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At least 19 recordsLinked to original sources

PubMed1977

Antigens on boar spermatozoa and the effect of antibodies against the spermatozoa and the accessory sexual gland fluids on boar spermatozoa.

Antigens on the acrosomes of boar spermatozoa could be detected by the anti-complement immunofluorescence test, using antisera against spermatozoa and against seminal vesicle and epididymis fluids. Weak fluorescence was observed even on the tail and no fluorescence in the postnuclear segment. Adequate absorption of the complement allowed the sperm-immobilizing test to be used with boar spermatozoa. Using this test, the specificity of the sperm-immobilizing effect on antispermatozoal antibodies was confirmed. The sperm-immobilizing effect and damage to the acrosomes was prevented by absorption of the sera with spermatozoa. Antibodies against the fluid of the boar prostate did not react with antigens on boar spermatozoa.

Agglutination Tests↗

PubMed1997

Cryopreservation of mouse spermatozoa. II. Relationship between survival after cryopreservation and osmotic tolerance of spermatozoa from three strains of mice.

A procedure to cryopreserve mouse spermatozoa has been derived to bank the genetics of valuable strains of mice in a practical way. The primary objective of this study was to apply the cryopreservation method developed for spermatozoa of strain B6D2F1 to those of strains 129/J and C57BL6/J. Using the capability of spermatozoa to fertilize oocytes in vitro as the criterion of survival, we found differences in survival after cryopreservation among the three strains. Blastocysts were obtained after in vitro fertilization of oocytes with frozen spermatozoa from B6D2F1 (51%) and 129/J (12%); none was obtained from C57BL6/J. Transfer of embryos into recipients resulted in the birth of 69 live pups from 164 embryos produced with frozen B6D2F1 spermatozoa and 11 pups from 35 embryos produced with 129/J spermatozoa. To seek an explanation of these differences among the three strains, spermatozoa were exposed to anisotonic solutions ranging from 5 to 3200 mOsm; viability of spermatozoa was assessed by a double stain using flow cytometry. Mouse spermatozoa tolerated exposure to solutions of osmolalities between 200 and 400 mOsm, but were damaged when exposed to solutions exceeding this range. Spermatozoa from C57BL6/J were the most sensitive: 20, 35, and 40% of C57BL6/J, 129/J, and B6D2F1 spermatozoa survived exposure to an 800 mOsm solution, respectively. This study suggests that there is a genetic basis for sensitivity of mouse spermatozoa to osmotic shock and freezing injury. Nevertheless, the birth of live pups produced with frozen spermatozoa from 129/J as well as with spermatozoa from B6D2F1 mice indicates that cryopreservation of spermatozoa can be used to preserve the genetics of valuable strains of mice.

Animals↗

PubMedDate not supplied

Fertilizing capacity of rat spermatozoa is correlated with decline in straight-line velocity measured by continuous computer-aided sperm analysis: epididymal rat spermatozoa from the proximal cauda have a greater fertilizing capacity in vitro than those from the distal cauda or vas deferens.

