Plasma membrane changes during the liquid storage of boar spermatozoa: A comparison of methods

2010 ◽  
Vol 58 (1) ◽  
pp. 105-116 ◽  
Author(s):  
Dariusz Gączarzewicz ◽  
Małgorzata Piasecka ◽  
Jan Udała ◽  
Barbara Błaszczyk ◽  
Tomasz Stankiewicz ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Semen Analysis ◽  
Membrane Integrity ◽  
Live Cells ◽  
Lower Percentage ◽  
Loss Of Function ◽  
Plasma Membrane Integrity ◽  
Sperm Plasma Membrane ◽  
Semen Preservation

Studies were performed on boar semen routinely used at the local artificial insemination (AI) centre. The semen was stored in a Safe Cell Plus commercial extender at 17 °C for nine days. The aim of our research was focused on changes in sperm plasma membrane integrity. The integrity of the sperm plasma membrane and acrosome as well as sperm motility decreased after dilution and during storage of the semen. The highest percentage of live sperm was identified by the eosin-nigrosin method, a lower percentage by the SYBR-14/PI test, and the lowest percentage of live cells was discovered by the hypoosmotic swelling (HOS) test (P < 0.01). There were significant differences between the results of staining methods and sperm motility (P < 0.01). No significant differences were found between the HOS test results and sperm motility. The plasma membrane integrity parameters positively correlated (P < 0.001) with each other and with sperm motility but negatively with aspartate aminotransferase activity. Our findings confirmed that the boar sperm aging changes, which increased during liquid semen preservation, were connected with the loss of function and integrity of the sperm plasma membrane. The employed complementary tests are comprehensive indicators of sperm membrane integrity during long-term semen preservation, and they can help establish the actual number of ‘healthy’ cells. The assays may be used in AI laboratories and should be incorporated into the routine of semen analysis.

Extender supplementation with catalase maintains the integrity of sperm plasma membrane after freezing–thawing of semen from capuchin monkey

Zygote ◽  
2017 ◽  
Vol 25 (2) ◽  
pp. 231-234 ◽  
Author(s):  
Danuza L. Leão ◽  
Adriel B. Brito ◽  
Stefânia A. Miranda ◽  
Karol G. Oliveira ◽  
Débora V.C. Almeida ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Capuchin Monkey ◽  
Plasma Membrane Integrity ◽  
Sapajus Apella ◽  
Sperm Plasma Membrane ◽  
Cryopreserved Sperm

SummaryWe aimed to evaluate the effect of supplementation of ACP-118® extender with the antioxidant catalase (10 and 50 µg/ml) on Sapajus apella sperm motility, vigour, and plasma membrane integrity during the processes of seminal liquefaction, cooling, and freezing. Catalase did not affect any of the evaluated parameters after semen dilution or cooling. Cryopreserved sperm in the presence of 50 µg/ml catalase presented a plasma membrane integrity similar to that fresh sperm, however.

scholarly journals Sheng Jing Decoction Can Promote Spermatogenesis and Increase Sperm Motility of the Oligozoospermia Mouse Model

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Guang Yan ◽  
Fang Tian ◽  
Peng Liu ◽  
Jianming Sun ◽  
Jianmin Mao ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Sperm Morphology ◽  
Sperm Concentration ◽  
Membrane Integrity ◽  
Testicular Tissue ◽  
Whole Body ◽  
Plasma Membrane Integrity ◽  
Normal Sperm ◽  
Sperm Plasma Membrane

