Analysis of hsp 30, hsp 70 and ubiquitin gene expression in Xenopus laevis tadpoles

Development ◽  
1988 ◽  
Vol 103 (1) ◽  
pp. 59-67
Author(s):  
P.H. Krone ◽  
J.J. Heikkila
Keyword(s):  
Heat Shock ◽  
Xenopus Laevis ◽  
Temporal Pattern ◽  
Size Range ◽  
Continuous Exposure ◽  
Hsp 70 ◽  
Blastula Stage ◽  
Tailbud Stage ◽  
Actin Mrna ◽  
Continuous Heat

Heat-induced accumulation of hsp 30 mRNA (1.1 kb) during early development of Xenopus laevis was first detectable at the tailbud stage (stage 30–34). This contrasts with heat-induced accumulation of hsp 70 mRNA (2.7 kb) and ubiquitin mRNA (size range = 1.7–3.1 kb), which was first detectable at the mid- to late-blastula stage. Continuous exposure of tadpoles to a 33 degrees C heat shock resulted in a coordinate, transient accumulation of hsp 30, hsp 70 and ubiquitin mRNA. A coordinate, temporal pattern was also observed for the decay of hsp 30, hsp 70 and ubiquitin mRNA in tadpoles recovering at 22 degrees C following a 1 h heat shock at 33 degrees C. Thus, while hsp 30 genes are regulated differently during development compared with hsp 70 and ubiquitin genes, these genes all exhibit a coordinate heat-inducible pattern of expression at the tadpole stage. Levels of alpha-cardiac actin mRNA remained unchanged during continuous heat shock and recovery experiments.

Heat shock gene expression in Xenopus laevis A6 cells in response to heat shock and sodium arsenite treatments

1988 ◽  
Vol 66 (8) ◽  
pp. 862-870 ◽  
Author(s):  
S. Darasch ◽  
D. D. Mosser ◽  
N. C. Bols ◽  
J. J. Heikkila
Keyword(s):  
Heat Shock ◽  
Xenopus Laevis ◽  
Sodium Arsenite ◽  
Polyacrylamide Gel Electrophoresis ◽  
Heat Shock Gene ◽  
Epithelial Cell Line ◽  
Continuous Exposure ◽  
Mrna Levels ◽  
Hsp 70 ◽  
Mrna Accumulation

Continuous exposure of a Xenopus laevis kidney epithelial cell line, A6, to either heat shock (33 °C) or sodium arsenite (50 μM) resulted in transient but markedly different temporal patterns of heat-shock protein (HSP) synthesis and HSP 70 and 30 mRNA accumulation. Heat-shock-induced synthesis of HSPs was detectable within 1 h and reached maximum levels by 2–3 h. While sodium arsenite induced the synthesis of some HSPs within 1 h, maximal HSP synthesis did not occur until 12 h. The pattern of HSP 70 and 30 mRNA accumulation was similar to the response observed at the protein level. During recovery from heat shock, a coordinate decline in HSPs and HSP 70 and 30 mRNA was observed. During recovery from sodium arsenite, a similar phenomenon occurred during the initial stages. However, after 6 h of recovery, HSP 70 mRNA levels persisted in contrast to the declining HSP 30 mRNA levels. Two-dimensional polyacrylamide gel electrophoresis revealed the presence of 5 HSPs in the HSP 70 family, of which two were constitutive, and 16 different stress-inducible proteins in the HSP 30 family. In conclusion, heat shock and sodium arsenite induce a similar set of HSPs but maximum synthesis of the HSP is temporally separated by 12–24 h.

Examination of heat shock protein mRNA accumulation in early Xenopus laevis embryos

1987 ◽  
Vol 65 (2) ◽  
pp. 87-94 ◽  
Author(s):  
J. J. Heikkila ◽  
N. Ovsenek ◽  
P. Krone
Keyword(s):  
Heat Shock ◽  
Xenopus Laevis ◽  
Heat Shock Protein ◽  
Incubation Temperature ◽  
Effect Of Temperature ◽  
Continuous Exposure ◽  
Hsp 70 ◽  
Mrna Accumulation ◽  
Mrna Induction ◽  
Xenopus Laevis Embryos

