Refinement of a technique for collecting and evaluating the osmolality of haemolymph from Drosophila larvae.

Ex vivo physiological experiments using small insect models such as Drosophila larvae have become increasingly useful to address fundamental biological questions. To perform such experiments, various artificial saline solutions have been developed, but their osmolality varies significantly from one to the next. Such a variation of osmolality stems, in part, from the difficulty of determining the true value of haemolymph osmolality in Drosophila larvae. Thus, there is a pressing need to refine protocols for collecting and measuring the osmolality of the larval haemolymph. Two major obstacles are thought to impede the accurate analysis of haemolymph collected from small insects: melanin formation and gut-derived contamination. Here, we greatly refined existing haemolymph collection methods, evaluated the purity of the collected haemolymph under melanin-free conditions, and concluded that the true value of haemolymph osmolality is close to 306.0 mOsm kg-1 in Drosophila larvae.

Belly roll, a GPI-anchored Ly6 protein, regulates Drosophila melanogaster escape behaviors by modulating the excitability of nociceptive peptidergic interneurons.

Appropriate modulation of escape behaviors in response to potentially damaging stimuli is essential for survival. Although nociceptive circuitry has been studied, it is poorly understood how genetic contexts affect relevant escape responses. Using an unbiased genome-wide association analysis, we identified an Ly6/α-neurotoxin family protein, Belly roll (Bero), which negatively regulates Drosophila nociceptive escape behavior. We show that Bero is expressed in abdominal leucokinin-producing neurons (ABLK neurons) and bero knockdown in ABLK neurons resulted in enhanced escape behavior. Furthermore, we demonstrated that ABLK neurons responded to activation of nociceptors and initiated the behavior. Notably, bero knockdown reduced persistent neuronal activity and increased evoked nociceptive responses in ABLK neurons. Our findings reveal that Bero modulates an escape response by regulating distinct neuronal activities in ABLK neurons.

Characterization of ferret Pit1 as a receptor of feline leukemia virus subgroup B.

Feline leukemia virus (FeLV) is a retrovirus that causes immune suppression and immunodeficiency, leading to opportunistic infections and leukemia/lymphoma in cats. Today, a variety of domestic mammals are kept in houses, and it is important to evaluate the possibility of interspecies transmission of FeLV. In this study, we assessed the infectivity of FeLV-B in ferrets that belong to Mustelidae. By pseudotype virus infection assay, we revealed that a ferret cell line, Mpf cells, is resistant to FeLV-B infection. The mRNA expression level of the FeLV-B receptor, Pit-1, was approximately half that of cat FEA cells in ferret Mpf cells. There was no significant difference in receptor usage between ferret's and cat's Pit1. These data may indicate the presence of the post-transcriptional modification and/or the restriction factor(s) against the FeLV-B infection in ferrets.

E-cadherin-dependent coordinated epithelial rotation on a two-dimensional discoidal pattern.

In vivo, cells collectively migrate in a variety of developmental and pathological contexts. Coordinated epithelial rotation represents a unique type of collective cell migrations, which has been modeled in vitro under spatially confined conditions. Although it is known that the coordinated rotation depends on intercellular interactions, the contribution of E-cadherin, a major cell-cell adhesion molecule, has not been directly addressed on two-dimensional (2D) confined substrates. Here, using well-controlled fibronectin-coated surfaces, we tracked and compared the migratory behaviors of MDCK cells expressing or lacking E-cadherin. We observed that wild-type MDCK II cells exhibited persistent and coordinated rotations on discoidal patterns, while E-cadherin knockout cells migrated in a less coordinated manner without large-scale rotation. Our comparison of the collective dynamics between these two cell types revealed a series of changes in migratory behavior caused by the loss of E-cadherin, including a decreased global migration speed, less regularity in quantified coordination, and increased average density of topological defects. Taken together, these data demonstrate that spontaneous initiation of collective epithelial rotations depends on E-cadherin under 2D discoidal confinements.