Rat spermatozoa recovered from different regions of the excurrent ducts of 10 adult males (proximal cauda epididymidis [PC], distal cauda epididymidis [DC], and vas deferens [VD]) were assessed by in vitro fertilization (LVF) using limited sperm numbers, and by continuous evaluation of motility parameters during 5 hours of incubation in vitro with automated computer-aided sperm analysis (CASA). Spermatozoa from the PC region fertilized (68 +/- 6%) a significantly greater (P < or = 0.005) number of oocytes than those from the DC (44 + 5%) or VD (47 +/- 7%). For pooled samples from all three regions, the mean fertilization rate (51 +/- 14%) was less tan for spermatozoa from the PC (P < 0.05) but was not significantly different from spermatozoa from the DC or VD. For each time point and sample, 1,592 +/- 428 sperm tracks were analyzed. CASA was verified by comparison with manual still-frame analysis of video recordings, by repeated analysis of the same or different samples of spermatozoa, and by examination of computer tracks. The coefficients of variation for various motion parameters suggested that the CASA obtained a high degree of precision. There were no significant differences in motility parameters for spermatozoa recovered from equivalent regions of the left or right tract or in motility parameters for spermatozoa from different regions of the tract immediately after recovery. However, during incubation in vitro, spermatozoa from the DC or VD regions exhibited a marked decline in straight-line velocity (VSL) compared with spermatozoa from the PC region. The reduction in VSL (combined values from right and left tract) for DC or VD spermatozoa compared with PC spermatozoa was significant at 2.5 hours of incubation (P < or = 0.05) and highly significant (P < or = 0.005) by the end of the incubation period. Differences in average path velocity (VAP) were also apparent after 4 hours (p < or = 0.05), but no significant differences were observed for measurements of curvilinear velocity (VCL) or lateral bead displacement (ALH). Overall, the decline in VSL over 5 hours was highly correlated (P < or = 0.001) with the outcome of fertilization in vitro. In contrast, initial VSL and changes in VCL of spermatozoa were not correlated with fertilization rate. These results indicate that the in vitro fertilizing capacity of rat spermatozoa is correlated with 1) the decline in straight-line velocity (VSL) as measured by repeated CASA during incubation in vitro and 2) with the site of recovery of mature rat spermatozoa from the distal excurrent duct. It is suggested that the deterioration of the quality of rat spermatozoa in the distal epididymidis and vas deferens during storage may occur sooner than previously realized, and therefore care must be taken when recovering samples for fertility assessment. In keeping with findings in other species, immediate "snapshot" analysis of rat motility was a poor predictor of sperm fertility. In contrast, continuous CASA provided significant information for determining sperm fertilizing capacity and will be a useful technique for reproductive toxicology.

Animals↗

PubMed2003

Effects of dead spermatozoa on motion characteristics and membrane integrity of live spermatozoa in fresh and cooled-stored equine semen.

The aim of this study was to determine if dead spermatozoa reduced motility or membrane integrity of live spermatozoa in fresh and cooled-stored equine semen. Three ejaculates from each of three stallions were centrifuged and virtually all seminal plasma was removed. Spermatozoa were resuspended to 25 x 10(6) spermatozoa/ml with EZ-Mixin CST extender and 10% autologous seminal plasma, then divided into aliquots to which 0 (control), 10, 25, 50, or 75% (v/v) dead spermatozoa were added. Dead spermatozoa preparations contained 25 x 10(6) spermatozoa/ml and 10% seminal plasma from pooled ejaculates of the three stallions, in EZ-Mixin CST extender. Spermatozoa were killed in the pooled ejaculates by repeated freezing and thawing, then stored at -20 degrees C until warmed to 37 degrees C and mixed with aliquots of fresh spermatozoa to be cooled and stored in an Equitainer for 24h. Motion characteristics (% total motility (MOT), % progressive motility (PMOT), and mean curvilinear velocity (VCL)) for fresh and 24h cooled samples were determined using a computerized spermatozoal motion analyzer. The presence of up to 75% dead spermatozoa did not adversely affect MOT or PMOT of live spermatozoa in either fresh or cooled-stored semen. However, VCL and the percentage of membrane-intact spermatozoa were reduced compared to control samples when 75% (v/v) dead spermatozoa were added. Membrane integrity, as assessed by staining with carboxyfluoresein diacetate-propidium iodide, was highly correlated (r>0.8; P<0.001) with MOT and PMOT in both fresh and cooled-stored semen samples. Results of this study have application to the processing of both cooled and frozen equine semen.

Animals↗

PubMed1996

Pregnancies achieved with testicular and ejaculated spermatozoa in combination with intracytoplasmic sperm injection in men with totally or initially immotile spermatozoa in the ejaculate.

