Coordination of tissue homeostasis and growth by the Scribble-α-Catenin-Septate junction complex

Conditional knockdown protocol for studying cellular communication using Drosophila melanogaster wing imaginal disc

“…It is worth noting that conditional KD of scrib alone exhibits oncogenic properties. 1 , 4 Nevertheless, our previous observations indicate that the likelihood of neoplasia formation strongly depends on the levels of scrib expression within the entire tissue. 1 For instance, neoplasia manifests in the wing discs after 3 days of scrib KD in animals with 2 copies of scrib .…”

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Figure 11 Verification of the phenotype of the wing disc epithelium using antibody staining (from Huang et al. 1 ) (A) Scribble antibody (magenta) is used to visualize the distribution pattern of Scrib in the wing disc. The loss of Scrib in adjacent cells from the Ptc-GFP domain (green) suggests a potential mechanism of cell-cell communication to disrupt ABP, ultimately leading to the neoplasia phenotype (4D ATS) Troubleshooting 3.

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“… Verification of the phenotype of the wing disc epithelium using antibody staining (from Huang et al. 1 ) …”

“…By implementing the protocol below, we observe changes in the distribution of Scrib, Hippo signaling executor Yorkie (Yki) signaling, and notable effects on tissue growth and cell migration, which we visualize through GFP expression in the Patched (Ptc) compartment in the wing imaginal discs. 1 , 4 These approaches assist in studying the spatiotemporal role of the gene of interest in tissue morphogenesis, as well as uncovering novel genes that contribute to tissue homeostasis and growth.…”

“…RNAi knockdown (KD) of scrib and clonal analysis of scrib alleles in Drosophila wing imaginal discs have been extensively employed to investigate scrib as a tumor suppressor gene. 1 , 3 , 4 , 6 While these approaches provide evidence that loss of scrib leads to varied phenotypes influenced by environmental conditions, the underlying mechanisms of cellular communication between sub-optimal (loss of scrib ) and optimal (wild type) cells, which give rise to diverse outcomes, remain largely unaddressed. Elucidating these mechanisms is particularly crucial for understanding the early phase of neoplasia formation mediated by loss of scrib .…”