Aging influences cellular and molecular responses of apoptosis to skeletal muscle unloading

2005 ◽  
Vol 288 (2) ◽  
pp. C338-C349 ◽  
Author(s):  
Parco M. Siu ◽  
Emidio E. Pistilli ◽  
David C. Butler ◽  
Stephen E. Alway
Keyword(s):  
Satellite Cell ◽  
Muscle Atrophy ◽  
Protein Level ◽  
Mrna Level ◽  
Age Groups ◽  
Experimental Period ◽  
Immunofluorescent Staining ◽  
Cell Nuclei ◽  
Protein Levels ◽  
Nuclear Apoptosis

The influence of aging on skeletal myocyte apoptosis is not well understood. In this study we examined apoptosis and apoptotic regulatory factor responses to muscle atrophy induced via limb unloading following loading-induced hypertrophy. Muscle hypertrophy was induced by attaching a weight to one wing of young and aged Japanese quails for 14 days. Removing the weight for 7 or 14 days after the initial 14 days of loading induced muscle atrophy. The contralateral wing served as the intra-animal control. A time-released bromodeoxyuridine (BrdU) pellet was implanted subcutaneously with wing weighting to identify activated satellite cells/muscle precursor cells throughout the experimental period. Bcl-2 mRNA and protein levels decreased after 7 days of unloading, but they were unchanged after 14 days of unloading in young muscles. Bcl-2 protein level but not mRNA level decreased after 7 days of unloading in muscles of aged birds. Seven days of unloading increased the mRNA level of Bax in muscles from both young and aged birds. Fourteen days of unloading increased mRNA and protein levels of Bcl-2, decreased protein levels of Bax, and decreased nuclear apoptosis-inducing factor (AIF) protein level in muscles of aged birds. BrdU-positive nuclei were found in all unloaded muscles from both age groups, but the number of BrdU-positive nuclei relative to the total nuclei decreased after 14 days of unloading compared with 7 days of unloading. The TdT-mediated dUTP nick end labeling (TUNEL) index was higher after 7 days of unloading in both young and aged muscles and after 14 days of unloading in aged muscles. Immunofluorescent staining revealed that almost all of the TUNEL-positive nuclei were also BrdU immunopositive, suggesting that activated satellite cell nuclei (both fused and nonfused) underwent nuclear apoptosis during unloading. There were significant correlations among levels of Bcl-2, Bax, and AIF and TUNEL index. Our data are consistent with the hypothesis that apoptosis regulates, at least in part, unloading-induced muscle atrophy and loss of activated satellite cell nuclei in previously loaded muscles. Moreover, these data suggest that aging influences the apoptotic responses to prolonged unloading following hypertrophy in skeletal myocytes.

Id2 and p53 participate in apoptosis during unloading-induced muscle atrophy

2005 ◽  
Vol 288 (5) ◽  
pp. C1058-C1073 ◽  
Author(s):  
Parco M. Siu ◽  
Stephen E. Alway
Keyword(s):  
Young Adult ◽  
Protein Content ◽  
Satellite Cell ◽  
Muscle Atrophy ◽  
Labeling Index ◽  
Immunofluorescent Staining ◽  
Age Group ◽  
Cell Nuclei ◽  
Regulatory Factors ◽  
And Control

Apoptotic signaling was examined in the patagialis (PAT) muscles of young adult and old quail. One wing was loaded for 14 days to induce hypertrophy and then unloaded for 7 or 14 days to induce muscle atrophy. Although the nuclear Id2 protein content was not different between unloaded and control muscles in either age group, cytoplasmic Id2 protein content of unloaded muscles was higher than that in contralateral control muscles after 7 days of unloading in young quails. Nuclear and cytoplasmic p53 contents and the p53 nuclear index of the unloaded muscles were higher than those in control muscles after 7 days of unloading in young quails, whereas in aged quails, the p53 and Id2 contents and p53 nuclear index of the unloaded muscles were not altered by unloading. Immunofluorescent staining indicated that myonuclei and activated satellite cell nuclei contributed to the increased number of p53-positive nuclei. Conversely, unloading in either young adult or aged PAT muscles did not alter c-Myc protein content. Although Cu-Zn-SOD content was not different in unloaded and control muscles, Mn-SOD content increased in PAT muscles after 7 days of unloading in young quails, suggesting that unloading induced an oxidative disturbance in these muscles. Moderate correlational relationships existed among Id2, p53, c-Myc, SOD, apoptosis-regulatory factors, and TdT-mediated dUTP nick end labeling index. These data indicate that Id2 and p53 are involved in the apoptotic responses during unloading-induced muscle atrophy after hypertrophy in young adult birds. Furthermore, our data suggest that there is an aging-dependent regulation of Id2 and p53 during unloading of previously hypertrophied muscles.

