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Number of results: 30
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Abstract

Electrophoretic methods were used to identify protein complexes formed between ostrich egg yolk lipoprotein fractions (LPFo) with seminal plasma (SP) of fractionated ejaculates, and to investigate the effect of these complexes on boar semen quality after cryopreservation. Chromatographic SP fractions (F1, F2 and F3), with or without LPFo solution, were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Comparative electrophoretic analyses of the SP revealed marked differences in the SDS-PAGE protein profiles among boars. Electrophoretic analyses showed that the interactions of LPFo with SP resulted in the appearance of high-intensity protein bands. Spermatozoa were exposed to SP chromatographic fractions originating from F1, F2 and F3, and the whole SP (wSP) before being frozen. Spermatozoa exposed to F1 and F2 exhibited significantly higher post-thaw motility compared to those treated with either F3 or wSP. In most of the boars the proportions of membrane-intact frozen-thawed spermatozoa differed among the treatments, being significantly lower in the wSP-treated samples. The incidence of frozen-thawed spermatozoa with DNA fragmentation was less prevalent in samples exposed to F3 or the wSP. The results of this study confirmed that the interactions of LPFo with fractionated SP during the cooling period contributed to alterations in the sperm membranes, rendering them less susceptible to temperature-related injury.

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Authors and Affiliations

L. Fraser
Ł. Zasiadczyk
J. Strzeżek
W. Kordan
A. Mańkowska
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Abstract

This is the first study to identify 23 protein spots corresponding to 13 proteins in mitochondria isolated from rabbit epididymal spermatozoa. In the group of protein spots identified in stress-induced samples, the abundance of 20 protein spots increased, whereas the abundance of three protein spots (GSTM3, CUNH9orf172, ODF1) decreased relative to the control. The results of this study provide valuable inputs for future research into the molecular mechanisms implicated in pathological processes during oxidative stress (OS).
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Authors and Affiliations

M. Lecewicz
1
W. Kordan
1
K. Górski
2
S. Kondracki
2

  1. Department of Animal Biochemistry and Biotechnology, University of Warmia and Mazury in Olsztyn, Oczapowskiego 5, 10-719 Olsztyn, Poland
  2. Institute of Animal Production and Fisheries, Siedlce University of Natural Sciences and Humanities, Prusa 14, 08-110 Siedlce, Poland
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Abstract

The number of spermatozoa in the ejaculate is important for its quality and that of the sperm contained in it. The number of ejaculated spermatozoa is also associated with sperm dimensions. The aim of this study was to assess the morphological structure of sperm and the frequency of morphological abnormalities in sperm on the ejaculation performance of boars, measured as the total number sperm per ejaculate. The study was conducted using 648 ejaculates collected from 31 Large White boars and 30 Landrace boars. All ejaculates were analysed for basic physical characteristics and the frequency of sperm with morphological abnormalities. In addition, morphometric measurements of the sperm were made and used to calculate their shape indexes. As a result of our study it was noted that sperm from ejaculates with the most spermatozoa have shorter heads with a smaller area than sperm from ejaculates with a small or intermediate number of spermatozoa. Landrace boars produce semen of better quality, with a smaller percentage of sperm with major abnormalities, and the differences between the breeds increase with the number of spermatozoa in the ejaculate. The sperm from Landrace boars have larger heads and longer flagella than the sperm from Large White boars. The differences in sperm dimensions between breeds decrease as the total number of spermatozoa in the ejaculate increases. The number of spermatozoa in the ejaculate was shown to influence the dimensions of the sperm. The effect of the number of ejaculated sperm on ejaculate characteristics and sperm morphology depends on the breed of the male.
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Bibliography

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Authors and Affiliations

S. Kondracki
1
K. Górski
1
M. Iwanina
1
W. Kordan
2
M. Lecewicz
2

  1. University of Siedlce, Faculty of Agricultural Sciences, Prusa 14, 08-110 Siedlce, Poland
  2. University of Warmia and Mazury in Olsztyn, Department of Animal Biochemistry and Biotechnology, Oczapowskiego 13, 10-719 Olsztyn, Poland
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Abstract

