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Caenorhabditis Elegans Mutants Predict Regulation of Fatty Acids and Endocannabinoids by the CYP-35A Gene Family.

Aarnio V, Lehtonen M, Storvik M, Callaway JC, Lakso M, Wong G - Front Pharmacol (2011)

Bottom Line: The overall fatty acid compositions of CYP mutants did not alter dramatically, although modest but significant changes were observed. cyp-35A1 and cyp-35A5 mutants had significantly higher levels of C18:1n7 and lower C18:2n6c.In the cyp-35A5 mutant, the levels of the endocannabinoid anandamide were found to be 4.6-fold higher than in wild-type.These results demonstrate a dynamic role for CYP-35A subfamily members in maintaining the diversity of fatty acid profiles in C. elegans, and more generally highlight the importance of CYPs in generating both structural and signaling fatty acid functions in other organisms.

View Article: PubMed Central - PubMed

Affiliation: Functional Genomics and Bioinformatics Laboratory, Department of Biosciences and Department of Neurobiology, A. I. Virtanen Institute, Biocenter Finland, University of Eastern Finland Kuopio, Finland.

ABSTRACT

Background: Cytochrome P450s (CYPs) are mono-oxygenases that metabolize endogenous compounds, such as fatty acids and lipid signaling molecules, and furthermore have a role in metabolism of xenobiotics. In order to investigate the role of CYP genes in fat metabolism at the molecular level, four Caenorhabditis elegans mutants lacking functional CYP-35A1, CYP-35A2, CYP-35A4, and CYP-35A5 were characterized. Relative amounts of fatty acids, as well as endocannabinoids, which regulate weight gain and accumulation of fats in mammals, were measured while fat contents in worms were visualized using Oil-Red-O staining.

Results: The cyp-35A1 and cyp-35A5 mutants had a significantly lower intestinal fat content than wild-type animals, whereas cyp-35A2 and cyp-35A4 mutants appeared normal. The overall fatty acid compositions of CYP mutants did not alter dramatically, although modest but significant changes were observed. cyp-35A1 and cyp-35A5 mutants had significantly higher levels of C18:1n7 and lower C18:2n6c. All four mutants had higher relative amounts of C18:1n7 than the wild-type. In the cyp-35A5 mutant, the levels of the endocannabinoid anandamide were found to be 4.6-fold higher than in wild-type. Several fatty acid synthesis genes were over-expressed in cyp-35A1 including fat-2. Feeding oleic or elaidic triglycerides to wild-type animals demonstrated that cyp-35A1 transcriptional levels are insensitive to environmental exposure of these fats, while cyp-35A2, cyp-35A4, and cyp-35A5 were significantly down regulated.

Conclusion: These results demonstrate a dynamic role for CYP-35A subfamily members in maintaining the diversity of fatty acid profiles in C. elegans, and more generally highlight the importance of CYPs in generating both structural and signaling fatty acid functions in other organisms.

No MeSH data available.


Related in: MedlinePlus

(A) Changes in the expression levels of fatty acid synthesis genes (fold change compared to wild-type ± SEM). *t-test p-value < 0.05, **t-test p-value < 0.01, ***t-test p-value < 0.001. (B) Schema of the synthesis of fatty acids and the enzymes involved.  = fatty acid percentage or gene expression increased in cyp-35A1 mutant,  = fatty acid percentage or gene expression decreased in cyp-35A1 mutant,  = fatty acid percentage or gene expression increased in cyp-35A5 mutant,  = fatty acid percentage or gene expression decreased in cyp-35A5 mutant.
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Figure 5: (A) Changes in the expression levels of fatty acid synthesis genes (fold change compared to wild-type ± SEM). *t-test p-value < 0.05, **t-test p-value < 0.01, ***t-test p-value < 0.001. (B) Schema of the synthesis of fatty acids and the enzymes involved.  = fatty acid percentage or gene expression increased in cyp-35A1 mutant,  = fatty acid percentage or gene expression decreased in cyp-35A1 mutant,  = fatty acid percentage or gene expression increased in cyp-35A5 mutant,  = fatty acid percentage or gene expression decreased in cyp-35A5 mutant.

Mentions: In the cyp-35A1(ok2306) mutant many genes required in the synthesis of long chain fatty acids appeared to be expressed in 2–3.5 times higher amounts than in wild-type (Figure 5A). The increases in fat-2, fat-3, fat-5, fat-6, fat-7, elo-1, and elo-6 were significant (p < 0.05). In a different cyp-35A1(ok1414) mutant allele strain, only fat-2 was significantly over-expressed, but all the measured genes were found in somewhat higher amounts than in the wild-type (data not shown). In the cyp-35A5 mutant many fatty acid synthesis genes seemed to be under-expressed, although only the changes in fat-1 and fat-3 were significant. The fatty acid beta-oxidation gene acs-2 was slightly (1.7-fold) over-expressed in the cyp-35A2 mutant and under-expressed (fold change 0.8) in the cyp-35A5 mutant (data not shown).