scholarly journals Status of the down-regulated canine testis using two different GNRH agonist implants in comparison with the juvenile testis

Reproduction ◽  
2013 ◽  
Vol 146 (6) ◽  
pp. 517-526 ◽  
Author(s):  
S Goericke-Pesch ◽  
M Gentil ◽  
A Spang ◽  
M P Kowalewski ◽  
K Failing ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Gnrh Agonist ◽  
Sertoli Cells ◽  
Protein Level ◽  
Limit Of Detection ◽  
Mrna Level ◽  
Cell Nuclei ◽  
Chain Cleavage ◽  
Cleavage Enzyme

Testicular function in the dog was down-regulated using two different GNRH agonist implants, with adult and juvenile testes serving as controls. Treatment resulted in an increased percentage of the interstitial area and decreased area of Leydig cell nuclei. Expression of StAR and the steroidogenic enzymes cytochrome P450 side-chain cleavage enzyme (P450scc, CYP11A1) and cytochrome P450 17α-hydroxylase-17,20-lyase (P450c17, CYP17A1) in Leydig cells was blocked at the mRNA and protein level, showing no differences between the two agonists. Staining for androgen receptor (AR) by immunohistochemistry was positive in Sertoli, Leydig and peritubular cells and some spermatogonia, with in situ hybridization confirming expression in Sertoli cells. At the mRNA level, expression of AR was not affected; however, translation was blocked (reduced percentage of AR-positive Sertoli cells), with the number of nuclei in basal position being decreased. In the juvenile testes, mRNA expression of StAR, CYP11A1 and CYP17A1 was higher compared with the other groups but distinctly lower for the AR. At the protein level, the expression was at the limit of detection for StAR; AR-positive Sertoli cells were not detected. Our observations show that the down-regulated testis is different from the juvenile one rather resembling the testicular status in seasonal breeders out of season.

scholarly journals Levels of G-proteins in liver and brain of lean and obese (ob/ob) mice

1992 ◽  
Vol 282 (1) ◽  
pp. 15-23 ◽  
Author(s):  
N McFarlane-Anderson ◽  
J Bailly ◽  
N Bégin-Heick
Keyword(s):  
Gene Expression ◽  
G Proteins ◽  
Protein Level ◽  
Mrna Level ◽  
Beta Subunits ◽  
Obese Mice ◽  
Protein Levels ◽  
Alpha Subunits ◽  
Brain Membranes ◽  
Liver Membranes

G-protein levels were assessed in liver and brain membranes of lean and obese mice. ADP-ribosylation and immunodetection studies revealed a decrease in the abundance of Gs and Gi alpha-subunits in the liver membranes of obese mice compared with lean mice. In contrast, in brain membranes, the abundance of these proteins was not significantly different between lean and obese mice. Studies at the mRNA level in both liver and brain revealed no difference in gene expression between lean and obese mice. Protein and mRNA studies both showed that Gs, Gi alpha 1, Gi alpha 2, Go alpha and G beta subunits are present in brain membranes, and Gi alpha 3 is barely detectable. In liver, Ga alpha, Gi alpha 2 and G beta subunits are the major constituents, whereas Gi alpha 1, Gi alpha 3 and Go alpha are barely detectable. It is possible that the differences observed at the protein level are due to different rates of translation of the mRNA. Different rates of release of the alpha-subunits from the membrane and/or different rates of degradation would also explain these results.