In our previous Genome-wise Association Study we found that Cystic Fibrosis Transmem- brane Conductance Regulator gene (CFTR) is a candidate gene for sperm motility in fresh semen of Holstein-Friesian bulls. Since in cows thawed semen is commonly used for the artificial insem- ination (AI) we have decided to find out whether functional polymorphism within CFTR gene coding sequence is associated with selected parameters of thawed sperm, including their motility evaluated by computer-assisted sperm analysis (CASA), the activity of three antioxidant enzymes: glutathione peroxidase (GPx) catalase (CAT), superoxide dismutase (SOD), ATP con- tent and integrity of sperm membranes. One hundred twenty Holstein Friesian bulls kept in uni- form environmental conditions (one AI company) were included in the study. Significant associ- ations between genotypes of missense mutation within exon 11 of the CFTR gene (Met468Leu) and the activity of antioxidant enzymes and sperm mitochondrial function were revealed. No effect of CFTR genotypes on sperm motility was observed. Significant differences in CAT and SOD activity were found between AA and TT homozygous individuals. Bulls with TT genotype had the lowest activity of both antioxidant enzymes. The same bulls also showed the lowest num- ber of sperm with active mitochondria. Our results demonstrate that missense mutation Met468Leu within CFTR gene is associated with antioxidant enzyme activity and mitochondrial function of bovine thawed sperm without affecting their motility.

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Authors and Affiliations

S. Kaminski
D.M. Hering
W. Kordan
M. Lecewicz
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Abstract

The aim of this study was to investigate the effect of different extenders on the post-thaw (PT) quality of sperm originating from the sperm-rich fraction (SRF) and post-sperm-rich fraction (PSRF) of boar ejaculate. Motility and velocity parameters, analyzed using a computer-assisted semen analysis (CASA) system, and membrane integrity parameters were markedly higher in frozen-thawed (FT) spermatozoa of the SRF in both the Belstville Thawing Solution (BTS) and Androhep Plus (AHP) extenders, irrespective of the post-thaw (PT) storage time. Furthermore, reduced cryo-survival was more marked in FT spermatozoa of the PSRF in both extenders following storage for 60 min. It was found that the SRF-stored samples in the AHP extender for 60 min exhibited significantly higher percentages of spermatozoa with total motility, mitochondrial function and acrosome integrity than those stored in the BTS extender. The findings of this study confirm that components of the ejaculate fractions and extender have varying effects on the cryo-survival of boar spermatozoa.
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Bibliography

1. Aquila S, Giordano F, Guido C, Rago V, Carpino A (2011) Nitric oxide involvement in the acrosome reaction triggered by leptin in pig sperm. Reprod Biol Endocrinol 9: 133.
2. Fraser L, Strzeżek J (2007) Effect of different procedures of ejaculate collection, extenders and packages on DNA integrity of boar sper-matozoa following freezing-thawing. Anim Reprod Sci 99: 317-329.
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6. Saravia F, Wallgren M, Johannisson A, Calvete JJ, Sanz L, Pena FJ, Roca J, Rodríguez-Martínez H (2009) Exposure to the seminal plasma of different portions of the boar ejaculate modulates the survival of spermatozoa cryopreserved in MiniFlatPacks. Theriogenology 71: 662-675.
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9. Wasilewska-Sakowska K, Zasiadczyk Ł, Fraser L (2019) Effect of fractionated seminal plasma on sperm characteristics following cryo-preservation of boar semen. Ann Anim Sci 19: 695-712.
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11. Yeste M (2016) Sperm cryopreservation update: cryodamage, markers, and factors affecting the sperm freezability in pigs. Theriogenol-ogy 85: 47-64.
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Authors and Affiliations

Ł. Zasiadczyk
1
K. Kurpanik
1
L. Fraser
1
W. Kordan
1

  1. Department of Animal Biochemistry and Biotechnology, Faculty of Animal Bioengineering, University of Warmia and Mazury in Olsztyn, Oczapowskiego 5, 10-719 Olsztyn, Poland
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Abstract

During the rutting season, stag semen is accompanied by a sticky, dense secretion called yellow fraction (YF). There is little information about the role, biology, physiology, and most importantly, the composition of this fluid. The aim of this study was to isolate and identify zinc ions (ZnBPs) and heparin binding proteins (HBPs) from YF of the red deer ( Cervus elaphus L.). Using liquid chromatography, the presence of 6 fractions of ZnBPs (71, 65, 55, 16, 14 and 12 kDa) and 22 fractions of HBPs (163, 140, 96, 78, 71, 65, 55, 49, 33, 31, 26, 25, 24, 22, 18, 16, 13, 12, 11, 10, 9 and 8 kDa) in YF proteome was demonstrated. By means of two-dimensional electrophoreses and MALDI-TOF/TOF mass spectrometry some of them were then identified. Amongst ZnBPs the following were identified: glutaminyl-peptide cyclotransferase, inhibitor of carbonic anhydrase-like, potassium voltage-gated channel subfamily E member 2, WD repeat-containing protein 38 isoform X4. Amongst the HBPs metalloproteinase inhibitor 2 (TIMP2), seminal plasma glycoprotein PSP-I and adseverin (scinderin) were identified. Identifying all ZnBPs and HBPs present in YF may broaden up-to-date knowledge concerning the biology, physiology and preservation of red deer semen.
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Authors and Affiliations

