Reducing endogenous estrogens during the neonatal and juvenile periods affects reproductive tract development and sperm production in postpuberal boars

Increased Sertoli cell proliferation during the neonatal period, transient negative effects on epididymal sperm maturation, larger postpuberal testis size reflective of increased Sertoli cell numbers, and increased testicular sperm production characterized boars subjected to continuous inhibition of endogenous estrogen production. The objective in the present experiment was to extend these previous observations to evaluate the effects of a shorter period of reduced estrogen production during the neonatal and juvenile periods on Sertoli cell proliferation, postpuberal testis size, sperm production and epididymal function. Experiments were designed to detect cumulative effects on accessory sex glands as well. Four pairs of littermate boars were assigned to the experiment with one member of each pair randomly selected to receive weekly oral treatment with the aromatase inhibitor, Letrozole, beginning at 1 week of age; the littermates received weekly oral treatment with the corn oil vehicle. Treatment stopped at 12 weeks of age and effects were examined at 10 months. Treated animals had approximately 25% larger testes (P<0.05) correlated with increased Sertoli cell numbers (P<0.05) and larger epididymides. Sperm quality was approximately equivalent in treated and control littermates. Accessory sex glands tended to be smaller in the treated boars. Sertoli cell proliferation during the neonatal and juvenile interval appears to be influenced by endogenous estrogen levels in the boar. A relatively short postnatal interval of Letrozole treatment effectively increased postpuberal testis size. Increased sperm production capacity in response to decreased endogenous estrogens has intriguing possibilities for animal agricultural production.     

Suppression of endogenous estrogen during development affects porcine epididymal sperm maturation

Estrogen plays an important role in male reproduction, critical for sustained fertility in some species. Reducing estrogen's interaction with its receptor(s) in monkey and mouse models is associated with reduced sperm motility and, in some cases, documented elimination of sperm fertilizing ability, suggesting that normal epididymal function may be estrogen dependent. The objective of these experiments was to evaluate the effects of reduced endogenous estrogen on development of epididymal function in the pig, a species in which males have very high levels of endogenous estrogen. Letrozole, a potent inhibitor of estrogen synthesis, was administered to neonatal boars from 1 week of age and markedly suppressed estrogen production. Epididymal function assessed as acquisition of sperm fertilizing ability (in vitro fertilization of zona-free oocytes) was reduced in Letrozole-treated animals at 24 and 28 weeks of age (23% and 30% fertilization, respectively compared with 37% and 54% in vehicle controls) but had recovered by 32 weeks of age. Cauda epididymal sperm numbers were reduced in treated animals (35% of control values at 20 weeks of age) but appeared to be recovering at 32 weeks of age. Reduction of endogenous estrogen had no effect on other aspects of epididymal function (percentage of motile sperm, sperm motion parameters, sperm head morphometrics, or ability of sperm to undergo an acrosome reaction). Reducing endogenous estrogen during postnatal development appears to have transient effects on porcine epididymal function. These transient effects suggest that the pig, with its high endogenous estrogen, may respond differently than other species to reduced estrogen synthesis.     

The effects of postnatal FSH substitution on Sertoli cell number and the sperm production capacity of the adult boar

The role of FSH for Sertoli cell establishment and sperm production in the boar is not definitely known. In order to elucidate its function FSH was substituted postnatally in male pigs and the resulting effects on testicular histological traits and sperm production capacity were investigated when the boars had reached maturity. Six male piglets received pFSH from 18 to 48 days postnatally. Another six piglets instead received saline and served as controls. Blood samples were drawn to measure FSH, LH, testosterone and estradiol. After 28 weeks, the boars were trained to mount a dummy so that the spermatogenic capacity was tested by increasing the frequency of semen collection at the age of 30 weeks. Libido (latency time) and ejaculate criteria (volume, motility, morphological abnormalities) were determined. Thereafter the boars were killed and their testes analyzed for morphology, number of Sertoli cells, germ cells and Leydig cells as well as the ratio between mitosis and apoptosis in the tubules.FSH concentrations were twofold due to FSH application when compared to the controls. LH was low during the first 2 weeks of FSH treatment. Thereafter concentrations increased in three of the six treated animals but not in controls. Testosterone increased slightly over the application period both in the controls and the treated piglets. Estradiol levels were similar in both groups. Increased ejaculation frequency reduced sperm concentrations and sperm motility in all boars and the percentage of morphologically abnormal sperm increased. Ejaculate volumes and the time of latency were not significantly altered. No differences were observed between the controls and the FSH treated boars. The testicular parameters of both FSH- and control boars were identical for morphology, number of spermatogenic and somatic cells as well as mitosis–apoptosis equilibrium. The data demonstrated that a prolonged postnatal period of FSH concentrations does not influence the sperm production of the adult boar.     