Elevation of the incubation temperature of Xenopus laevis neurulae from 22 to 33–35 °C induced the accumulation of heat shock protein (hsp) 70 mRNA (2.7 kilobases (kb)) and a putative hsp 87 mRNA (3.2 kb). While constitutive levels of both hsp mRNAs were detectable in unfertilized eggs and cleavage-stage embryos, heat-induced accumulation was not observed until after the mid-blastula stage. Exposure of Xenopus laevis embryos to other stressors, such as sodium arsenite or ethanol, also induced a developmental stage-dependent accumulation of hsp 70 mRNA. To characterize the effect of temperature on hsp 70 mRNA induction, neurulae were exposed to a range of temperatures (27–37 °C) for 1 h. Heat-induced hsp 70 mRNA accumulation was first detectable at 27 °C, with relatively greater levels at 30–35 °C and lower levels at 37 °C. A more complex effect of temperature on hsp 70 mRNA accumulation was observed in a series of time course experiments. While continuous exposure of neurulae to heat shock (27–35 °C) induced a transient accumulation of hsp 70 mRNA, the temporal pattern of hsp 70 mRNA accumulation was temperature dependent. Exposure of embryos to 33–35 °C induced maximum relative levels of hsp 70 mRNA within 1–1.5 h, while at 30 and 27 °C peak hsp 70 mRNA accumulation occurred at 3 and 12 h, respectively. Finally, placement of Xenopus neurulae at 22 °C after a 1-h heat shock at 33 °C produced an initial decrease in hsp 70 mRNA within 15–30 min, followed by a transient increase in hsp 70 mRNA at 1–2 h before decaying to background levels by 7 h.

Analysis of the temperature-dependent temporal pattern of heat-shock-protein synthesis in fish cells

1983 ◽  
Vol 3 (7) ◽  
pp. 647-658 ◽  
Author(s):  
Lashitew Gedamu ◽  
Beverly Culham ◽  
John J. Heikkila
Keyword(s):  
Protein Synthesis ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Temporal Pattern ◽  
Stress Proteins ◽  
Incubation Temperature ◽  
Stress Protein ◽  
In Vitro Translation ◽  
Continuous Exposure ◽  
Temperature Dependent

Continuous exposure of Chinook salmon embryo cells to an elevated incubation temperature of 24°C induces the transient expression of a set of heat-shock or stress proteins whereas maintenance of the cells at a higher incubation temperature of 28°C produces a continuous synthesis of these stress proteins. In vitro translation studies suggest that the temperature-dependent temporal pattern of stress-protein synthesis is correlated with the levels of stress-protein mRNA. This was verified using a recombinant-DNA probe complementary to the 70K heat-shock-protein mRNA. A transient increase in the level of the fish heat-shock 70K mRNA was observed in RNA samples isolated from cells continuously exposed at 24°C However, a constant increase in the level of this specific mRNA was found in RNA preparations obtained from cells maintained at 28°C Therefore, the temperature-dependent pattern of fish heat-shockprotein synthesis appears to be directly related to the level of heat-shock-protein mRNA.

Expression of microinjected hsp 70/CAT and hsp 30/CAT chimeric genes in developing Xenopus laevis embryos

Development ◽  
1989 ◽  
Vol 106 (2) ◽  
pp. 271-281
Author(s):  
P.H. Krone ◽  
J.J. Heikkila
Keyword(s):  
Xenopus Laevis ◽  
Fusion Gene ◽  
Developmental Regulation ◽  
Hsp 70 ◽  
Inhibitory System ◽  
Developmentally Regulated ◽  
Promoter Sequences ◽  
Chimeric Genes ◽  
Stage Of Development ◽  
Tailbud Stage

The expression of microinjected chimeric genes containing Drosophila hsp 70 and Xenopus hsp 70 and hsp 30 promoters linked to the reporter gene coding for bacterial chloramphenicol acetyltransferase (CAT) was examined during early development of Xenopus laevis. Heat-inducible expression of fusion genes containing either the Drosophila hsp 70 promoter (1100 bp) or the Xenopus hsp 70 promoter (750 bp) was first detectable after the midblastula stage of development. This coincides with the embryonic stage at which the endogenous hsp 70 gene is first heat-inducible. A Xenopus hsp 30/CAT fusion gene containing 350 bp of promoter sequences was also heat-inducible after the midblastula stage unlike the endogenous hsp 30 genes which were not heat-inducible until the early tailbud stage (stage 23–24). Sequences that are present within either the coding or 3′ region of the hsp 30 clone do not cause the microinjected hsp 30 gene to be developmentally regulated in a normal manner. Additionally, microinjected hsp 30 gene sequences have no effect on the developmental regulation of endogenous hsp 30 genes which continue to be activated at the tailbud stage of development. Our data suggest, that an inhibitory system, which may control the expression of the endogenous hsp 30 gene during development, does not regulate the expression of the injected hsp 30 gene.

