Table of Contents

• Coverage
• Dataset Description
  • Methods & Sampling
  • Data Processing Description
• Data Files
• Supplemental Files
• Parameters
• Instruments
• Project Information
• Funding

Embryos collected with a rake from small boat on 5 dates across three months and kept in a cooler with ambient seawater for 1-5 hours until returned to the lab.  1-5 embryos flash frozen and stored cyogenically for 3-6 months until processing.

Embryos were physically broken with a micropestle in a microcentriguge tube, then processed 3x 30 seconds with stainless steel beads in a bullet blender.  Homogenate was removed from the bead then sonicated 3x. and digested using trypsin and a standard s-trap protocol.  Tubes were dried using a speed vap and reconstituted in water/ACN. 

Samples were spiked with Pierce retention time standards and with yeast enolase as a standard in some cases.  A 90 minute gradient was run on the LC serving the Lumos. 

NSF Award Abstract:
Pacific Herring represent a critical link in marine food webs along the West Coast of the United States, connecting the plankton they eat with larger predators (fish, sea birds, and marine mammals). Temperature strongly influences the development and success of herring. This investigation targets the underlying pathways that drive their temperature response by examining seasonal differences in protein expression. The outcome will be a better understanding of the processes most influenced by temperature, such as specific metabolic processes or stress responses. The project supports training for the investigator in new proteomics techniques and for undergraduate students at a Primarily Undergraduate Institution. Outreach includes engagement with local stakeholders and coastal indigenous communities. Societal benefits include a better understanding of population differences to inform conservation and recovery efforts for a culturally, economically, and ecologically important species.

Pacific Herring are ecologically important forage fish; fluctuations in their biomass drive far reaching food web responses. Climate variability is suspected to be a major driver of population trends, but the underlying mechanisms driving physiological responses remain unknown. Protein expression is a sensitive indicator of sub-lethal differences in stress response and metabolic state; therefore, comparisons across seasons unveil the cellular processes driving organismal responses to climate factors. Project goals are 1) a deeper understanding of the mechanisms driving the response of a key forage fish species to temperature and 2) workforce development, bringing cutting-edge molecular capabilities to faculty and students at a primarily undergraduate institution. Through a comparison between the robust Semiahmoo Bay (SB) population and the genetically and behaviorally distinct, and much depleted, Cherry Point (CP) herring population, the research team is detecting biomarker molecules of key physiological differences. Investigators are profiling SB and CP embryos collected from January through June using proteomic analyses, then developing targeted assays for peptides of interest, with total lipids and relevant environmental variables (T, Salinity, pH) providing meaningful context. Cohorts of embryos are also being reared to hatch from each collection date for comparison of protein biomarkers associated with survival or morphometric differences in the hatched larvae. This project provides the first large-scale survey of proteins present in early life stage Pacific Herring under different temperature regimes, advancing our understanding of herring response to environmental conditions associated with global change and ocean/atmosphere cycles.

This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.