Abstract
We have recently characterized three different, alternatively spliced, partial turkey LH receptor (tLH-R) cDNA isoforms by the combination of reverse transcription-polymerase chain reaction (RT-PCR) and 5'- and 3'-rapid amplification of cDNA ends. The first cDNA (intact form: tLH-R(intact)) showed 98% and 72-75% similarity with chicken and mammalian LH receptor sequences, respectively. The other two cloned cDNA isoforms (insertion and truncated forms: tLH-R(insert) and tLH-R(trunc)) could encode truncated soluble protein isoforms that lack the transmembrane region. Northern blot analysis detected two transcripts of 3.0 kilobases (kb) (tLH-R(intact)) and 1.5 kb (tLH-R(trunc)) in the turkey ovary but could not discriminate a third alternatively spliced transcript (tLH-R(insert)) due to the small 86-base pair difference in the size range of approximately 3.0-kb mRNAs. But with the combination of RNase protection assay, RT-PCR, and Northern blot analysis, three different alternatively spliced tLH-R mRNA isoforms were quantified. Differential expression of the tLH-R mRNA isoforms was demonstrated in ovarian stromal tissue during various reproductive stages and in the theca and granulosa layer through follicular development. To gain a better understanding of the physiological significance of the three different tLH-R isoforms, total RNA from the theca layer through follicular development after prolactin (PRL) treatment was analyzed by RT-PCR. PRL treatment for 8-14 days significantly increased the steady-state levels of total tLH-R mRNAs, including tLH-R(insert) and tLH-R(trunc) mRNAs, compared to those in nontreated controls. In contrast, the steady-state levels of tLH-R(intact) mRNA during the same period was not significantly changed when compared to that in nontreated controls. The present study shows that tLH-R transcripts are alternatively spliced in a tissue-specific manner in the turkey and that the mechanism may, in part, be controlled hormonally.
| Original language | English |
|---|---|
| Pages (from-to) | 117-124 |
| Number of pages | 8 |
| Journal | Biology of Reproduction |
| Volume | 62 |
| Issue number | 1 |
| Publication status | Published - 2000 Jan 10 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Cell Biology
- Developmental Biology
- Embryology
Cite this
Three different Turkey luteinizing hormone receptor (tLH-R) isoforms II : Characterization of differentially regulated tLH-R messenger ribonucleic acid isoforms in the ovary. / You, Seungkwon; Kim, Hyunggee; El Halawani, Mohamed E.; Foster, Douglas N.
In: Biology of Reproduction, Vol. 62, No. 1, 10.01.2000, p. 117-124.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Three different Turkey luteinizing hormone receptor (tLH-R) isoforms II
T2 - Characterization of differentially regulated tLH-R messenger ribonucleic acid isoforms in the ovary
AU - You, Seungkwon
AU - Kim, Hyunggee
AU - El Halawani, Mohamed E.
AU - Foster, Douglas N.
PY - 2000/1/10
Y1 - 2000/1/10
N2 - We have recently characterized three different, alternatively spliced, partial turkey LH receptor (tLH-R) cDNA isoforms by the combination of reverse transcription-polymerase chain reaction (RT-PCR) and 5'- and 3'-rapid amplification of cDNA ends. The first cDNA (intact form: tLH-R(intact)) showed 98% and 72-75% similarity with chicken and mammalian LH receptor sequences, respectively. The other two cloned cDNA isoforms (insertion and truncated forms: tLH-R(insert) and tLH-R(trunc)) could encode truncated soluble protein isoforms that lack the transmembrane region. Northern blot analysis detected two transcripts of 3.0 kilobases (kb) (tLH-R(intact)) and 1.5 kb (tLH-R(trunc)) in the turkey ovary but could not discriminate a third alternatively spliced transcript (tLH-R(insert)) due to the small 86-base pair difference in the size range of approximately 3.0-kb mRNAs. But with the combination of RNase protection assay, RT-PCR, and Northern blot analysis, three different alternatively spliced tLH-R mRNA isoforms were quantified. Differential expression of the tLH-R mRNA isoforms was demonstrated in ovarian stromal tissue during various reproductive stages and in the theca and granulosa layer through follicular development. To gain a better understanding of the physiological significance of the three different tLH-R isoforms, total RNA from the theca layer through follicular development after prolactin (PRL) treatment was analyzed by RT-PCR. PRL treatment for 8-14 days significantly increased the steady-state levels of total tLH-R mRNAs, including tLH-R(insert) and tLH-R(trunc) mRNAs, compared to those in nontreated controls. In contrast, the steady-state levels of tLH-R(intact) mRNA during the same period was not significantly changed when compared to that in nontreated controls. The present study shows that tLH-R transcripts are alternatively spliced in a tissue-specific manner in the turkey and that the mechanism may, in part, be controlled hormonally.
AB - We have recently characterized three different, alternatively spliced, partial turkey LH receptor (tLH-R) cDNA isoforms by the combination of reverse transcription-polymerase chain reaction (RT-PCR) and 5'- and 3'-rapid amplification of cDNA ends. The first cDNA (intact form: tLH-R(intact)) showed 98% and 72-75% similarity with chicken and mammalian LH receptor sequences, respectively. The other two cloned cDNA isoforms (insertion and truncated forms: tLH-R(insert) and tLH-R(trunc)) could encode truncated soluble protein isoforms that lack the transmembrane region. Northern blot analysis detected two transcripts of 3.0 kilobases (kb) (tLH-R(intact)) and 1.5 kb (tLH-R(trunc)) in the turkey ovary but could not discriminate a third alternatively spliced transcript (tLH-R(insert)) due to the small 86-base pair difference in the size range of approximately 3.0-kb mRNAs. But with the combination of RNase protection assay, RT-PCR, and Northern blot analysis, three different alternatively spliced tLH-R mRNA isoforms were quantified. Differential expression of the tLH-R mRNA isoforms was demonstrated in ovarian stromal tissue during various reproductive stages and in the theca and granulosa layer through follicular development. To gain a better understanding of the physiological significance of the three different tLH-R isoforms, total RNA from the theca layer through follicular development after prolactin (PRL) treatment was analyzed by RT-PCR. PRL treatment for 8-14 days significantly increased the steady-state levels of total tLH-R mRNAs, including tLH-R(insert) and tLH-R(trunc) mRNAs, compared to those in nontreated controls. In contrast, the steady-state levels of tLH-R(intact) mRNA during the same period was not significantly changed when compared to that in nontreated controls. The present study shows that tLH-R transcripts are alternatively spliced in a tissue-specific manner in the turkey and that the mechanism may, in part, be controlled hormonally.
UR - http://www.scopus.com/inward/record.url?scp=0033971096&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0033971096&partnerID=8YFLogxK
M3 - Article
C2 - 10611075
AN - SCOPUS:0033971096
VL - 62
SP - 117
EP - 124
JO - Biology of Reproduction
JF - Biology of Reproduction
SN - 0006-3363
IS - 1
ER -