Molecular Response to Osmotic Shock

Sharfstein, Susan T.; Shen, Duan; Kiehl, Thomas R.; Zhou, Ruihao

doi:10.1007/1-4020-5252-9_7

Cited by 6 publications

(4 citation statements)

References 102 publications

(119 reference statements)

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“…Evidence for this conclusion is that various compounds, such as glucose, sucrose, mannitol, or sorbitol, can reproduce the effect observed with NaCl. 4 , 20 , 27 , 30 Second, the yield increase described in our study was notably higher than the rate of increase reported for mAb production, with most studies reporting a relatively moderate mAb rate increase between 2- and 3-fold. 20 , 23 , 26 This is likely the result of critical differences between mAbs and rAAV production: mAb production relies essentially on transcription and translation in cells that are selected for this performance; on the contrary, rAAV production involves genome rescue and replication, gene expression, and genome packaging, all steps heavily controlled both by cellular and helper virus pathways.…”

Section: Discussioncontrasting

confidence: 70%

NaCl and KCl mediate log increase in AAV vector particles and infectious titers in a specific/timely manner with the HSV platform

Trivedi

Chaudhuri

et al. 2021

Molecular Therapy - Methods & Clinical Development

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The increasing demand for adeno-associated virus (AAV) vectors, a result from the surging interest for their potential to cure human genetic diseases by gene transfer, tumbled on low-performing production systems. Innovative improvements to increase both yield and quality of the vector produced have become a priority undertaking in the field. In a previous study, we showed that adding a specific concentration of sodium chloride (NaCl) to the production medium resulted in a dramatic increase of AAV vector particle and infectious titers when using the herpes simplex virus (HSV) production system, both in adherent or suspension platforms. In this work, we studied additional salts and their impact on AAV vector production. We found that potassium chloride (KCl), or a combination of KCl and NaCl, resulted in the highest increase in AAV vector production. We determined that the salt-mediated effect was the most impactful when the salt was present between 8 and approximately 16 h post-infection, with the highest rate increase occurring within the first 24 h of the production cycle. We showed that the AAV vector yield increase did not result from an increase in cell growth, size, or viability. Furthermore, we demonstrated that the impact on AAV vector production was specifically mediated by NaCl and KCl independently of their impact on the osmolality of the production media. Our findings convincingly showed that NaCl and KCl were uniquely efficacious to promote up to a 10-fold increase in the production of highly infectious AAV vectors when produced in the presence of HSV. We think that this study will provide unique and important new insights in AAV biology toward the establishment of more successful production protocols.

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Section: Discussioncontrasting

confidence: 70%

NaCl and KCl mediate log increase in AAV vector particles and infectious titers in a specific/timely manner with the HSV platform

Trivedi

Chaudhuri

et al. 2021

Molecular Therapy - Methods & Clinical Development

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“…The hypo‐osmotic stress response of C. cryptica shared features of “general” environmental stress responses seen in other eukaryotes (Gasch et al, 2000; Mager & De Kruijff, 1995), including repression of genes involved in cell growth (e.g., ribosome biogenesis and RNA metabolism) and induction of genes canonically involved in stress defence such as ROS scavengers and HSP chaperones. Like other prokaryotic and eukaryotic microbes (Rojas et al, 2017; Sharfstein et al, 2007; Warner, 1999), diatoms halt cell division in the early stages of osmotic shock until ionic and osmotic equilibria are restored. Under such conditions, energy is probably redirected to cellular processes essential for acute stress management, rather than energetically expensive processes related to cell growth such as ribosome biogenesis (Albert et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

“…Like other prokaryotic and eukaryotic microbes (Rojas et al, 2017;Sharfstein et al, 2007;Warner, 1999), diatoms halt cell division in the early stages of osmotic shock until ionic and osmotic equilibria are restored. Under such conditions, energy is probably redirected to cellular processes essential for acute stress management, rather than energetically expensive processes related to cell growth such as ribosome biogenesis (Albert et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