Influence of supplemental lighting on serum LH,testosterone and semen quality in prepubertal and postpubertal boars

Thirty-six prepubertal boars received either natural lighting or supplemental lighting from incandescent or fluorescent sources between 8 and 35 weeks of age. Serum concentrations of luteinizing hormone (LH) and testosterone were similar among the treatment groups (P > 0.05). Even though libido scores were higher at 20–24 weeks of age in boars that received supplemental lighting, the libido scores in control boars and supplemental-lighted boars were similar by 30 weeks of age. From 44 to 83 weeks of age, 20 postpubertal boars were exposed to natural or supplemental lighting and semen was collected 3 times in one week per month. No differences in spermatozoa concentration, forward motility, or total motile spermatozoa were found among the treatments. Semen volume of boars maintained on supplemental lighting was lower (P < 0.05) than that of control boars switched to supplemental lighting and lower than that of boars exposed to supplemental lighting that were switched to control. No differences in serum LH or testosterone were observed among treatments in boars from 42 to 82 weeks of age. When boars were bilaterally castrated at 83 weeks of age, no differences in testes weight, epididymal weight, sperm per gram testes, relative testes weight, or relative epididymal weight were found. Data from the present experiment indicate that supplemental lighting does not enhance semen quality of postpubertal boars.     

Effects of exposing boars to different artificial light regimens on semen plasma markers and "in vivo" fertilizing capacity

This study was designed to assess the effects of exposing boars to an artificial photoperiod on semen quality in terms of sperm concentration, sperm vitality, sperm motility and acrosome integrity. We also determined biochemical semen plasma variables, such as total protein concentration, phosphorylated tyrosine residues and fructose, glucose and sorbitol contents, along with their effects on the fertility, prolificacy and libido of the boars. Three groups of 10 males were kept for 3 months under experimental conditions of 24, 12 and 0 h of artificial light, and a constant temperature of 21 +/- 1 degrees C and 60-75% humidity. The animals were fed a nutritious diet and subjected to semen collection twice per week. Semen samples were analyzed throughout the entire experimental period. Our results indicate that, while the extreme photoperiods (0 and 24 h of light) affected semen quality in terms of sperm concentration, acrosome integrity and semen volume, its fertilizing capacity was only significantly reduced under conditions of absolute darkness. Sperm motility was found to be a poor indicator of fertilizing ability, while other sperm factors, such as acrosome integrity or other functional variables seemed to behave better. The photoperiod was found to affect the production of accessory sex gland secretions more than their composition. In addition, light effects on fertility, prolificacy and libido seemed to be achieved through independent mechanisms.     

Dietary l-arginine supplementation improves semen quality and libido of boars under high ambient temperature

