scholarly journals Butaphosphan and Cyanocobalamin Supplementation in Semen Extender on Chilled Boar Sperm Quality and Life Span

2020 ◽  
Vol 7 ◽  
Author(s):  
J. Suwimonteerabutr ◽  
S. Chumsri ◽  
P. Tummaruk ◽  
Morakot Nuntapaitoon
Keyword(s):  
Plasma Membrane ◽  
Life Span ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Control Group ◽  
Plasma Membrane Integrity ◽  
Progressive Motility ◽  
Boar Sperm ◽  
Acrosome Integrity ◽  
Semen Extender

The objective of the present study was to determine the effect of butaphosphan and cyanocobalamin supplementation in semen extender on chilled boar sperm quality and life span. A total of 35 ejaculates of boar semen were included. The semen was diluted with Beltsville thawing solution extender supplemented with different concentrations of butaphosphan and cyanocobalamin [0 (control), 0.1, 0.2, 0.3, 0.4, and 0.5%] in the diluted semen. The semen samples were evaluated using a computer-assisted sperm analysis system to determine sperm motility and sperm kinetic parameters (i.e., the curvilinear velocity, VCL; straight line velocity, VSL; average path velocity, VAP; linearity, LIN; straightness, STR; amplitude of lateral head, ALH; wobble, WOB; and beat cross frequency, BCF). Additionally, sperm viability, acrosome integrity, mitochondrial activity, and plasma membrane integrity were evaluated after 4 (day 0), 72 (day 3), 120 (day 5), and 168 (day 7) h of storage using SYBR-14–ethidium homodimer-1 (EthD-1), EthD-1, JC-1, and the short hypo-osmotic swelling test, respectively. The analyses were carried out by using the general linear mixed model (MIXED) procedure of SAS. The statistical models for each data set included group, day after storage, and interaction between group and day after storage. The boar was included as a random effect. On day 0 after storage, progressive motility, VCL, VSL, VAP, and plasma membrane integrity of boar sperm in 0.3% of butaphosphan and cyanocobalamin supplementation were greater than those in the 0.4 and 0.5% groups (P < 0.05). On day 3 after storage, total motility and progressive motility, VCL, VSL, VAP, LIN, WOB, BCF, and plasma membrane integrity in 0.3% of butaphosphan and cyanocobalamin supplementation were significantly greater than those in the control group (P < 0.05). The total motility and progressive motility, VAP, and WOB in 0.3% of butaphosphan and cyanocobalamin supplementation were greater than those in the control group on day 5 after storage (P < 0.05). No effects of butaphosphan and cyanocobalamin supplementation on acrosome integrity and mitochondria activity were found on days 3, 5, and 7 after storage. However, the motility and progressive motility and the values for all sperm kinetic parameters except ALH in 0.3% of butaphosphan and cyanocobalamin supplementation were greater than those in the control group on day 7 after storage (P < 0.05). In conclusion, 0.3% of butaphosphan and cyanocobalamin supplementation in semen extender improved sperm motility, sperm activity, morphology, and life span in chilled boar sperm.

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.

Effect of platelet activating factor (PAF) supplementation in semen extender on viability and ATP content of cryopreserved canine spermatozoa

2010 ◽  
Vol 13 (4) ◽  
pp. 571-579 ◽  
Author(s):  
W. Kordan ◽  
M. Lecewicz ◽  
R. Strzeżek ◽  
A. Dziekońska ◽  
L. Fraser
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Mitochondrial Function ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Platelet Activating Factor ◽  
Computer Assisted ◽  
Atp Content ◽  
Plasma Membrane Integrity ◽  
Semen Extender

Effect of platelet activating factor (PAF) supplementation in semen extender on viability and ATP content of cryopreserved canine spermatozoa The aim of this study was to investigate the effect of platelet activating factor (PAF) on the quality characteristics of cryopreserved canine spermatozoa. Cryopreserved semen of 5 mixed-breed dogs was treated with different concentrations of exogenous PAF (1 × 10-3M, 1 × 10-4M, 1 × 10-5M and 1 × 10-6M) and examined at different time intervals (0, 30, 60 and 120 min). Cryopreserved semen treated without PAF was used as the control. Sperm quality was evaluated for motility (computer-assisted semen analysis, CASA), mitochondrial function (JC-1/PI assay) and plasma membrane integrity (SYBR-14/PI assay and Hoechst 33258). Also, ATP content of spermatozoa was determined using a bioluminescence assay. Treatment of cryopreserved semen with 1 × 10-3 M PAF at 120 min of incubation resulted in significantly higher total sperm motility compared with the control. It was observed that PAF-improved total sperm motility was concurrent with enhanced sperm motility patterns after treatment of cryopreserved semen. Treatment of cryopreserved semen with PAF did not improve either sperm mitochondrial function or plasma membrane integrity, as monitored by different fluorescent membrane markers. Furthermore, ATP content of cryopreserved spermatozoa was significantly higher when PAF was used at a concentration of 1 × 10-3 M compared with the control and other PAF treatments, regardless of the incubation time. The findings of this study indicated that treatment with 1 × 10-3 M PAF at 120 min of incubation rendered better quality of cryopreserved canine semen, which was associated with improved sperm motility parameters and ATP content. It can be suggested that exogenous PAF addition is beneficial as a supplement for canine semen extender used for.

