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Changes in Northern Elephant Seal Skeletal Muscle Following Thirty Days of Fasting and Reduced Activity

ABSTRACT: Changes in Northern Elephant Seal Skeletal Muscle Following Thirty Days of Fasting and Reduced Activity

PROVIDER: PRJNA636169 | ENA |

REPOSITORIES: ENA

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Changes in Northern Elephant Seal Skeletal Muscle Following Thirty Days of Fasting and Reduced Activity

Project description:Northern elephant seals (NES, Mirounga angustirostris) undergo an annual molt during which they spend ~40 days fasting on land with reduced activity and lose approximately one-quarter of their body mass. Reduced activity and muscle load in stereotypic terrestrial mammalian models results in decreased muscle mass and capacity for force production and aerobic metabolism. However, the majority of lost mass in fasting female NES is from fat while muscle mass is largely preserved. Although muscle mass is preserved, potential changes to the metabolic and contractile capacity are unknown. To assess potential changes in NES skeletal muscle during molt, we collected muscle biopsies from 6 adult female NES at the beginning of the molt and after ~30 days at the end of the molt. Skeletal muscle was assessed for respiratory capacity using high resolution respirometry, and RNA was extracted to assess changes in gene expression. Despite a month of reduced activity, fasting, and weight loss, skeletal muscle respiratory capacity was preserved with no change in OXPHOS respiratory capacity. Molt was associated with 162 upregulated genes including genes favoring lipid metabolism and regulating cell cycles. We identified 172 downregulated genes including those coding for ribosomal proteins and genes associated with skeletal muscle force transduction and glucose metabolism. Following ~30 days of molt, NES skeletal muscle metabolic capacity appears largely preserved although mechanotransduction may be compromised. In the absence of exercise stimulus, fasting-induced shifts in skeletal muscle lipid metabolism may stimulate lipid signaling pathways associated with preserving the mass and metabolic capacity of slow oxidative muscle.

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Mirounga angustirostris strain:northern elephant seal

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Project description:Northern elephant seal skeletal muscle, blubber, and plasma were collected from 5 adult females early and late in a 5-week molting fast (mean 33 days apart; see Sample Key in supplementary files). Proteins were isolated from muscle using the Trizol method and blubber using Qiazol with the Qiagen RNeasy Lipid Mini kit (see Khudyakov et al., 2018, doi: 10.1242/bio.036731). Plasma samples were used raw (without depletion of highly abundant proteins). Muscle, blubber, and plasma proteins were denatured using 1% sodium deoxycholate, 8 M urea, and 5 mM DTT in 50 mM ammonium bicarbonate and digested in-solution using trypsin (1:50 trypsin to protein ratio).

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Project description:Proteins were extracted from the outer (closer to skin) blubber layer of 4 northern elephant seals (Mirounga angustirostris) using either 1) A standard detergent lysis buffer containing 1% sodium deoxycholate, 8 M urea, 5 mM DTT in 50 mM ammonium bicarbonate (SDC method) or 2) the phenol and guanidine thiocyanate reagent Qiazol (QIA method). Mass spectrometry analysis was performed using an Orbitrap Fusion Tribrid mass spectrometer in DDA mode. Briefly, full MS1 scans were resolved by the orbitrap, and ions were selected for MS2 using DDA. These precursor ions were quadrupole filtered and subsequently fragmented using stepped collision HCD. MS2 product ions were resolved by the orbitrap. MS/MS data was analyzed using Proteome Discoverer v2.2.

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Project description:The identity of most functional elements in the mammalian genome and the phenotypes they impact are unclear. Here, we perform a genome-wide comparative analysis of patterns of accelerated evolution in species with highly distinctive traits to discover candidate functional elements for clinically important phenotypes. We identify accelerated regions (ARs) in the elephant, hibernating bat, orca, dolphin, naked mole rat and thirteen-lined ground squirrel lineages in mammalian conserved regions, uncovering ~33,000 elements that bind hundreds of different regulatory proteins in humans and mice. ARs in the elephant, the largest land mammal, are uniquely enriched at elephant DNA damage response genes and changed conserved regulatory sites. The genomic hotspot for elephant ARs is the E3 ligase subunit of the Fanconi Anemia Complex, a master regulator of DNA repair. Additionally, ARs in the six species are associated with specific human clinical phenotypes that have apparent concordance with overt traits in each species.

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