Mechanism of the reduction in DNA content in sperm cells preserved in vitro

1969 ◽  
Vol 67 (4) ◽  
pp. 383-385 ◽  
Author(s):  
I. I. Ivanov ◽  
N. V. Korban ◽  
V. I. Sharobaiko
Keyword(s):  
Dna Content ◽  
Sperm Cells

scholarly journals DECREASE IN NUCLEAR FEULGEN-POSITIVE MATERIAL (DNA) UPON AGING IN IN VITRO STORAGE OF BOVINE SPERMATOZOA

1961 ◽  
Vol 10 (3) ◽  
pp. 353-359 ◽  
Author(s):  
G. W. Salisbury ◽  
W. J. Birge ◽  
L. de la Torre ◽  
J. R. Lodge
Keyword(s):  
Dna Content ◽  
Artificial Insemination ◽  
Embryonic Mortality ◽  
Sperm Cells ◽  
Progressive Decrease ◽  
Bovine Spermatozoa ◽  
The Individual ◽  
Feulgen Technique ◽  
Absorption Measurements

The Feulgen-DNA content of sperm cells from 5 bulls was studied by means of microspectrophotometry after storage at 5°C for 2, 3, 5, and 10 days in a yolk-citrate diluent permitting slow aerobic metabolism. A subsample of sperm cells from each bull was subjected to the Feulgen technique on each of the storage days selected. The cells sampled on each of these days received a standard 12 minute, 60°C hydrolysis. Absorption measurements at 546 mµof the individual cells indicated a marked progressive decrease in the Feulgen-DNA content of the stored spermatozoa. The loss of 30 per cent of the initial DNA at the end of 5 days' storage was highly significant statistically. This decrease approximately parallels the known decrease in fertility of stored sperm cells, as well as the increase in apparent embryonic mortality resulting from the use of similarly aged spermatozoa for artificial insemination.

scholarly journals Cooling of equine semen at 16°C for 36 hours with addition of different glutathione concentrations

2015 ◽  
Vol 36 (6) ◽  
pp. 3699
Author(s):  
Rodrigo Arruda de Oliveira ◽  
Marco Antônio De Oliveira Viu ◽  
Maria Lúcia Gambarini
Keyword(s):  
Lipid Peroxidation ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Membrane Lipid ◽  
Sperm Cells ◽  
Sperm Viability ◽  
Membrane Lipid Peroxidation ◽  
Acrosomal Membrane ◽  
In Vitro Effects

Handling equine semen during the refrigeration process reduces sperm viability, and consequently causes membrane lipid peroxidation, among other challenges. The present study aimed to evaluate the in vitro effects of glutathione (control, 1. 0, 1. 5, and 2. 5 mM) on equine semen in a refrigeration protocol of 16ºC for 36 hours. The following variables were evaluated after 0, 12, 24, and 36 hours refrigeration: total sperm motility, vigor, viability, and plasma and acrosomal membrane integrity. Motility was higher with 2. 5mM of glutathione (57. 8 ± 7. 3) after 12 hours of refrigeration compared to the control (53. 2 ± 8. 3) (P < 0. 05). After 36 hours of refrigeration, motility was higher with 1. 5 mM (43. 4 ± 12. 7) and 2. 5mM glutathione (45. 5 ± 6. 2), than it was with 1mM glutathione (38. 2 ± 9) and the control (35. 5 ± 18. 4) (P < 0. 05), respectively. Vigor was highest with 1. 5mM glutathione (3. 7 ± 0. 3) after 36 hours compared to the control (3. 2 ± 1. 1), (P < 0. 05). Viability differed between control and 1mM treatments (79. 5 ± 1. 8) only after 24 hours (75. 5 ± 9. 7) (P < 0. 05). Throughout the investigation, no significant differences were noted in plasma and acrosomal membrane integrity (P > 0. 05). The 1. 5 and 2. 5mM glutathione levels were more efficient in protecting sperm cells and yielded higher total motility values after 36 hours of refrigeration.