l-Arginine is a nutritionally essential amino acid for spermatogenesis and plays versatile roles in animal health and can be utilized as a potential agent to improve reproductive performance of boars under high ambient temperature. The present study aimed to determine whether dietary l-arginine could alleviate heat stress-induced infertility in boars. In all, 20 boars (PIC 1040; 248.59±3.84 kg BW and 407.65±6.40 days of age) were selected and randomly assigned to four groups (group 0.0%, basal diet; group 0.6%, 0.8% or 1.0%, basal diet added with 0.6%, 0.8% or 1.0% l-arginine (wt:wt), respectively.) The four diets were made isonitrogenous by addition of appropriate amounts of l-alanine. Boars were pre-fed the corresponding experimental diet for 42 days. Then, the semen characteristics and libido were accessed for 6 weeks during the hot summer period (25.5° to 33.0°C). Results show that dietary l-arginine remarkably improved sperm motility, normality, total sperm number and effective total sperm number. Also, dietary l-arginine improved semen antioxidant capacity, such as decrease of malondialdehyde and 8-Hydroxy-2'-deoxyguanosine content in sperm (P<0.05), increase of the ratio of glutathione and oxidized glutathione, total antioxidant capacity, glutathione peroxidase and catalase activities in seminal plasma (P<0.05). Most of mitochondria contained intact ultrastructure in l-arginine-supplemented group which also accompany with higher ATP content than the 0.0% group. The boars fed 0.8% l-arginine show increased levels of estradiol-17β and testosterone and exhibit improved libido performance than boars in the 0.0% group. Adding dietary l-arginine linearly increased (P=0.002) nitric oxide content (as l-arginine increased). The scrotal surface temperature in the 0.6%, 0.8% and 1.0% group were decreased by 0.9°C, 0.9°C and 0.4°C, respectively, compared with the 0.0% group. l-Arginine levels caused linear effect on semen quality and antioxidant capacity, also caused quadratic effect on libido performance. During the hot summer months, the predicted optimal l-arginine levels for best semen quality and antioxidant capacity was 0.8% to 1.0% and for best libido performance was 0.8%. It can be concluded that l-arginine can be used as an effective agent to alleviate heat stress-induced infertility of boar, and that 0.8% to 1.0% can be considered as the optimum dosage.     

Prediction of porcine semen fertility by homologous in vitro penetration (hIVP) assay

A field trial was conducted to compare the fertility predicting capacity of different sperm assays applying classical semen analysis, sperm function and the homologous in vitro penetration test (hIVP) to 60 ejaculates from four boars collected over a period of 15 weeks. No differences were found between the groups of fertility (Low Fertility: <20%; Intermediate: 40–60% and High: >80%) for sperm-rich fraction volume collection, sperm concentration, total sperm number, cationic contents in seminal plasma and ATP concentration. Partial differences were found in the parameters of motility, normal morphology, normal apical ridge (NAR), viability with eosin–nigrosin stain, hypo-osmotic swelling test (HOS), osmotic resistance test (ORT) and functional membrane integrity (with carboxyfluorescein diacetate, DCF). These parameters would be useful for detecting sperm with poor fertility, but they are not precise enough to discriminate an ejaculate with higher fertility than the herd median. Only the penetration percentage (10.24±1.45 vs. 55.13±3.35 vs. 84.72±1.73) and sperm number per oocyte (1.29±0.07 vs. 11.29±1.79 vs. 25.86±1.43) in a hIVP system were parameters with a predictive capacity to discriminate between the three fertility groups. Consequently, hIVP was found to be the best seminal assay and it may improve the in vitro assessment of sperm fertilizing ability.     

Reducing the time rabbit sperm are held at 5 °C negatively affects their fertilizing ability after cryopreservation

Cooling sperm to and equilibrating the sperm at 5 °C require the most time in any sperm cryopreservation protocol. Reducing the time required for these phases would simplify sperm freezing protocols and allow greater number of ejaculates to be processed and frozen in a given time. This study determined how holding rabbit sperm at 5 °C for different lengths of time (0, 10, 15, 20, 30, or 45 minutes) affected the quality of rabbit sperm, measured by in vitro assays, and if reducing the cooling time to only 10 minutes affected the fertilizing ability of the sperm. Reducing the time sperm were held at 5 °C to 10 minutes did not affect the in vitro quality of the sperm (percent motile and with intact plasma membranes), although eliminating the cooling phase completely (directly freezing the sperm from room temperature) decreased in vitro assessed sperm quality (P < 0.01). However, reducing the time sperm were held at 5 °C, from 45 to 10 minutes, negatively affected the fertilizing ability of sperm in vivo (P < 0.05). In conclusion, completely eliminating cooling rabbit sperm to 5 °C before freezing is detrimental for rabbit sperm cryosurvival, and although shortening the time sperm are held at 5 °C to 10 minutes does not reduce in vitro sperm quality, it does reduce the fertility of rabbit sperm. Therefore, the length of time rabbit sperm equilibrate at 5 °C is crucial to the fertilizing ability of rabbit sperm and must be longer than 10 minutes. Currently, it is not known if holding rabbit sperm at 5 °C for less than 45 minutes will affect sperm fertilizing ability.     