Cryoprotective Effect of Glycerol Concentrations on Indian Red Jungle Fowl (Gallus Gallus Murghi) Spermatozoa

2018 ◽  
Vol 11 (2) ◽  
pp. 80-88 ◽  
Author(s):  
Bushra Allah Rakha ◽  
Muhammad Sajjad Ansari ◽  
Shamim Akhter ◽  
Elisabeth Blesbois
Keyword(s):  
Plasma Membrane ◽  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Gallus Gallus ◽  
Recovery Rates ◽  
Plasma Membrane Integrity ◽  
Red Jungle Fowl ◽  
Frozen Semen ◽  
Acrosome Integrity

Semen cryopreservation protocols for wild avian species need to be optimised in order to achieve optimum post-thaw sperm quality and fertility. The present study was designed to evaluate the cryoprotective effect of different glycerol concentrations (11%, 15% and 20%) on post-thaw quality, recovery rates, absolute livability index and fertility of Indian Red Jungle Fowl (Gallus gallus murghi) semen. Semen was collected from eight mature cocks and cryopreserved for storage at −196 °C. Frozen semen was thawed at 37 °C for 30 s and assessed for motility, plasma membrane integrity, viability and acrosome integrity at 0, 2 and 4 h incubation at 37 °C. Percentages of motility, plasma membrane integrity, viability and acrosome integrity were recorded higher (P<0.05) post-thaw at 0, 2 and 4 h at 37 °C with 20% glycerol compared to 15% and 11% glycerol. Likewise, recovery rates (%) of aforementioned parameters after cryopreservation and absolute livability index were observed highest (P<0.05) with 20% glycerol. By comparing values of R2 after multivariate regression analysis, least negative effects of hours of incubation were observed on semen quality in extenders with 20% glycerol followed by 15% and 11% glycerol. The fertility outcomes (number of fertile eggs, fertility [%], number of hatched chicks, percent hatch and hatchability of fertilised eggs) were recorded higher (P<0.05) with 20% glycerol followed by 15% and 11% glycerol. It is concluded that the concentration of 20% glycerol gives the best cryoprotection for quality and fertility of Indian Red Jungle Fowl semen.

scholarly journals Effect of warming temperatures on donkey sperm vitrification in 0.5 mL straws in comparison to conventional freezing

2019 ◽  
Vol 17 (3) ◽  
pp. e0406 ◽  
Author(s):  
Maria Diaz-Jimenez ◽  
Jesus Dorado ◽  
Cesar Consuegra ◽  
Blasa Pereira ◽  
Isabel Ortiz ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Slow Freezing ◽  
Kinematic Parameters ◽  
Plasma Membrane Integrity ◽  
Progressive Motility ◽  
Acrosome Integrity ◽  
Sperm Freezing ◽  
Conventional Freezing

Aim of study: There is little information about vitrification of sperm in large volumes (up to 0.5 mL). This study aimed to develop the vitrification technique in 0.5 mL straws in donkey sperm, evaluating the effect of three warming temperatures.Area of study: Cordoba, Spain.Material and methods: Ejaculates from five donkeys were divided in four groups: one control subjected to conventional slow freezing (C) and three vitrified in 0.5 mL straws and warmed using different protocols (W1: 37ºC/30s, W2: 43ºC/20s and W3: 70ºC/8s+37ºC/52s). Sperm motility, kinematic parameters, plasma membrane and acrosome integrity were evaluated. Conventional freezing resulted in significantly higher values for total (42.7±19.6%), and progressive motility (30.3±16.7%), plasma membrane (49.1±10.4%) and acrosome integrity (39.6±14.5%) respect to vitrification method.Main results: Values after warming ranged between 0.2-2.8% for total motility; 0.2-2.1% for progressive motility; 5.5-20.0% for plasma membrane integrity and 14.5-29.8% for acrosome integrity in all warming protocols after sperm vitrification. However, no differences were found between W3 and C for kinematic parameters; and W3 resulted in significantly higher values for membrane integrity (20.0±11.0%) in comparison to W1 (5.5±3.6%) and W2 (9.3±8.4%).Research highlights: High warming rates seem to be better for donkey sperm vitrification in large volumes; but this methodology is still not an alternative to conventional sperm freezing.

