TY - JOUR
T1 - Quantitative analysis of thyroid-stimulating hormone messenger RNA and heterogeneous nuclear RNA in hypothyroid rats
AU - Sugimoto, Koreaki
AU - Mori, Kouki
AU - Uchida, Katsuya
AU - Kobayashi, Daisuke
AU - Itoi, Keiichi
N1 - Funding Information:
This work was partly supported by the Saito Ho-On Kai Foundation and the Research Grant for Scientific Research (C) 18592066 from the Ministry of Education, Culture, Sports, Science and Technology.
Funding Information:
The authors declare no conflict of interest. The study was not funded by Roche Applied Science Inc. The authors have no financial interest related to the material described in the manuscript.
PY - 2007/9/14
Y1 - 2007/9/14
N2 - Thyroid-stimulating hormone (TSH) stimulates the synthesis and release of thyroid hormones including triiodothyronine (T3) and thyroxine (T4). Semiquantitative analyses using northern blot and in situ hybridization suggested that TSH gene transcription is upregulated under conditions of hypothyroidism. However, no quantitative analysis of TSH gene expression using real-time polymerase chain reaction (PCR) has been reported. In this study, we quantitated the TSHβ messenger ribonucleic acid (mRNA) level as well as the TSHβ heterogeneous nuclear ribonucleic acid (hnRNA) level in the anterior pituitary of hypothyroid rats, by real-time PCR using the LightCycler® system. The hnRNA is the primary deoxyribonucleic acid (DNA) transcript, which reflects the transcription rate more reliably than the mRNA because of its short half-life. In the anterior pituitary of rats with methimazol-induced chronic hypothyroidism, both mRNA and hnRNA expression of TSHβ were upregulated fourfold relative to normal rats (n = 4). Our method provides a rapid and accurate measure of gene transcription. In the present report, we described a technique for accurate measurement of TSHβ hnRNA level.
AB - Thyroid-stimulating hormone (TSH) stimulates the synthesis and release of thyroid hormones including triiodothyronine (T3) and thyroxine (T4). Semiquantitative analyses using northern blot and in situ hybridization suggested that TSH gene transcription is upregulated under conditions of hypothyroidism. However, no quantitative analysis of TSH gene expression using real-time polymerase chain reaction (PCR) has been reported. In this study, we quantitated the TSHβ messenger ribonucleic acid (mRNA) level as well as the TSHβ heterogeneous nuclear ribonucleic acid (hnRNA) level in the anterior pituitary of hypothyroid rats, by real-time PCR using the LightCycler® system. The hnRNA is the primary deoxyribonucleic acid (DNA) transcript, which reflects the transcription rate more reliably than the mRNA because of its short half-life. In the anterior pituitary of rats with methimazol-induced chronic hypothyroidism, both mRNA and hnRNA expression of TSHβ were upregulated fourfold relative to normal rats (n = 4). Our method provides a rapid and accurate measure of gene transcription. In the present report, we described a technique for accurate measurement of TSHβ hnRNA level.
KW - Heterogeneous nuclear RNA (hnRNA)
KW - LightCycler
KW - Real-time PCR
KW - Thyroid-stimulating hormone (TSH
KW - Thyrotropin-releasing hormone (TRH)
KW - thyrotropin)
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U2 - 10.1016/j.brainresbull.2007.06.004
DO - 10.1016/j.brainresbull.2007.06.004
M3 - Article
C2 - 17683800
AN - SCOPUS:34547507537
VL - 74
SP - 142
EP - 146
JO - Journal of Electrophysiological Techniques
JF - Journal of Electrophysiological Techniques
SN - 0361-9230
IS - 1-3
ER -