Immunological castration temporarily reduces testis size and function without long-term effects on libido and sperm quality in boars

The objective was to determine the effects of immunization against gonadotropin-releasing hormone on reproductive characteristics in boars. A total of 72 boars were used in a randomized design with three treatments: single immunization (SI) (10 weeks of age) or double immunization (DI) (10 and 15 weeks of age) with Improvest® and intact controls (no Improvest®; CNT) (n=24/group). At 10, 15, 20, 25 and 40 weeks of age, blood was collected and serum harvested to evaluate testosterone concentrations. Testosterone concentrations were less for DI boars compared with CNT boars and SI boars at 20 and 25 weeks (P<0.001), but not at 40 weeks of age. At week 25, 18 pigs (n=6/group) were sacrificed and testes were removed, weighed and measured, and seminiferous tubules were examined and scored using histological slides of testes parenchyma. A sample of neck fat was assessed for boar taint aroma. All testicular measurements and weights and seminiferous tubule scores were less for DI boars compared with SI and CNT boars (P<0.001). More (P<0.05) SI and CNT boars had detectable boar taint aroma than DI boars. Libido was assessed at 32, 36, 47, 60 and 63 weeks of age and semen collected at 60 weeks of age was analyzed for indicators of quality. There were no effects of treatment (P=0.41) or treatment by week (P=0.71) on libido. Semen volume, gel weight and total number of sperm cells, determined in a subset of boars (n=3/treatment), were not different among treatments. Sperm concentration was greater for DI than SI (P=0.01), and tended to be greater for DI compared with CNT (P=0.10). Sperm motility tended to be greater for DI boars compared with CNT boars (P=0.066). In conclusion, our results show that there are no long-term effects of immunocastration on reproductive characteristics in boars.     

Season and social environment influence the membrane integrity of ejaculated boar spermatozoa as assessed by ouabain sensitivity

Eight adult Landrace boars were housed for 12 months in one of two social environments. Socially nonrestricted boars were penned near estrual females and socially restricted boars were penned behind solid walls to eliminate visual and physical contact with other pigs. All animals were subjected to natural changes in day length. The sensitivity of ejaculated spermatozoa to ouabain (in inhibitor of Na+-K+ ATPase) was determined on 4 consecutive weeks in November, March-April, and July-August. Semen was diluted in Tyrode's solution (pH 7.4) with and without 10(-3) M ouabain. Duplicate samples of control and ouabain-treated spermatozoa were incubated at 37 degrees C for 4 h, and percent motile sperm, motility type, and motility index (combination of percent and type) were determined at hourly intervals. Ouabain-induced decreases in most motility parameters varied with season (season X treatment, P less than 0.05). At hour 4, induced decreases in percent motile sperm were more pronounced in November and July-August than in March-April for socially nonrestricted boars. Decreases in motility type were greater (P less than 0.05) in November and July-August than in March-April for socially nonrestricted boars and were greater (P less than 0.01) in November than in July-August for restricted boars. In March-April motility type decreased (P less than 0.01) to a greater extent for socially restricted vs. nonrestricted boars. Similar season and social environment differences were observed for motility index values. Given the interrelationships between ouabain sensitivity, the functional integrity of sperm cells, and fertilizing capacity, season and social environment differences in ouabain-induced motility depression probably reflect qualitative changes in boar spermatozoa.     

Arylsulfatase A deficiency causes seminolipid accumulation and a lysosomal storage disorder in Sertoli cells

Sulfogalactosylglycerolipid (SGG) is the major sulfoglycolipid of male germ cells. During spermatogenesis, apoptosis occurs in >50% of total germ cells. Sertoli cells phagocytose these apoptotic germ cells and degrade their components using lysosomal enzymes. Here we demonstrated that SGG was a physiological substrate of Sertoli lysosomal arylsulfatase A (ARSA). SGG accumulated in Sertoli cells of Arsa(-/-) mice, and at 8 months of age, this buildup led to lysosomal swelling and other cellular abnormalities typical of a lysosomal storage disorder. This disorder likely compromised Sertoli cell functions, manifesting as impaired spermatogenesis and production of sperm with near-zero fertilizing ability in vitro. Fecundity of Arsa(-/-) males was thus reduced when they were older than 5 months. Sperm SGG is known for its roles in fertilization. Therefore, the minimal sperm fertilizing ability of 8-month-old Arsa(-/-) males may be explained by the 50% reduction of their sperm SGG levels, a result that was also observed in testicular germ cells. These unexpected decreases in SGG levels might be partly due to depletion of the backbone lipid palmitylpalmitoylglycerol that is generated from the SGG degradation pathway in Sertoli cells and normally recycled to new generations of primary spermatocytes for SGG synthesis.     