Chlortetracycline staining patterns of frozen-thawed bull spermatozoa treated with β-cyclodextrins, dibutyryl cAMP and progesterone

Zygote ◽  
2000 ◽  
Vol 8 (3) ◽  
pp. 245-256 ◽  
Author(s):  
Michael B. Dinkins ◽  
Benjamin G. Brackett
Keyword(s):  
Intestinal Mucosa ◽  
Sperm Cells ◽  
Fluorescent Staining ◽  
Sperm Capacitation ◽  
Dibutyryl Camp ◽  
Bovine Spermatozoa ◽  
Differential Responses ◽  
Definition Of

Efforts to achieve complete chemical definition of media used for in vitro capacitation of bovine spermatozoa including removal of heparin purified from porcine intestinal mucosa are presented. Fluorescent staining with chlortetracycline (CTC), known to reflect changes coincident with sperm capacitation in certain species, was studied following treatments of frozen-thawed bull spermatozoa with β-cyclodextrins, dibutyryl cAMP (dbcAMP) and progesterone in comparison with heparin. The CTC staining patterns (F, B and AR) were confirmed to correlate with known conditions that effectively prepare cryopreserved bull spermatozoa for fertilisation in vitro. In the absence of glucose, the routinely employed heparin-containing capacitating medium caused an increase in spermatozoa displaying the AR pattern. Both progesterone (100 μM) and dbcAMP (0.01–0.1 mM) were able to increase the proportion of B pattern stained sperm cells more than after exposure to control (mDM) conditions without a significant reduction in motility. Exposure to either dbcAMP or β-cyclodextrins was accompanied by an increase in proportions of spermatozoa displaying the AR pattern over those seen in controls. Exposure to β-cyclodextrins did not increase the proportion of B pattern stained spermatozoa. Comparison of spermatozoa from two bulls revealed differential responses of spermatozoa from different males to treatments with heparin and progesterone. In vitro fertilisation results demonstrated that previously cryopreserved bull spermatozoa could be capacitated in chemically defined conditions devoid of heparin or other biological components.

scholarly journals In vitro propagation, DNA content and essential oil composition of Teucrium scorodonia L. ssp. scorodonia

2016 ◽  
Vol 127 (1) ◽  
pp. 1-13 ◽  
Author(s):  
Joanna Makowczyńska ◽  
Elwira Sliwinska ◽  
Danuta Kalemba ◽  
Ewelina Piątczak ◽  
Halina Wysokińska
Keyword(s):  
Essential Oil ◽  
Dna Content ◽  
In Vitro Propagation ◽  
Essential Oil Composition ◽  
Oil Composition

scholarly journals Spermatozoa Obtained From Alpaca vas deferens. Effects of Seminal Plasma Added at Post-thawing

2021 ◽  
Vol 8 ◽  
Author(s):  
Eduardo G. Aisen ◽  
Wilfredo Huanca López ◽  
Manuel G. Pérez Durand ◽  
Edita Torres Mamani ◽  
Juan C. Villanueva Mori ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Artificial Insemination ◽  
Reproductive Tract ◽  
Vas Deferens ◽  
Female Reproductive Tract ◽  
Low Fertility ◽  
Sperm Cells ◽  
South American Camelids ◽  
Thawing Process

The viscous seminal plasma (SP) is currently a major impediment to the handling of ejaculate and the development of some biotechnologies in South American camelids. The vas deferens-collected spermatozoa of alpacas is a useful technique to avoid this problem. On the other hand, SP contains a large protein component that has been implicated in the function of spermatozoa within the female reproductive tract. In this sense, the low fertility achieved using transcervical insemination with frozen-thawed spermatozoa in alpacas could be improved by adding SP. This study aimed to evaluate the effect of the whole SP on some in vitro parameters of alpaca spermatozoa after the freezing-thawing-process and the fertility after artificial insemination. It would contribute to a better understanding of the interaction between thawed sperm cells and SP. Spermatozoa were obtained by surgically diverted vas deferens. The samples were diluted with a Tris-based extender, packaged in straws, and frozen. At thawing, each straw was divided into two post-thawing conditions: with the addition of 10% of PBS (control) or with 10% SP (treatment). The sperm cells were evaluated using dynamic parameters, sperm cell morphology, and morphometry. Fertility was assessed by an artificial insemination trial. All in vitro parameters were analyzed by ANOVA. A heterogeneity test was scheduled for the fertility trial. After the freezing-thawing process, motility and plasma membrane functionality was improved when SP was added. No differences were found for post-thaw viability between the control and treatment samples. The percentage of normal cells was higher with SP at post-thawing, and a decrease of the presence of bent tailed spermatozoa with a droplet in the SP group was observed. The length of the head spermatozoa was 3.4% higher in the samples with PBS compared to those in which SP was added. Females pregnant at day 25 post-insemination were 0/12 (with SP inside the straw) and 1/10 (without SP inside the straw). In conclusion, the presence of 10% SP at post-thawing improves sperm cells' motility, functionality, and morphology, indicating that it would be beneficial to improve the frozen-thawed alpaca's physiology spermatozoa. More fertility trials must be developed to increase this knowledge.

