Functionally conserved effects of rapamycin exposure on zebrafish

Sucularli, Ceren; Shehwana, Huma; Kuscu, Cem; Dungul, Dilay Ciglidag; Özdağ, Hilal; Konu, Özlen

doi:10.3892/mmr.2016.5059

Cited by 13 publications

(17 citation statements)

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“…S2C). Thus, a one-hour pulse of heat shock treatment introduces ectopic β-SNAP1 expression for at least 24 h. Next, we injected the DNA construct into coa mutants, and repeated the heat shock treatment at 12-h intervals from 36 to 72 hpf at four different starting points (36,48, 60, 72 hpf). Cone and rod survival was evaluated at 84 hpf by labeling with zpr1 antibody and fluorescent signals of Tg[XlaRho:XP-GFP], respectively (Fig.…”

Section: Resultsmentioning

confidence: 99%

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β-SNAP activity in the outer segment growth period is critical for preventing BNip1-dependent apoptosis in zebrafish photoreceptors

Nishiwaki

Masai

2020

Sci Rep

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BNip1, which functions as a t-SNARE component of the syntaxin18 complex, is localized on the ER membrane and regulates retrograde transport from Golgi to the ER. BNip1 also has a BH3 domain, which generally releases pro-apoptotic proteins from Bcl2-mediated inhibition. Previously we reported that retinal photoreceptors undergo BNip1-dependent apoptosis in zebrafish β-snap1 mutants. Here, we investigated physiological roles of BNip1-dependent photoreceptor apoptosis. First, we examined the spatio-temporal profile of photoreceptor apoptosis in β-snap1 mutants, and found that apoptosis occurs only during a small developmental window, 2–4 days-post-fertilization (dpf), in which an apical photoreceptive membrane structure, called the outer segment (OS), grows rapidly. Transient expression of β-SNAP1 during this OS growing period prevents photoreceptor apoptosis in β-snap1 mutants, enabling cone to survive until at least 21 dpf. These observations suggest that BNip1-mediated apoptosis is linked to excessive activation of vesicular transport associated with rapid growth of the OS. Consistently, knockdown of Ift88 and Kif3b, which inhibits protein transport to the OS, rescued photoreceptor apoptosis in β-snap1 mutants. Treatment with rapamycin, which inhibits protein synthesis via the mTOR pathway, also rescued photoreceptor apoptosis in β-snap1 mutants. These data suggest that BNip1 performs risk assessment to detect excessive vesicular transport in photoreceptors.

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Section: Resultsmentioning

confidence: 99%

“…In accordance with a previous report 36 , wild-type and coa mutant embryos were treated with rapamycin (LC laboratories, R-5000) at 10 μM from 34 to 97 hpf, and fixed in 4% PFA. Photoreceptor survival was evaluated with rhodamine-conjugated phalloidin labeling.…”

Section: Methodsmentioning

confidence: 95%

β-SNAP activity in the outer segment growth period is critical for preventing BNip1-dependent apoptosis in zebrafish photoreceptors

Nishiwaki

Masai

2020

Sci Rep

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“…After yolk depletion, liver provides glucose for all tissues of the body by breaking down its own stores of glycogen via the process of glycolysis (23, 24). At the same time, liver replaces the consumed glycogen through gluconeogenesis and/or lipolysis that includes the formation of alternative glycogen from non carbohydrate sources, such as lactate, pyruvate, glycerol, and alanine (25, 26).…”

Section: Resultsmentioning

confidence: 99%

“…Rapamycin might be effective in age-related pathologies via activation of autophagy by enhancing the clearance of aggregate-prone proteins in vitro (55–57) However, Stimulation of autophagy by rapamycin in knocked-out models is still poorly understood. Previously, morphometric analysis of rapamycin-treated zebrafish has been established that revealed a great reduction in the epithelial cells number, size and proliferation hence lower yolk consumption till 9dpf (26, 58). In our study, rapamycin increasing the longevity of atg7 - and beclin1 -mutated through slowing metabolism and dietary restriction which reduced cellular food demand without malnutrition resulted in metabolic equilibrium.…”

Section: Discussionmentioning

confidence: 99%

Autophagy-related genesatg7andbeclin1are essential for energy metabolism and survival during the larval-to-juvenile transition stage of zebrafish

