Fixation Temperature Affects DNA Integrity in the Testis as Measured by the TUNEL Assay

2012 ◽  
Vol 40 (4) ◽  
pp. 667-674 ◽  
Author(s):  
Dibyendu Dutta ◽  
In Park ◽  
Nathaniel C. Mills
Keyword(s):  
Tunel Assay ◽  
Dna Integrity ◽  
Fixation Temperature

scholarly journals Relationships between sperm DNA fragmentation, sperm apoptotic markers and serum levels of CB-153 and p,p′-DDE in European and Inuit populations

Reproduction ◽  
2006 ◽  
Vol 132 (6) ◽  
pp. 949-958 ◽  
Author(s):  
A Stronati ◽  
G C Manicardi ◽  
M Cecati ◽  
M Bordicchia ◽  
L Ferrante ◽  
...  
Keyword(s):  
Dna Fragmentation ◽  
Serum Levels ◽  
Study Groups ◽  
Tunel Assay ◽  
Dna Integrity ◽  
Sperm Dna Fragmentation ◽  
Adult Males ◽  
Apoptotic Markers ◽  
Sperm Dna

Persistent organochlorine pollutants (POPs) are suspected to interfere with hormone activity and the normal homeostasis of spermatogenesis. We investigated the relationships between sperm DNA fragmentation, apoptotic markers identified on ejaculated spermatozoa and POP levels in the blood of 652 adult males (200 Inuits from Greenland, 166 Swedish, 134 Polish and 152 Ukrainian). Serum levels of 2, 2′, 4, 4′, 5, 5′-hexachlorobiphenyl (CB-153), as a proxy of the total POP burden, and of 1,1-dichloro-2,2-bis(p-chlorophenyl)-ethylene (p,p′-DDE), as a proxy of the total DDT exposure were determined. Sperm DNA fragmentation was measured by using the TUNEL assay, whereas immunofluorescence methods were utilized for detecting pro-apoptotic (Fas) and anti-apoptotic (Bcl-xL) markers. Both TUNEL assay and apoptotic markers were statistically differed across the four populations. No correlation between neither sperm DNA fragmentation nor apoptotic sperm parameters and the large variations in POPs exposure was observed for the separate study groups. However, considering the European populations taken together, we showed that both %TUNEL positivity and Bcl-xL were related to CB-153 serum levels, whereas our study failed to demonstrate any relations between DDE and %TUNEL positivity and apoptotic sperm biomarkers (Fas and Bcl-xL) in any region or overall regions. These results suggest that CB-153 and related chemicals might alter sperm DNA integrity and Bcl-xL levels in European adult males, but not in the highly exposed Inuit men. Additional issues (genetic background, lifestyle habits and characterization of total xeno-hormonal activities) need to be investigated in order to fully assess the population variations observed.

Assessment of DNA damage in goat preantral follicles after vitrification of the ovarian cortex

2015 ◽  
Vol 27 (3) ◽  
pp. 440 ◽  
Author(s):  
Luciana R. Faustino ◽  
Adeline A. Carvalho ◽  
Cleidson M. G. Silva ◽  
Rafael Rossetto ◽  
Cláudio A. P. Lopes ◽  
...  
Keyword(s):  
Dna Damage ◽  
Ovarian Tissue ◽  
Tunel Assay ◽  
Lower Percentage ◽  
Dna Integrity ◽  
Preantral Follicles ◽  
Histone H2ax ◽  
Γh2ax Foci ◽  
Control Samples

Effective methods for gamete preservation should have low impact on DNA integrity. The present study investigated the effects of vitrification of goat ovarian tissues on the occurrence of DNA fragmentation and DNA double-stand breaks using the terminal deoxyribonucleotidyl transferase-mediated dUTP–digoxigenin nick end-labelling (TUNEL) assay and detection of phosphorylated histone H2AX (γH2AX), respectively. Goat ovaries were collected at a local abattoir and 12 tissue fragments were prepared from each ovarian pair. Tissue fragments were used as fresh control samples or were cultured in vitro, vitrified or vitrified and cultured. Vitrification was performed using the Ovarian Tissue Cryosystem. Fragments from all groups (control and treatments) were processed for histology, transmission electron microscopy, TUNEL assay and immunofluorescence. Compared with fresh control samples, a lower percentage of morphologically normal follicles was detected in the vitrification followed by culture treatment group (P < 0.05). Normal follicular ultrastructure was observed in all groups. Immunofluorescence revealed the presence of γH2AX foci in few oocytes and ovarian stromal cells. TUNEL-positive follicles were found in samples without significant differences among groups (P > 0.05). In conclusion, the vitrification protocol used in the present study did not increase DNA damage in preantral follicles enclosed in goat ovarian tissues.

