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Cloning and characterization of prostaglandin endoperoxide synthase-1 and -2 from the brook trout ovary
S B Roberts1, D M Langenau, F W Goetz
1University of Notre Dame, Department of Biological Sciences, P.O. Box 369, Notre Dame, IN 46556-0369, USA.
Abstract:
Using a combination of reverse transcription-PCR and library screening, the cDNAs for prostaglandin endoperoxide synthase-1 (PGS-1) and 2 (PGS-2) were isolated from the brook trout ovary. The brook trout PGS-1 cDNA encodes for a 598 amino acid protein that is 69% identical to mammalian PGS-1. PGS-1 transcripts were observed in the ovary, spleen, gills, head kidney, trunk kidney, intestine, stomach, skin and heart. To our knowledge, this is the first characterization of a non-mammalian PGS-1 cDNA. The brook trout PGS-2 encodes for a 607 amino acid protein that is 69% identical to mammalian PGS-2 and was observed in the skin, gills, stomach and heart. PGS-2 transcripts were highly upregulated in the ovaries by the phorbol ester, phorbol-12-myristate-13-acetate, in combination with the calcium ionophore, A23187. However, PGS-2 was not observed in the ovary of brook trout undergoing natural oocyte maturation and ovulation.
Insights
This study characterizes prostaglandin endoperoxide synthase-1 (PGS-1) and -2 (PGS-2) cDNAs in brook trout, revealing their presence and regulation, particularly PGS-2
Area of Science:
- Comparative genomics
- Fish reproductive biology
- Prostaglandin synthesis pathways
Background:
- Prostaglandin endoperoxide synthases (PGS) are key enzymes in prostaglandin synthesis.
- Understanding PGS function in non-mammalian vertebrates is crucial for comparative physiology.
- This research focuses on brook trout (Salvelinus fontinalis) as a model organism.
Purpose of the Study:
- To isolate and characterize prostaglandin endoperoxide synthase-1 (PGS-1) and -2 (PGS-2) cDNAs from brook trout.
- To investigate the tissue distribution of PGS-1 and PGS-2 transcripts in brook trout.
- To examine the regulation of PGS-2 expression in brook trout ovaries.
Main Methods:
- cDNA isolation using reverse transcription-PCR and library screening.
- Sequence analysis to determine protein identity and homology to mammalian PGS.
- Transcript distribution analysis via detection of PGS-1 and PGS-2 mRNA in various tissues.
- Hormonal induction studies using phorbol ester and calcium ionophore to assess PGS-2 regulation in ovaries.
Main Results:
- Brook trout PGS-1 cDNA encodes a 598 amino acid protein, 69% identical to mammalian PGS-1, with widespread tissue distribution.
- Brook trout PGS-2 cDNA encodes a 607 amino acid protein, 69% identical to mammalian PGS-2, found in skin, gills, stomach, and heart.
- PGS-2 transcripts were significantly upregulated in ovaries upon stimulation with phorbol-12-myristate-13-acetate and A23187.
- PGS-2 was notably absent in ovaries of brook trout undergoing natural oocyte maturation and ovulation.
Conclusions:
- This is the first characterization of non-mammalian PGS-1 and PGS-2 cDNAs, providing insights into fish prostaglandin synthesis.
- Differential expression and regulation of PGS-2 in brook trout ovaries suggest a role in reproductive processes.
- The absence of PGS-2 during natural ovulation warrants further investigation into its specific role in fish reproduction.