Disruption of spermatogenesis in boars sub-clinically infected with Trypanosoma brucei brucei

Data from 14 crossbred (Landreace x Large white) boars aged 10-12 months were used to investigate specific germ cells and to what extent Sertoli cells are prone to sub-clinical infection with strain Y58/98 Trypanosome brucei brucei and effects on spermatogenesis. Boars were divided into three groups, A, B and C of 5, 5 and 4 animals, respectively. Groups A and B were infected intraperitoneally with 2.8x10(6) trypanosomes per animal. Group C consisted of intact controls. At stable sub-clinical trypanosomiasis, boars in groups A and B together with two from the controls were weighed, scrotal circumferences were measured and animals were castrated on days 56 and 84 post infection, respectively. Testes were weighed. A portion of a testis was processed for histomorphometric assessment and another portion was used to determine gonadal sperm reserves by haemocytometry. Crude cells were converted to true cells.Sub-clinical trypanosomiasis was characterised by low live and testes weights, reduced scrotal circumference, scanty parasitemia peaks at long intervals and decreased libido. Histomorphometry of animals infected with T. brucei brucei revealed somniferous tubular distortion, denudation and or degeneration of germ cells and Sertoli cells leading to distortion of spermatogenesis. Spermatids and young primary spermatocytes were most prone to, while Sertoli cells and spermatogonia were least affected by sub-clinical trypanosomiasis. There was evidence of regeneration of germ cells from precursor stem cells, resulting in slightly increased gonodal sperm reserves as the post infection period increased. Infected boars may not attain original fertility levels consequently. It was concluded that boars in tropical regions that harbour endemic disease should be maintained under prophylactic conditions.     

Egg water from the amphibian Bufo arenarum induces capacitation-like changes in homologous spermatozoa

Mammalian sperm acquire fertilizing capacity after residing in the female tract, where physiological changes named capacitation take place. In animals with external fertilization as amphibians, gamete interactions are first established between sperm and molecules of the egg jelly coat released into the medium. Since dejellied oocytes are not normally fertilized, the aim of this study was to determine if the jelly coat of the toad Bufo arenarum promotes a “capacitating” activity on homologous sperm. We found that sperm incubation in diffusible substances of the jelly coat (egg water) for 90-180 s is sufficient to render sperm transiently capable of fertilizing dejellied oocytes. The fertilizing state was correlated with an increase of protein tyrosine phosphorylation and a decrease of sperm cholesterol content. Inhibition of either the increase in tyrosine phosphorylation or cholesterol efflux affected the acquisition of fertilizing capacity. Phosphorylation and fertilization could be promoted with NaHCO₃ and also by addition of beta cyclodextrin. Moreover, sperm could gain the ability to fertilize dejellied oocytes in the presence of these compounds. These data indicate that sperm should undergo a series of molecular changes to gain fertilizing capacity; these changes are reminiscent of mammalian sperm capacitation and take place before the acrosome reaction.     

Development of Apical Blebbing in the Boar Epididymis

Microvesicles are of increasing interest in biology as part of normal function of numerous systems; from the immune system (T cell activation) to implantation of the embryo (invasion of the trophoblasts) and sperm maturation (protein transfer in the epididymis). Yet, the mechanisms involved in the appearance of apical blebbing from healthy cells as part of their normal function remain understudied. Microvesicles are produced via one of two pathways: exocytosis or apical blebbing also termed ectocytosis. This work quantifies the histological appearance of apical blebbing in the porcine epididymis during development and examines the role of endogenous estrogens in regulating this blebbing. Apical blebbing appears at puberty and increases in a linear manner into sexual maturity suggesting that this blebbing is a mature phenotype. Endogenous estrogen levels were reduced with an aromatase inhibitor but such a reduction did not affect apical blebbing in treated animals compared with their vehicle-treated littermates. Epididymal production of apical blebs is a secretion mechanism of functionally mature principal cells regulated by factors other than estradiol.     

