scholarly journals Induction of expression and phosphorylation of heat shock protein B5 (CRYAB) in rat myometrium during pregnancy and labour

Reproduction ◽  
2016 ◽  
Vol 152 (1) ◽  
pp. 69-79 ◽  
Author(s):  
J G Nicoletti ◽  
B G White ◽  
E I Miskiewicz ◽  
D J MacPhee
Keyword(s):  
Heat Shock ◽  
Protein Expression ◽  
Post Partum ◽  
Focal Adhesions ◽  
Late Pregnancy ◽  
Immunoblot Analysis ◽  
Small Molecular Weight ◽  
Pregnant Rats ◽  
Immune System Activation ◽  
Cryab Protein

During pregnancy the myometrium undergoes a programme of differentiation induced by endocrine, cellular, and biophysical inputs. Small heat shock proteins (HSPs) are a family of ten (B1–B10) small-molecular-weight proteins that not only act as chaperones, but also assist in processes such as cytoskeleton rearrangements and immune system activation. Thus, it was hypothesized that HSPB5 (CRYAB) would be highly expressed in the rat myometrium during the contractile and labour phases of myometrial differentiation when such processes are prominent. Immunoblot analysis revealed that myometrial CRYAB protein expression significantly increased from day (D) 15 to D23 (labour;P<0.05). In correlation with these findings, serine 59-phosphorylated (pSer59) CRYAB protein expression significantly increased from D15 to D23, and was also elevated 1-day post-partum (P<0.05). pSer59-CRYAB was detected in the cytoplasm of myocytes within both uterine muscle layers mid- to late-pregnancy. In unilaterally pregnant rats, pSer59-CRYAB protein expression was significantly elevated in the gravid uterine horns at both D19 and D23 of gestation compared with non-gravid horns. Co-immunolocalization experiments using the hTERT-human myometrial cell line and confocal microscopy demonstrated that pSer59-CRYAB co-localized with the focal adhesion protein FERMT2 at the ends of actin filaments as well as with the exosomal marker CD63. Overall, pSer59-CRYAB is highly expressed in myometrium during late pregnancy and labour and its expression appears to be regulated by uterine distension. CRYAB may be involved in the regulation of actin filament dynamics at focal adhesions and could be secreted by exosomes as a prelude to involvement in immune activation in the myometrium.

scholarly journals Expression of small heat shock-related protein 20 (HSP20) in rat myometrium is markedly decreased during late pregnancy and labour

Reproduction ◽  
2007 ◽  
Vol 133 (4) ◽  
pp. 807-817 ◽  
Author(s):  
B E Cross ◽  
H M O’Dea ◽  
D J MacPhee
Keyword(s):  
Smooth Muscle ◽  
Heat Shock ◽  
Longitudinal Muscle ◽  
Post Partum ◽  
Muscle Relaxation ◽  
Late Pregnancy ◽  
Smooth Muscle Relaxation ◽  
Pregnant Rats ◽  
Longitudinal Muscle Layer ◽  
Mrna Detection

The underlying mechanisms regulating uterine contractions during labour are still poorly understood. Heat shock protein 20 (HSP20) is known to be present at high levels in smooth muscle and implicated in muscle relaxation, but HSP20 expression in the myometrium is completely undetermined. Since HSP20 has been implicated in smooth muscle relaxation, we hypothesized that HSP20 would be highly expressed in the rat myometrium during early and mid-pregnancy when the myometrium is relatively quiescent. Northern blot analysis particularly demonstrated that HSP20 mRNA detection was significantly decreased from day (d) 22 of pregnancy to 1-daypost-partum(PP) compared with d6 (P< 0.05). HSP20 mRNA detection was also significantly decreased from d22 to d23 of gestation compared with non-pregnant (NP) samples. Immunoblot analysis showed that detection of HSP20 was significantly decreased at d23 compared with d12 and d15 (P< 0.05). HSP20 detection also significantly decreased at PP compared with d15 (P< 0.05). Immunofluorescence analysis demonstrated that after d15, plasma membrane-associated localization of HSP20 decreased markedly in both circular and longitudinal muscle layers. In addition, HSP20 was detectable near cell membranes at much higher levels in the longitudinal muscle layer of progesterone-treated pregnant rats (delayed labour) at all gestational time points examined, compared with controls. Our results demonstrate that HSP20 mRNA and protein are highly expressed during early and mid-pregnancy and then the expression markedly decreases during late pregnancy and labour. The observed patterns of HSP20 expression are consistent with a potential role for HSP20 in facilitating myometrium quiescence during early and mid-pregnancy.