Differential regulation of G protein expression in rat hearts exposed to chronic hypoxia

1995 ◽  
Vol 269 (6) ◽  
pp. H1865-H1873 ◽  
Author(s):  
R. Kacimi ◽  
J. M. Moalic ◽  
A. Aldashev ◽  
D. E. Vatner ◽  
J. P. Richalet ◽  
...  
Keyword(s):  
Gene Expression ◽  
G Protein ◽  
Functional Activity ◽  
Protein Level ◽  
Chronic Hypoxia ◽  
Mrna Level ◽  
Mrna Levels ◽  
Protein Levels ◽  
Rat Hearts ◽  
And Function

Chronic hypoxia impairs adrenergic responsiveness. A modulation of Gs and/or G1 protein alpha-subunits may be associated with the downregulation of the beta-adrenergic receptors previously found in chronic hypoxia. G protein gene expression and protein level and function in rat hearts exposed to a 30-day hypobaric chronic hypoxia were compared with control rat hearts. No change was observed in G alpha s mRNA levels in either right or left ventricles. In right ventricles, mRNA levels of G alpha i-2 increased by 40% (P < 0.05), but not in left ventricles. In both left and right ventricles, chronic hypoxia did not modify G alpha i-2 and G alpha s protein amounts, but significantly decreased functional activity of G alpha s. In conclusion, gene expression, protein levels of G alpha s and G alpha i-2, and activity of G alpha s do not change in parallel fashion with chronic hypoxia. In chronic hypoxic right ventricles, although the mRNA level of G alpha i-2 is increased, the protein level is unchanged. One potential mechanism of desensitization to catecholamines in chronic hypoxia appears to involve a decreased functional activity of G alpha s in spite of normal mRNA and protein levels

scholarly journals Losartan Protects the Heart Against Ischemia Reperfusion Injury: Sirtuin3 Involvement

2015 ◽  
Vol 18 (1) ◽  
pp. 112 ◽  
Author(s):  
Mohsen Sharifi Klishadi ◽  
Farideh Zarei ◽  
Seyyed Hassan Hejazian ◽  
Ali Moradi ◽  
Mahdieh Hemati ◽  
...  
Keyword(s):  
Protein Level ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Mrna Level ◽  
Experimental Studies ◽  
Control Group ◽  
Signaling Mechanism ◽  
Survival Factors ◽  
Protein Levels ◽  
Ischemic Tissue

PURPOSE: Sirtuin-3 (SIRT3) deacetylase protects the heart against oxidative stress via survival factors upregulation. Clinical and experimental studies have demonstrated that activation of systemic and local renin-angiotensin system (RAS) is implicated in ischemia-induced cardiac injury. However, the relation between RAS and SIRT3 in pathophysiology of myocardial ischemia reperfusion is unknown. In this study, the cardiac transcription and expression of SIRT3 levels was examined in response to ischemia reperfusion in untreated and losartan treated rats. METHODS: Rats were divided into control group, losartan group (L), and ischemia reperfusion (IR) groups with (L+IR) or without losatran pretreatment. Some rats were included as sham-operated and saline groups. IR was induced by left anterior descending artery occlusion. SIRT3 protein levels were determined by Western blot technique. The genes expression was specified by real-time RT-PCR. Arrhythmias were assessed according to the Lambeth conventions. RESULTS: In L+IR group a significant reduction was noted in the number of ventricular ectopic beats (VEBs) and episodes of ventricular tachycardia (VT) (VEBs: P<0.001; VT: P<0.01 vs. IR). In IR group, SIRT3 protein level was decreased in the ischemic tissue by 26.7±5.9% (P<0.01 vs. Control). However, in the non-ischemic tissue the changes of SIRT3 protein content were not significant. In L+IR group SIRT3 protein levels in the ischemic part of Left ventricle were significantly different from IR group (P<0.001). SIRT3 mRNA level did not change significantly among the experimental groups. Thioredoxin-1 and catalase transcription level was increased in L+IR group compared to IR group (P<0.01). CONCLUSION: A decreased SIRT3 protein levels subsequent to IR might be a novel signaling mechanism involved in IR injury. Losartan at non–hypotensive dose exerts anti-ischemic effects in part by normalizing the SIRT3 protein level and upregulating the survival factors encoding genes transcription in ischemic tissue of the heart. This article is open to POST-PUBLICATION REVIEW. Registered readers (see “For Readers”) may comment by clicking on ABSTRACT on the issue’s contents page.

scholarly journals CRUDE PROTEIN REQUIREMENTS OF GUINEA FOWL KEETS (NUMIDA MELIAGRIS GALEATA, PALLAS) IN THE TROPICS

2021 ◽  
Vol 14 ◽  
pp. 88-94
Author(s):  
M. C. Njike ◽  
A. S. Ahmed ◽  
E. S. Haruna
Keyword(s):  
Growth Rate ◽  
Protein Level ◽  
Crude Protein ◽  
Experimental Period ◽  
Guinea Fowl ◽  
Feed Consumption ◽  
Energy Concentration ◽  
Protein Requirements ◽  
Protein Levels ◽  
The Tropics