Intra- and interboar variability in flow cytometric sperm sex sorting

To improve the efficiency of porcine sperm sex sorting using flow cytometry, the aims of the present study were to determine the relevance of inter- and intraboar variability in sperm sortability and to evaluate the significance of ejaculate semen characteristics in such variability. In addition, the variability among boars in the ability of sex-sorted spermatozoa to survive liquid storage at 15 °C to 17 °C was also evaluated. In total, 132 ejaculates collected from 67 boars of different breeds that were housed at an artificial insemination center were used in three experiments. X- and Y-chromosome–bearing sperm were simultaneously separated according to the Beltsville sperm-sorting technology using a high-speed flow cytometer. In the first experiment, interboar variability in the ability of the ejaculated spermatozoa to undergo the flow-based sex-sorting procedure was observed; the ejaculates of nearly 15% of the boars (n = 67) did not exhibit well-defined X- and Y-chromosome–bearing spermatozoa peaks in the histogram, and the ejaculate sperm concentration demonstrated good predictive value for explaining this variation, as indicated by the area under the receiver operating characteristics curve (0.88, P < 0.001). In the second experiment, a certain degree of intraboar variability was observed only in the boars that showed poor sperm sortability (measured according to the presence or not a well-defined split together with sperm sortability parameters) in the first ejaculate (n = 3). In contrast, boars classified as having good sperm sortability in the first ejaculate (n = 5) maintained this condition in five ejaculates collected over the subsequent 5 months. In the third experiment, sex-sorted spermatozoa from boars with good sperm sortability (n = 5) remained viable and motile (above 70% in all boars) after 48 hours of storage at 15 °C to 17 °C, which may facilitate the commercial application of sex-sorted spermatozoa in swine artificial insemination programs.     

Sertoli cells in the boar testis: changes during development and compensatory hypertrophy after hemicastration at different ages

Changes in Sertoli cell numbers and testicular structure during normal development and compensatory hypertrophy were assessed in crossbred Meishan x White Composite males. Boars were assigned at birth to unilateral castration at 1, 10, 56, or 112 days or to remain as intact controls through 220 days. The first testes removed were compared to assess testicular development. At 220 days, testicular structure was evaluated in boars representing the 25% with the largest (Lg) testis and the 25% with the smallest (Sm) testis in each treatment group. The number of Sertoli cells per testis reached a maximum by Day 56 in Sm testis but not until Day 112 in Lg testis boars, indicating a longer duration of Sertoli cell proliferation in Lg testis boars. Unilateral castration of Lg testis boars on Days 1, 10, 56, and 112 caused the weight of the remaining testis to hypertrophy by 149%, 135%, 119%, and 120%, respectively, and total sperm production to increase to 127%, 128%, 97%, and 106%, respectively. However, Sertoli cell numbers changed little in hemicastrate boars. In Lg testis boars, compensatory hypertrophy primarily involved proliferation of Leydig cells and expansion of existing Sertoli cells with little increase in Sertoli cell numbers, but in Sm testis boars, it involved expansion of existing Leydig and Sertoli cells without increase in cell numbers. These results indicate that Lg and Sm testis boars display intriguing differences during both development and compensatory hypertrophy, and they identify a unique animal model for further studies of factors that program and control Sertoli cell proliferation.     

Cellular localization of estrogen receptor-alpha (ERα) and -beta (ERβ) mRNA in the boar testis

Boar testes synthesize high amounts of estrogens which are known to stimulate several male sexual functions in a variety of extragonadal target tissues. Possible effects within the testis depend on the existence of the estrogen receptor subtypes α and β (ERα, ERβ). The precise cellular localization of these subtypes within the testis was, so far, based mainly on protein expression studies using different antibodies in several species including boars shows contradictory results. Therefore, we investigated the ERα and ERβ gene expression using RT-PCR of testis homogenates and RT-PCR after UV-single cell microdissection combined with in-situ hybridization of four fertile boars with an average age of 32 weeks. Both ERα and ERβ mRNA were found in testis homogenates. Using in-situ hybridization and UV-single cell microdissection ERα mRNA was present in type A and type B spermatogonia up to mid-pachytene primary spermatocytes in stage V–VIII and stage I of the seminiferous epithelial cycle, but not in other cells. ERβ mRNA was found only in Sertoli cells. Interstitial Leydig cells revealed neither ERα nor ERβ mRNA. The data suggest a direct impact of estrogen in the boar on Sertoli cell function via ERβ and germ cell formation via ERα.     