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 Inducible heat shock protein A1A (HSPA1A) is markedly expressed in rat myometrium by labour and secreted via myometrial cell-derived extracellular vesicles

2021 ◽  
Vol 33 (4) ◽  
pp. 279
Author(s):  
M. F. Russell ◽  
G. C. Bailey ◽  
E. I. Miskiewicz ◽  
D. J. MacPhee
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Extracellular Vesicles ◽  
Susceptibility Gene ◽  
Protein A ◽  
Circular Muscle ◽  
Late Pregnancy ◽  
Immunoblot Analysis ◽  
Interacting Protein ◽  
Myometrial Cell

The myometrium goes through physiological, cellular and molecular alterations during gestation that necessitate effective cellular proteostasis. Inducible heat shock protein A1A (HSPA1A) is a member of the 70-kDa heat shock protein A (HSPA) family, which acts as a chaperone to regulate proteostasis; however, HSPA1A also participates as a cytokine in inflammatory regulation, leading to its designation as a chaperokine. This study examined the spatiotemporal expression of HSPA1A protein in the rat myometrium throughout gestation and assessed whether it is secreted as cargo of myometrial cell-derived extracellular vesicles (EVs). Immunoblot analysis demonstrated that HSPA1A expression was markedly elevated during late pregnancy and labour and increased by uterine distension. Myometrial HSPA1A expression insitu increased in myocytes of longitudinal and circular muscle layers from Day 19 through to postpartum, specifically in the cytoplasm and nuclei of myocytes from both muscle layers, but frequently detectable just outside myocyte membranes. Scanning electron microscopy examination of samples isolated from hTERT-HM cell-conditioned culture medium, using EV isolation spin columns, confirmed the presence of EVs. EV lysates contained HSPA8, HSPA1A and the EV markers apoptosis-linked gene 2-interacting protein X (Alix), the tetraspanin cluster of differentiation 63 (CD63), tumour susceptibility gene 101 (TSG101) and HSP90, but not the endoplasmic reticulum protein calnexin. These results indicate that HSPA1A may act as a chaperokine in the myometrium during pregnancy.

scholarly journals Small heat shock protein 27 (Hsp27) expression is highly induced in rat myometrium during late pregnancy and labour

Reproduction ◽  
2005 ◽  
Vol 129 (1) ◽  
pp. 115-126 ◽  
Author(s):  
B G White ◽  
S J Williams ◽  
K Highmore ◽  
D J MacPhee
Keyword(s):  
Heat Shock ◽  
Heat Shock Protein ◽  
Regulatory Protein ◽  
Circular Muscle ◽  
Small Heat Shock Protein ◽  
Late Pregnancy ◽  
Immunoblot Analysis ◽  
Heat Shock Protein 27 ◽  
Potential Candidate ◽  
Hsp27 Expression

The underlying mechanisms that regulate uterine contractions during labour are still poorly understood. A candidate regulatory protein is heat shock protein 27 (Hsp27). It belongs to the small heat shock protein family and can regulate actin cytoskeleton dynamics, act as a chaperone, and may regulate contractile protein activation. As a result, we hypothesized that Hsp27 expression would be highly induced during late pregnancy and labour. Hsp27 mRNA expression was significantly elevated (P< 0.05) on days 17 to 22 of gestation. In addition, immunoblot analysis demonstrated that detection of total Hsp27 increased (P< 0.05) between day 21 and 1 day post-partum (PP) inclusive. Since phosphorylation of Hsp27 has been reported to be a prerequisite for smooth muscle contraction, we examined the temporal and spatial expression of Ser-15 phosphorylated Hsp27. Immunoblot analysis showed that the detection of Ser-15 phosphorylated Hsp27 significantly increased (P< 0.05) between days 19 and 23 (active labour) inclusive, in parallel with detection of total Hsp27. Immunocytochemical analysis of Ser-15 phosphorylated Hsp27 expressionin situdemonstrated that phosphorylated Hsp27 in circular muscle became detectable in peri-nuclear and membrane regions on days 19 to 22, but was primarily restricted to the cytoplasm on days 23 to PP. In contrast, phosphorylated Hsp27 in longitudinal muscle was primarily detected in myocyte membranes on days 15 to 22, and then also became detectable in the cytoplasm of myocytes on days 23 and PP. Our results demonstrate that Hsp27 expression is highly upregulated during late pregnancy and labour and suggest that Hsp27 is a potential candidate contraction-associated protein.

