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Published on: January 6, 2023
Discovery and characterization of endometrial epithelial messenger ribonucleic acids using the ovine uterine gland
T E Spencer1, A G Stagg, M M Joyce
1Department of Animal Science, Albert B. Alkek Institute of Biosciences, Texas A&M University, College Station 77843-2471, USA. tspencer@ansc.tamu.edu
Abstract:
Prolonged exposure of the developing neonatal ovine uterus to a progestin from birth prevents uterine gland development and creates an adult endometrial phenotype characterized by the absence of glandular epithelium, the uterine gland knockout (UGKO) phenotype. This study used endometrium from normal and UGKO sheep to identify messenger RNAs (mRNAs) expressed differentially in the endometrial epithelium using the molecular techniques of mRNA differential display PCR (DD-PCR) and suppression subtractive complementary DNA (cDNA) hybridization (SSH). Sequence analyses of DD- and SSH-identified and cloned cDNAs indicated similarity of some to known mRNAs, including beta-lactoglobulin, alkaline phosphatase, type B and D endogenous sheep retroviruses, gp330/megalin, matrix Gla protein, and others. Other cDNAs were not similar to any known sequences and are considered novel, although some of these match human expressed sequence tags. In situ hybridization analyses of uteri from cyclic and pregnant ewes indicated that all DD-PCR- and SSH-identified mRNAs were expressed in either the endometrial lumenal and/or glandular epithelium, although some were also expressed in other uterine cell types. Northern and in situ hybridization analyses revealed that patterns of mRNA expression for most clones were affected by the day of the estrous cycle and pregnancy in a manner consistent with regulation by progesterone. Studies demonstrate the utility of the ovine UGKO model as a tool with which to identify known and novel uterine epithelial-specific genes. Cloned cDNAs identified here are expressed sequence tags useful for comparative and physical genetic mapping and may be used to reveal new factors and pathways regulating endometrial function.
Insights
Neonatal progestin exposure creates a uterine gland knockout (UGKO) sheep model. This study identified novel and known endometrial genes in UGKO sheep, aiding research into uterine function regulation.
Area of Science:
- Reproductive Biology
- Genomics
- Molecular Endocrinology
Background:
- Neonatal exposure to progestins disrupts ovine uterine development, leading to the uterine gland knockout (UGKO) phenotype.
- The UGKO phenotype is characterized by an absence of glandular epithelium in the adult endometrium.
- Understanding gene expression in the UGKO model provides insights into endometrial development and function.
Purpose of the Study:
- To identify differentially expressed messenger RNAs (mRNAs) in the endometrial epithelium of normal versus UGKO sheep.
- To characterize known and novel genes involved in uterine function using molecular techniques.
- To validate the ovine UGKO model for identifying uterine epithelial-specific genes.
Main Methods:
- Differential display PCR (DD-PCR) and suppression subtractive complementary DNA (cDNA) hybridization (SSH) were employed to identify unique cDNAs.
- Sequence analysis was performed on cloned cDNAs to identify similarities to known genes and novel sequences.
- In situ and Northern hybridization were used to determine mRNA expression patterns in uterine tissues from cyclic and pregnant ewes.
Main Results:
- Several known mRNAs, including beta-lactoglobulin and alkaline phosphatase, were identified, alongside novel sequences, some matching human expressed sequence tags.
- All identified mRNAs were localized to the endometrial epithelium, with some also found in other uterine cell types.
- mRNA expression patterns varied with the estrous cycle and pregnancy, suggesting progesterone regulation.
Conclusions:
- The ovine UGKO model is effective for identifying both known and novel uterine epithelial-specific genes.
- The identified cDNAs serve as expressed sequence tags for genetic mapping and understanding endometrial regulation.
- This research provides valuable tools and data for investigating factors and pathways governing endometrial function.