The efficacy of intracytoplasmic sperm injection (ICSI) employing testicular and ejaculated spermatozoa was assessed in 24 couples with totally or initially immotile spermatozoa. No criteria were employed in selecting which patients would be treated with testicular or ejaculated spermatozoa. The men were chosen at random. Testicular spermatozoa obtained by testicular sperm extraction were used in 14 and ejaculated spermatozoa were used in 10 of these couples. In all cases. asthenozoospermia was total in their basal semen sample. In 12 male partners, spermatozoa were totally immotile before and after Percoll gradient fractionation (totally immotile). In the remaining 12 men, spermatozoa initially showed a total absence of motility; however, some of the spermatozoa had showed very poor motility (0. 1%) after Percoll gradient fractionation and a 1.5-2.0 h incubation period (initially immotile). Of these 24 total asthenozoospermic males, 14 also had total teratozoospermia. The fertilization and cleavage rates in the testicular and ejaculated sperm groups were 53. 5 and 96.3 and 54.5 and 94.4% respectively. One cycle resulted in complete fertilization failure, and in 23 embryo transfer cycles a total of 10 pregnancies were obtained (41.6%). Eight pregnancies were achieved in the testicular sperm group, while only two pregnancies were obtained in the ejaculated sperm group. Four pregnancies, two from the ejaculated sperm group and two from the testicular sperm group, resulted in clinical abortions in the first trimester. Of the remaining six pregnancies, two have already resulted in healthy births and four pregnancies are now in the second or third trimester in the testicular sperm group. Using testicular spermatozoa in combination with ICSI can be an alternative mode of treatment in cases with totally or initially immotile spermatozoa in the ejaculate. Very low pregnancy rates have been obtained and no ongoing pregnancy has been achieved using ejaculated spermatozoa in these cases.

Adult↗

PubMed1997

Cryopreservation of mouse spermatozoa. I. Effect of seeding on fertilizing ability of cryopreserved spermatozoa.

To examine the effect of seeding to induce ice formation during cryopreservation on their survival, spermatozoa from B6D2F1 mice were cooled to and held at -4 degrees C for 30 min in phosphate-buffered saline (PBS) alone, in egg yolk-supplemented PBS, or in PBS with raffinose + glycerol as cryoprotective additives (CPAs). Seeding and holding spermatozoa at -4 degrees C did not affect their viability as judged by vital staining. Egg yolk protected spermatozoa against chilling injury, as cooling them to -4 degrees C in the presence of egg yolk yielded higher survivals than those cooled without egg yolk (34.4 +/- 3.4 v 9.0 +/- 1.3% in three replicates of >400 spermatozoa/replicate). To study effects of seeding on their fertilizing ability, spermatozoa in the raffinose-glycerol-egg yolk solution were frozen to -196 degrees C either without seeding or after seeding at -4 degrees C. Development of 222 oocytes into two-cell embryos after in vitro fertilization (IVF) with spermatozoa frozen without seeding was 43%; development rates of 186, 186, and 207 oocytes after IVF with spermatozoa frozen after seeding and being held at -4 degrees C for 5, 10, or 30 min were 46, 44, and 9%, respectively. In a direct comparison, after IVF with seeded or unseeded spermatozoa the respective cleavage rates into two-cell embryos were 83% of 275 oocytes and 69% of 304 oocytes, a difference that was small but significant by chi2 analysis. An additional 925 oocytes were fertilized with spermatozoa after being seeded and frozen to -196 degrees C in four separate batches of CPA solutions. Overall, after IVF with frozen sperm, 68% of those oocytes cleaved into two-cell embryos and 59% developed into 544 blastocysts. Based on these results, we concluded that embryo production by IVF seemed to be improved using spermatozoa frozen after being seeded. Mouse spermatozoa cryopreserved by the method described here are capable of fertilizing oocytes at a rather high rate.

Animals↗

PubMed1981

[Studies of fertilizing capacity of spermatozoa from infertile men with oligozoospermia. AIH by washed and concentrated spermatozoa and in vitro fertilizing capacity to zona free hamster eggs (author's transl)].

The fertilizing abilities of spermatozoa from infertile men with oligozoospermia were evaluated by examining pregnancy rates after AIH using washed and concentrated spermatozoa, and also by in vitro fertilizing rates to zona free hamster eggs. The spermatozoa from ten oligozoospermic men with less than 20 x 10(6)/ml of sperm counts were used in the experiments. After the procedures of washing and concentrating spermatozoa from oligozoospermic men, the sperm concentration was increased from two to six times and the sperm motility was also increased up to two times. However none of the patients' wives conceived after several AIHs with washed and concentrated spermatozoa. When the in vitro fertilization of zona free hamster eggs by the spermatozoa was used, the fertilization rates were null in case of 7 oligozoospermic patients, and 5, 10 and 33.3% in case of another 3 oligozoospermic patients respectively in contrast with 73.3% in use of spermatozoa from control fertile men. The fertilizing abilities of spermatozoa from husbands of 4 sterile couples with an unknown cause were examined by in vitro fertilization test and 7.1, 37.5, 50.0 and 55.6% fertilization rates were obtained. Among these men, the spermatozoa from one man indicated a very low fertilization rate without any other pathological findings. This suggests that the impairment of fertilizing ability of spermatozoa might be present in the case of sterile men with an unknown cause.

