Nanocarried antioxidants in freezing extenders for boar spermatozoa

Andrologia ◽  
2021 ◽  
Author(s):  
S. L. Soares ◽  
Camila R. C. Brito ◽  
Andreia Nobre Anciuti ◽  
Norton C. Gatti ◽  
Carine Dahl Corcini ◽  
...  
Keyword(s):  
Author(s):  
N. Seki ◽  
Y. Toyama ◽  
T. Nagano

It is believed that i ntramembra.nous sterols play an essential role in membrane stability and permeability. To investigate the distribution changes of sterols in sperm membrane during epididymal maturation and capacitation, filipin has been used as a cytochemical probe for the detection for membrane sterols. Using this technique in combination with freeze fracturing, we examined the boar spermatozoa under various physiological conditions.The spermatozoa were collected from: 1) caput, corpus and cauda epididymides, 2) sperm rich fraction of ejaculates, and 3)the uterus 2hr after natural coition. They were fixed with 2.5% glutaraldehyde in 0.05M cacodylate buffer (pH 7.4), and treated with the filipin solution (final concentration : 0.02.0.05%) for 24hr at 4°C with constant agitation. After the filipin treatment, replicas were made by conventional freeze-fracture technique. The density of filipin-sterol complexes (FSCs) was determined in the E face of the plasma membrane of head regions.


Reproduction ◽  
2000 ◽  
pp. 325-335 ◽  
Author(s):  
A Calvo ◽  
LM Pastor ◽  
S Bonet ◽  
E Pinart ◽  
M Ventura

Lectin histochemistry was used to perform in situ characterization of the glycoconjugates present in boar testis and epididymis. Thirteen horseradish peroxidase- or digoxigenin-labelled lectins were used in samples obtained from healthy fertile boars. The acrosomes of the spermatids were stained intensely by lectins with affinity for galactose and N-acetyl-galactosamine residues, these being soybean, peanut and Ricinus communis agglutinins. Sertoli cells were stained selectively by Maackia ammurensis agglutinin. The lamina propria of seminiferous tubules showed the most intense staining with fucose-binding lectins. The Golgi area and the apical part of the principal cells of the epididymis were stained intensely with many lectins and their distribution was similar in the three zones of the epididymis. On the basis of lectin affinity, both testis and epididymis appear to have N- and O-linked glycoconjugates. Spermatozoa from different epididymal regions showed different expression of terminal galactose and N-acetyl-galactosamine. Sialic acid (specifically alpha2,3 neuraminic-5 acid) was probably incorporated into spermatozoa along the extratesticular ducts. These findings indicate that the development and maturation of boar spermatozoa are accompanied by changes in glycoconjugates. As some lectins stain cellular or extracellular compartments specifically, these lectins could be useful markers in histopathological evaluation of diseases of boar testis and epididymis.


Reproduction ◽  
2000 ◽  
pp. 145-152 ◽  
Author(s):  
B Pintado ◽  
J de la Fuente ◽  
ER Roldan

This study was designed to assess whether nucleic acid stains such as propidium iodide and Hoechst 33258 and the cytosolic stain eosin identified equivalent proportions of non-viable cells. Sub-samples of boar spermatozoa stored for up to 72 h, and frozen bull spermatozoa stored in straws and thawed before staining, were exposed to either propidium iodide or Hoechst 33258 alone or in combination. Additional sub-samples were stained with eosin-nigrosin and subsequently with Giemsa. The proportion of non-viable cells identified by propidium iodide alone was equivalent to that observed when it was used in combination with the other fluorescent probe. Similar results were observed for Hoechst 33258. However, direct microscopic examination of sub-samples exposed to both stains revealed that a proportion of spermatozoa stained with propidium iodide did not incorporate Hoechst 33258. This was found consistently in boar and bull spermatozoa under the different experimental conditions used. Quantification showed that the proportion of propidium iodide-positive cells was significantly higher than Hoechst 33258-positive cells. Furthermore, the proportion of propidium iodide-positive cells was higher than cells stained with eosin, but no differences were found between the number of cells stained with Hoechst 33258 or eosin. The proportion of cells stained with propidium iodide was positively correlated with the proportion stained with either Hoechst 33258 or eosin, despite the observation that more cells incorporated propidium iodide. Taken together, these results indicate that there are differences in the ability of fluorescent probes to identify non-viable sperm cells and that this should be considered when staining protocols are used to analyse sperm viability, or when viability is used as a discriminating factor in functional studies, such as those related to acrosomal exocytosis.