A. Orzołek
1
K. Dyrda
1
K. Rafalska
1
P. Wysocki
1
W. Kordan
1
W. Giżejewski
2

  1. Department of Animal Biochemistry and Biotechnology, University of Warmia and Mazury, Oczapowskiego 5, 10-719 Olsztyn, Poland
  2. Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland
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Abstract

Mature males of a wild boar-pig crossbreed, during the long and short day season, were used for the study which demonstrates that the chemical light carrier CO regulates the expression of biological clock genes in the hypothalamus via humoral pathways. Autologous blood with experimentally elevated concentrations of endogenous CO (using lamps with white light-emitting diodes) was infused into the ophthalmic venous sinus via the right dorsal nasal vein. Molecular biology methods: qPCR and Western Blot were used to determine the expression of genes and biological clock proteins. The results showed that elevated endogenous CO levels, through blood irradiation, induces changes in genes expression involved in the functioning of the main biological clock located in suprachiasmatic nuclei. Changes in the expression of the transcription factors Bmal1, Clock and Npas2 have a similar pattern in both structures, where a very large decrease in gene expression was shown after exposure to elevated endogenous CO levels. The changes in the gene expression of PER 1-2, CRY 1-2, and REV-ERB α-β and ROR β are not the same for both POA and DH hypothalamic structures, indicating that both structures respond differently to the humoral signal received.
The results indicate that CO is a chemical light molecule whose production in an organism depends on the amount of light. An adequate amount of light is an essential factor for the proper functioning of the main biological clock.
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Authors and Affiliations

P. Gilun
1
M. Koziorowska-Gilun
2
B. Wąsowska
1
M. Sowa-Kućma
3
K. Kozioł
4
M. Romerowicz-Misielak
4
W. Kordan
2
M. Koziorowski
4

  1. Department of Local Physiological Regulations, Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences in Olsztyn, Tuwima 10, 10-748, Olsztyn, Poland
  2. Department of Animal Biochemistry and Biotechnology, Faculty of Animal Bioengineering, University of Warmia and Mazury, Oczapowskiego 5, 10-719 Olsztyn, Poland
  3. Department of Human Physiology, Institute of Medical Sciences, Medical College of Rzeszow University, Kopisto 2a, Rzeszow, 35-959, Poland
  4. Department of Animal Physiology and Reproduction, Institute of Biology and Biotechnology, Collegium Scientarium Naturalium, University of Rzeszow, Werynia 502, 36-100 Kolbuszowa, Poland
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Abstract

The aim of this study was to identify the proteoforms of albumin and kallikrein in stallion seminal plasma (SP), and to determine their correlations with sperm motility parameters. The experimental material consisted of ejaculates from 8 stallions, which were collected during the breeding and non-breeding seasons (BS and NBS, respectively). SP proteins were identified by 2-D PAGE and mass spectrometry (MALDI TOT/TOF MS). Sperm motility parameters were analyzed using the CASA system. Protein expression (integrated optical density-IOD) of albumin proteoforms 1 (ALB 1) and 2 (ALB 2) and kallikrein proteoforms 1 (KAL 1) and 2 (KAL 2) was correlated (p<0.05) with sperm motility parameters (total motility and progressive motility) during the BS. No significant correlations were found between the expression of albumin or kallikrein and sperm motility parameters during the NBS. The presence of correlations between the expression of ALB 1, ALB 2, KAL 1, KAL 2 and selected sperm motility parameters could suggest that the analyzed components of the SP belong to the group of fertility-associated pro- teins (FAPs).

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Authors and Affiliations

M. Mogielnicka-Brzozowska
L. Fraser
A. Dziekońska
K. Gackowska
M. Sobiewska
A. Kuzborska
A.M. Majewska
K. Filipowicz
W. Kordan
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Abstract

The aim of this study was to measure the NO level in boar semen held in a liquid state and to determine its putative relation to spermatozoa motility, plasma membrane integrity, mitochondrial membrane potential and ATP content. Generally, the percentage of spermatozoa which generated nitric oxide gradually increased, while NO level in the surrounding medium declined during the liquid preservation. NO generation in semen preserved in BTS was higher as compared to those in Androhep®Plus. We demonstrated the positive correlation between the NO level in fresh spermatozoa and their quality. We also showed negative correlation between nitric oxide level in spermatozoa preserved in BTS and sperm cells motility as well as plasma membrane integrity. Results obtained in this study confirm that NO may affect sperm physiology in a dualistic manner.
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Authors and Affiliations

A. Orzołek
Ł. Zasiadczyk
P. Wysocki
W. Kordan
P. Krysztofiak

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