Mawed

Zhang

Ren

et al. 2019

Preprint

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High mortality is usually observed during the transition from larvae to juvenile in teleost which is related to the transition from endogenous to exogenous feeding. Autophagy is an evolutionary regulated cellular mechanism highly conserved in eukaryotic organisms to maintain energy homeostasis against stress including starvation. To investigate whether autophagy plays a role during the larval-juvenile transition, we generated atg7 and beclin1 zebrafish mutant lines using CRISPR/Cas9 technology. In this study, both atg7 and beclin1 null zebrafish exhibited a normal body confirmation; nevertheless, they completely died around 15 dpf and 9 dpf respectively. During larval-juvenile transition period, atg7 and beclin1 mutants were unable to cope with the metabolic stress after yolk absorption at 5 dpf and fail to activate autophagy in response to nutrient restriction, and without external feeding, all mutants died nearly at 8 dpf. Dramatic defects in the intestine architecture and metabolic functions in the liver were observed even though providing larvae with an external food supply, suggesting that autophagy isn’t only important during yolk depletion but also within food plenty. Treatment with rapamycin, an activator of autophagy, could effectively extend the survival time of both atg7 and beclin1 null zebrafish through lowering the metabolic rate while it couldn’t activate autophagy in mutants via the canonical pathway. Our findings provided a molecular evidence for the physiological, histological and metabolic changes that occur during the transition process from the larval to the juvenile stages and the chief role of autophagy on the body metabolism during these turning milestones.Author summaryZebrafish Danio rerio has emergrd one of the most powerful research models for studying genes expression during early embryogenesis and postnatal development. On the basis of the cell mechanisms, Macroautphagy, a natural regulated pathway disassembles unnecessary or dysfunctional components orchestrated by more than 36 autophagy related-genes conserved from yeast to mammals. Among those genes are atg7 and beclin1 which have been proved to play an important role in regulating post natal development in some mammals however their roles during zebrafish development still unedited. During this research, CRISPER/CAS9 were adopted to know atg7 and beclin1 knockout effects on the mutants’ metabolism during shifting from maternal yolk acquisition to exogenous feeding and the role of autophagy during the larvae to pre-juvenile development. Herein, we found out that larvae couldn’t abandon autophagy in both fasting and feeding conditions as larvae died earlier before pre-juvenile development despite feeding declaring the importance of autophagy not only to provide the cell with essential nutrients during starvation but also to get rid of cargos inside the eukaryotic cells. Briefly, if the larvae didn’t recycle those cargos due to autophagy perturbations, they will die despite providing suitable conditions including food and acclimatization.

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“…This technique has been used to explore the molecular mechanisms of various diseases, such as different cancers (16,17), osteoarthritis (18) and degenerative diseases (19). Furthermore, metaanalysis and the comparison of gene expression profiles of different species enables researchers to discover conserved molecular mechanisms (20).…”

Section: Introductionmentioning

confidence: 99%

Investigations of Microtubule-associated Protein 2 Gene Expression in Spinal Muscular Atrophy

Bora¹,

Sucularli²,

Hensel³

et al. 2019

jpr

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Spinal muscular atrophy (SMA) is a devastating genetic disease in childhood andff is caused by the absence of functional survival motor neuron (SMN) protein, which leads to impairments of the cytoskeleton, especially in neurons. Dysregulation of actin dynamics have been linked to SMA patho mechanisms, however involvement of altered microtubule dynamics is largely unknown. In this study, we investigated differentially expressed microtubule-related genes using in vitro and in vivo SMA model systems. Materials and Methods: By focusing on microtubule-related genes, we re-analyzed publically available gene expression arrays, which were previously performed with induced pluripotent stem cell-derived motor neurons of SMA patients and the spinal cords of SMA mice. We found altered expressions of microtubule-associated protein 2 (MAP2), which was validated by real time reverse-transcription polymerase chain reaction using the SMN knock-down NSC34 cell line and the severe SMA mouse model. Results: We showed that the expression of MAP2 gene was significantly upregulated in both expression arrays. Upregulation was also detected in the brain and spinal cord tissues of severe SMA mice at different developmental stages. Conclusion: Our findings suggest that microtubule regulatory proteins may be altered in SMN depleted cells and further research is needed to elucidate the contribution of dysregulated microtubule dynamics towards SMA.

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Functionally conserved effects of rapamycin exposure on zebrafish

Cited by 13 publications

References 50 publications

β-SNAP activity in the outer segment growth period is critical for preventing BNip1-dependent apoptosis in zebrafish photoreceptors

β-SNAP activity in the outer segment growth period is critical for preventing BNip1-dependent apoptosis in zebrafish photoreceptors

Autophagy-related genesatg7andbeclin1are essential for energy metabolism and survival during the larval-to-juvenile transition stage of zebrafish

Investigations of Microtubule-associated Protein 2 Gene Expression in Spinal Muscular Atrophy

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