a nuclear receptor most closely related to two C. elegans receptors, NHR-8 and NHR-48, Drosophila DHR96, and vertebrate vitamin D and pregnane-X receptors. daf-12 has three predicted protein isoforms, two of which contain DNA- and ligand-binding domains, and one of which contains the ligand-binding domain only. Mutations cluster in DNA- and ligand-binding domains, but correspond to distinct phenotypic classes. DAF-12 is expressed widely in target tissues from embryo to adult, but is upregulated during midlarval stages. In the adult, expression persists in nervous system and somatic gonad, two tissues that regulate adult longevity. We propose that DAF-12 integrates hormonal signals in cellular targets to coordinate major life history traits.
    Type of Medium: Online Resource
    ISSN: 0890-9369 , 1549-5477
    URL: Article
    DOI: 10.1101/gad.14.12.1512
    RVK:
    WA 15000
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2000
    detail.hit.zdb_id: 1467414-2
    SSG: 12
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  • 4
    Online Resource
    Online Resource
    Nonlethal Blood Sampling from the Killifish Nothobranchius furzeri
    Cold Spring Harbor Laboratory ; 2023
    In:  Cold Spring Harbor Protocols Vol. 2023, No. 8 ( 2023-08), p. pdb.prot107745-
    Details
    In: Cold Spring Harbor Protocols, Cold Spring Harbor Laboratory, Vol. 2023, No. 8 ( 2023-08), p. pdb.prot107745-
    Abstract: Blood withdrawal is a common procedure performed on laboratory animals to monitor key processes and indicators of fish health and physiology, such as hematopoiesis, hemostasis, and lipid and glucose metabolism. Moreover, the ability to extract blood with minimal invasiveness and without sacrificing animals enables repeated sampling, allowing both longitudinal studies of individual animals, as well as reducing the number of experimental animals needed in a study. The African turquoise killifish is an emerging animal model that is progressively being adopted worldwide for aging studies because of its naturally short life span. However, because of the small body size of this species, nonlethal blood collection is a challenging procedure. Here we present a detailed protocol enabling repeated blood sampling from the same individual fish. This method, if correctly executed, is minimally invasive and does not cause any lasting damage. The protocol has been tested on animals spanning from 6 to 24 wk of age and the amount of blood that could be extracted varied from 0.5 to 8 µL, greatly depending on specimen age, sex, and size. This volume is sufficient to perform analyses such as blood glucose measurement, blood cell counts, or histological stains on blood smears.
    Type of Medium: Online Resource
    ISSN: 1940-3402 , 1559-6095
    URL: Article
    DOI: 10.1101/pdb.prot107745
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2023
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  • 5
    Online Resource
    Online Resource
    Identification of C. elegans DAF-12-binding sites, response elements, and target genes
    Cold Spring Harbor Laboratory ; 2004
    In:  Genes & Development Vol. 18, No. 20 ( 2004-10-15), p. 2529-2544
    Details
    In: Genes & Development, Cold Spring Harbor Laboratory, Vol. 18, No. 20 ( 2004-10-15), p. 2529-2544
    Abstract: Intracellular receptor DAF-12 regulates dauer formation and developmental age and affects Caenorhabditis elegans lifespan. Genetic analyses place DAF-12 at the convergence of several signal transduction pathways; however, the downstream effectors and the molecular basis for the receptor's multiple physiological outputs are unknown. Beginning with C. elegans genomic DNA, we devised a procedure for multiple rounds of selection and amplification that yielded fragments bearing DAF-12-binding sites. These genomic fragments mediated DAF-12-dependent transcriptional regulation both in Saccharomyces cerevisiae and in C. elegans ; that is, they served as functional DAF-12 response elements. We determined that most of the genomic fragments that displayed DAF-12 response element activity in yeast were linked to genes that were regulated by DAF-12 in C. elegans ; indeed, the response element-containing fragments typically resided within clusters of DAF-12-regulated genes. DAF-12 target gene regulation was developmental program and stage specific, potentially predicting a fit of these targets into regulatory networks governing aspects of C. elegans reproductive development and dauer formation.
    Type of Medium: Online Resource
    ISSN: 0890-9369 , 1549-5477
    URL: Article
    DOI: 10.1101/gad.1218504
    RVK:
    WA 15000
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2004
    detail.hit.zdb_id: 1467414-2
    SSG: 12
    Location Call Number Limitation Availability
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  • 6
    Online Resource
    Online Resource
    C. elegans dauer formation and the molecular basis of plasticity
    Cold Spring Harbor Laboratory ; 2008
    In:  Genes & Development Vol. 22, No. 16 ( 2008-08-15), p. 2149-2165
    Details
    In: Genes & Development, Cold Spring Harbor Laboratory, Vol. 22, No. 16 ( 2008-08-15), p. 2149-2165
    Abstract: Because life is often unpredictable, dynamic, and complex, all animals have evolved remarkable abilities to cope with changes in their external environment and internal physiology. This regulatory plasticity leads to shifts in behavior and metabolism, as well as to changes in development, growth, and reproduction, which is thought to improve the chances of survival and reproductive success. In favorable environments, the nematode Caenorhabditis elegans develops rapidly to reproductive maturity, but in adverse environments, animals arrest at the dauer diapause, a long-lived stress resistant stage. A molecular and genetic analysis of dauer formation has revealed key insights into how sensory and dietary cues are coupled to conserved endocrine pathways, including insulin/IGF, TGF-β, serotonergic, and steroid hormone signal transduction, which govern the choice between reproduction and survival. These and other pathways reveal a molecular basis for metazoan plasticity in response to extrinsic and intrinsic signals.
    Type of Medium: Online Resource
    ISSN: 0890-9369 , 1549-5477
    URL: Article
    DOI: 10.1101/gad.1701508
    RVK:
    WA 15000
    Language: English
    Publisher: Cold Spring Harbor Laboratory
    Publication Date: 2008
    detail.hit.zdb_id: 1467414-2
    SSG: 12
    Location Call Number Limitation Availability
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