KLF regulation of insulin pathway genes

Huan Wang^{1

2}, Christopher W Brey³, Yi Wang⁴, Randy Gaugler⁴, Sarwar Hashmi^{4

2}

Affiliations

¹ College of Bioscience and Biotechnology, Shenyang Agriculture University, Shanyang, 110866 China.
² Rutgers Center for Lipid Research, New Jersey Institute for Food, Nutrition, and Health, Rutgers University, New Brunswick, USA.
³ Science Department, Marywood University, Scranton, PA 18505 USA.
⁴ Laboratory of Developmental Biology, Center for Vector Biology, Rutgers University, 180 Jones Avenue, New Brunswick, NJ 08901 USA.

PMID: 36816753
PMCID: PMC9935763
DOI: 10.1007/s13205-023-03502-5

KLF regulation of insulin pathway genes

Huan Wang et al. 3 Biotech. 2023 Mar.

. 2023 Mar;13(3):87.

doi: 10.1007/s13205-023-03502-5. Epub 2023 Feb 16.

Authors

Huan Wang^{1

2}, Christopher W Brey³, Yi Wang⁴, Randy Gaugler⁴, Sarwar Hashmi^{4

2}

Affiliations

¹ College of Bioscience and Biotechnology, Shenyang Agriculture University, Shanyang, 110866 China.
² Rutgers Center for Lipid Research, New Jersey Institute for Food, Nutrition, and Health, Rutgers University, New Brunswick, USA.
³ Science Department, Marywood University, Scranton, PA 18505 USA.
⁴ Laboratory of Developmental Biology, Center for Vector Biology, Rutgers University, 180 Jones Avenue, New Brunswick, NJ 08901 USA.

PMID: 36816753
PMCID: PMC9935763
DOI: 10.1007/s13205-023-03502-5

Abstract

Alteration in lipid metabolism can result in fat accumulation in adipose tissues, which may lead to two most important human diseases, obesity and diabetes. A shift in lipid metabolism deregulates signaling pathways which regulates obesity and/or diabetes. In this study, we examined the components of insulin/ TGF-β pathways and their genetic interaction with Krüppel-like transcription factors (KLFs). Their role in energy homeostasis were discussed. We separately created klf/daf genes double mutants by carrying out klfs RNAi on daf-2 (e1391), daf-4 (e1364), daf-7 (e1372); dpy-1 (e1), daf-14 (m77), daf-16 (mgDf50) mutants. And then conducted Oil O Red staining to assay the klf/daf RNAi worms for fat deposits and examine genetic interaction between klfs and daf genes. The results showed that worms bearing klf-1, 2, or 3 and daf-2, or daf-4 mutations deposit large, but similar fat levels as individual mutants. The results suggested that they target the same molecular pathway of fat storage. klf-1, 2 or 3 RNAi /daf-7 worms showed higher fat deposits in klf-1, 2, or 3 RNAi/daf-7 worms than klf-1, 2, or 3 RNAi or daf-7 mutants alone, which showed a functional interaction between klfs and daf-7 in perhaps TGF-β-like pathway. Altogether our study suggests a direct role of klfs in insulin signaling pathway.

Keywords: Caenorhabditis elegans; Insulin pathway; Krüppel-like transcription factors; TGF beta pathway; daf; klf.

© King Abdulaziz City for Science and Technology 2023, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

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Conflict of interest statement

Conflict of interestAll the authors declare that they are no conflict of interest.

Figures

**Fig. 1**
Genetic interaction between *klf-1, klf-2, klf-3* and *daf-*2, and *daf-16*: Genetic interactions between *klf-1, 2, 3* and *daf-2* and *daf-16* genes were assessed by the measurement of fat mass in set of RNAi animals and their F1 progenies. *Klf-1, 2, or 3 RNAi* were separately performed on *daf-2* (*e1391*) and *daf-16* (*mgDF50*) L4 worms, then RNAi worms and their F1 progenies were stained with Oil Red O staining solution. The quantitation of fat mass based on Oil-Red staining intensity. Multiple representative images from each treatment were chosen for image quantitation as described in Materials and Methods. A wild-type animal showing low fat mass; B high fat mass in the intestine of *klf-1 RNAi* worm; C high fat mass in the intestine of *daf-2* (*e1391*) worm; D high fat mass in *klf-1 RNAi/ daf-2* worm; E high fat mass in *klf-2* (*ok1043*) worm; F high fat mass in *klf-2 RNAi/ daf-2* (*e1391*) worm; G high fat mass in *klf-3* (*ok1975*) worm; H high fat mass in *klf-3 RNAi/ daf-2* (*e1391*) worm; I low fat mass in *daf-16* (*mgDf50*) worm; J high fat mass in *klf-1 RNAi/ daf-16* (*mgDF50*) worm; K high fat mass in *klf-2 RNAi/ daf-16* (*mgDF50*) worm; L high fat mass in *klf-3 RNAi/ daf-16* (*mgDF50*) worm. Worms were observed under Olympus U-Tr0.63Xc optics attached to Axioplan Zeiss microscope and photographed using a digital camera Prog Res CF scan (magnification: 200X). Images are the representative of approximately 100 animals