Adult↗

PubMed1995

Epididymal spermatozoa: recovery and subsequent improvements of mouse epididymal spermatozoa via the SpermPrep filtration method.

This study was designed to determine the effects of Sephadex filtration (SpermPrep method) on the separation of viable motile, morphologically normal mouse epididymal spermatozoa and to study the viability of the recovered spermatozoa over a 3 hr incubation period. Spermatozoa were harvested from the caudae epididymides (5 animals per run or replication; n = 10) following bilateral testicular excision and incubated in 2-ml of Test-Yolk buffer (TYB) at 37 degrees C for 15 min. The specimen was then split into 2 aliquots, with Alquot 1 as the control and Aliquot 2 used for filtration. SpermPrep I column was employed according to the manufacturer's specifications (ZBL, Inc., Lexington, KY, USA) using TYB. During filtration (10 min), different fractions were obtained: first 5 min (Sample 1) and second 5 min (Sample 2). The filtered fractions were evaluated and incubated at 37 degrees C and assessed for percentage and grade of motility (0-4) every 30 min for 3 hr. Filtration resulted in a significant improvement in percentage and grade of motility (91.5% and 3.0 vs. 76.5% and 2.5, respectively). The results point out very clearly that the filtration via the SpermPrep method improved the percentage and grade of motility (p < 0.05) but not the percentage normal morphology of the spermatozoa. Also the SpermPrep I enabled the recovery of 45% (8.3 x 10(6) spermatozoa) of the total spermatozoa processed in the control aliquot (18.4 x 10(6) spermatozoa) which is consistent with previous observations. Most importantly, filtered spermatozoa incubated for 3 hr showed greater percentage and grade of motility than the control spermatozoa (63% and 1.66 vs. 39% and 0.82, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1982

Studies on prolonged spermatozoa survival in Chiroptera: a morphological examination of storage and clearance of intrauterine and cauda epididymal spermatozoa in the bats Myotis lucifugus and M. velifer.

The cauda epididymidis, uterine corpus, and cornua and uterotubal junction of Myotis function to retain and preserve normal spermatozoa throughout hibernation. In none of the sites do spermatozoa show features that might account for their extended viability. Spermatozoa stored in the uterus and epididymis show no special orientation toward the epithelium lining these sites, whereas an intimate relationship is established between some sperm and the epithelial cells of the uterotubal junction which might either account for extended postcoital sperm survival or forecast their removal from further participation. Transmission and scanning electron microscopic observations do not disclose any morphological changes in stored luminal spermatozoa. A low rate of phagocytosis of sperm is evident in the female tract during hibernation. However, spermatozoa are evidently not vulnerable to being removed from the storage sites until spring arousal when ovulation occurs. Both uterotubal epithelial cells and phagocytes appear to be involved in the disposal of spermatozoa in the female, whereas epididymal spermatozoa apparently are primarily voided during urination. A mechanism that delays capacitation must underlie the ability of spermatozoa to survive in the female reproductive tract of the hibernating bat.

Animals↗

PubMed2005

Morphometry characterisation of European eel spermatozoa with computer-assisted spermatozoa analysis and scanning electron microscopy.

The aim of the present study was to characterise European eel spermatozoa morphometrically, as a basis for future studies on the morphological effects of methods for sperm cryopreservation and sperm quality. This characterisation was carried out measuring several spermatozoa morphology parameters (head length, width, area and perimeter) by scanning electron microscopy (SEM), in comparison with measurements developed in European eel spermatozoa with computer-assisted morphology analysis (ASMA). Spermatozoa head morphology showed differences in width (1.15+/-0.01 microm versus 1.12+/-0.01 microm), perimeter (14.68+/-0.13 microm versus 13.72+/-0.19 microm) and area (5.36+/-0.06 microm2 versus 1.12+/-0.01 microm2) for ASMA and SEM, respectively. When head length was evaluated, significant differences were found, being higher for SEM methodology (5.09+/-0.04 microm versus 4.29+/-0.03 microm). The curved and elongated spermatozoa head in eels means a problem for the ASMA system (Sperm Class Analyser), Morfo Version 1.1, Imagesp, Barcelona, Spain), causing an error in the length measurements. However, similar results were obtained by both techniques when spermatozoa head length was considered as the greater length between two points within the object (4.29+/-0.03 microm versus 4.31+/-0.04 microm for ASMA and SEM, respectively). In conclusion, this is one of the first applications of ASMA in fish and the first in this species, and confirms this system as a useful tool with wide applications in future fish spermatozoa studies. Width, perimeter and area could be used as parameters for the spermatozoa morphology evaluation, whereas the length requires a new programming of the Imagesp software.