A quantitative study of rat uterine sympathetic innervation during pregnancy and post partum

2006 ◽  
Vol 18 (5) ◽  
pp. 525 ◽  
Author(s):  
R. Chávez-Genaro ◽  
P. Lombide ◽  
G. Anesetti
Keyword(s):  
Quantitative Study ◽  
Post Partum ◽  
Sympathetic Innervation ◽  
Glyoxylic Acid ◽  
Late Pregnancy ◽  
Uterine Horn ◽  
Pregnant Rats ◽  
Tissue Sections ◽  
Hormonal Milieu

In mammals, pregnancy induces a transient and extensive degeneration of uterine sympathetic innervation. We used the models of unilateral oviduct ligation and in oculo myometrium transplant in pregnant rats to address the role of stretching forces and/or hormone milieu in the loss of sympathetic innervation. The sympathetic fibres of the uterine horn and in oculo myometrial transplants were quantified on tissue sections processed by the glyoxylic acid technique. In normal pregnant rats, the density of uterine horn innervation was significantly reduced at late pregnancy and recovery took place during post partum. The empty horn of pregnant rats showed no significant changes in density of myometrial innervation during pregnancy or post partum. In oculo myometrial transplants were organotypically reinnervated in virgin animals. When the transplants were exposed to gestational hormonal milieu, few or no fibres were observed to the end of pregnancy; however, a significant increase at post partum was observed. Results showed that both the effects of stretching and the hormone milieu derived from the fetus–placenta complex play a role as inductors of changes on sympathetic myometrial innervation during pregnancy and support the idea that immature muscular uterine fibres are more susceptible to the effects of pregnancy than those originating from adult animals.

Serum concentrations of testosterone throughout pregnancy in rats

1982 ◽  
Vol 94 (1) ◽  
pp. 21-27 ◽  
Author(s):  
R. S. Bridges ◽  
R. B. Todd ◽  
C. M. Logue
Keyword(s):  
Post Partum ◽  
Late Pregnancy ◽  
Serum Testosterone ◽  
Serum Levels ◽  
Diurnal Variations ◽  
Serum Prolactin ◽  
Serum Concentrations ◽  
Pregnant Rats ◽  
Pregnant Rat ◽  
Microcolumn Chromatography

Testosterone concentrations in serum of rats bled throughout pregnancy and post partum were measured using Celite microcolumn chromatography and a radioimmunoassay for testosterone. Mean serum levels of testosterone ranged from about 170 to 340 pmol/l during the first 10 days of pregnancy. Significant increases in concentrations of testosterone in serum of pregnant rats were found on days 12, 15 and 18 of gestation. The highest testosterone concentrations occurred on days 18 and 20 of pregnancy when mean levels were 3228 and 3685 pmol/l respectively. Testosterone levels declined before parturition on day 22 (mean = 1449 pmol/l and declined further after parturition (mean = 315 pmol/l). In order to determine whether serum testosterone concentrations varied during the day in the pregnant rat, samples were collected at 6-h intervals on days 6–7 and 14–15 of gestation. Diurnal variations in serum testosterone concentrations were not evident during early or late pregnancy, unlike the rhythmic changes in serum prolactin levels found at these times during early pregnancy. The possible sources of the increased titres of serum testosterone during the second part of gestation in rats are discussed.

Distension of the uterus induces HspB1 expression in rat uterine smooth muscle

2011 ◽  
Vol 301 (5) ◽  
pp. R1418-R1426 ◽  
Author(s):  
B. G. White ◽  
D. J. MacPhee
Keyword(s):  
Protein Expression ◽  
Stress Protein ◽  
Late Pregnancy ◽  
Cytoplasmic Localization ◽  
Pregnant Rats ◽  
Pregnant Rat ◽  
Myometrial Cells ◽  
Uterine Smooth Muscle ◽  
Stimulation Of

The uterine musculature, or myometrium, demonstrates tremendous plasticity during pregnancy under the influences of the endocrine environment and mechanical stresses. Expression of the small stress protein heat shock protein B1 (HspB1) has been reported to increase dramatically during late pregnancy, a period marked by myometrial hypertrophy caused by fetal growth-induced uterine distension. Thus, using unilaterally pregnant rat models and ovariectomized nonpregnant rats with uteri containing laminaria tents to induce uterine distension, we examined the effect of uterine distension on myometrial HspB1 expression. In unilaterally pregnant rats, HspB1 mRNA and Ser15-phosphorylated HspB1 (pSer15 HspB1) protein expression were significantly elevated in distended gravid uterine horns at days 19 and 23 (labor) of gestation compared with nongravid horns. Similarly, pSer15 HspB1 protein in situ was only readily detectable in the distended horns compared with the nongravid horns at days 19 and 23; however, pSer15 HspB1 was primarily detectable in situ at day 19 in membrane-associated regions, while it had primarily a cytoplasmic localization in myometrial cells at day 23. HspB1 mRNA and pSer15 HspB1 protein expression were also markedly increased in ovariectomized nonpregnant rat myometrium distended for 24 h with laminaria tents compared with empty horns. Therefore, uterine distension plays a major role in the stimulation of myometrial HspB1 expression, and increased expression of this small stress protein could be a mechanoadaptive response to the increasing uterine distension that occurs during pregnancy.