Sheng Jing Decoction (SJD), as a traditional Chinese medicine prescription, is mainly be used to treat male infertility. However, the pharmacological functions and molecular mechanisms of SJD are poorly understood. In this study, we investigated the functions of SJD on spermatogenesis and sperm motility and explored the potential mechanisms involved. Here, we demonstrated that high, medium, and low doses of SJD are effective in restoring the impairments of the whole body and testicular tissue by cyclophosphamide inducing and to rescue the damage of testicular tissue cells including Sertoli cells and germ cells. SJD can partly restore the decrease in sperm concentration, sperm vitality, sperm motility, and normal sperm morphology rate in oligozoospermic mouse models. Ki67 staining analyses confirm SJD can promote testicular tissue cell proliferation. Real-time RT-PCR analyses also reveal that SJD can upregulate the expression of proliferation-associated gene Lin28a and differentiation-associated genes Kit, Sohlh2, and Stra8. SJD can also reduce the impairment of mitochondrial membrane potential (MMP) and sperm plasma membrane integrity by cyclophosphamide inducing. Our results reveal that SJD is effective in improving both sperm quantity and quality by increasing the sperm concentration, sperm vitality, sperm motility, and normal sperm morphology rate. SJD can promote spermatogenesis by upregulating the expression of the proliferation-associated gene Lin28a and the differentiation-associated genes (Kit, Sohlh2, and Stra8). SJD can sustain MMP and sperm plasma membrane integrity to increase sperm motility.

118 Semen quality and fertilization ability of myostatin-knockout boars

2020 ◽  
Vol 32 (2) ◽  
pp. 186
Author(s):  
M.-F. Xuan ◽  
S.-Z. Han ◽  
B.-H. Quan ◽  
X.-J. Yin ◽  
J.-D. Kang
Keyword(s):  
Plasma Membrane ◽  
Semen Quality ◽  
Semen Analysis ◽  
Membrane Integrity ◽  
Mitochondrial Activity ◽  
Human Consumption ◽  
Computer Assisted ◽  
Plasma Membrane Integrity ◽  
Sperm Plasma Membrane ◽  
Student’S T

Myostatin-knockout (MSTN−/−) pigs may provide a source of healthy lean protein for human consumption. However, little is known about the effect of this knockout on semen quality, which is important if these pigs are used for production. The purpose of this study is to evaluate the semen quality and fertility of MSTN−/− boars. We generated MSTN−/− boars from Duroc-Landrace-Yorkshire hybrid pig cell lines by somatic cell nuclear transfer, and all 12 boars showed sexual maturation with an obvious “double muscling” phenotype. Semen was collected randomly from three MSTN−/− boars using the gloved-hand technique by one technician and then tested by computer-assisted semen analysis. Semen acrosomal integrity and deformity were measured using Coomassie blue- and eosin-stained smears, respectively. Sperm plasma membrane integrity and mitochondrial activity were evaluated by Hoechst 33342, propidium iodide, and JC-1 multiple staining. The reproductive performance of MSTN−/− boars was evaluated by IVF and by AI. All data were analysed by Student's t-tests. The results showed that the semen color, odor, and pH had no abnormalities. The concentration, motility, plasma membrane integrity, deformity, acrosome integrity, and mitochondrial activity of the semen presented no significant differences from those of the control semen (Duroc). The ejaculation volume of the MSTN−/− boars was significantly lower than that of the control (168.78±6.70 and 223.11±21.21mL, respectively), although the total sperm number was not significantly different. The rate of cleavage and blastocyst formation (247 to 254 oocytes per boar) was not significantly different from those of the control (69.1±0.7 vs. 65.2±1.6%, and 20.2±1.2 vs. 22.8±1.4%, respectively). Seventeen healthy offspring were successfully produced from three sows through AI using semen from one MSTN−/− boar. However, the genotype of piglets has not been tested at present. Thus, MSTN−/− boar may be used as sires, and these pigs are expected to be developed to provide new super-lean meat varieties in the future.

scholarly journals Sheng Jing Decoction, as a Traditional Chinese Medicine Prescription, Can Promote Spermatogenesis and Increase Sperm Motility