Animals↗

PubMed2000

Expression of the platelet-activating factor receptor in human spermatozoa: differences in messenger ribonucleic acid content and protein distribution between normal and abnormal spermatozoa.

OBJECTIVE: To determine the expression and distribution of the platelet-activating factor (PAF) receptor in normal and abnormal specimens of human spermatozoa. DESIGN: Prospective analysis of membrane-bound PAF receptors by immunofluorescence and PAF receptor messenger RNA by quantitated reverse transcription-polymerase chain reaction in normal and abnormal spermatozoa. SETTING: University-based reproductive genetics laboratory. PATIENT(S): Men undergoing routine semen analysis. INTERVENTION(S): Normal and abnormal spermatozoa were exposed to rabbit anti-PAF receptor antibody, fluorescein isothiocyanate-conjugated goat anti-rabbit antibody, and fluorescent microscopy or subjected to RNA isolation by acid-phenol extraction and quantitated (MIMIC Construction Kit [Clontech Laboratories, Inc., Palo Alto, CA]) reverse transcription-polymerase chain reaction. MAIN OUTCOME MEASURE(S): Fluorescent intensities at six locations along spermatozoa (end piece, principal tail, midpiece, neck, proximal head, and acrosomal region) and PAF receptor expression (messenger RNA) levels. RESULT(S): Immunofluorescence demonstrated a significant difference in PAF receptor distribution between normal and abnormal human spermatozoa, specifically at the neck region. Additionally, abnormal spermatozoa were found to have statistically significantly more PAF receptor messenger RNA than normal spermatozoa. CONCLUSION(S): Platelet-activating factor receptor expression and distribution are significantly altered in abnormal spermatozoa and this may be the result of some defect in gene transcription.

Animals↗

PubMed1998

Verification of flow cytometorically-sorted X- and Y-bearing porcine spermatozoa and reanalysis of spermatozoa for DNA content using the fluorescence in situ hybridization (FISH) technique.

Flow cytometric sperm sorting based on X and Y sperm DNA difference has been established as the only effective method for sexing the spermatozoa of mammals. The standard method for verifying the purity of sorted X and Y spermatozoa has been to reanalyze sorted sperm aliquots. We verified the purity of flow-sorted porcine X and Y spermatozoa and accuracy of DNA reanalysis by fluorescence in situ hybridization (FISH) using chromosome Y and 1 DNA probe. Eight ejaculates from 4 boars were sorted according to the Beltsville Sperm Sexing method. Porcine chromosome Y- and chromosome 1-specific DNA probes were used on sorted sperm populations in combination with FISH. Aliquots of the sorted sperm samples were reanalyzed for DNA content by flow cytometry. The purity of the sorted X-bearing spermatozoa was 87.4% for FISH and 87.0% for flow cytometric reanalysis; purity for the sorted Y-bearing spermatozoa was 85.9% for FISH and 84.8% for flow cytometric reanalysis. A total of 4,424 X sperm cells and 4,256 Y sperm cells was examined by FISH across the 8 ejaculates. For flow cytometry, 5,000 sorted X spermatozoa and 5,000 Y spermatozoa were reanalyzed for DNA content for each ejaculate. These results confirm the high purity of flow sorted porcine X and Y sperm cells and the validity of reanalysis of DNA in determining the proportions of X- and Y-sorted spermatozoa from viewing thousands of individual sperm chromosomes directly using FISH.

Animals↗

PubMed2002

Comparison of intrabursal transfer of spermatozoa, a new method for artificial insemination in mice, with intraoviductal transfer of spermatozoa.

