Limits...
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

Fat content visualized by Oil-Red-O staining. N2, starved N2, daf-2, cyp-35A1, cyp-35A2, cyp-35A4, cyp-35A5. Animals were grown and treated as described in Materials and Methods. Staining was performed on populations of day-1 adult animals and photographed using transmitted light microscopy at 200 magnification. Panels shown are three typical fields per mutant or treatment. (A-C), wild-type; (D-F), wild-type starved; (G-I), daf-2 mutant; (J-L), cyp-35A1 mutant; (M-O), cyp-35A2 mutant; (P-R), cyp-35A4 mutant; (S-U), cyp-35A5 mutant.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3108558&req=5

Figure 2: Fat content visualized by Oil-Red-O staining. N2, starved N2, daf-2, cyp-35A1, cyp-35A2, cyp-35A4, cyp-35A5. Animals were grown and treated as described in Materials and Methods. Staining was performed on populations of day-1 adult animals and photographed using transmitted light microscopy at 200 magnification. Panels shown are three typical fields per mutant or treatment. (A-C), wild-type; (D-F), wild-type starved; (G-I), daf-2 mutant; (J-L), cyp-35A1 mutant; (M-O), cyp-35A2 mutant; (P-R), cyp-35A4 mutant; (S-U), cyp-35A5 mutant.

Mentions: Wild-type N2 strain animals feeding on bacteria show abundant Oil-Red-O staining, particularly in the intestine and mature oocytes (Figures 2A–C). Animals starved for a day show loss of staining, in the intestine and less in oocytes (Figures 2D–F). The daf-2 mutant, as a positive control, showed heavy fat staining in the intestine (Figures 2G–I). In contrast, the cyp-35A1 mutant showed light Oil-Red-O staining in the intestine, mainly in the anterior part, and oocytes (Figures 2J–L), resembling the starved wild-type that also had staining in the eggs but not visible amounts in the intestine. The cyp-35A2 and cyp-35A4 mutants had about the same level of Oil-Red-O staining as the wild-type (Figures 2M–R). The cyp-35A5 had some Oil-Red-O staining in the intestine but visibly less than wild-type (Figures 2S–U).


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)

Fat content visualized by Oil-Red-O staining. N2, starved N2, daf-2, cyp-35A1, cyp-35A2, cyp-35A4, cyp-35A5. Animals were grown and treated as described in Materials and Methods. Staining was performed on populations of day-1 adult animals and photographed using transmitted light microscopy at 200 magnification. Panels shown are three typical fields per mutant or treatment. (A-C), wild-type; (D-F), wild-type starved; (G-I), daf-2 mutant; (J-L), cyp-35A1 mutant; (M-O), cyp-35A2 mutant; (P-R), cyp-35A4 mutant; (S-U), cyp-35A5 mutant.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 2: Fat content visualized by Oil-Red-O staining. N2, starved N2, daf-2, cyp-35A1, cyp-35A2, cyp-35A4, cyp-35A5. Animals were grown and treated as described in Materials and Methods. Staining was performed on populations of day-1 adult animals and photographed using transmitted light microscopy at 200 magnification. Panels shown are three typical fields per mutant or treatment. (A-C), wild-type; (D-F), wild-type starved; (G-I), daf-2 mutant; (J-L), cyp-35A1 mutant; (M-O), cyp-35A2 mutant; (P-R), cyp-35A4 mutant; (S-U), cyp-35A5 mutant.
Mentions: Wild-type N2 strain animals feeding on bacteria show abundant Oil-Red-O staining, particularly in the intestine and mature oocytes (Figures 2A–C). Animals starved for a day show loss of staining, in the intestine and less in oocytes (Figures 2D–F). The daf-2 mutant, as a positive control, showed heavy fat staining in the intestine (Figures 2G–I). In contrast, the cyp-35A1 mutant showed light Oil-Red-O staining in the intestine, mainly in the anterior part, and oocytes (Figures 2J–L), resembling the starved wild-type that also had staining in the eggs but not visible amounts in the intestine. The cyp-35A2 and cyp-35A4 mutants had about the same level of Oil-Red-O staining as the wild-type (Figures 2M–R). The cyp-35A5 had some Oil-Red-O staining in the intestine but visibly less than wild-type (Figures 2S–U).

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