Premature ovarian ageing following heterozygous loss of Senataxin

2020 ◽  
Author(s):  
G N Subramanian ◽  
M Lavin ◽  
H A Homer
Keyword(s):  
Dna Damage ◽  
Neurodegenerative Disorder ◽  
Dna Helicase ◽  
Dna Integrity ◽  
Ovarian Activity ◽  
Homozygous Mutation ◽  
Female Mice ◽  
Mating Trial ◽  
High Prevalence

Abstract Premature loss of ovarian activity before 40 years of age is known as primary ovarian insufficiency (POI) and occurs in ∼1% of women. A more subtle decline in ovarian activity, known as premature ovarian ageing (POA), occurs in ∼10% of women. Despite the high prevalence of POA, very little is known regarding its genetic causation. Senataxin (SETX) is an RNA/DNA helicase involved in repair of oxidative stress-induced DNA damage. Homozygous mutation of SETX leads to the neurodegenerative disorder, ataxia oculomotor apraxia type 2 (AOA2). There have been reports of POI in AOA2 females suggesting a link between SETX and ovarian ageing. Here, we studied female mice lacking either one (Setx+/−) or both (Setx−/−) copies of SETX over a 12- to 14-month period. We find that DNA damage is increased in oocytes from 8-month-old Setx+/− and Setx−/− females compared with Setx+/+ oocytes leading to a marked reduction in all classes of ovarian follicles at least 4 months earlier than typically occurs in female mice. Furthermore, during a 12-month long mating trial, Setx+/− and Setx−/− females produced significantly fewer pups than Setx+/+ females from 7 months of age onwards. These data show that SETX is critical for preventing POA in mice, likely by preserving DNA integrity in oocytes. Intriguingly, heterozygous Setx loss causes an equally severe impact on ovarian ageing as homozygous Setx loss. Because heterozygous SETX disruption is less likely to produce systemic effects, SETX compromise could underpin some cases of insidious POA.

DNA integrity and ecophysiological responses of Spanish populations of Ulmus glabra to increasing ozone levels

Ecotoxicology ◽  
2021 ◽  
Author(s):  
Ambra Dell’Orso ◽  
Elena Kuzminsky ◽  
Victoria Bermejo-Bermejo ◽  
Raquel Ruiz-Checa ◽  
Rocío Alonso-Del Amo ◽  
...  
Keyword(s):  
Dna Integrity ◽  
Ulmus Glabra ◽  
Spanish Populations ◽  
Ecophysiological Responses ◽  
Ozone Levels

scholarly journals TUNEL Assay: A Powerful Tool for Kidney Injury Evaluation

2021 ◽  
Vol 22 (1) ◽  
pp. 412
Author(s):  
Christopher L. Moore ◽  
Alena V. Savenka ◽  
Alexei G. Basnakian
Keyword(s):  
Cell Death ◽  
Dna Fragmentation ◽  
Cell Injury ◽  
Kidney Injury ◽  
Cell Types ◽  
Tunel Assay ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Hypoxic Injury ◽  
Additional Information ◽  
Tissue Compartments

Terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay is a long-established assay used to detect cell death-associated DNA fragmentation (3’-OH DNA termini) by endonucleases. Because these enzymes are particularly active in the kidney, TUNEL is widely used to identify and quantify DNA fragmentation and cell death in cultured kidney cells and animal and human kidneys resulting from toxic or hypoxic injury. The early characterization of TUNEL as an apoptotic assay has led to numerous misinterpretations of the mechanisms of kidney cell injury. Nevertheless, TUNEL is becoming increasingly popular for kidney injury assessment because it can be used universally in cultured and tissue cells and for all mechanisms of cell death. Furthermore, it is sensitive, accurate, quantitative, easily linked to particular cells or tissue compartments, and can be combined with immunohistochemistry to allow reliable identification of cell types or likely mechanisms of cell death. Traditionally, TUNEL analysis has been limited to the presence or absence of a TUNEL signal. However, additional information on the mechanism of cell death can be obtained from the analysis of TUNEL patterns.

scholarly journals Toxicity and chemical transformation of silver nanoparticles in A549 lung cells: dose-rate-dependent genotoxic impact

2021 ◽  
Author(s):  
Laure Bobyk ◽  
Adeline Tarantini ◽  
David Beal ◽  
Giulia Veronesi ◽  
Isabelle Kieffer ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Silver Nanoparticles ◽  
Dose Rate ◽  
Cell Metabolism ◽  
A549 Cells ◽  
Dna Integrity ◽  
Lung Cells ◽  
Ag Nps ◽  
Rate Dependent ◽  
A549 Lung

Acute exposure of A549 cells to Ag-NPs induces stronger effects on DNA integrity, ROS level, cell metabolism and cell cycle than repeated exposure. Ag-NPs dissolves in both exposure conditions and Ag ions recombine with thiolated proteins.

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