Atlantic Salmon mucus from skin, pyloric caeca and distal intestine

Announcement date

2019/04/10

Responsible:

Sara Lindén . [no affiliation]

Description

Diseases cause ethical concerns and economic losses in the Salmonid industry. The mucus layer comprised of highly O-glycosylated mucins is the first contact between pathogens and fish. Mucin glycans govern pathogen adhesion, growth and virulence. The Atlantic salmon O-glycome from a single location has been characterized and the interindividual variation was low. Because interindividual variation is considered a population-based defense, hindering the entire population from being wiped out by a single infection, low interindividual variation among Atlantic salmon may be a concern. Here, we analyzed the O-glycome of 25 Atlantic salmon from six cohorts grown under various conditions from Sweden, Norway and Australia (Tasmania) using mass spectrometry. This expanded the known Atlantic salmon O-glycome by 60% to 169 identified structures. The mucin O-glycosylation was relatively stable over time within a geographical region, but the size of the fish affected skin mucin glycosylation. The skin mucin glycan repertoires from Swedish and Norwegian Atlantic salmon populations were closely related compared with Tasmanian ones, regardless of size and salinity, with differences in glycan size and composition. The internal mucin glycan repertoire also clustered based on geographical origin and into pyloric cecal and distal intestinal groups, regardless of cohort and fish size. Fucosylated structures were more abundant in Tasmanian pyloric caeca and distal intestine mucins compared with Swedish ones. Overall, Tasmanian Atlantic salmon mucins have more O-glycan structures in skin but less in the gastrointestinal tract compared with Swedish fish. Low interindividual variation was confirmed within each cohort. The results can serve as a library for identifying structures of importance for host-pathogen interactions, understanding population differences of salmon mucin glycosylation in resistance to diseases and during breeding and selection of strains. The results could make it possible to predict potential vulnerabilities to diseases and suggest that inter-region breeding may increase the glycan diversity.

Sample preparation

---

1. Sample Origin

General information:
Atlantic Salmon mucus from skin, pyloric caeca and distal intestine.

1.1 Biologically derived material

Biologically derived material - Recombinantly produced material

Cell type:
N/A

Growth/harvest conditions for recombinantly produced material:
N/A

Biologically derived material - Biological origin of Material

Origin (biological fluid, tissue, etc):
Mucus

Species:
Salmo Salar

Treatments and/or storage conditions:
Stored in -80 C

Glycoprotein:
N/A

Biologically derived material - Purchased from commercial manufacturer

Vendor and applicable item information:
N/A

1.2 Chemically derived material

Synthesis steps or specify where the equivalent reaction protocol is available:
N/A

Description of starting material:
N/A

2. Sample Processing

2.1 Sample Processing - Isolation

Chemical treatments

Define the technique for oligosaccharide release or other chemical modifications:
Release method-REDUCTIVE BETA ELIMINATION

Reaction conditions (temperature, duration, volume and chemical concentrations):
1 M NaBH4 + 50 mM NaOH

2.2 Sample Processing - Modification

2.3 Sample Processing - Purification

Purification steps:
Crude mucin fraction obtained after dissolved in GuHCl extraction buffer and centrifugation to remove insoluble material.

3. Defined Sample

Sample name:
O-linked oligosaccharides

Liquid chromatography

---

N/A

MS

---

1. General features

(a) Global descriptors

Instrument manufacturer:
Thermo Fisher

Instrument model:
LTQ Linear Ion Trap

Customizations:
N/A

Ion mode:
Negative

(b) Control and analysis software

2. Ion sources

(a) Electrospray Ionisation (ESI)

Supply type (static, or fed):
Fed

Interface name:
N/A

Catalog number, vendor, and any modifications made to the standard specification:
N/A

Sprayer name:
IonMax standard ESI source

Sprayer type, coating, manufacturer, model and catalog number (where available):
Equipped with a stainless steel needle

Relevant voltages where appropriate (tip, cone, acceleration):
Electrospray voltage of 3.5 kV and capillary voltage of −33.0 V..

Degree of prompt fragmentation evaluated:
Yes

Whether in-source dissociation performed:
Yes

Other parameters if discriminant for the experiment (such as nebulizing gas and pressure):
N/A

(b) MALDI

Plate composition (or type):
N/A

Matrix composition (if applicable):
N/A

Deposition technique:
N/A

Relevant voltages where appropriate:
N/A

Degree of prompt fragmentation evaluated:
N/A

PSD (or LID/ISD) summary, if performed:
N/A

Operation with or without delayed extraction:
N/A

Laser type (e.g., nitrogen) and wavelength (nm):
N/A

Other laser related parameters, if discriminating for the experiment:
N/A

3. Ion transfer optics

Hardware options:
N/A

(a) Post-source componentry - Collision cell

Collision-Induced Dissociation (CID)

Gas composition:
He

Gas pressure:
N/A

Collision energy CID/function:
Collision energy 35%

Electron Transfer Dissociation (ETD)

Reagent gas:
N/A

Pressure:
N/A

Reaction time:
N/A

Number of reagent atoms:
N/A

Electron Capture Dissociation (ECD)

Emitter type:
N/A

Voltage:
N/A

Current:
N/A

(b) Post-source componentry - TOF drift tube

Reflectron status (on, off, none):
N/A

(c) Post-source componentry - Ion trap

Final MS stage achieved:
N/A

(d) Post-source componentry - Ion mobility

Gas:
N/A

Pressure:
N/A

Instrument-specific parameters:
N/A

(e) Post-source componentry - FT-ICR

Peak selection:
N/A

Pulse:
N/A

Width:
N/A

Voltage:
N/A

Decay time:
N/A

IR:
N/A

Other parameters:
N/A

(f) Post-source componentry - Detectors

Detector type:
N/A