**Fig. 2**
Genetic interaction between *klf-1, klf-2, klf-3* and *daf-4* (*e1364*): Genetic interactions between *klf-1, 2, 3* and *daf-4* (*e1364*) genes were assessed by the measurement of fat mass in set of RNAi animals and their F1 progenies. *klf-1, 2, or 3 RNAi* were separately performed on *daf-4* (*e1364*) L4 worms, then RNAi worms and their F1 progenies were stained with Oil Red O staining solution. The quantitation of fat mass based on Oil-Red staining intensity. Multiple representative images from each treatment were chosen for image quantitation. A wild-type animal showing low fat mass; B high fat mass deposit in the intestine of *klf-1 RNAi* worm; C high fat mass deposit in the intestine of *daf-4* (*e1364*) worm; D high fat mass deposit in *klf-1 RNAi/ daf-4* (*e1364*) worm; E high fat mass deposit in *klf-2* (*ok1043*) worms; F high fat mass deposit in *klf-2 RNAi/ daf-4* (*e1364*) worms; G high fat mass deposit in *klf-3* (*ok1975*) worm; H high fat mass deposit in *klf-3 RNAi/ daf-4* (*e1364*) worm. Worms were observed under Olympus U-Tr0.63Xc optics attached to Axioplan Zeiss microscope and photographed using a digital camera Prog Res CF scan (magnification: 200X). Images are the representative of approximately 100 animals

**Fig. 3**
Genetic interaction between *klf-1, klf-2, klf-3* and *daf-7*, and *daf-14* genes: Genetic interactions between *klf-1, 2, 3* and *daf-7* and 14 genes were assessed by the measurement of fat mass in set of RNAi animals and their F1 progenies. *Klf-1, 2, or 3 RNAi* were separately performed on *daf-7* (*e1372*) and *daf-14* (*m77*) L4 worms, then RNAi worms and their F1 progenies were stained with Oil Red O staining solution. The quantitation of fat mass based on Oil-Red staining intensity. Multiple representative images from each treatment were chosen for image quantitation as described in Materials and Methods. A wild-type animal showing low fat mass; B high fat mass in *klf-1 RNAi* worm; C *daf-7* (*e1372*) worm showing intense fat staining; D more fat deposit in *klf-1 RNAi/daf-7* (*e1372*) worm than either *klf-1 RNAi* or *daf-7*(*e1372*) worm; E high fat mass in *klf-2* (*ok1043*) worms; F more fat mass in *klf-2 RNAi/daf-7* (*e1372*) worms than either *klf-2* (*ok1043*) or *daf-7* (*e1372*) worm; G high fat mass in *klf-3* (*ok1975*) worm; H more fat deposit in *klf-3 RNAi/ daf-7* (*e1372*) worm than either *klf-3* (*ok1975*) or *daf-7* (*e1372*) worm; I low fat mass in *daf-14* (*m77*) worm; J high fat mass in *klf-1 RNAi/daf-14(m77)* worm; K high fat mass in *klf-2 RNAi/ daf-14* (*m77*) worm; L high fat mass in *klf-3 RNAi/ daf-14* (*m77*) worm. Worms were observed under Olympus U-Tr0.63Xc optics attached to Axioplan Zeiss microscope and photographed using a digital camera Prog Res CF scan (magnification: 200X). Images are the representative of approximately 100 animals

See this image and copyright information in PMC

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KLF regulation of insulin pathway genes

Affiliations

KLF regulation of insulin pathway genes

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

References

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