Expression of prostaglandin G/H synthase (PGHS) and heat shock protein-70 (HSP-70) in the corpus luteum (CL) of prostaglandin F2α-treated immature superovulated rats

2004 ◽  
Vol 82 (6) ◽  
pp. 363-371 ◽  
Author(s):  
R M Narayansingh ◽  
M Senchyna ◽  
M M Vijayan ◽  
J C Carlson
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Protein Expression ◽  
Corpus Luteum ◽  
Heat Shock Protein 70 ◽  
Prostaglandin F2α ◽  
Sampling Period ◽  
Immunoblot Analysis ◽  
Hsp 70 ◽  
Prostaglandin F

In this study we examined the mechanism of corpus luteum (CL) regression by measuring changes in expression of prostaglandin G/H synthase-1 (PGHS-1) and -2 (PGHS-2) in day 4 CL and inducible heat shock protein 70 (HSP-70) in day 4 and day 9 CL of immature superovulated rats. The rats were superovulated and treated with 500 µg of prostaglandin F2α (PGF2α) on day 4 or day 9 after CL formation. Ovaries and serial blood samples were removed during the 24-hour period following treatment. Plasma progesterone was determined by radioimmunoassay while mRNA abundance and protein expression were assessed by semiquantitative RT-PCR and immunoblot analysis, respectively. One hour after PGF2α, both day 4 and day 9 rats exhibited a significant decrease in progesterone secretion; however, there was a greater decrease in day 9 rats. In ovarian samples removed on day 4, there was a significant increase in mRNA for PGHS-2 at 1 hour after PGF2α. PGHS-1 mRNA content remained unchanged. Immunoblot analyses showed an increase in PGHS-2 protein expression only at 8 h. There were no changes in PGHS-1 protein expression. In day 9 rats, ovarian HSP-70 protein levels increased by 50% after PGF2α injection; however, on day 4 there was no change in expression of this protein over the sampling period. These results suggest that expression of PGHS-2 may be involved in inhibiting progesterone production and that expression of HSP-70 may be required for complete CL regression in the rat.Key words: rat, prostaglandin F2α, corpus luteum, prostaglandin G/H synthase, heat shock protein-70.

scholarly journals Differential activation of heat-shock and oxidation-specific stress genes in chemically induced oxidative stress

1995 ◽  
Vol 309 (2) ◽  
pp. 453-459 ◽  
Author(s):  
L Tacchini ◽  
G Pogliaghi ◽  
L Radice ◽  
E Anzon ◽  
A Bernelli-Zazzera
Keyword(s):  
Oxidative Stress ◽  
Heat Shock ◽  
Time Course ◽  
Consensus Sequence ◽  
Single Agent ◽  
Stress Protein ◽  
Hsp 70 ◽  
Activator Protein ◽  
Chemically Induced ◽  
Haem Oxygenase

Post-ischaemic reperfusion increases the level of the major heat-shock (stress) protein hsp 70 and of its mRNA by transcriptional mechanisms, and activates the binding of the heat-shock factor HSF to the consensus sequence HSE. In common with CoCl2 treatment, post-ischaemic reperfusion increases the level of haem oxygenase mRNA, an indicator of oxidative stress, but CoCl2 does not seem to induce the expression of the hsp 70 gene [Tacchini, Schiaffonati, Pappalardo, Gatti and Bernelli-Zazzera (1993) Lab. Invest. 68, 465-471]. Starting from these observations, we have now studied the expression of two genes of the hsp 70 family and of other possibly related genes under conditions of oxidative stress. Three different chemicals, which cause oxidative stress by various mechanisms and induce haem oxygenase, enhance the expression of the cognate hsc 73 gene, but do not activate the inducible hsp 70 gene. Expression of the other genes that have been studied seems to vary in intensity and/or time course, in relation to the particular mechanism of action of any single agent. The pattern of induction of the early-immediate response genes c-fos and c-jun observed during oxidative stress differs from that found in post-ischaemic reperfused livers. Oxidative-stress-inducing agents do not promote the binding of HSF to its consensus sequence HSE, such as occurs in heat-shock and post-ischaemic reperfusion, and fail to activate AP-1 (activator protein 1). With the possible exception of Phorone, the oxidative stress chemically induced in rat liver activates NFkB (nuclear factor kB) and AP-2 (activator protein 2) transcription factors.

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