scholarly journals A Comparison of Freezing Methods and Diluents Types on Post-Thaw Sperm Quality of Ram Sperm

2020 ◽  
Vol 7 (2) ◽  
pp. 235-241
Author(s):  
Pankaj Kumar Jha ◽  
M Golam Shahi Alam ◽  
Farida Yeasmin Bari
Keyword(s):  
Plasma Membrane ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Sperm Parameters ◽  
Plasma Membrane Integrity ◽  
One Step ◽  
Acrosome Integrity ◽  
Ram Sperm

The effect of freezing methods and diluents types on post-thaw sperm quality of Bangladeshi ram semen was studied. Two freezing methods and three diluents was tested as pooling effects (freezing methods or diluents) on post-thaw sperm parameters; sperm motility (SM), viability (SV), plasma membrane integrity (SPMI) and acrosome integrity (SAI), respectively. From selected ten rams, eight ejaculates were used for each freezing group (freezing methods × diluents). Semen samples were diluted by using two-steps for hand-made tris-based diluents (20% egg yolk): D1 (7% glycerol) and D2 (5% glycerol), and one-step dilution for commercial diluents: D3 (Triladyl®) at 35°C. After 4h of equilibration of temperature at 5°C, diluted semen samples was aspirated into 0.25 mL straws, and sealed. Straws were frozen in liquid nitrogen (LN2) vapour using two methods: F1 (manually in Styrofoam box, using three-steps method; +5°C to -80°C at -11.33°C/min, -80°C to -120°C at -26.66°C/min, and -120°C to -140°C at - 13.33°C/min) and F2 (programmable bio-freezer, using two-steps method; +5°C to -100°C at - 20°C/min and -100°C to -140°C at -10°C/min). Two semen straws from each batch were evaluated (37°C for 20 sec) for sperm parameters. In pool effects between freezing methods; SAI differed significantly (P < 0.001). The SM (56%) and SV (72%) were observed competitive. However, SPMI (67.58 ± 2.02%) and SAI (76.13 ± 1.42%) were higher in F1. Among diluents, SM (P < 0.006), SV (P < 0.008), SPMI (P < 0.012) and SAI (P < 0.019) differed significantly. The SM (61.25 ± 1.80%), SV (77.13 ± 1.47%), SPMI (68.31 ± 1.91%) and SAI (74.75 ± 1.64%) were highest in D3. In conclusion, the combination of manual freezing (three-steps) and handmade tris-based diluents (20% egg yolk, 5% glycerol) is suitable and sustainable method for cryopreservation of ram semen. Res. Agric., Livest. Fish.7(2): 235-241,  August 2020

scholarly journals PENGARUH METODE PENCUCIAN SPERMATOZOA SAPI ACEH TERHADAP MOTILITAS, PERSENTASE HIDUP, DAN INTEGRITAS MEMBRAN PLASMA UTUH SPERMATOZOA (The Infuence of Aceh Bull Spermatozoa Washing Method on Spermatozoa Motility and Plasma Membrane Integrity of Intact Spermatozoa)

2015 ◽  
Vol 9 (2) ◽  
Author(s):  
Listin Handayani ◽  
Dasrul Dasrul ◽  
Muslim Akmal ◽  
Cut Nila Thasmi ◽  
Hamdan Hamdan ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Quality Parameters ◽  
Control Group ◽  
Plasma Membrane Integrity ◽  
Spermatozoa Motility ◽  
Sperm Washing ◽  
Fresh Semen

This study aimed to determine the effect of sperm washing by swim up and centrifugation in isotonic medium on sperm quality of aceh bull. In this study, fresh semen from healthy male aceh bull aged 3-4 months was collected using artificial vagina. Immediately after semen collection, fresh semen quality was examined macroscopically and microscopically. Subsequently, sperm washing was performed by centrifugation and swim up in sperm washing medium. Group 1 (P0) as control group, cement washed with isotonic solution (andromed medium: saline solution) with ratio of 1:8. 2. Group 2 (P1), cement was separated by centrifugation method, group 3 (P2), all cement was separated by swim up method then examined the sperm quality sperm washing results. Each treatment was repeated 5 times. Quality parameters measured were the percentage of spermatozoa motility, sperm viability, and plasma membrane integrity intact spermatozoa. Data were analyzed with analysis of variance one-way pattern, followed by Duncan's multiple test. The results showed the mean ± SD percentage of sperm motility of each treatment group (P0; P1; P2) respectively amounted to 72.00±3.74, 66.40±4.77, and 73.60±3.29%. The percentage of viability was 72.00 ±3.74%, 66.40±2.88%, 71.80±2.17%. The percentage of plasma membrane integrity is intact spermatozoa was 68.20±1.79%, 57.20±3.77%, 69.00±2.00%. Results of this study showed that the percentage of motility, live spermatozoa and plasma membrane integrity intact after separation by swim-up method were significantly different (P <0.05) compared with no separation.Key words: spermatozoa quality, aceh bulls, centrifugation, swim up

