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author Hildebrand, Jasper
Stumpp, Meike
author_facet Hildebrand, Jasper
Stumpp, Meike
collection Datos científicos de ciencias marinas y ambientales
contents Strongylocentrotus purpuratus (Stimpson 1857) originally collected in November 2022 from La Jolla, USA (Lat: 32.842674; Long: -117.257767) were held in flow-through tanks which were filled with water from Kiel Fjord adjusted to 31.5 psu in the Christian-Albrechts-Universität zu Kiel at 10 °C. For experimental characterization of carbohydrate digestion between September and November 2023 S. purpuratus larvae were generated by gently shaking adult males and females and held in climate chambers at 15°C and 31.5 psu up to 10 days under two different food conditions. For genetic analysis of carbohydrate metabolism RT-qPCR was conducted. In brief, mRNA of samples from nine different time points (2 days post fertilization (dpf) – 10 dpf, n=4) was isolated and reverse transcripted into cDNA. Primer pairs were designed through NCBI primer blast, specifically targeting exon junctions of the selected carbohydrates enzyme genes for which the mRNA sequence is accessible in the S. purpuratus genome database echinobase.org. Genes were selected based on transcription patterns of the genes during early development. Primer specificity was validated through standard PCR, confirming the amplification of a single, specific product within the anticipated size range. Gene expression utilized the ∆∆CT method, and levels were normalized to the stable housekeeping gene EF1a (SPU_000595).
format Dataset Open Access
id pangaea_https___doi_org_10_1594_PANGAEA_968978
institution PANGAEA
language en
publishDate 2025
publisher PANGAEA
record_format pangaea
spellingShingle Genetic analysis of carbohydrate metabolism in Strongylocentrotus purpuratus larvae during laboratory experiments with different food treatments
Hildebrand, Jasper
Stumpp, Meike
algae; Carbohydrate microarrays; carbohydrates; culture experiment; DIVER; energetics; Experiment; La_Jolla_S_purpuratus; Laboratory; Laboratory experiment; La Jolla, USA; laminarin; Microscope, Zeiss, Axio Imager Z; measured with Hemocytometer; Real-time-PCR-system, ThermoFisher Scientific, QuantStudio 1; Sample ID; Sampling by diver; Sampling date/time, experiment; sea urchin; Strongylocentrotus purpuratus; Strongylocentrotus purpuratus, larvae, alpha-amylase, relative gene expression; Strongylocentrotus purpuratus, larvae, beta-1,3-glucanase, relative gene expression; Strongylocentrotus purpuratus, larvae, endo-beta-1,4-glucanase D-like, relative gene expression; Strongylocentrotus purpuratus, larvae, trehalase, relative gene expression; Strongylocentrotus purpuratus, larvae, xylanase/beta-glucanase-like, relative gene expression; Strongylocentrotus purpuratus, larval age; Treatment: food; Type of study
Strongylocentrotus purpuratus (Stimpson 1857) originally collected in November 2022 from La Jolla, USA (Lat: 32.842674; Long: -117.257767) were held in flow-through tanks which were filled with water from Kiel Fjord adjusted to 31.5 psu in the Christian-Albrechts-Universität zu Kiel at 10 °C. For experimental characterization of carbohydrate digestion between September and November 2023 S. purpuratus larvae were generated by gently shaking adult males and females and held in climate chambers at 15°C and 31.5 psu up to 10 days under two different food conditions. For genetic analysis of carbohydrate metabolism RT-qPCR was conducted. In brief, mRNA of samples from nine different time points (2 days post fertilization (dpf) – 10 dpf, n=4) was isolated and reverse transcripted into cDNA. Primer pairs were designed through NCBI primer blast, specifically targeting exon junctions of the selected carbohydrates enzyme genes for which the mRNA sequence is accessible in the S. purpuratus genome database echinobase.org. Genes were selected based on transcription patterns of the genes during early development. Primer specificity was validated through standard PCR, confirming the amplification of a single, specific product within the anticipated size range. Gene expression utilized the ∆∆CT method, and levels were normalized to the stable housekeeping gene EF1a (SPU_000595).
title Genetic analysis of carbohydrate metabolism in Strongylocentrotus purpuratus larvae during laboratory experiments with different food treatments
topic algae; Carbohydrate microarrays; carbohydrates; culture experiment; DIVER; energetics; Experiment; La_Jolla_S_purpuratus; Laboratory; Laboratory experiment; La Jolla, USA; laminarin; Microscope, Zeiss, Axio Imager Z; measured with Hemocytometer; Real-time-PCR-system, ThermoFisher Scientific, QuantStudio 1; Sample ID; Sampling by diver; Sampling date/time, experiment; sea urchin; Strongylocentrotus purpuratus; Strongylocentrotus purpuratus, larvae, alpha-amylase, relative gene expression; Strongylocentrotus purpuratus, larvae, beta-1,3-glucanase, relative gene expression; Strongylocentrotus purpuratus, larvae, endo-beta-1,4-glucanase D-like, relative gene expression; Strongylocentrotus purpuratus, larvae, trehalase, relative gene expression; Strongylocentrotus purpuratus, larvae, xylanase/beta-glucanase-like, relative gene expression; Strongylocentrotus purpuratus, larval age; Treatment: food; Type of study
url https://doi.org/10.1594/PANGAEA.968978