The dynamic response to hypo‐osmotic stress reveals distinct stages of freshwater acclimation by a euryhaline diatom

et al. 2022

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The salinity gradient separating marine and freshwater environments is a major ecological divide, and the mechanisms by which most organisms adapt to new salinity environments are poorly understood. Diatoms are a lineage of ancestrally marine microalgae that have repeatedly colonized and diversified in freshwaters. Cyclotella cryptica is a euryhaline diatom that naturally tolerates a broad range of salinities, thus providing a powerful system for understanding the genomic mechanisms for mitigating and acclimating to low salinity. To understand how diatoms mitigate acute hypoosmotic stress, we abruptly shifted C. cryptica from seawater to freshwater and performed transcriptional profiling during the first 10 hours. Freshwater shock dramatically remodeled the transcriptome, with ~50% of the genome differentially expressed in at least one time point. The peak response occurred within 1 hour, with strong repression of genes involved in cell growth and osmolyte production, and strong induction of specific stress defense genes. Transcripts largely returned to baseline levels within 4-10 hours, with growth resuming shortly thereafter, suggesting that gene expression dynamics may be useful for predicting acclimation. Moreover, comparison to a transcriptomics study of C. cryptica following months-long acclimation to freshwater revealed little overlap between the genes and processes differentially expressed in cells exposed to acute stress versus fully acclimated conditions. Altogether, this study highlights the power of time-resolved transcriptomics to reveal fundamental insights into how cells dynamically respond to an acute environmental shift and provides new insights into how diatoms mitigate natural salinity fluctuations and have successfully diversified across freshwater habitats worldwide..

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“…These molecules are the physical basis for regulation of the neuroendocrine and immune systems in organisms. Metabolites are the final products that generated during living activities, and metabolomics analysis is study of these downstream molecules ( Sharfstein et al, 2007 ). Metabolomics is an important tool for systems biology, as it provides the ability to measure the concentrations of endogenous small molecules in biofluids and characterize the quantitative changes of the metabolites that occur in specific physiological states of organisms ( Nicholson et al, 1999 ; Sun et al, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

The Effect of Melatonin on Locomotor Behavior and Muscle Physiology in the Sea Cucumber Apostichopus japonicus

Ding

Zhang

et al. 2019

Front. Physiol.

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Melatonin is a highly conserved hormone in evolutionary history. It occurs in numerous organisms and plays a role in the endocrine and immune systems. Locomotor behavior is a basic behavior in animals and is an important indicator of circadian rhythms, which are coordinated by the nervous and endocrine systems. To date, the effect of melatonin on locomotor behavior has been studied in vertebrates, including syrian hamsters, sparrows, rats, zebrafish, goldfish, and flatworms. However, there have been few studies of the effects of melatonin on locomotor behavior in marine invertebrates. The goals of present study were to show the existence of melatonin in the sea cucumber Apostichopus japonicus and to evaluate its effect on locomotor activity. In addition, muscle tissues from control and melatonin-treated sea cucumbers were tested using ultra performance liquid chromatography and quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) to determine the changes of metabolic activity in muscle. Melatonin was present in the coelomic fluid of A. japonicus at a concentration of ∼135.0 ng/L. The total distance traveled and number steps taken over 9 h after melatonin administration decreased with increasing concentration of the melatonin dose. Mean and maximum velocity of movement and stride length and stride frequency also decreased, but their differences were not statistically significant. Overall, these results suggest that melatonin administration had a sedative effect on A. japonicus. The levels of 22 different metabolites were altered in the muscle tissues of melatonin-treated sea cucumbers. Serotonin, 9-cis retinoic acid, all-trans retinoic acid, flavin mononucleotide in muscles were downregulated after melatonin administration. Moreover, a high free fatty acid (FFA) concentration and a decrease in the adenosine 5′-triphosphate (ATP) concentration in the muscle tissues of the melatonin-treated group were detected as well. These results suggest that the sedative effect of melatonin involves some other metabolic pathways, and the reduced locomotor modulator—serotonin, inhibited fatty acid oxidation and disturbed oxidative phosphorylation are potential physiological mechanisms that result in the inhibitory effect of melatonin on locomotion in sea cucumbers.

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Molecular Response to Osmotic Shock

Cited by 6 publications

References 102 publications

NaCl and KCl mediate log increase in AAV vector particles and infectious titers in a specific/timely manner with the HSV platform

NaCl and KCl mediate log increase in AAV vector particles and infectious titers in a specific/timely manner with the HSV platform

The dynamic response to hypo‐osmotic stress reveals distinct stages of freshwater acclimation by a euryhaline diatom

The Effect of Melatonin on Locomotor Behavior and Muscle Physiology in the Sea Cucumber Apostichopus japonicus

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