1968 ◽  
Vol 27 (4) ◽  
pp. 1032-1036 ◽  
Author(s):  
N. L. First ◽  
R. E. Short ◽  
J. B. Peters ◽  
F. W. Stratman
Keyword(s):  

1966 ◽  
Vol 25 (2) ◽  
pp. 406-409 ◽  
Author(s):  
G. A. Schul ◽  
C. W. Foley ◽  
C. D. Heinze ◽  
R. E. Erb ◽  
R. B. Harrington

Author(s):  
Svitlana Lobchenko ◽  
Tetiana Husar ◽  
Viktor Lobchenko

The results of studies of the viability of spermatozoa with different incubation time at different concentrations and using different diluents are highlighted in the article. (Un) concentrated spermatozoa were diluented: 1) with their native plasma; 2) medium 199; 3) a mixture of equal volumes of plasma and medium 199. The experiment was designed to generate experimental samples with spermatozoa concentrations prepared according to the method, namely: 0.2; 0.1; 0.05; 0.025 billion / ml. The sperm was evaluated after 2, 4, 6 and 8 hours. The perspective of such a study is significant and makes it possible to research various aspects of the subject in a wide range. In this regard, a series of experiments were conducted in this area. The data obtained are statistically processed and allow us to highlight the results that relate to each stage of the study. In particular, in this article it was found out some regularities between the viability of sperm, the type of diluent and the rate of rarefaction, as evidenced by the data presented in the tables. As a result of sperm incubation, the viability of spermatozoa remains at least the highest trend when sperm are diluted to a concentration of 0.1 billion / ml, regardless of the type of diluent used. To maintain the viability of sperm using this concentration of medium 199 is not better than its native plasma, and its mixture with an equal volume of plasma through any length of time incubation of such sperm. Most often it is at this concentration of sperm that their viability is characterized by the lowest coefficient of variation, regardless of the type of diluent used, which may indicate the greatest stability of the result under these conditions. The viability of spermatozoa with a concentration of 0.1 billion / ml is statistically significantly reduced only after 6 or even 8 hours of incubation. If the sperm are incubated for only 2 hours, regardless of the type of diluent used, the sperm concentrations tested do not affect the viability of the sperm. Key words: boar, spermatozoa, sperm plasma, concentration, incubation, medium 199, activity, viability, rarefaction.


2021 ◽  
Author(s):  
Helen Jäkel ◽  
Heiko Henning ◽  
Anne‐Marie Luther ◽  
Karl Rohn ◽  
Dagmar Waberski

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
M. Schulze ◽  
F. Schröter ◽  
M. Jung ◽  
U. Jakop

AbstractThe increase of fertility performance in sows is one of the biggest achievements in pig production over the last 30 years. Nevertheless, pig farms using artificial insemination (AI) repeatedly experienced in recent year’s fertility problems with dramatic consequences due to toxic compounds from plastic semen bags. In particular, bisphenol A diglycidyl-ether (BADGE) present in multilayer plastic bags can leach into the semen and could affect the functionality of the spermatozoa. Former studies could not find any alterations in spermatozoa based on the exposure to BADGE. The aim of the study was to evaluate effects of BADGE on boar spermatozoa using an extended panel of spermatological methods. In spring 2019, a large drop in farrowing rates from 92.6 ± 2.3% to 63.7 ± 11.1% in four sow farms in Croatia was detected. In migration studies, BADGE could be identified as a causal toxic compound and leached into the extended semen in concentration of 0.37 ± 0.05 mg/L. Detailed spermatological studies showed that significant predictors for effects on spermatozoa were different levels of motility and kinematic data after a prolonged storage time, thermo-resistance test (prolonged incubation time), mitochondrial activity, membrane integrity and fluidity. No serious effects were observed for sperm morphology and DNA fragmentation. These results provide new insights into the development of a new quality assurance concept for a detailed spermatological examination during testing of plastic materials for boar semen preservation. It could be shown that boar spermatozoa are an excellent biosensor to detect potential toxicity and fertility-relevant compounds.


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