50 EFFECTS OF VITAMINS ON THE QUALITY AND FERTILITY OF BOAR SEMEN AFTER LIQUID PRESERVATION AT 5°C

2012 ◽  
Vol 24 (1) ◽  
pp. 137
Author(s):  
Z. Namula ◽  
V. V. Luu ◽  
Y. Kaedei ◽  
R. Kodama ◽  
T. Otoi
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Control Group ◽  
Computer Assisted ◽  
Control Groups ◽  
Plasma Membrane Integrity ◽  
Boar Semen ◽  
Acrosome Integrity ◽  
And Control

Liquid preservation can be used as an alternative to freeze-thawing for preserving semen for AI. The efficiency of some boar semen extenders has been studied over storage periods of 5 to 7 days. The objective of this study was to evaluate the viability and penetrability of boar spermatozoa preserved at 5°C in a modified Modena-based extender supplemented with either 100 μM vitamin C (Vc), 100 μM vitamin E (Ve), or 100 μM Vc + 100 μM Ve (Vc + e). The final sperm concentration was adjusted to cells mL–1 and the semen was then stored at 5°C for 4 weeks. In Experiment 1, the semen samples were assessed every week during the 4-week storage in each extender for the following factors: motility, by using computer-assisted semen analysis (CASA); viability, by using the Live/Dead fluorescence viability assay; plasma membrane integrity, by using the hypoosmotic swelling test (HOST); and acrosome integrity, by using fluorescein isothiocyanate (FITC)-labelled peanut agglutinin staining. In Experiment 2, we examined the penetrability of spermatozoa that had been stored in each extender for 4 weeks and the development of fertilized oocytes. Data were analysed using ANOVA. In Experiment 1, when the semen was stored for 2 weeks, the mean percentage values of total sperm motility and viability for semen stored with Ve were significantly higher than those for semen stored without Vc and Ve (control group) (84.3 vs 67.9% and 59.8 vs 51.2%, respectively; P < 0.05). Moreover, the percentage sperm motility for semen stored for 4 weeks tended to be higher in the Ve group than in the control group (44.2 vs 32.7%; P < 0.1). Storage with Vc or Vc + e did not improve sperm motility and viability of semen. The plasma membrane integrity and acrosome integrity of semen did not significantly differ among the groups during the 4-week storage. In Experiment 2, the rates of sperm penetration and of development to blastocysts of fertilized oocytes did not differ between the Ve and control groups (33.0 vs 28.5% and 14.9 vs 10.1%, respectively; P > 0.05). However, storage with Vc reduced the rate of oocyte development compared with the Ve and control groups (1.1%; P < 0.05). In conclusion, adding Ve to the semen extender may improve the motility and fertility of boar semen stored at 5°C. However, adding Vc has a harmful effect on the quality and fertility of stored boar semen.

Effects of fulvic acids on goat sperm

Zygote ◽  
2018 ◽  
Vol 26 (3) ◽  
pp. 220-223
Author(s):  
Yu Xiao ◽  
Zhengmu Wu ◽  
Min Wang
Keyword(s):  
Superoxide Dismutase ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Fulvic Acids ◽  
Quality Parameters ◽  
Control Group ◽  
Progressive Motility ◽  
Acrosome Integrity

SummaryThe effects of adding fulvic acids (FAs) to semen extenders on the quality parameters of frozen–thawed goat buck spermatozoa remain undetermined. Buck semen samples collected from six mature goat bucks once a week were diluted with Tris–egg yolk-based extenders. The diluted semen samples were supplemented with FAs (0.2, 0.4 and 0.6%, w/w), cryopreserved, and evaluated for sperm-quality parameters. Addition of FAs to the extender increased progressive motility, acrosome integrity, membrane integrity, and superoxide dismutase and catalase activities and decreased percentage abnormality and sperm malondialdehyde level compared with the control group. However, excessive FA addition (>0.4%, w/w) to semen extenders did not improve the efficiency. The results indicated that FAs could be a promising cryoprotectant for goat buck sperm.

