Evaluating the Death and Recovery of Lateral Line Hair Cells Following Repeated Neomycin Treatments

Acute chemical ablation of lateral line hair cells is an important tool to understand lateral line-mediated behaviors in free-swimming fish larvae and adults. However, lateral line-mediated behaviors have not been described in fish larvae prior to swim bladder inflation, possibly because single doses of ototoxin do not effectively silence lateral line function at early developmental stages. To determine whether ototoxins can disrupt lateral line hair cells during early development, we repeatedly exposed zebrafish larvae to the ototoxin neomycin during a 36 h period from 3 to 4 days post-fertilization (dpf). We use simultaneous transgenic and vital dye labeling of hair cells to compare 6-h and 12-h repeated treatment timelines and neomycin concentrations between 0 and 400 µM in terms of larval survival, hair cell death, regeneration, and functional recovery. Following exposure to neomycin, we find that the emergence of newly functional hair cells outpaces cellular regeneration, likely due to the maturation of ototoxin-resistant hair cells that survive treatment. Furthermore, hair cells of 4 dpf larvae exhibit faster recovery compared to 3 dpf larvae. Our data suggest that the rapid functional maturation of ototoxin-resistant hair cells limits the effectiveness of chemical-based methods to disrupt lateral line function. Furthermore, we show that repeated neomycin treatments can continually ablate functional lateral line hair cells between 3 and 4 dpf in larval zebrafish.

Evaluating the Death and Recovery of Lateral Line Hair Cells Following Repeated Neomycin Treatments

Acute chemical ablation of lateral line hair cells is an important tool to understand lateral line-mediated behaviors in free-swimming fish larvae and adults. However, lateral line-mediated behaviors have not been described in fish larvae prior to swim bladder inflation, possibly because single doses of ototoxin do not effectively silence lateral line function at early developmental stages. To determine if ototoxins can effectively silence the lateral line during early development, we repeatedly expose zebrafish larvae to the ototoxin neomycin during a 36-hour period from 3-4 days post-fertilization (dpf). We use simultaneous transgenic and vital dye labeling of hair cells to compare 6- hour and 12-hour repeated treatment timelines and neomycin concentrations between 0–400 µM in terms of larval survival, hair cell death, regeneration, and functional recovery. Following exposure to neomycin, we find that the emergence of newly functional hair cells outpaces cellular regeneration, likely due to the maturation of ototoxin-resistant hair cells that survive treatment. Furthermore, hair cells of 4 dpf larvae exhibit faster recovery compared to 3 dpf larvae. Our data suggest that the rapid functional maturation of ototoxin-resistant hair cells limits the effectiveness of chemical-based methods to disrupt lateral line function. Furthermore, we show that repeated neomycin treatments can continually ablate lateral line hair cells between 3–4 dpf in larval zebrafish.

Intracellular Interactions Between Arboviruses and Wolbachia in Aedes aegypti

Aedes aegypti is inherently susceptible to arboviruses. The geographical expansion of this vector host species has led to the persistence of Dengue, Zika, and Chikungunya human infections. These viruses take advantage of the mosquito’s cell to create an environment conducive for their growth. Arboviral infection triggers transcriptomic and protein dysregulation in Ae. aegypti and in effect, host antiviral mechanisms are compromised. Currently, there are no existing vaccines able to protect human hosts from these infections and thus, vector control strategies such as Wolbachia mass release program is regarded as a viable option. Considerable evidence demonstrates how the presence of Wolbachia interferes with arboviruses by decreasing host cytoskeletal proteins and lipids essential for arboviral infection. Also, Wolbachia strengthens host immunity, cellular regeneration and causes the expression of microRNAs which could potentially be involved in virus inhibition. However, variation in the magnitude of Wolbachia’s pathogen blocking effect that is not due to the endosymbiont’s density has been recently reported. Furthermore, the cellular mechanisms involved in this phenotype differs depending on Wolbachia strain and host species. This prompts the need to explore the cellular interactions between Ae. aegypti-arboviruses-Wolbachia and how different Wolbachia strains overall affect the mosquito’s cell. Understanding what happens at the cellular and molecular level will provide evidence on the sustainability of Wolbachia vector control.

Immunomodulatory and antimicrobial non-mulberry Antheraea mylitta silk fibroin accelerates in vitro fibroblast repair and regeneration by protecting oxidative stress

Initially SF accelerated pro-inflammatory cytokines, restricted anti-inflammatory cytokines; later it regulated in reverse order. SF potentially eradicated ROS and promoted Ki-67 cellular regeneration whereas pristine PU could not.

Role of hemocytes in the regeneration of germline stem cells in Drosophila

Cellular regeneration, which relies on extensive restructuring of cytoplasmic materials, is an essential process to restore tissues and organs lost during aging, degenerative diseases and injury. At early stages of Drosophila spermatogenesis, when cellular constituents are intensely remodeled, there are two different populations of stem cells, the somatic stem cells and the germline stem cells (GSCs). GSCs divide by asymmetric division to give rise two distinct daughter cells. One of them will leave the stem cells’ niche and differentiate into spermatogonial cells (SCs). Both aging and cellular stress can lead to the loss of GSCs. Lost GSCs can be restored by dedifferentiation of SCs into functional GSCs. In other tissues, macrophages provide specific conditions for cellular transformation. Here we examined the potential role of immune surveillance cells called hemocytes during dedifferentiation of SGs into GSCs. We found an elevated number of hemocytes during this dedifferentiation process. Immune depletion of hemocytes decreased the regeneration capacity of germline. We also show that autophagy, which plays a pivotal role in cellular differentiation by eliminating unwanted, superfluous parts of the cytoplasm, becomes upregulated in dedifferentiating SCs upon JAK-STAT signaling emitted by hemocytes. Furthermore, these immune cells regulate expression of Omi/HtrA2, a key regulator of apoptosis in early spermatogenesis. Together, we suggest that hemocytes have important functions in the dedifferentiation process of GSCs.

Selective Chemonucleolysis With Condoliase in Cynomolgus Monkeys

Selective chemonucleolytic effects of condoliase, a glycosaminoglycan degrading enzyme, was investigated histopathologically in cynomolgus monkeys. Condoliase was administered once into the lumber intervertebral disc (IVD), and as a comparative control, chymopapain, a proteolytic enzyme, was administered in a similar manner. Histopathological changes of the IVD and the adjacent vertebral body (VB) were examined at 1 to 26 weeks after administration. Major changes induced by condoliase in the IVD were degenerative and necrotic changes in the nucleus pulposus, annulus fibrosus, cartilaginous endplate (CEP), and epiphyseal growth plate (EGP); focal disappearance of the EGP; and neovascularization and ossification of the CEP. Decreased/necrosis of bone marrow cells with new bone formation was observed in the VB. Cellular regeneration in the IVD was observed as a recovery changes on and after week 4. The changes in the IVD and VB subsided at week 26. Chymopapain induced qualitatively similar but more widely extended changes. The degrees of the changes in the IVD and VB were more severe than those of condoliase, and the changes were exacerbated even at week 26. These results indicated that histopathological changes caused by condoliase were less severe and more selective than those by chymopapain.

Cellular regeneration and proliferation on polymeric 3D inverse-space substrates and the effect of doxorubicin

2D substrates promote cell attachment with lateral compression; 3DIS scaffolding restores the 3D cell structure allowing more realistic cellular-drug responses.