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Expression of growth-related genes in muscle during fasting and refeeding of juvenile Atlantic halibut, Hippoglossus hippoglossus L
Ist Teil von
Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 2009, Vol.152 (1), p.47-53
Ort / Verlag
England: Elsevier Inc
Erscheinungsjahr
2009
Quelle
ScienceDirect
Beschreibungen/Notizen
The aim of this study was to establish a fasting–refeeding protocol to investigate the expression of growth-related genes during the transition between catabolic and anabolic states in Atlantic halibut (
Hippoglossus hippoglossus L
.). Juveniles of ~
950 g were maintained at ambient temperature (5–8 °C) until the 1st of May, then fasted for two months and refed for two months at 7.7–8.0 °C under continuous low light. Fast epaxial myotomal muscle was sampled at −
64 d (days), −
38 d, 0 d (start of refeeding), 3 d, 7 d, 14 d, 30 d and 60 d. Average body mass was unchanged over the fasting period but increased by 24.4% following 60 d refeeding. qPCR was used to analyse the stability of expression of five potential reference genes (
Eef2,
Fau,
18S
rRNA,
Actb and
Tubb2) with
GeNorm and
Normfinder. Expression of the growth-related genes,
cathepsin
B (
ctsb),
cathepsin
D (
ctsd),
insulin-like
growth
factor-I
and
II (
IGF-I and
II) and
insulin-like
growth
factor-I
receptor
1a (
IGF-IRa) was normalised using the geometric average of the two most stable housekeeping genes,
Fau and
18S
rRNA.
IGF-I mRNA showed a transient 2.6-fold increase in abundance with refeeding at 7 d whilst transcripts for
IGF-II and
IGF-IRa were elevated during fasting and decreased 3.8-fold and 3-fold between the 0 d and 3 d samples respectively.
Ctsb expression increased between −
64 d and 0 d and then decreased ~
10-fold by 14 d refeeding. In contrast,
ctsd was relatively unaffected by the fasting–refeeding cycle, showing a modest (~
35%) transient decrease in expression between the 0 d and 30 d refeeding samples. It was concluded that the experimental protocol adopted and housekeeping genes identified were suitable for investigating the catabolic–anabolic transition in halibut skeletal muscle.