Measurement of PTHrP, PTHR1, and CaSR Expression Levels in Tissues of Sea Bream (Sparus aurata) Using Quantitative PCR

: A quantitative PCR (Q‐PCR) method has been established to measure the mRNA expression levels of parathyroid hormone‐related protein (PTHrP), parathyroid hormone receptor type 1 (PTHR1), and calcium‐sensing receptor (CaSR) in sea bream (Sparus aurata), using the housekeeping gene, β‐actin, as endog...

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Bibliographic Details
Published in:Annals of the New York Academy of Sciences 2005-04, Vol.1040 (1), p.340-344
Main Authors: HANG, X M, POWER, D, FLIK, G, BALMENT, R J
Format: Article
Language:English
Subjects:
Animals
Bream
CaSR
Gene expression
Gene Expression Regulation - physiology
Genes
Gonads
Hormones
Kidneys
Marine
Parathyroid Hormone-Related Protein - biosynthesis
Parathyroid Hormone-Related Protein - genetics
Polymerase Chain Reaction - methods
PTHR1
PTHrP
Q-PCR
Receptor, Parathyroid Hormone, Type 1 - biosynthesis
Receptor, Parathyroid Hormone, Type 1 - genetics
Receptors
Receptors, Calcium-Sensing - biosynthesis
Receptors, Calcium-Sensing - genetics
sea bream
Sea Bream - genetics
Sea Bream - metabolism
Sea water
Sparus aurata
Tissue Distribution - genetics
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Summary:: A quantitative PCR (Q‐PCR) method has been established to measure the mRNA expression levels of parathyroid hormone‐related protein (PTHrP), parathyroid hormone receptor type 1 (PTHR1), and calcium‐sensing receptor (CaSR) in sea bream (Sparus aurata), using the housekeeping gene, β‐actin, as endogenous control. TaqMan primers and probes were designed using the Primer Express program, according to the published/unpublished sequences of the three target genes and β‐actin of sea bream. Different tissues including gill, kidney, duodenum, hindgut, rectum, liver, heart, brain, pituitary, skin, muscle, and gonad were removed and immediately snap‐frozen from three juvenile sea bream (100‐150 g) cultured in sea water. The mRNAs were extracted and reverse‐transcribed into cDNAs, which were subsequently examined by the ABI 5700 system using an optimized Q‐PCR method. Triplicate measures of each sample indicated consistency of the technique. However, the mRNA expression levels for each transcript in these tissues were variable between fish and also relatively low. Nevertheless, this methodology can be used in the future studies of factors that may alter gene expression in these tissues.
ISSN:0077-8923
1749-6632
DOI:10.1196/annals.1327.056