Real-time PCR quantification of the in vitro effects of crustacean immunostimulants on gene expression in lobster ( Homarus gammarus ) granular haemocytes

This paper presents data from an investigation of the mode of action of five different crustacean immunostimulants presented to European lobster (Homarus gammarus) granulocytes cultured in vitro. The experiments were designed to test whether or not the immunostimulants could cause the short-term up-...

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Bibliographic Details
Published in:Developmental & Comparative Immunology
Main Authors: Hauton, Christopher, Hammond, JA, Smith, Valerie Jane
Format: Article in Journal/Newspaper
Language:English
Published: 2005
Subjects:
real-time PCR
immunostimulants
prophenoloxidase
beta-1,3-glucan binding protein
lobster
Homarus gammarus
granulocytes
PROPHENOLOXIDASE-ACTIVATING SYSTEM
CRAYFISH PACIFASTACUS-LENIUSCULUS
BETA-1,3-GLUCAN BINDING-PROTEIN
FRESH-WATER CRAYFISH
MESSENGER-RNA LEVELS
BLACK TIGER SHRIMP
PENAEUS-MONODON
RT-PCR
LIPOPOLYSACCHARIDE
AQUACULTURE
European lobster
Online Access:https://research-portal.st-andrews.ac.uk/en/researchoutput/realtime-pcr-quantification-of-the-in-vitro-effects-of-crustacean-immunostimulants-on-gene-expression-in-lobster-homarus-gammarus-granular-haemocytes(543a77a1-1d7e-45a7-a6b0-41dbcd23e113).html
https://doi.org/10.1016/j.dci.2004.05.006
http://www.scopus.com/inward/record.url?scp=4344591431&partnerID=8YFLogxK
Description
Summary:This paper presents data from an investigation of the mode of action of five different crustacean immunostimulants presented to European lobster (Homarus gammarus) granulocytes cultured in vitro. The experiments were designed to test whether or not the immunostimulants could cause the short-term up-regulation of genes coding for immune proteins without causing the cells to degranulate. Quantitative measurements of mRNA transcript abundance were made using real-time PCR and it was first necessary to isolate the complete gene sequences coding for the proteins prophenoloxidase (proPO), beta-1,3-glucan binding protein (betaGBP) and beta-actin (beta-act) in the lobster. These sequences were used to design TaqMan(TM) primer and fluorescent probe sets. The presented data indicated that the majority of the tested immunostimulants did not induce the up-regulation of immune-related gene expression in the granulocytes in isolation. Alternative modes of action, including the in vivo up-regulation of gene expression in haemopoetic tissues, are discussed. (C) 2004 Elsevier Ltd. All rights reserved.

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