Gonadal sperm reserve in purebred Landrace and Large White boars of high average daily gain

The present study investigated the effects of average growth rate (AGR) levels and age on the number of sperm cells per gram of testis parenchyma and on the gonadal reserve in Landrace (LD) and Large White (LW) boars. In Experiment 1, the effects of breed (LD, LW), level of AGR from birth up to 90 days of age (Level 1: 384 +/- 32 g/day; Level 2: 512 +/- 22 g/day; Level 3: 624 +/- 41 g/day), and age (13, 15, 17, 19 and 21 weeks) on testicular cell concentration were evaluated. Data were analyzed under a 2 x 3 x 4 factorial design. There were significant effects associated with breed (P < 0.001) and age (P < 0.001) but not with AGR (P > 0.05) on sperm cell number per gram of testicular parenchyma. The number of cells increased with age and was greater in LW than in LD young boars, mainly those up to 19 weeks of age. In Experiment 2, the effect of two AGR levels (Level 1: 649-694 g/day; Level 2: 813-885 g/day) from birth up to 100 kg body weight on the number of sperm cells per gram of testis parenchyma and on the gonadal reserve was investigated using 59 purebred LD and LW boars. The boars were castrated at 23, 25, 29 and 33 weeks of age. Age of boars significantly affected gonadal sperm reserve and the number of sperm cells per gram of testicular tissue (P < 0.001). Breed of boars and AGR Levels did not significantly affect number of sperm cells and gonadal sperm reserve (P > 0.05). It was concluded that the number of sperm cells in the testicular tissue of young boars is influenced by their breed and age, but not by the level of their AGR.     

Effect of estrogens on boar sperm capacitation <Emphasis Type="Italic">in vitro</Emphasis>

Background  

Mammalian sperm must undergo a series of controlled molecular processes in the female reproductive tract called capacitation before they are capable of penetrating and fertilizing the egg. Capacitation, as a complex biological process, is influenced by many molecular factors, among which steroidal hormone estrogens play their role. Estrogens, present in a high concentration in the female reproductive tract are generally considered as primarily female hormones. However, there is increasing evidence of their important impact on male reproductive parameters. The purpose of this study is to investigate the effect of three natural estrogens such as estrone (E1), 17beta-estradiol (E2) and estriol (E3) as well as the synthetical one, 17alpha-ethynylestradiol (EE2) on boar sperm capacitation in vitro.     

In vitro fertilization of porcine oocytes with fresh and frozen-thawed ejaculated or frozen-thawed epididymal semen obtained from identical boars

The objective of this study was to compare the in vitro fertilizing capacity of porcine spermatozoa from fresh and frozen-thawed semen and frozen-thawed epididymal spermatozoa obtained from identical boars. Prior to IVF, fresh spermatozoa were capacitated in TCM 199. Frozen semen samples were stored in 0.25-ml plastic straws using a lactose/glycerol/orvus-es-paste extender. Cumulus-oocyte-complexes (COC) obtained from superovulated prepuberal gilts were fertilized in vitro within 2 h after aspiration with one of the semen samples. After final dilution for IVF, frozen-thawed epididymal semen samples showed motility rates (72.2 +/- 5.6%) similar to those of spermatozoa in fresh semen (76.4 +/- 4.5%), while sperm motility decreased in frozen-thawed ejaculated semen (40.2 +/- 9.4%). Considerable individual differences in sperm motility between boars were observed for ejaculated semen but not for epididymal semen. Enhanced fertilizing capacity of frozen-thawed epididymal spermatozoa was confirmed by pronucleus formation and cleavage rates, with significantly more embryos developing to the 2- and 4-cell stages compared with the groups fertilized with fresh or with frozen-thawed ejaculated semen (59.7 vs 14.6 and 16%). In conclusion, consistent in vitro fertilization rates with minimal semen variability are obtained using frozen-thawed epididymal semen. Following a modified freezing protocol, epididymal spermatozoa can easily be frozen in small containers for IVF, with higher resultant motility and fertilization rates than with ejaculated semen.