Caenorhabditis Elegans Mutants Predict Regulation of Fatty Acids and Endocannabinoids by the CYP-35A Gene Family.

Aarnio V, Lehtonen M, Storvik M, Callaway JC, Lakso M, Wong G - Front Pharmacol (2011)

(A) Changes in the expression levels of fatty acid synthesis genes (fold change compared to wild-type ± SEM). *t-test p-value < 0.05, **t-test p-value < 0.01, ***t-test p-value < 0.001. (B) Schema of the synthesis of fatty acids and the enzymes involved.  = fatty acid percentage or gene expression increased in cyp-35A1 mutant,  = fatty acid percentage or gene expression decreased in cyp-35A1 mutant,  = fatty acid percentage or gene expression increased in cyp-35A5 mutant,  = fatty acid percentage or gene expression decreased in cyp-35A5 mutant.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3108558&req=5

Figure 5: (A) Changes in the expression levels of fatty acid synthesis genes (fold change compared to wild-type ± SEM). *t-test p-value < 0.05, **t-test p-value < 0.01, ***t-test p-value < 0.001. (B) Schema of the synthesis of fatty acids and the enzymes involved.  = fatty acid percentage or gene expression increased in cyp-35A1 mutant,  = fatty acid percentage or gene expression decreased in cyp-35A1 mutant,  = fatty acid percentage or gene expression increased in cyp-35A5 mutant,  = fatty acid percentage or gene expression decreased in cyp-35A5 mutant.
Mentions: In the cyp-35A1(ok2306) mutant many genes required in the synthesis of long chain fatty acids appeared to be expressed in 2–3.5 times higher amounts than in wild-type (Figure 5A). The increases in fat-2, fat-3, fat-5, fat-6, fat-7, elo-1, and elo-6 were significant (p < 0.05). In a different cyp-35A1(ok1414) mutant allele strain, only fat-2 was significantly over-expressed, but all the measured genes were found in somewhat higher amounts than in the wild-type (data not shown). In the cyp-35A5 mutant many fatty acid synthesis genes seemed to be under-expressed, although only the changes in fat-1 and fat-3 were significant. The fatty acid beta-oxidation gene acs-2 was slightly (1.7-fold) over-expressed in the cyp-35A2 mutant and under-expressed (fold change 0.8) in the cyp-35A5 mutant (data not shown).

Bottom Line: The overall fatty acid compositions of CYP mutants did not alter dramatically, although modest but significant changes were observed. cyp-35A1 and cyp-35A5 mutants had significantly higher levels of C18:1n7 and lower C18:2n6c.In the cyp-35A5 mutant, the levels of the endocannabinoid anandamide were found to be 4.6-fold higher than in wild-type.These results demonstrate a dynamic role for CYP-35A subfamily members in maintaining the diversity of fatty acid profiles in C. elegans, and more generally highlight the importance of CYPs in generating both structural and signaling fatty acid functions in other organisms.

View Article: PubMed Central - PubMed

Affiliation: Functional Genomics and Bioinformatics Laboratory, Department of Biosciences and Department of Neurobiology, A. I. Virtanen Institute, Biocenter Finland, University of Eastern Finland Kuopio, Finland.

ABSTRACT

Background: Cytochrome P450s (CYPs) are mono-oxygenases that metabolize endogenous compounds, such as fatty acids and lipid signaling molecules, and furthermore have a role in metabolism of xenobiotics. In order to investigate the role of CYP genes in fat metabolism at the molecular level, four Caenorhabditis elegans mutants lacking functional CYP-35A1, CYP-35A2, CYP-35A4, and CYP-35A5 were characterized. Relative amounts of fatty acids, as well as endocannabinoids, which regulate weight gain and accumulation of fats in mammals, were measured while fat contents in worms were visualized using Oil-Red-O staining.

Results: The cyp-35A1 and cyp-35A5 mutants had a significantly lower intestinal fat content than wild-type animals, whereas cyp-35A2 and cyp-35A4 mutants appeared normal. The overall fatty acid compositions of CYP mutants did not alter dramatically, although modest but significant changes were observed. cyp-35A1 and cyp-35A5 mutants had significantly higher levels of C18:1n7 and lower C18:2n6c. All four mutants had higher relative amounts of C18:1n7 than the wild-type. In the cyp-35A5 mutant, the levels of the endocannabinoid anandamide were found to be 4.6-fold higher than in wild-type. Several fatty acid synthesis genes were over-expressed in cyp-35A1 including fat-2. Feeding oleic or elaidic triglycerides to wild-type animals demonstrated that cyp-35A1 transcriptional levels are insensitive to environmental exposure of these fats, while cyp-35A2, cyp-35A4, and cyp-35A5 were significantly down regulated.

Conclusion: These results demonstrate a dynamic role for CYP-35A subfamily members in maintaining the diversity of fatty acid profiles in C. elegans, and more generally highlight the importance of CYPs in generating both structural and signaling fatty acid functions in other organisms.

No MeSH data available.


Related in: MedlinePlus