DNA content distribution of in vivo and in vitro lines of Lewis lung carcinoma

1982 ◽  
Vol 18 (10) ◽  
pp. 973-978 ◽  
Author(s):  
G. Starace ◽  
G. Badaracco ◽  
C. Greco ◽  
A. Sacchi ◽  
G. Zupi
Keyword(s):  
Lung Carcinoma ◽  
Dna Content ◽  
Lewis Lung Carcinoma ◽  
Content Distribution ◽  
Lewis Lung

scholarly journals The in vitro toxicity of atrazine on kinematics and DNA fragmentation in common carp (Cyprinus carpio) sperm cells

2019 ◽  
Vol 70 (3) ◽  
pp. 1639
Author(s):  
M.E. ÖZGÜR
Keyword(s):  
Dna Fragmentation ◽  
Common Carp ◽  
Cyprinus Carpio ◽  
In Vitro Toxicity ◽  
Computer Assisted ◽  
Sperm Cells ◽  
Common Carp Cyprinus Carpio ◽  
Significant Difference ◽  
Carp Cyprinus Carpio

This study investigated the in vitro effects of different concentrations of Atrazine (0.001, 0.01, 0.1, 0.5 mg/L) added to motile and immotile solutions on kinematics quality of sperm cells of common carp, Cyprinus carpio, which is a fish of economic significance. The kinematics of the sperm cells was analyzed by a computer-assisted sperm analysis system (CASA). As a result of the study, while there was a significant difference (P < 0.05) between the groups in terms of the VSL (μm/s) and VCL (μm/s) values after the Atrazine-added immotile solution’s (IMS) and incubation for 3 hours, there was a significant difference (P < 0.05) in only the VSL values directly activated by the Atrazine-added motile solution (MS). DNA fragmentation was evident but not in higher numbers in the 0.1 mg/L atrazine group. Finally, it was determined the effective concentration (EC50) values of the VSL value of the motile and immotile solution as 0.34 mg/L and 0.03 mg/L, respectively.

Role of membrane phosphotyrosine proteins in human spermatozoal function

1991 ◽  
Vol 99 (1) ◽  
pp. 157-165
Author(s):  
R.K. Naz ◽  
K. Ahmad ◽  
R. Kumar
Keyword(s):  
Physiological State ◽  
Human Sperm ◽  
Kinase Assay ◽  
Relative Molecular Mass ◽  
Sperm Cells ◽  
Western Blots ◽  
Tyrosine Residues ◽  
Sperm Extract ◽  
32P Incorporation

The monoclonal anti-phosphotyrosine antibody (PTA) recognized proteins related to relative molecular mass regions of 94,000 +/− 3000 and 46,000 +/− 3000 Mr on Western blots of detergent-solubilized non-capacitated human sperm extract (HSE). The pattern of phosphorylation at tyrosine residues depended upon the physiological state of the sperm cells. At least six protein bands corresponding to four molecular regions of 94,000 +/− 3000, 46,000 +/− 3000, 25,000 +/− 7000 and 12,000 +/− 2000 Mr, respectively, were labeled with 32P when human sperm were capacitated in vitro; the proteins belonging to the former three regions were phosphotyrosine proteins as they were precipitable by PTA. In vitro kinase assay performed directly on HSE indicated autophosphorylation of proteins of the same four molecular regions, with the capacitated sperm preparations having 30% higher 32P incorporation into 94,000 +/− 3000 Mr proteins and 17% less incorporation into 12,000 +/− 2000 Mr proteins as compared to the non-capacitated sperm preparations. Both of these protein regions were also autophosphorylated at tyrosine residues when immunoprecipitated phosphotyrosine proteins were used for the kinase assay. Phosphorylation of tyrosine residues of 94,000 +/− 3000 Mr proteins was further stimulated by 1.38- to 1.46-fold in response to exposure to zona pellucida proteins, namely the porcine ZP3 and human zona proteins (HZP); the HZP induced the highest response. Immunofluorescence observations on fixed human sperm demonstrated that capacitation as well as exposure to zona proteins increased the degree of tyrosine-specific fluorescence per sperm cell as well as the number of sperm cells that showed fluorescence at the acrosomal region of the spermhead.(ABSTRACT TRUNCATED AT 250 WORDS)

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