Guinea fowl of both sexes were fed from 0 to 8 weeks on protein level ranging from 18 to 26% using constant energy concentration of 3000 kilocalories/kg diet. At the end of the experimental period, keats on 24 and 26% protein levels with liveweights of 854 and 867.3g respectively were significantly heavier than keets on the other diets. Keets on these two rations showed no significant differences in the final liveweights, liveweight gains and feed consumption. However, it appeared from the result that feed consumption was related to growth rate. The feed/gain ratio was significantly better for keets that received 24% protein level than for those on 26% protein. But feed/gain ratios generally tended to be inversely related to growth rate. On the basis of this study it is recommended that the diet of guinea fowl keets in the tropics should not contain less than 24% crude protein for optimal results.   

scholarly journals Reduction of serotonin receptor 2A protein level in peripheral blood lymphocytes during antipsychotic medication is biomarker of therapy safety

2020 ◽  
pp. 58-59
Author(s):  
А.М. Заботина ◽  
М.Н. Грунина ◽  
Е.В. Волкова ◽  
Р.Ф. Насырова ◽  
А.Е. Тараскина
Keyword(s):  
Peripheral Blood ◽  
Protein Level ◽  
Serotonin Receptor ◽  
Mrna Level ◽  
Peripheral Blood Lymphocytes ◽  
Antipsychotic Therapy ◽  
Blood Lymphocytes ◽  
Protein Levels ◽  
Potential Biomarker ◽  
Serotonin Receptor 2A

В лимфоцитах периферической крови (ЛПК) в группе пациентов с коморбидным течением шизофрении с синдромом алкогольной зависимости показаны более низкие значения количества белка рецептора 5-НТ2А и уровня его мРНК по сравнению с группой с неотягощенной психической патологией (р<0.001). Безопасная фармакотерапия ассоциирована со снижением количества рецептора 5-НТ2А в ЛПК на 28-й день без изменения уровня транскрипции гена, возможно вызванного интернализацией рецептора. We studied mRNA and protein levels of serotonin receptor 2A (5-НТ2А) in lymphocytes of schizophrenia patients both before and after 28 days of antipsychotic therapy in order to estimate biomarkers of efficacy and safety of the treatment. We found lower mRNA and protein levels of 5-НТ2А in peripheral blood lymphocytes (PBL) of schizophrenia patients with alcohol addiction syndrome than in PBL of schizophrenia patients without addictions (р<0.001). Absence of common adverse side effects (antipsychotic-induced weight gain, parkinsonism, akathisia) during antipsychotic therapy was associated with reduction of 5-НТ2А protein level in PBL by 28th day of the treatment, while HTR2A mRNA level was not changed. Reduction of 5-НТ2А protein level in PBL by 28th day may be a potential biomarker of the antipsychotic therapy safety.

Transcripts and protein levels of CSN1S1 and CSN3 genes in dairy cattle mammary gland secretory tissue during chronic staphylococcal infection

2021 ◽  
Vol 88 (1) ◽  
pp. 73-77
Author(s):  
Ewelina Kawecka-Grochocka ◽  
Magdalena Zalewska ◽  
Aleksandra Kapusta ◽  
Tomasz Ząbek ◽  
Magdalena Rzewuska ◽  
...  
Keyword(s):  
Protein Level ◽  
Mrna Level ◽  
Milk Proteins ◽  
Staphylococcal Infection ◽  
Coagulase Negative Staphylococci ◽  
Post Translational Modifications ◽  
Protein Levels ◽  
Secretory Tissue ◽  
Cheese Curd ◽  
Coagulase Positive Staphylococci

AbstractOur objective was to determine the influence of chronic coagulase-positive staphylococci (CoPS) or coagulase-negative staphylococci (CoNS) infection on the mRNA and protein levels of two main milk proteins responsible for cheese curd quantity and quality, alpha-S1-casein (CSN1S1) and kappa-casein (CSN3). Measurements were made in cow mammary parenchyma with a prevalence of secretory tissue (MGST). Samples of MGST were collected from the separate quarters and divided into CoPS, CoNS and bacteria-free (H) groups according to the microbiological status of the quarter milk. No differences in CSN1S1 and CSN3 mRNA level were found between groups, however, CSN1S1 protein level was significantly higher in the H group than the CoNS group, and CSN3 protein level was significantly higher in H than CoPS group. Hence, while the CSN1S1 and CSN3 genes appear to be constitutively expressed at the mRNA level in dairy cow MGST during mastitis, CoNS infection negatively affected CSN1S1 protein level, and CoPS infection negatively affected CSN3 protein level. The lack of change at the mRNA level suggests that staphylococcal infection may affect the post-transcriptional or post-translational modifications.

