NDLI: Expression and cellular pattern of relaxin mRNA in porcine corpora lutea during pregnancy

Content Provider	Springer Nature Link
Author	Kohsaka, Tetsuya Singh, Udai Pratap Yogo, Keiichiro Sasada, Hiroshi Taya, Kazuyoshi Hashizume, Kazuyoshi
Copyright Year	2007
Abstract	We developed an in situ hybridization method for detecting relaxin mRNA in the porcine corpus luteum (CL) by employing a non-radioactive probe and microwave fixation. We subsequently examined the expression and cellular patterns of relaxin mRNA in the CL during pregnancy and then evaluated whether relaxin mRNA was a factor limiting hormone production by the CL. Digoxigenin (DIG)-labeled RNA probes complementary to porcine relaxin mRNA were produced by in vitro transcription. The specificity was validated by showing, by Northern analysis, that the anti-sense probe hybridized to a 1.0-kb relaxin transcript in the CL. Microwave fixation (2-min irradiation in a conventional microwave oven) combined with DIG-labeled cRNA probes allowed precise and reliable analysis of relaxin mRNA, with superior retention of the mRNA and a higher resolving power. Application of this method to the porcine CL during pregnancy demonstrated that the relaxin mRNA level per cell and the percentage of mRNA-expressing cells increased as gestation progressed, with a marked decline near term. Northern analysis revealed the cellular pattern of relaxin mRNA localization, showing that the increase of relaxin mRNA with advancing pregnancy was attributable to an increase of both the cellular mRNA level and the percentage of mRNA-expressing cells. The present findings, taken together with known relaxin levels in the CL, reveal that changes of relaxin mRNA are correlated with changes of the hormone in the CL during pregnancy, suggesting that the relaxin level is determined by the amount of mRNA available for translation.
Starting Page	303
Ending Page	312
Page Count	10
File Format	PDF
ISSN	0302766X
Journal	Cell and Tissue Research
Volume Number	330
Issue Number	2
e-ISSN	14320878
Language	English
Publisher	Springer-Verlag
Publisher Date	2007-09-13
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Relaxin Gene expression In situ hybridization Northern hybridization Pregnancy Corpora lutea Pig Molecular Medicine Proteomics Human Genetics
Content Type	Text
Resource Type	Article
Subject	Cell Biology Histology Pathology and Forensic Medicine

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