scholarly journals Effects of Acute Heat Stress on a Newly Established Chicken Hepatocyte—Nonparenchymal Cell Co-Culture Model

Animals ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 409 ◽  
Author(s):  
Máté Mackei ◽  
Andor Molnár ◽  
Szabolcs Nagy ◽  
László Pál ◽  
Csaba Kővágó ◽  
...  
Keyword(s):  
Cell Culture ◽  
Heat Stress ◽  
Stress Response ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Cultured Cells ◽  
Heat Exposure ◽  
Liver Cells ◽  
Culture Models ◽  
Cell Culture Models

Heat stress is one of the most important issues in broiler flocks impairing animal health and productivity. On a cellular level, excess heat exposure can trigger heat shock response acting for the restoration of cell homeostasis by several mechanisms, such as affecting heat shock protein synthesis, redox homeostasis and pro-inflammatory cytokine production. The major aim of this study was to establish a novel avian hepatocyte—nonparenchymal cell co-culture as a model for investigating the cellular effects of heat stress and its interaction with inflammation in chicken liver. Cell fractions were isolated by differential centrifugation from a freshly perfused chicken liver, and hepatocyte mono-cultures as well as hepatocyte–nonparenchymal cell co-cultures (with cell ratio 6:1, hepatocytes to nonparenchymal cells, mimicking a milder hepatic inflammation) were prepared. Isolated and cultured cells were characterized by flow cytometry and immunocytochemistry applying hepatocyte- and macrophage-specific antibodies. Confluent cell cultures were exposed to 43 °C temperature for 1 or 2 h, while controls were cultured at 38.5 °C. The metabolic activity, LDH enzyme activity, reactive oxygen species (H2O2) production, extracellular concentration of heat shock protein 70 (HSP70), and that of the pro-inflammatory cytokines interleukin (IL-)6 and IL-8 were assessed. Shorter heat stress applied for 1 h could strongly influence liver cell function by significantly increasing catabolic metabolism and extracellular H2O2 release, and by significantly decreasing HSP70, IL-6, and IL-8 production on both cell culture models. However, all these alterations were restored after 2 h heat exposure, indicating a fast recovery of liver cells. Hepatocyte mono-cultures and hepatocyte—nonparenchymal cell co-cultures responded to heat stress in a similar manner, but the higher metabolic rate of co-cultured cells may have contributed to a better capability of inflamed liver cells for accommodation to stress conditions. In conclusion, the established new primary cell culture models provide suitable tools for studying the hepatic inflammatory and stress response. The results of this study highlight the impact of short-term heat stress on the liver in chickens, underline the mediatory role of oxidative stress in acute stress response, and suggest a fast cellular adaptation potential in liver cells.

scholarly journals The Effect of β-Alanine Supplementation on the Heat Shock Protein, Inflammatory and Neurotrophin Response in Animals Exposed to an Acute Heat Stress

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 1210-1210
Author(s):  
Jay Hoffman ◽  
Tal Belety ◽  
Michal Horowitz ◽  
Yoram Epstein ◽  
Yaron Bruchim ◽  
...  
Keyword(s):  
Heat Stress ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Heat Exposure ◽  
Normal Diet ◽  
Hsp70 Expression ◽  
Bdnf Expression ◽  
Funding Sources ◽  
Supplementation Period ◽  
Acute Heat Stress

Abstract Objectives The effect of 30-days of β-alanine (BA) supplementation on heat shock protein 70 (HSP70), inflammatory and neurotrophin responses in the hippocampus and hypothalamus of rats exposed to an acute heat stress was investigated. Methods Animals were randomized to either a control (CTL) group or BA supplementation (100mg·kg−1) group. All animal were fed a normal diet and only differed regarding supplementation. Following supplementation animals were either exposed to the heat stress (120 min at 40–41°C) or were unexposed. Following the acute heat stress, or at the end of the supplementation period, animals were harvested and their brains removed. Immunohistochemical technique was used to detect expression of HSP70, brain-derived neurotrophic factor (BDNF), cyclooxygenase-2 (COX2) and neuropeptide Y (NPY) in the hippocampus subregions and paraventricular nuclear (PVN) region of the hypothalamus. Results Three animals in CTL and one in BA did not survive the heat stress. Significant attenuation (P's < 0.005) in BDNF expression was noted in animals exposed to the heat stress compared to unexposed in all subregions (CA1, CA3 and DG) of the hippocampus and PVN. A significant elevation in BDNF expression in the CA3 subregion of rats fed BA and exposed to the heat stress was observed compared to exposed CTL animals. Significant elevations in COX2 was also noted in the CA1 and CA3 subregions in exposed compared to unexposed animals. COX2 expression was significantly greater (P ≤ 0.0065) in CTL compared with BA during heat exposure in the CA1 subregion of the hippocampus. Animals supplemented with BA also realized significantly higher HSP70 expression (P = 0.02) in the CA3 subregion of the hippocampus compared to CTL. Significant differences (P’s < 0.05) in NPY expression in all subregions of the hippocampus and PVN were noted between exposed and unexposed animals. However, NPY expression was significantly higher (P ≤ 0.03) for BA compared to CTL in exposed animals in the PVN. Conclusions Results suggested that BA supplementation appeared to increase resiliency to an acute heat stress and reduced the inflammatory response, while increasing HSP70 and neurotrophins expression. Funding Sources Natural Alternatives International Inc., Carlsbad, CA, USA.