scholarly journals The Effect of Adding Different Levels of Curcumin and Its Nanoparticles to Extender on Post-Thaw Quality of Cryopreserved Rabbit Sperm

Animals ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1508 ◽  
Author(s):  
Sameh Abdelnour ◽  
Mahmoud Hassan ◽  
Amer Mohammed ◽  
Ahmad Alhimaidi ◽  
Naif Al-Gabri ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Integrity ◽  
Positive Influence ◽  
Control Group ◽  
Curcumin Nanoparticles ◽  
Total Antioxidant ◽  
Semen Extender ◽  
Apoptosis Process ◽  
Structural Levels

The cryopreservation process adversely affects sperm function and quality traits, causing some changes at biochemical and structural levels, due to mechanical, thermal, osmotic, and oxidative damage. Supplementation with curcumin nanoparticles could prevent and even revert this effect and could enhance the post/thawed sperm quality in the rabbit. The study amid to explore the effect of curcumin (CU) and curcumin nanoparticles (CUNPs) supplementation in semen extender on post/thawed rabbit sperm quality. Twelve fertile, healthy rabbit bucks were included, and the ejaculates were collected using artificial vaginas. Rabbit pooled semen was cryopreserved in tris-yolk fructose (TYF) extender without any supplement (control group) or extender supplemented with CU at levels of 0.5, 1 or 1.5 µg/mL (CU0.5, CU1.0, and CU1.5, respectively) or CUNPs at levels of 0.5, 1, 1.5 (CUNPs0.5, CUNPs1.0, and CUNPs1.5, respectively) and was packed in straws (0.25 mL) and stored in liquid nitrogen (−196 °C). Results revealed that CUNPs1.5 had a positive influence (p < 0.05) on post-thawing sperm progressive motility, viability, and membrane integrity as compared with the other groups. Percentages of dead sperm, abnormalities, early apoptotic, apoptotic, and necrotic sperm cells reduced (p < 0.05) in CUNPs1.5 as compared to other treatments. Using 1.5 µg/mL of CUNPs significantly improved total antioxidant capacity (TAC), GPx, while MDA and POC reduced (p < 0.05) in CU1.5 in comparison with other groups. SOD values were enhanced (p < 0.05) in CUNPs1.0 and CUNPs1.5 in relation with other treatments. Conclusively, the addition of curcumin and its nanoparticles to the extender can improve the post-thawed quality of rabbit sperm via redox signaling and reduce the apoptosis process.

Immunolocalisation and expression of oxytocin receptors and sex hormone-binding globulin in the testis and epididymis of dogs: correlation with sperm function

2019 ◽  
Vol 31 (9) ◽  
pp. 1434
Author(s):  
Andressa Dalmazzo ◽  
João D. A. Losano ◽  
Daniel S. R. Angrimani ◽  
Isabel V. A. Pereira ◽  
Marcelo D. Goissis ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Protein Expression ◽  
Membrane Integrity ◽  
Sex Hormone ◽  
Sex Hormone Binding Globulin ◽  
Mitochondrial Activity ◽  
Hormone Binding ◽  
Plasma Membrane Integrity ◽  
Gene And Protein Expression ◽  
Acrosome Integrity

The aim of this study was to confirm gene and protein expression of oxytocin receptor (OTR) and sex hormone-binding globulin (SHBG) in the testis and epididymis of dogs, correlating these data with sperm quality and production and testosterone concentrations. Positive correlations were found between OTR and SHBG expression in both the testis and epididymis. Testicular OTR expression was positively associated with plasma membrane and acrosome integrity in canine spermatozoa, whereas SHBG expression in the testis was positively correlated with various sperm characteristics, such as sperm concentration, total and progressive motility, plasma membrane integrity and acrosome integrity. Testicular expression of both OTR and SHBG was negatively correlated with low sperm mitochondrial activity. In the epididymis, SHBG expression was only positively correlated with plasma membrane integrity. Analysis of protein expression revealed that testicular OTR was positively correlated with testosterone concentrations and negatively correlated with the absence of sperm mitochondrial activity. In addition, SHBG expression in the testes was associated with epididymis SHBG expression and morphologically normal cells. Immunohistochemical (IHC) analysis revealed the presence of both OTR and SHBG in testicular smooth muscles and Leydig cells. However, in the epididymis, OTR was only located in smooth muscle cells, whereas neither IHC nor western blotting detected SHBG. Together, the results of this study suggest that OTR and SHBG play key roles in spermatogenesis and sperm maturation, being essential for male reproductive success.

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