PURPOSE: The objective of this paper was to compare the in vivo fertilizing abilities of fresh epididymal spermatozoa with a new method of artificial insemination in mice, so-called "intrabursal transfer of spermatozoa (ITS)," which requires transfer of spermatozoa into a space near the infundibulum between the ovary and ovarian bursa of superovulated females, and the previous method, so-called "intraoviductal transfer of spermatozoa (IOTS)," especially as regards sperm number and capacitation. METHODS: Spermatozoa freshly isolated from B6C3F1 males were injected into superovulated B6C3F1 females on E 0.4 (10:00 AM) by the IOTS or ITS method. Embryos at two-cell stage were collected from the females 1 day after injection and their morphology was scored. Some females were allowed to survive at midgestational stages and inspected for development of normal fetuses. RESULTS: When 1 microL of a sperm suspension containing uncapacitated 1 x 10(5) spermatozoa freshly isolated from B6C3F1 males was injected by the IOTS or ITS method, normal two-cell embryos were recovered from the females at rates ranging from 14 to 23% with each method. This rate was much lower than that (93% on average) for embryos obtained by natural mating. Neither injection of 1 x 10(3) or 1 x 10(4) spermatozoa nor induction of capacitation improved in vivo fertilization rate. In both cases, females given spermatozoa exogenously yielded midgestational fetuses (E 12.5-13.5) with average litter sizes between 2.5 and 2.8. CONCLUSION: ITS was comparable to IOTS with the conditions used. These two methods will be valuable for artificial insemination in mice for propagation of offspring from particular transgenic or mutant lines that are difficult to breed, although further attempts to improve in vivo fertilization ability are required.

Animals↗

PubMed1982

Site of aromatic L-amino acid oxidase in dead bovine spermatozoa and determination of between-bull differences in the percentage of dead spermatozoa by oxidase activity.

Most (94%) of the aromatic L-amino acid oxidase activity in dead bovine spermatozoa was recovered in tail preparations. The enzyme was released from the cell in sodium citrate but not in sodium phosphate or sodium chloride solutions but oxidase activity was not significantly different in sodium phosphate or sodium citrate buffers (9.6 microliters O2/h and 11.2 microliters O2/h). The activity in ejaculated spermatozoa was correlated with the percentage of dead spermatozoa (r = 0.954, P less than 0.01) but could not be detected in freshly collected epididymal spermatozoa. Killed epididymal spermatozoa showed oxidase activity (10.1 microliters O2/h) similar to that of killed ejaculated spermatozoa (9.2 microliters O2/h). It is concluded that death of spermatozoa occurs in the ampulla and/or at ejaculation and that between-bull differences in the percentage of dead spermatozoa are a consequence of differences between bulls in conditions in the ampulla and/or at ejaculation.

Amino Acid Oxidoreductases↗

PubMed1995

Increased zona-binding ability after incubation of spermatozoa with proteins extracted from spermatozoa of fertile semen.

The involvement of proteins extracted from spermatozoa of fertile semen in sperm-zona binding was examined under hemizona assay conditions. One droplet of a suspension of spermatozoa was exposed to sperm proteins and then tested for zona binding, while a parallel semen suspension droplet incubated with culture medium served as a control. The reliability of the test was increased by relating the number of spermatozoa bound to each inseminated hemizona to the surface area of the hemizona and expressed as the binding index. For spermatozoa incubated with extracted proteins, the binding index was greater than (P = 0.001) that of controls (125.2 +/- 45.1 versus 63.6 +/- 29.2, respectively). As a first control, two other protein sources (fetal calf serum and human follicular fluid) were tested in the hemizona assay. No significant differences were found in zona binding for other protein-exposed spermatozoa compared with controls. As a second and reverse control, exposure of one hemizona to sperm proteins before insemination with untreated spermatozoa induced a marked decrease (P = 0.0003) in sperm binding, compared with that of the matched hemizona not exposed to sperm proteins (control) (3.4 +/- 1.4 versus 74.5 +/- 6.8, respectively). Taken together, these findings confirm the involvement of extracted sperm proteins in sperm-zona interactions. Therefore, in the cases in which fertilization in vitro fails because of a lack of sperm-zona binding, incubation of deficient spermatozoa with proteins extracted from spermatozoa of fertile ejaculates should restore their ability to interact with the oocyte and, thus, should enhance the prognosis for in vitro fertilization.

