2006 - Progress

• Time-course analysis of spiggin mRNA expression in kidney was performed using MT exposure fish.
• Cloning of full-length ARb and partial ARa.
• Immunohistochmical analysis of the sticleback AR protein in kidney using human AR-antibody.
• Continued transfer of knowledge on stickleback breeding practices from UK to Japan, in order to achieve efficient maturation in the laboratory.

2006 - Objectives

• Application of spiggin mRNA quantification to in vivo evaluation system using female
  stickleback.
• Molecular cloning of stickleback ARs (continued)
• Preparation of reporter gene assay of ARs.
• Histological localization of AR (using antibody and RNA probe)
• Transfer of knowledge on stickleback breeding practices from UK to Japan, in order to
  achieve efficient maturation in the laboratory.
• Characterisation of physiologically relevant androgen(s) during male sexual maturation
• Differential protein expression in male brain after exposure to EDCs (2D gels)
• Evaluate effects of anti-androgens on physiology and behaviour (continued) 

 

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Collaborative Achievements up to 2005

• Cloning of two types of spiggin genes (SPG-I and SPG-II)
• Expression data for spiggin genes (SPG-I and SPG-II) in kidney
• Fundamental studies establishing primary cultivation of stickleback cells (kidney and liver)
• Purification of vitellogenin and lipovitellin in stickleback

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2005 - Objectives

• Development of method for detecting androgenic/anti-androgenic effects of chemicals in vivo, using spiggin.
• Development of stickleback AR reporter gene assays and thier use for assessing (anti-) androgenic activity of chemicals and environmental samples.
• Establishment of a full life-cycle test for stickleback test

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2005 - Progress against Objectives

 

Development of method for detecting androgenic/anti-androgenic effects of chemicals in vivo, using spiggin

• Cloning of spiggin I and II, development of QPCR assays, and assessments of androgenic chemicals in those assays.
• Method (in vivo) established to assess the potencies of anti-androgens, including linuron and fenitrothione

Development of stickleback AR reporter gene assays and assessment of chemicals and environmental samples through these receptors

• Cloning of partial fragment of stickleback AR alpha and beta

Establishment of stickleback breeding method for life-cycle test

• Literature review and recommended approach reported to DEFRA
• Good development of a pair breeding assay. Integration of behavioural traits to compliment physiological responses to EDCs
• Establishment of breeding system for stickleback in Japan

Additional deliverables

• Cloning and developmental expression of stickleback ER alpha and ER beta, and demonstration of an up-regulation of ER alpha on exposure to oestrogen 

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