OSMO_LUS Osmoregulation in Salmon Lice
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1 OSMO_LUS Osmoregulation in Salmon Lice Anna Z Komisarczuk
2 Content 1. Aim 2. Background: 1. Osmoregulation 2. Osmoregulation in salmon louse 3. Fresh water effects on salmon louse: 1. Survival 2. Osmotic stress response 3. Ion exchangers/transporters 4. Ion transporters not only osmoregulation
3 Aim Fresh water treatment is one of the methods currently used to fight salmon lice infections OSMO_LUS project: è gain more knowledge about fresh water tolerance in salmon louse; è physiological changes in salmon louse triggered by fresh water exposure; è identification of the main players (genes/proteins) involved in osmoregulation in salmon louse ; è identification of anatomical structures associated with osmoregulation;
4 Osmoregulation OSMOREGULATION: Maintaining cell/organism homeostasis with respect to solute concentration and balance of water content controlled movement of solutes and water between internal fluids and the external environment; narrow cellular limits; OSMOLARITY: the solute concentration in a solution (solute: mainly salts), determines movement of water across a selectively permeable membrane: Isosmotic solution: equal concentrations of water and solute the movement of water is equal in both directions Hypoosmotic solution: lower solute and higher free water concentration water movement to cell; Hyperosmotic solution: higher solute and lower free water concentration water movement from cell;
5 Osmoregulation in Salmon Louse Regulation of water transport through body/cell surface is crucial for organisms living in water. Environmental solution has very high or very low solute content, often different than solute concentration of body and tissue fluids: Sea water (hyperosmotic solution): 34 ppt salinity = 1000 mosm/l Fresh water (hypoosmotic solution): < 3 ppt salinity = 1 10 mosm/l Salmon louse (Lepeophtheirus salmonis) is osmoconformer it is isoosmotic with the marine environment and in normal conditions does not regulate its osmolarity (the internal osmolarity of the salmon louse 1000mOsm). Osmoconformers have low tolerance of osmolarity change of their environment Without the host: è salmon louse cannot tolerate substantial changes in external osmolarity and die within hours. When attached to its host: è salmon louse is able to survive large fluctuations in external osmolarity, and control water influx in hypoosmotic environment by uptake of osmolytes from its host.
6 Ion Exchange/Transport Structures Salmon louse lacks obvious osmoregulatory structures. è Ion exchange/transport structures were identified in many locations on the animal body Adult female Copepodid Crusalis organs
7 Fresh Water Effects on Adult Lice Treatment time 6h 2 groups: Fresh water and Seawater Sampling: Lost lice every 10 minutes; Attached lice every hour; Seawater Fresh water 2 weeks acclimatization 6h fresh water Seawater Treatment 0h hours
8 Fresh Water Effects on Adult Lice Lice Loss Lice Egg strings
9 Fresh Water Effects on Adult Lice Osmotic Stress Heat Shock Proteins (HSP): Stressful conditions è cell response è the upregulation of the HSPs many HSPs perform chaperone function - help to refold proteins that were damaged by the cell stress Adult male
10 Fresh Water Effects on Adult Lice Ion Exchangers/ Transporters Ion Exchangers/Transporters a family of membrane bound proteins pump ions to/from cell with use od energy (ATP) involved in number of physiological processes including osmoregulation Adult male
11 Fresh Water Effects on Copepodids Treatment time 24h 11 groups 100% 0 sea water content. Evaluation: 30 min, 1h, 2h, 4h, 8h, 24h Movement; Staining; Result: 100% Sea water 100% 100% - 40% sea water 100% survival after 24h; 30% - 20% sea water 100% - 90% survival after 24h; 20% - 24h 90% 80% alive 70% 60% 50% 40% 30% CONTROL dead 20% 10% 100% Fresh water 10% 30 min 1h 4h
12 Fresh Water Effects on Copepodids Osmotic Stress Heat Shock Proteins 70 (HSP70): Copepodids 100%, 75%, 50% and 25% sea water Borchel et al. 2017
13 Fresh Water Effects on Copepodids Ion Exchangers/Transporters Ion Exchangers/Transporters: Copepodids 25% sea water
14 LsalNa + /K + -ATPase Osmoregulation Sodium-Potassium ATPase Na + ions out of the cell and K + to the cell; osmoregulation cell volume regulation; membrane excitability; transport of nutrients;
15 LsalNa + /K + -ATPase Gene Expression copepodid adults
16 LsalNa + /K + -ATPase RNAi Copepodids Movement impairment: è Defects in muscle development control dsnka
17 LsalNa + /K + -ATPase RNAi Adult Females dsnka
18 LsalNa + /K + -ATPase RNAi Adult Females Defects in reproductions Defects in glands dsnka control
19 Conclusions Gill-like structures (ion exchange structures) spread around the whole animal body Adult lice can deal with decreased salinity when attached to the fish Copepodids are able to survive in the environment with very low salinity è Osmotic stress response genes and ion exchangers/transporters are upregulated in order to counteract negative effects of fresh water Na + /K + -ATPase has number of functions in salmon louse: è osmoregulation; è reproduction and secretion of fluids;
20 OSMO_LUS FHF project: Acknowledgements Frank Nilsen Heidi Kongshaug Per Gunnar Espedal Lars Hamre
OSMO_LUS Osmoregulation in Salmon Lice
OSMO_LUS Osmoregulation in Salmon Lice Anna Z Komisarczuk 23.01.2018 Aims Fresh water treatment is one of the methods currently used to fight salmon lice infections OSMO_LUS project: gain more knowledge
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