Pengaruh Lama Pengasinan Terhadap Kadar Protein Putih Telur Itik

2014 ◽  
Vol 1 (1) ◽  
pp. 36 ◽  
Author(s):  
Siti Fatimah ◽  
Muji Rahayu ◽  
Siti Aminah
Keyword(s):  
Protein Level ◽  
Egg White ◽  
Special Treatment ◽  
Egg White Protein ◽  
Protein Levels ◽  
Duck Egg ◽  
Egg Period ◽  
Graph Presentation ◽  
And Control ◽  
The Impact

Background : Egg  is one of the animal protein source, which has delicious taste, easy to digest and highly nutritious. Besides its affordable price, its supply availability is unquestionable as well. However, due to its short storability, it requires special treatment, such as preserving, to store it for long period. One way to preserve the egg is by pickling egg, which generally requires seven to ten days of marinating. During the process of marinating, there will be a visual change of egg white and yolk. Their structures  will be more solid (the occurrence of thickening process) because salinization will lead to protein denaturalization. Consequently, it has an influence as well towards the content of egg white protein of duck egg. This study is aimed to explore the impact of various time of pickling egg towards egg white protein of duck egg. Method  : The study where takes place in a laboratories, is a true experimental study for the reason that the researcher must provide intervention, hence all of potentially confounding variables are manageable. Samples that had been used in this study are duck eggs which were bought from North Brebes. This study is expected to generate data from four various time of pickling egg and control (no treatment). Since there are four samples, accordingly the number of data resulted are twenty. The resulted data will be descriptively presented in table, graph, presentation, and narration. Result  : Protein level examination within duck white egg shows changes  in protein levels that occurs in every variation of pickling egg time, where the average results of the assay of duck egg white protein is 14.94% without treatment (control), in five days of pickling time is 13.68%, in seven days of pickling time is 13.29%, in nine days of pickling time is 12.87% and eleven days of pickling time is 12.78%. Conclusion  : There is a significant impact among the period of pickling time to the protein level degradation of duck white egg. Keywords : Duck egg, period of pickling time, level protein of duck white egg.

scholarly journals Functional Comparison between VP64-dCas9-VP64 and dCas9-VP192 CRISPR Activators in Human Embryonic Kidney Cells

2021 ◽  
Vol 22 (1) ◽  
pp. 397
Author(s):  
Nasir Javaid ◽  
Thuong L. H. Pham ◽  
Sangdun Choi
Keyword(s):  
Protein Level ◽  
Reference Genes ◽  
Mrna Level ◽  
High Specificity ◽  
Kidney Cells ◽  
Human Embryonic Kidney ◽  
Human Embryonic Kidney Cells ◽  
Sox2 Expression

Reversal in the transcriptional status of desired genes has been exploited for multiple research, therapeutic, and biotechnological purposes. CRISPR/dCas9-based activators can activate transcriptionally silenced genes after being guided by gene-specific gRNA(s). Here, we performed a functional comparison between two such activators, VP64-dCas9-VP64 and dCas9-VP192, in human embryonic kidney cells by the concomitant targeting of POU5F1 and SOX2. We found 22- and 6-fold upregulations in the mRNA level of POU5F1 by dCas9-VP192 and VP64-dCas9-VP64, respectively. Likewise, SOX2 was up-regulated 4- and 2-fold using dCas9-VP192 and VP64dCas9VP64, respectively. For the POU5F1 protein level, we observed 3.7- and 2.2-fold increases with dCas9-VP192 and VP64-dCas9-VP64, respectively. Similarly, the SOX2 expression was 2.4- and 2-fold higher with dCas9-VP192 and VP64-dCas9-VP64, respectively. We also confirmed that activation only happened upon co-transfecting an activator plasmid with multiplex gRNA plasmid with a high specificity to the reference genes. Our data revealed that dCas9-VP192 is more efficient than VP64-dCas9-VP64 for activating reference genes.

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