OESTRADIOL CONTENT OF OVARIAN VENOUS BLOOD AND OVARIAN TISSUE IN HYPOPHYSECTOMIZED RATS DURING LATE PREGNANCY

1972 ◽  
Vol 54 (1) ◽  
pp. 79-85 ◽  
Author(s):  
H. B. WAYNFORTH ◽  
D. M. ROBERTSON
Keyword(s):  
Corpus Luteum ◽  
Post Partum ◽  
Ovarian Tissue ◽  
Venous Blood ◽  
Late Pregnancy ◽  
Corpora Lutea ◽  
Pregnant Rats ◽  
Hypophysectomized Rats ◽  
Competitive Protein Binding ◽  
Intact Rats

SUMMARY Oestradiol-17β in ovarian venous blood and ovarian tissue was assayed by a competitive protein-binding method. Oestradiol was found in similar amounts in the ovarian vein blood of pregnant rats hypophysectomized on Day 12 and killed on Days 16 and 21 and in pregnant rats sham-hypophysectomized on Day 12 and killed on Day 16. The pituitary therefore plays no part in oestrogen production after mid-pregnancy until some time between Day 16 and Day 21, when it gives rise to an increased ovarian venous blood oestradiol content just before parturition, in intact sham-hypophysectomized rats. It is suggested that this increase is associated with the advent of the post-partum ovulation. The corpus luteum and the extraluteal component of the ovary in hypophysectomized rats autopsied on Days 16 and 21 and in sham-hypophysectomized rats autopsied on Day 16, contain similar amounts of oestradiol within each group. The extraluteal component contains about five times more oestradiol than corpora lutea in sham-hypophysectomized intact rats autopsied on Day 21. The ovaries of these animals also show an increased amount of oestradiol over that of the ovaries in the other three groups. It is suggested that secretion of oestradiol after mid-pregnancy in rats involves concurrently both the corpus luteum and the extraluteal component of the ovary.

Effect of pregnancy on rat myometrial β2-adrenoceptor mRNA and isoproterenol-induced relaxation of isolated uterine strips

1997 ◽  
Vol 153 (3) ◽  
pp. 393-399 ◽  
Author(s):  
T Engstrøm ◽  
P Bratholm ◽  
H Vilhardt ◽  
N J Christensen
Keyword(s):  
De Novo ◽  
Contractile Activity ◽  
Post Partum ◽  
Late Pregnancy ◽  
Maximal Effect ◽  
Adrenergic Stimulation ◽  
Pregnant Rats ◽  
Binding Characteristics ◽  
Near Term ◽  
One Way Anova

Abstract The altered myometrial contractile activity near term of pregnancy is partly due to changes in the responsiveness to catecholamines. Previous experiments have basically been concerned with uterine adrenoceptor binding characteristics. In the present study we have evaluated total myometrial DNA, β2-adrenoceptor mRNA and isoproterenol-induced relaxation of rat isolated uterine strips pre-contracted with potassium on days 0, 7, 14 and 21 of pregnancy and on day 5 post-partum. Total myometrial DNA expressed per milligram wet tissue peaked at day 14 of pregnancy followed by a decrease at the end of gestation. This suggests that hyperplasia predominates in the growth of the uterus in early gestation, whereas hypertrophy may be more marked in late pregnancy. The concentration of β2-adrenoceptor mRNA decreased linearly throughout the gestational period (0·73 ± 0·20 amol/mg wet tissue on day 0 vs 0·34 ± 0·09 amol/mg wet tissue on day 21, P<0·05). Five days after parturition, at which time the uterus had returned to its pre-pregnant weight, β2-adrenoceptor mRNA was found to have increased 8-fold (2·79 ± 0·14 amol/mg wet tissue, P<0·05) as compared with day 21. The maximal effect of isoproterenol on pre-contracted uterine strips in which α-receptors were blocked by phentolamine showed a similar decrease which on day 21 reached 67% of the day 0 level (P<0·001). EC50 values were unchanged in all groups except day 21 pregnant rats in which an increase was observed. One-way ANOVA with Bonferroni's t-test showed statistically significant differences only between the day 21 group and either the day 5 post-partum group or the day 14 pregnant group (P<0·05). The observed alteration in EC50 prior to the end of gestation indicates that the system becomes less sensitive to β2-adrenergic stimulation at this time. We conclude that a reduction of de novo synthesis of β2-adrenoceptors may play a role in contributing to the increased myometrial activity at term. We further suggest that the dramatic up-regulation of β2-adrenoceptor mRNA postpartum may protect the fully involuted uterus against excessive contractions induced by oxytocin secreted during lactation. Journal of Endocrinology (1997) 153, 393–399

Sign in / Sign up

Export Citation Format

Share Document