2021 ◽  
Author(s):  
Guang Yan ◽  
Fang Tian ◽  
Peng Liu ◽  
Jianmin Mao ◽  
Jianming Sun ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Sperm Morphology ◽  
Sperm Concentration ◽  
Membrane Integrity ◽  
Testicular Tissue ◽  
Plasma Membrane Integrity ◽  
Medicine Prescription ◽  
Sperm Plasma Membrane ◽  
Chinese Medicine Prescription

Abstract BackgroundSheng Jing Decoction (SJD), as a traditional Chinese medicine prescription, is mainly be used to treat male infertility. It has been applied in many hospitals in China, and the clinical effect of patients' reaction is satisfactory. However, pharmacological function and molecular mechanism of SJD are poorly understood. In this study, mainly investigated the function of SJD on spermatogenesis and sperm motility, and explored the potential mechanisms.MethodsThe oligozoospermia model of ICR mice was induced by injecting intraperitoneally (ip) with 60 mg/kg dose of cyclophosphamide. At the same time of modeling, high, medium and low doses of SJD were given orally for treatment respectively. Sperm vitality and motility were detected and analyzed by CASA, and sperm morphology was tested by Papanicolaou staining. Sperm mitochondrial membrane potential (MMP) was tested by JC-1 staining. Sperm plasma membrane integrity was determined by SYBR-14/PI double staining. Histopathological changes of testis were analyzed by HE staining and Immunohistochemistry. Genes expression related to spermatogenesis and sperm development were detected by real-time RT-PCR.Results High, medium and low doses of SJD are effective to recover from the impairment of the whole body and testicular tissue by cyclophosphamide inducing, and to rescue the damage of testicular tissue cells including sertoli cells and germ cells by cyclophosphamide inducing. SJD can all partly restore the decrease in sperm concentration, sperm vitality, sperm motility and normal sperm morphology rate by cyclophosphamide inducing. Ki67 staining analyses confirm SJD can promoted testicular tissue cells proliferation. Real-time RT-PCR analyses reveal SJD can up-regulates the expression of proliferation-associated gene Lin28a, and differentiation-associated genes Kit, Sohlh2 and Stra8. SJD can also reduce the impairment of MMP and sperm plasma membrane integrity by cyclophosphamide inducing.ConclusionSJD is effective to support sperm quantity and quality by increasing sperm concentration, sperm vitality, sperm motility and normal sperm morphology rate. SJD can promote spermatogenesis by up-regulating the expression of the proliferation-associated gene Lin28a, and the differentiation-associated genes (Kit, Sohlh2 and Stra8). SJD can sustain MMP and sperm plasma membrane integrity to increase sperm motility.

87 OSMOTIC SENSITIVITY OF OKAPI SPERMATOZOA AND DEVELOPMENT OF CRYOPRESERVATION PROTOCOLS USING CRYOMICROSCOPY

2008 ◽  
Vol 20 (1) ◽  
pp. 124 ◽  
Author(s):  
L. M. Penfold
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Repeated Measures ◽  
Membrane Integrity ◽  
Latin Square ◽  
Crystal Formation ◽  
Plasma Membrane Integrity ◽  
Power Of The Test ◽  
Before And After ◽  
Cooling Effects