Role of heat stress response in the tolerance of immature renal tubules to anoxia

1998 ◽  
Vol 274 (6) ◽  
pp. F1029-F1036 ◽  
Author(s):  
Karen M. Gaudio ◽  
Gunilla Thulin ◽  
Andrea Mann ◽  
Michael Kashgarian ◽  
Norman J. Siegel
Keyword(s):  
Transcription Factor ◽  
Heat Stress ◽  
Stress Response ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Metabolic Response ◽  
Renal Tubules ◽  
Heat Stress Response

The stress response was studied in suspensions of tubules from immature (IT) and mature (MT) rats after noninjury, heat, oxygen, and anoxia. Under all conditions, IT exhibited more exuberant activation of heat shock transcription factor (HSF) than MT. Characterization of activated HSF in immature cortex revealed HSF1. Also, 2 h after each condition, heat shock protein-72 (HSP-72) mRNA was twofold in IT. As the metabolic response to 45 min of anoxia, 20-min reoxygenation was assessed by measuring O2 consumption (O2C). Basal O2C was manipulated with ouabain, nystatin, and carbonylcyanide p-chloromethyoxyphenylhydrazone (CCCP). Basal O2C in IT were one-half the value of MT. After anoxia, basal O2C was reduced by a greater degree in MT. Ouabain reduced O2C to half the basal value in both noninjured and anoxic groups. Basal O2C was significantly stimulated by nystatin but not to the same level following anoxia in MT and IT. Basal O2C was also stimulated by CCCP, but after anoxia, CCCP O2C was significantly less in MT with no decrease in IT, suggesting mitochondria are better preserved in IT. Also, O2C devoted to nontransport activity was better maintained in IT.

Multiscale iterative voting for differential analysis of stress response for 2D and 3D cell culture models

2010 ◽  
Vol 241 (3) ◽  
pp. 315-326 ◽  
Author(s):  
J. HAN ◽  
H. CHANG ◽  
Q. YANG ◽  
G. FONTENAY ◽  
T. GROESSER ◽  
...  
Keyword(s):  
Cell Culture ◽  
Stress Response ◽  
3D Cell Culture ◽  
Differential Analysis ◽  
Culture Models ◽  
2D And 3D ◽  
Cell Culture Models ◽  
Iterative Voting

scholarly journals Genome-wide identification and analysis of the heat shock transcription factor family in moso bamboo (Phyllostachys edulis)

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Bin Huang ◽  
Zhinuo Huang ◽  
Ruifang Ma ◽  
Jialu Chen ◽  
Zhijun Zhang ◽  
...  
Keyword(s):  
Heat Stress ◽  
Stress Response ◽  
Heat Shock ◽  
Gene Family ◽  
Regulatory Elements ◽  
Moso Bamboo ◽  
Naphthalene Acetic Acid ◽  
Plant Responses ◽  
Heat Stress Response ◽  
Regulatory Pathways

AbstractHeat shock transcription factors (HSFs) are central elements in the regulatory network that controls plant heat stress response. They are involved in multiple transcriptional regulatory pathways and play important roles in heat stress signaling and responses to a variety of other stresses. We identified 41 members of the HSF gene family in moso bamboo, which were distributed non-uniformly across its 19 chromosomes. Phylogenetic analysis showed that the moso bamboo HSF genes could be divided into three major subfamilies; HSFs from the same subfamily shared relatively conserved gene structures and sequences and encoded similar amino acids. All HSF genes contained HSF signature domains. Subcellular localization prediction indicated that about 80% of the HSF proteins were located in the nucleus, consistent with the results of GO enrichment analysis. A large number of stress response–associated cis-regulatory elements were identified in the HSF upstream promoter sequences. Synteny analysis indicated that the HSFs in the moso bamboo genome had greater collinearity with those of rice and maize than with those of Arabidopsis and pepper. Numerous segmental duplicates were found in the moso bamboo HSF gene family. Transcriptome data indicated that the expression of a number of PeHsfs differed in response to exogenous gibberellin (GA) and naphthalene acetic acid (NAA). A number of HSF genes were highly expressed in the panicles and in young shoots, suggesting that they may have functions in reproductive growth and the early development of rapidly-growing shoots. This study provides fundamental information on members of the bamboo HSF gene family and lays a foundation for further study of their biological functions in the regulation of plant responses to adversity.

scholarly journals Factors to consider when interrogating 3D culture models with plate readers or automated microscopes

2021 ◽  
Author(s):  
Terry Riss ◽  
O. Joseph Trask
Keyword(s):  
Cell Culture ◽  
Three Dimensional ◽  
3D Culture ◽  
Fluorescent Imaging ◽  
Culture Models ◽  
Assay Methods ◽  
Diffusion Rates ◽  
Cell Culture Models ◽  
Dimensional Cell ◽  
Cells Cultured

AbstractAlong with the increased use of more physiologically relevant three-dimensional cell culture models comes the responsibility of researchers to validate new assay methods that measure events in structures that are physically larger and more complex compared to monolayers of cells. It should not be assumed that assays designed using monolayers of cells will work for cells cultured as larger three-dimensional masses. The size and barriers for penetration of molecules through the layers of cells result in a different microenvironment for the cells in the outer layer compared to the center of three-dimensional structures. Diffusion rates for nutrients and oxygen may limit metabolic activity which is often measured as a marker for cell viability. For assays that lyse cells, the penetration of reagents to achieve uniform cell lysis must be considered. For live cell fluorescent imaging assays, the diffusion of fluorescent probes and penetration of photons of light for probe excitation and fluorescent emission must be considered. This review will provide an overview of factors to consider when implementing assays to interrogate three dimensional cell culture models.

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