Dissection, Fixation, and Immunostaining of the Drosophila Midgut

doi:10.1007/978-1-0716-2035-9_20

. 2022:2438:309-321.

doi: 10.1007/978-1-0716-2035-9_20.

Dissection, Fixation, and Immunostaining of the Drosophila Midgut

Jia Chen¹, Daniel St Johnston²

Affiliations

¹ The Gurdon Institute and the Department of Genetics, University of Cambridge, Cambridge, UK.
² The Gurdon Institute and the Department of Genetics, University of Cambridge, Cambridge, UK. d.stjohnston@gurdon.cam.ac.uk.

PMID: 35147951
DOI: 10.1007/978-1-0716-2035-9_20

Dissection, Fixation, and Immunostaining of the Drosophila Midgut

Jia Chen et al. Methods Mol Biol. 2022.

. 2022:2438:309-321.

doi: 10.1007/978-1-0716-2035-9_20.

Authors

Jia Chen¹, Daniel St Johnston²

Affiliations

¹ The Gurdon Institute and the Department of Genetics, University of Cambridge, Cambridge, UK.
² The Gurdon Institute and the Department of Genetics, University of Cambridge, Cambridge, UK. d.stjohnston@gurdon.cam.ac.uk.

PMID: 35147951
DOI: 10.1007/978-1-0716-2035-9_20

Abstract

The Drosophila midgut is mainly composed of highly polarized epithelial cells called enterocytes that establish their apical-basal polarity in a fundamentally different way from other Drosophila epithelia. The roles of polarity factors in the midgut can be studied by generating clones of homozygous mutant cells in the background of wild-type tissue. In this chapter, we will introduce and discuss the procedures for producing positively marked mutant clones in the midgut and describe specific protocols for dissecting, fixing, and immunostaining this tissue.

Keywords: Dissection; Drosophila midgut; Heat fixation; Immunostaining; MARCM.

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References

1. Miguel-Aliaga I, Jasper H, Lemaitre B (2018) Anatomy and physiology of the digestive tract of Drosophila melanogaster. Genetics 210(2):357–396. https://doi.org/10.1534/genetics.118.300224 - DOI - PubMed - PMC
1. O'Brien LE, Soliman SS, Li X, Bilder D (2011) Altered modes of stem cell division drive adaptive intestinal growth. Cell 147(3):603–614. https://doi.org/10.1016/j.cell.2011.08.048 - DOI - PubMed - PMC
1. Chen J, Sayadian AC (2018) An alternative mode of epithelial polarity in the drosophila midgut. PLoS Biol 16(10):e3000041. https://doi.org/10.1371/journal.pbio.3000041 - DOI - PubMed - PMC
1. Micchelli CA, Perrimon N (2006) Evidence that stem cells reside in the adult drosophila midgut epithelium. Nature 439(7075):475–479. https://doi.org/10.1038/nature04371 - DOI - PubMed
1. Ohlstein B, Spradling A (2006) The adult drosophila posterior midgut is maintained by pluripotent stem cells. Nature 439(7075):470–474. https://doi.org/10.1038/nature04333 - DOI - PubMed

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[1] Miguel-Aliaga I, Jasper H, Lemaitre B (2018) Anatomy and physiology of the digestive tract of Drosophila melanogaster. Genetics 210(2):357–396. https://doi.org/10.1534/genetics.118.300224 - DOI - PubMed - PMC

[2] Miguel-Aliaga I, Jasper H, Lemaitre B (2018) Anatomy and physiology of the digestive tract of Drosophila melanogaster. Genetics 210(2):357–396. https://doi.org/10.1534/genetics.118.300224 - DOI - PubMed - PMC

[3] O'Brien LE, Soliman SS, Li X, Bilder D (2011) Altered modes of stem cell division drive adaptive intestinal growth. Cell 147(3):603–614. https://doi.org/10.1016/j.cell.2011.08.048 - DOI - PubMed - PMC

[4] O'Brien LE, Soliman SS, Li X, Bilder D (2011) Altered modes of stem cell division drive adaptive intestinal growth. Cell 147(3):603–614. https://doi.org/10.1016/j.cell.2011.08.048 - DOI - PubMed - PMC

[5] Chen J, Sayadian AC (2018) An alternative mode of epithelial polarity in the drosophila midgut. PLoS Biol 16(10):e3000041. https://doi.org/10.1371/journal.pbio.3000041 - DOI - PubMed - PMC

[6] Chen J, Sayadian AC (2018) An alternative mode of epithelial polarity in the drosophila midgut. PLoS Biol 16(10):e3000041. https://doi.org/10.1371/journal.pbio.3000041 - DOI - PubMed - PMC

[7] Micchelli CA, Perrimon N (2006) Evidence that stem cells reside in the adult drosophila midgut epithelium. Nature 439(7075):475–479. https://doi.org/10.1038/nature04371 - DOI - PubMed

[8] Micchelli CA, Perrimon N (2006) Evidence that stem cells reside in the adult drosophila midgut epithelium. Nature 439(7075):475–479. https://doi.org/10.1038/nature04371 - DOI - PubMed

[9] Ohlstein B, Spradling A (2006) The adult drosophila posterior midgut is maintained by pluripotent stem cells. Nature 439(7075):470–474. https://doi.org/10.1038/nature04333 - DOI - PubMed

[10] Ohlstein B, Spradling A (2006) The adult drosophila posterior midgut is maintained by pluripotent stem cells. Nature 439(7075):470–474. https://doi.org/10.1038/nature04333 - DOI - PubMed

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Dissection, Fixation, and Immunostaining of the Drosophila Midgut

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Dissection, Fixation, and Immunostaining of the Drosophila Midgut

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