Identification and initial characterization of calcyclin and phospholipase A2 in equine conceptuses.
Abstract: For development to proceed normally, the appropriate genes must be expressed in the correct tissues and in the correct time frame. Knowledge of gene expression during development provides information about the changes taking place within the conceptus as well as possible reasons for pregnancy failure. However, little is known about gene expression during development in the equine conceptus. In this study, we examined differences in gene expression between day 12 and day 15 equine conceptuses by suppression subtractive hybridization. This technique was used to isolate transcripts that are more abundantly expressed in day 15 conceptuses compared to day 12 conceptuses. Between day 12 and 15 of pregnancy in horses, maternal recognition of pregnancy occurs, gastrulation is taking place, and mesoderm is beginning to form. Fifty cDNA clones were isolated, sequenced, and compared to known sequences in the GenBank database. Two cDNA clones identified that were of primary interest were calcyclin and phospholipase A2. Calcyclin is a calcium-binding protein of the S-100 protein family that has been found in mouse decidua and trophoblast. Calcyclin was found to be expressed in both day 12 and 15 equine conceptuses, with approximately a 30-fold increase in transcript abundance between days 12 and 15. Phospholipase A2 is an enzyme that cleaves phospholipids to release fatty acids and is involved in arachidonic acid release needed for prostaglandin, thromboxane, and leukotriene synthesis. Multiple forms of PLA2, that appear to be differentially regulated in day 12 and 15 conceptuses, were detected by northern blotting.
Publication Date: 1999-05-20 PubMed ID: 10331456DOI: 10.1002/(SICI)1098-2795(199906)53:2<179::AID-MRD7>3.0.CO;2-PGoogle Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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The research discusses the gene expression in equine (horse) conceptuses and identifies the presence of calcyclin and phospholipase A2. Their expression increases between the crucial developmental stages of day 12 and day 15, suggesting potential roles in equine development and pregnancy.
Study Objective and Process
- The focus of the study was to understand the gene expression during the development of equine conceptuses. Specifically, the researchers aimed to explore the differences in gene expression between day 12 and day 15 of the development stage.
- To achieve this, a technique called suppression subtractive hybridization was employed. This process allows for the isolation of transcripts (segments of RNA) that are more abundantly expressed in day 15 conceptuses compared to those on day 12.
Significance of Days 12 and 15 in Equine Pregnancy
- Between day 12 and day 15 of pregnancy in horses, several key developmental processes occur. These include the maternal recognition of pregnancy, gastrulation (the formation of germ layers that will develop into specialized body tissue), and the initial formation of the mesoderm (a layer that results from gastrulation).
Identification of Key Genes
- The researchers sequenced and compared fifty cDNA clones derived from the gene expression analysis to known sequences in the GenBank database.
- Two cDNA clones identified were calcyclin and phospholipase A2, which seemed to play significant roles considering their increased expression between day 12 and day 15.
Role of Calcyclin and Phospholipase A2
- Calcyclin is a calcium-binding protein of the S-100 protein family, previously found in mouse decidua and trophoblast, tissues essential to embryo implantation and placenta formation. Its presence and 30-fold increase in expression suggest importance for equine pregnancy and development.
- Phospholipase A2 acts as an enzyme that splits phospholipids to release fatty acids, playing a part in the synthesis of essential substances like prostaglandins, thromboxanes, and leukotrienes. Multiple forms of phospholipase A2 were detected in day 12 and 15 conceptuses, suggesting differential regulation during these stages.
Cite This Article
APA
Simpson KS, Adams MH, Behrendt-Adam CY, Baker CB, McDowell KJ.
(1999).
Identification and initial characterization of calcyclin and phospholipase A2 in equine conceptuses.
Mol Reprod Dev, 53(2), 179-187.
https://doi.org/10.1002/(SICI)1098-2795(199906)53:2<179::AID-MRD7>3.0.CO;2-P Publication
Researcher Affiliations
- Gluck Equine Research Center, Department of Veterinary Science, University of Kentucky, Lexington 40546-0099, USA. kss@pop.uky.edu
MeSH Terms
- Amino Acid Sequence
- Animals
- Base Sequence
- Cell Cycle Proteins
- DNA, Complementary
- Embryo Implantation
- Embryonic and Fetal Development
- Female
- Gene Expression Regulation, Developmental
- Gene Library
- Horses / embryology
- Horses / genetics
- Humans
- Male
- Mice
- Molecular Sequence Data
- Nucleic Acid Hybridization
- Phospholipases A / genetics
- Phospholipases A2
- Pregnancy
- S100 Calcium Binding Protein A6
- S100 Proteins / genetics
Grant Funding
- NIH T32HD07346 / NICHD NIH HHS
Citations
This article has been cited 1 times.- Gadgil C, Rink A, Beattie C, Hu WS. A mathematical model for suppression subtractive hybridization.. Comp Funct Genomics 2002;3(5):405-22.
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