Culture Media, Conditioned↗

PubMedDate not supplied

Cryopreservation of a small number of fresh human testicular spermatozoa and testicular spermatozoa cultured in vitro for 3 days in an empty zona pellucida.

It is known that the motility of human testicular sperm can be improved when they are cultured in vitro for a few days. The purpose of this study was to determine whether it is better to freeze human testicular spermatozoa on the day of biopsy (fresh) or after they were cultured for 3 days. A modified, single-sperm freezing technique was used in this study. The study consisted of two parts: (1) ejaculated spermatozoa were used to examine the influence of different concentrations of glycerol and synthetic serum substitute (SSS) on the survival rate after cryopreservation, and (2) the survival rates between cryopreserved fresh testicular spermatozoa (Group 1) and testicular spermatozoa that were cultured for 3 days before freezing (Group 2) were compared. Empty zonae pellucidae were obtained from mouse eggs. Five to 10 motile spermatozoa were selected and injected into an empty zona pellucida. For freezing, the zona pellucida with spermatozoa was transferred into a HEPES-buffered human tubal fluid containing different concentrations of glycerol and kept at room temperature for 10 to 15 minutes, and then loaded into a 0.25-ml-plastic straw. The straws were exposed to liquid nitrogen vapor for 2 hours and then plunged into liquid nitrogen. For thawing, the straws were taken out of liquid nitrogen and placed into a 37 degrees C waterbath for 25 to 30 seconds. There was no statistically significant difference in survival rates between 3% and 10% SSS with different glycerol concentrations. There was no statistically significant difference in the survival rates of spermatozoa between Group 1 and Group 2 after cryopreservation. It appears that in vitro culture of testicular spermatozoa before freezing does not increase survival rate.

Animals↗

PubMed1980

Heavy metals and spermatozoa. 1. Inhibition of the motility and metabolism of spermatozoa by metals related to copper.

The toxicity to human spermatozoa of seven metals (nickel, palladium, platinum, silver, gold, zinc, and cadmium) and one alloy (brass: 80% copper, 20% zinc) related to copper was assessed in vitro. Only brass and cadmium significantly reduced the percentage of motile unwashed spermatozoa; however, washing the spermatozoa increased the spermicidal effectiveness of both brass and cadmium and also resulted in a significant reduction in motility caused by zinc and silver. Oxygen consumption by once-washed spermatozoa was apparently increased by zinc and brass, but the high rate of oxidation of these metals confounds interpretation of their effect. Silver caused a decline in the oxygen uptake of spermatozoa. Silver, zinc, brass, and, to a lesser extent, cadmium decreased the quantity of glucose utilized by spermatozoa and also decreased the glucose oxidized. Accumulation of lactate by washed spermatozoa was impaired severely by zinc and less severely by brass and cadmium.

Adult↗

PubMed1998

Morphology of spermatozoa in the cauda epididymidis before and after electroejaculation and a comparison with ejaculated spermatozoa in the domestic cat.

When 2 ejaculates are collected by electroejaculation from the domestic cat within a period of 10 min the first ejaculate has a higher proportion of abnormal spermatozoa than the second. The reason for this difference is not known for the domestic cat, but in other species long-term epididymal storage results in a higher proportion of abnormal spermatozoa. The aims of this study were to determine the proportions of abnormal spermatozoa in the cauda epididymidis and to ascertain if electroejaculation affects this proportion. Therefore the proportions of spermatozoa in the cauda epididymidis with different morphological abnormalities were compared before and after ejaculation. In addition, the proportion of morphologically abnormal spermatozoa in the epididymis was compared with that in the ejaculate. Nine privately-owned domestic cats were anesthetized, and one testicle was surgically removed. An ejaculate was collected by electroejaculation, after which the remaining testicle was ectomized. There were no significant differences in the proportions of different sperm abnormalities between the cauda epididymidis removed before ejaculation and the one removed after ejaculation. A significantly (P = 0.009) higher proportion of spermatozoa with tail abnormalities was found in the ejaculates compared with the cauda epididymides (11.1 and 1.6%, respectively), while, as expected, there was a lower proportion of spermatozoa with distal droplets in the ejaculates than in the cauda epididymides (35.1 and 75.9%, respectively). This new information contributes to the understanding of the etiology of sperm defects in the domestic cat, and is of importance when evaluating a semen sample in this species.

Animals↗