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Identification of candidate protein biomarkers associated with domoic acid toxicosis in cerebrospinal fluid of California sea lions (Zalophus californianus)

ABSTRACT: Since 1998, California sea lion stranding events associated with domoic acid toxicosis (DAT) have consistently increased and there are no practical non-lethal clinical tests for the diagnosis of domoic acid toxicosis that can be utilized in a large-scale rehabilitation facility. Proteomic analysis was conducted to discover candidate protein markers of DAT using cerebrospinal fluid (CSF) from stranded sea lions with acute DAT, chronic DAT, or without DAT. A total of 2005 protein families were identified across 40 CSF samples (FDR<0.01) using the annotated California sea lion genome. Of these proteins, 83 were significantly different in abundance across the three groups (p<0.05). Comparisons between all sea lions with DAT versus those without DAT indicated that 119 proteins were significantly different between both groups (p<0.05); whereas, 47 proteins were significantly different between acute DAT and chronic DAT (p<0.05). Significant proteins were assessed as classifiers using ROC curves. Compared to sea lions with non-DAT, those with either acute or chronic DAT displayed higher levels of 14-3-3 proteins and malate dehydrogenase, and lower levels of 5’-3’ exonuclease PLD3, neurosecretory protein VGF, disintegrin and metalloproteinase domain-containing protein, and calsyntenin-1. When comparing acute DAT versus chronic DAT, 4 proteins were identified as good classifiers. Elevated levels of beta-synuclein was detected in acute DAT, and was identified as a high classifier for both comparisons. Many of these proteins have been implicated in a variety of neurodegenerative diseases. These proteins should be considered potential markers for DAT in California sea lions, as well as markers to discriminate between acute or chronic DAT, and should be considered priority for future validation studies as biomarkers.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Zalophus Californianus (california Sealion)

TISSUE(S): Cerebrospinal Fluid

SUBMITTER: Benjamin Neely

LAB HEAD: Gautam Ghosh, Alison M. Bland, Michael G. Janech, Benjamin A. Neely

PROVIDER: PXD041356 | Pride | 2023-04-05

REPOSITORIES: Pride

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			Action	DRS
	2022-8-30_CSF-01-F003230.dat	Other
	2022-8-30_CSF-01-F003230.mzid.gz	Mzid
	2022-8-30_CSF-01.mgf	Mgf
	2022-8-30_CSF-01.raw	Raw
	2022-8-30_CSF-02-F003231.dat	Other

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Project description:Urinary proteins provide valuable insights into renal health, with implications spanning human and domestic animal veterinary medical research. However, the field of marine mammal medicine lacks comprehensive studies on urine protein composition. This research aimed to fill this gap by 1) selecting an optimal search strategy that yields the highest number of protein families in the bottlenose dolphin urine based on post-translational modifications (PTMs), 2) describing the urine proteome of wild bottlenose dolphins (Tursiops truncatus) in the Gulf of the Mexico, USA, considering sex (females vs. males), site (Barataria Bay, LA vs. Sarasota Bay, FL), and comparing with California sea lions (Zalophus californianus). Ten urine samples (Barataria Bay, LA: N= 6; Sarasota Bay, FL: N=4) collected during 2023 catch-and-release heath assessment were proteolytically digested and analyzed using an UltiMate 3000 Nano LC and Fusion Lumos Orbitrap mass spectrometer. A 2-step search strategy, incorporating dehydroalanine formation and semi-trypsin on the list of unassigned spectra, significantly increased the number of identified protein families by an average of 6.2% compared to the 1-step strategy (P < 0.001, t = -8.32). The top 30 proteins in bottlenose dolphin urine were ranked according to an exponentially modified protein abundance index for comparison based on sex, site, and species. There were no significant differences in urine proteins between sexes or sites (padj > 0.05), although there was sperm contribution in two of the male bottlenose dolphin urine samples. Two putative antimicrobial proteins (cathelicidin and lysozyme) were identified and found to be abundant in bottlenose dolphin urine, similar to California sea lions. The study also identified 27 potential markers of acute kidney injury and 12 regulators of kidney stone formation. This study established a reference database of urinary proteins from bottlenose dolphins, aiding future research in monitoring and evaluating renal health in marine mammals.

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Identification of candidate protein biomarkers associated with domoic acid toxicosis in cerebrospinal fluid of California sea lions (Zalophus californianus)

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