Identification of stable and oestrus cycle-independent housekeeping genes in the rat mammary gland and other tissues
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Identification of stable and oestrus cycle-independent housekeeping genes in the rat mammary gland and other tissues. / Hvid, Henning; Ekstrøm, Claus Thorn; Vienberg, Sara; Oleksiewicz, Martin B.; Klopfleisch, Robert.
In: Veterinary Journal, Vol. 190, No. 1, 2011, p. 103-108.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Identification of stable and oestrus cycle-independent housekeeping genes in the rat mammary gland and other tissues
AU - Hvid, Henning
AU - Ekstrøm, Claus Thorn
AU - Vienberg, Sara
AU - Oleksiewicz, Martin B.
AU - Klopfleisch, Robert
N1 - Copyright © 2010 Elsevier Ltd. All rights reserved.
PY - 2011
Y1 - 2011
N2 - The function and development of the rat mammary gland is dependent on the oestrus cycle. Normalization of gene expression in mammary gland samples assessed by quantitative RT-PCR therefore requires housekeeping genes (HKGs) which are stably expressed during the oestrus cycle. mRNA expression of 10 HKGs was measured in the rat mammary gland at different phases of the oestrus cycle. In addition, mRNA expression of the HKGs was measured in a panel of other rat tissues comprising laser microdissected mammary gland alveolar lobules and interlobular connective tissue and macrodissected mammary gland, liver, skeletal muscle, colon and ovary samples. Expression and ranking of HKGs varied between tissues and oestrus cycle phases and several HKGs were necessary for normalization between samples. In the mammary gland samples, three HKGs (Sdha, Tbp, and Atp5b) were identified as the optimal combination of stably expressed genes across oestrus cycle phases. For normalization between samples from the entire panel of rat tissues, eight HKGs (Rps18, Eef1a1, B2m, Actb, Tbp, Hprt, Pgk1, and Sdha) were identified as the optimal combination. These HKGs are of general relevance for studies comparing gene expression between different rat tissues.
AB - The function and development of the rat mammary gland is dependent on the oestrus cycle. Normalization of gene expression in mammary gland samples assessed by quantitative RT-PCR therefore requires housekeeping genes (HKGs) which are stably expressed during the oestrus cycle. mRNA expression of 10 HKGs was measured in the rat mammary gland at different phases of the oestrus cycle. In addition, mRNA expression of the HKGs was measured in a panel of other rat tissues comprising laser microdissected mammary gland alveolar lobules and interlobular connective tissue and macrodissected mammary gland, liver, skeletal muscle, colon and ovary samples. Expression and ranking of HKGs varied between tissues and oestrus cycle phases and several HKGs were necessary for normalization between samples. In the mammary gland samples, three HKGs (Sdha, Tbp, and Atp5b) were identified as the optimal combination of stably expressed genes across oestrus cycle phases. For normalization between samples from the entire panel of rat tissues, eight HKGs (Rps18, Eef1a1, B2m, Actb, Tbp, Hprt, Pgk1, and Sdha) were identified as the optimal combination. These HKGs are of general relevance for studies comparing gene expression between different rat tissues.
KW - Animals
KW - Biological Markers
KW - Female
KW - Gene Expression Profiling
KW - Genes, Essential
KW - Mammary Glands, Animal
KW - Microdissection
KW - Polymerase Chain Reaction
KW - RNA, Messenger
KW - Rats
KW - Rats, Sprague-Dawley
KW - Reference Standards
KW - Tissue Distribution
U2 - 10.1016/j.tvjl.2010.09.002
DO - 10.1016/j.tvjl.2010.09.002
M3 - Journal article
C2 - 20952231
VL - 190
SP - 103
EP - 108
JO - The Veterinary Journal
JF - The Veterinary Journal
SN - 1090-0233
IS - 1
ER -
ID: 32350424