Okapi would benefit from artificial insemination with frozen-thawed sperm in cases where aggression prevents mating or where individuals are geographically disparate. Effective sperm cryopreservation is a prerequisite to this goal. Ejaculates (n = 20) were collected from 7 anesthetized adult male okapi housed individually, or with a female for breeding, throughout the year by electroejaculation, and semen and sperm parameters were assessed. Semen aliquots were centrifuged; resuspended in 500 µL of PBS with the osmolarity adjusted to 35, 75, 150, 600, 1200, and 2400 mOsm; and incubated for 30 min before returning to isosmotic conditions. Semen was extended in TEST containing 1%, 2%, or 4% glycerol with or without 0.5% Equex (Minitube, Verona, WI, USA); 5-µL aliquots were cooled in a Latin square design on a Linkam BCS 196 cryomicrostage (Linkam Scientific, Tadworth, Surrey, UK) at 20�C min–1 to –6� –12�C at which point ice crystal formation was induced (seeded), and cooled further to –70�C before warming at 50�C min–1 to 35�C (okapi body temperature). To investigate cooling effects only, raw ejaculate was cooled to –6�C without seeding and warmed to 35�C. Percent sperm motility and plasma membrane integrity (PMI) were recorded before and after treatments. Differences were examined using one-way repeated measures analysis of variance. No differences in motility, total sperm numbers, or percent normal morphology were observed throughout the year (P > 0.05), although the power of the test was low so that negative findings should be interpreted cautiously. Mean semen volume was 1.3 � 0.19 mL, sperm motility was 29 � 3.2%, with a PMI of 39 � 6.8%; 48 � 2.8% were morphologically normal. High proportions of non-motile, plasma membrane-damaged cells were noted in every ejaculate, and whiplash motility, possibly indicating spontaneous capacitation, was observed in several ejaculates 1 h after collection. Motility was dramatically reduced on either side of isosmotic conditions and was more sensitive to osmotic pressure than was plasma membrane integrity. Cooling of raw ejaculate to sub-zero temperatures without freezing did not result in any loss of motility or PMI, indicating cold tolerence. Superior results were obtained when sperm were frozen-thawed in TEST containing 4% glycerol with 0.5% Equex. Findings indicate that okapi semen collected by electroejaculation routinely contain high numbers of non-motile and plasma membrane-damaged spermatozoa, apparently unrelated to season or the length of time since the male was housed with a breeding female. Okapi spermatozoa are remarkably intolerant of departures from isosmotic conditions, indicating a lack of ability to regulate or withstand volume excursions during osmotic stress events; however, cooling to sub-zero temperatures in the absence of cryoprotectant did not reduce percent sperm motility or PMI, indicating resistance to cold shock. Increasing and maintaining proportions of motile, membrane-intact spermatozoa prior to and during cryopreservation will be critical for development of freezing protocols for this species.

The effects of chronic administration of ghrelin on rat sperm quality and membrane integrity

2009 ◽  
Vol 59 (2) ◽  
pp. 159-168 ◽  
Author(s):  
Arash Kheradmand ◽  
Majid Taati ◽  
Homayoon Babaei
Keyword(s):  
Plasma Membrane ◽  
Treatment Group ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Membrane Integrity ◽  
Chronic Administration ◽  
Control Group ◽  
Plasma Membrane Integrity ◽  
Significant Difference

AbstractAlthough ghrelin acts as a modulator of feeding behavior and energy metabolism in the central nervous system, recent studies have implicated the peripheral actions of ghrelin in reproductive tissues. Here, we investigated the effects of chronic administration of ghrelin on the motility, plasma membrane integrity and concentration of rat spermatozoa. 45-d male Wistar rats were scheduled for the study and were divided into control and treatment groups. In the treatment group, 1 nmol of ghrelin was administered as sc injection for 10 consecutive days or vehicle (physiological saline) to the control rats. Sperm collection was achieved by killing of the rats on days 15, 25 and 50 after first injection. Total sperm motility and forward progressive movement did not exhibit significant difference during the experiment, although, there was a tendency for greater motion rate on d 15 and 25 in the treated rats compared to the control group. Plasma membrane integrity (HOS-reacted spermatozoa) was significantly higher in the treated animals, especially on day 15 as well as day 25, because of possible antioxidant properties of ghrelin. This value was statistically higher on day 15 than that of day 25 (P <0.05). Likewise, there was a significant correlation between the FPM (P <0.0001, r = 0.79) and TSM (P <0.01, r = 0.52) with the HOS test percentage in the treatment group. It was not observed statistically difference in the sperm concentration between groups during all of the experimental days. In conclusion, chronic administration of ghrelin (similar to induced by energy deficiency such as fasting) increased the integrity of sperm membrane, however, the sperm motility and concentration did not display any alterations.

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