A cautionary tale on genetic screens based on a gain-of-expression approach: The case of LanB1

Celis, José F. de; Molnar, Cristina

doi:10.4161/fly.4.1.10720

Cited by 5 publications

(6 citation statements)

References 12 publications

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“…To test the role of laminin in the peripheral glia, we used RNAi transgenes to knock down laminin expression in all glia using repo-GAL4. Unfortunately, quantification of laminin deposition in the neural lamella by glia is not possible as hemocytes and fat bodies contribute extensively to the ECM surrounding peripheral nerves (Bunt et al, 2010;Pastor-Pareja and Xu, 2011). To overcome this limitation, we used a range of RNAi lines to knock down the ␣ subunit, LanA, or the LanB1 and LanB2 subunits.…”

Section: Knockdown Of Laminin Subunits In Glia Results In Severe Morpmentioning

confidence: 99%

“…In Drosophila, collagen IV is not observed within the peripheral glia (Xie and Auld, 2011; Fig. 5 A, B), and is deposited by circulating hemocytes and fat bodies (Bunt et al, 2010;Pastor-Pareja and Xu, 2011). We examined the effect of Tango1 knockdown on Viking::GFP, the Drosophila homolog of collagen IV endogenously tagged with GFP (Morin et al, 2001).…”

Section: Tango1 Mediates Laminin Secretion Via a Collagenindependent mentioning

confidence: 99%

“…Loss of integrins and metalloproteinase-mediated degradation of the surrounding ECM does not lead to glial swelling or formation of vacuoles, but rather triggers a loss of perineurial glia wrapping of the axon (Xie and Auld, 2011). In Drosophila, the fat body and hemocytes contribute to the deposition of ECM proteins in the basement membrane surrounding the nervous system (Bunt et al, 2010;Pastor-Pareja and Xu, 2011), explaining why the loss of laminin secretion from glia in Tango1 does not lead to the same phenotypes as integrin knockdown.…”

Section: Figurementioning

confidence: 99%

“…Mutations in LanB1 result in embryonic lethality, absent basement membranes, and accumulation of other ECM proteins in mutant tissues (Urbano et al, 2009;Wolfstetter and Holz, 2012). The embryonic lethality of laminin mutations is likely because many tissues secrete laminin during development, including hemocytes and fat bodies, both of which contribute to the ECM surrounding tissues, such as the nervous system (Bunt et al, 2010;Pastor-Pareja and Xu, 2011). Muscle cells also secrete laminin and secretion of LanA regulates larval neuromuscular junction growth (Tsai et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

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Accumulation of Laminin Monomers inDrosophilaGlia Leads to Glial Endoplasmic Reticulum Stress and Disrupted Larval Locomotion

et al. 2016

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The nervous system is surrounded by an extracellular matrix composed of large glycoproteins, including perlecan, collagens, and laminins. Glial cells in many organisms secrete laminin, a large heterotrimeric protein consisting of an ␣, ␤, and ␥ subunit. Prior studies have found that loss of laminin subunits from vertebrate Schwann cells causes loss of myelination and neuropathies, results attributed to loss of laminin-receptor signaling. We demonstrate that loss of the laminin ␥ subunit (LanB2) in the peripheral glia of Drosophila melanogaster results in the disruption of glial morphology due to disruption of laminin secretion. Specifically, knockdown of LanB2 in peripheral glia results in accumulation of the ␤ subunit (LanB1), leading to distended endoplasmic reticulum (ER), ER stress, and glial swelling. The physiological consequences of disruption of laminin secretion in glia included decreased larval locomotion and ultimately lethality. Loss of the ␥ subunit from wrapping glia resulted in a disruption in the glial ensheathment of axons but surprisingly did not affect animal locomotion. We found that Tango1, a protein thought to exclusively mediate collagen secretion, is also important for laminin secretion in glia via a collagen-independent mechanism. However loss of secretion of the laminin trimer does not disrupt animal locomotion. Rather, it is the loss of one subunit that leads to deleterious consequences through the accumulation of the remaining subunits.

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Section: Knockdown Of Laminin Subunits In Glia Results In Severe Morpmentioning

confidence: 99%

Section: Tango1 Mediates Laminin Secretion Via a Collagenindependent mentioning

confidence: 99%

Section: Figurementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Accumulation of Laminin Monomers inDrosophilaGlia Leads to Glial Endoplasmic Reticulum Stress and Disrupted Larval Locomotion

et al. 2016

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“…First is the possibility of unspecific effects of the misexpression, which could result in interesting phenotypes unrelated to the normal function of the gene (de Celis and Molnar 2010). Although this is likely the case for an unknown fraction of overexpressed genes, it is interesting to point out that the overlap of genes selected comparing different screens is low, suggesting some degree of specificity relative to the cell type or process under examination.…”

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confidence: 99%

Genetic Annotation of Gain-Of-Function Screens Using RNA Interference and in Situ Hybridization of Candidate Genes in the Drosophila Wing

et al. 2012

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Gain-of-function screens in Drosophila are an effective method with which to identify genes that affect the development of particular structures or cell types. It has been found that a fraction of 2-10% of the genes tested, depending on the particularities of the screen, results in a discernible phenotype when overexpressed. However, it is not clear to what extent a gain-of-function phenotype generated by overexpression is informative about the normal function of the gene. Thus, very few reports attempt to correlate the loss-and overexpression phenotype for collections of genes identified in gain-of-function screens. In this work we use RNA interference and in situ hybridization to annotate a collection of 123 P-GS insertions that in combination with different Gal4 drivers affect the size and/or patterning of the wing. We identify the gene causing the overexpression phenotype by expressing, in a background of overexpression, RNA interference for the genes affected by each P-GS insertion. Then, we compare the loss and gainof-function phenotypes obtained for each gene and relate them to its expression pattern in the wing disc. We find that 52% of genes identified by their overexpression phenotype are required during normal development. However, only in 9% of the cases analyzed was there some complementarity between the gain-and loss-of-function phenotype, suggesting that, in general, the overexpression phenotypes would not be indicative of the normal requirements of the gene.

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Protein retention in the endoplasmic reticulum rescues Aβ toxicity in Drosophila

Catterson,

Minkley,

Aspe

et al. 2023

Neurobiology of Aging

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A cautionary tale on genetic screens based on a gain-of-expression approach: The case of LanB1

Cited by 5 publications

References 12 publications

Accumulation of Laminin Monomers inDrosophilaGlia Leads to Glial Endoplasmic Reticulum Stress and Disrupted Larval Locomotion

Accumulation of Laminin Monomers inDrosophilaGlia Leads to Glial Endoplasmic Reticulum Stress and Disrupted Larval Locomotion

Genetic Annotation of Gain-Of-Function Screens Using RNA Interference and in Situ Hybridization of Candidate Genes in the Drosophila Wing

Protein retention in the endoplasmic reticulum rescues Aβ toxicity in Drosophila

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