- Semen collection, semen analysis and artificial insemination in Columbian sharp-tailed grouse (Tympanuchus phasianellus columbianus) as part of a species conservation project. [Journal Article]
- TTheriogenology 2019 Jul 01; 132:128-137
- Columbian sharp-tailed grouse (Tympanuchus phasianellus columbianus; hereafter CSTG) have experienced substantial decreases in population numbers and geographic range during the early 20th century, p…
Columbian sharp-tailed grouse (Tympanuchus phasianellus columbianus; hereafter CSTG) have experienced substantial decreases in population numbers and geographic range during the early 20th century, primarily due to habitat loss. The conservation aim of this project was to re-establish a self-sustaining population of CSTG within an unoccupied portion of their historic range in northeastern Nevada via reintroduction from source populations in Idaho, USA. Female nest initiation rates post-translocation due to low fertilization rates are believed to be one limiting factor in the establishment of some translocated CSTG populations. However, studies on semen collection and artificial insemination in this species are absent. Assisted reproduction was evaluated as an additional tool in this species conservation project in order to gain knowledge on the reproductive status of yearling and adult male CSTG, establish orientation values for semen parameters and evaluate artificial insemination procedures on female CSTG. In two consecutive breeding seasons, semen collection was attempted 51 times in 47 males using the established massage method, and a novel electro-stimulation technique. Semen collection was successful in all attempts, even in yearling grouse, which represents a novel confirmation that yearling male CSTG can produce live spermatozoa in their first breeding season. Volume, color, consistency, contamination, pH of semen, and the motility, concentration, viability and morphology of spermatozoa were analyzed. Extracted semen volume ranged between 6 and 74 μl and the mean pH was 6.9 ± 0.5 (x¯ ± SD). Morphology analysis revealed that on average, 42.8% of sperm was morphologically normal, but 34.8% had malformed tails. Additionally, artificial insemination was practiced in 17 females (sham-insemination group; insemination lacking spermatozoa) and performed in 17 females. Intravenous catheters G20 1.0 mm × 32 mm enabled safe intravaginal insemination under visual control. Significant (p ≤ 0.05) differences in semen parameters between adult and yearling birds were detected. It is well established that adult males receive a majority of copulations during lekking, but our novel findings also indicate that they produce significantly more spermatozoa which is of higher quality than yearling males. This finding offers insights into male reproductive biology in a lekking grouse species.
- The incorporation of cystine by the soluble carrier family 7 member 11 (SLC7A11) is a component of the redox regulatory mechanism in stallion spermatozoa. [Journal Article]
- BRBiol Reprod 2019 Apr 18
- Oxidative stress is considered a major mechanism causing sperm damage during cryopreservation and storage, and underlies male factor infertility. Currently, oxidative stress is no longer believed to …
Oxidative stress is considered a major mechanism causing sperm damage during cryopreservation and storage, and underlies male factor infertility. Currently, oxidative stress is no longer believed to be caused only by the overproduction of reactive oxygen species, but rather by the deregulation of redox signaling and control mechanisms. With this concept in mind, here, we describe for the first time the presence of the soluble carrier family 7 member 11 (SLC7A11) antiporter, which exchanges extracellular cystine (Cyss) for intracellular glutamate, in stallion spermatozoa, as well as its impact on sperm function using the specific inhibitor sulfasalazine. Spermatozoa incubated with cystine exhibited an increased intracellular GSH content compared with controls (P < 0.01): 50% in fresh extended stallion spermatozoa and 30% in frozen-thawed spermatozoa. This effect was prevented by the addition of sulfasalazine to the media. Cyss supplementation also reduced the oxidation-reduction potential of spermatozoa, with sulfasalazine only preventing this effect on fresh spermatozoa that were incubated for three hours at 37°C, but not in frozen-thawed spermatozoa. While sulfasalazine reduced the motility of frozen-thawed spermatozoa, it increased motility in fresh samples. The present findings provide new and relevant data on the mechanism regulating the redox status of spermatozoa and suggest that a different redox regulatory mechanism exists in cryopreserved spermatozoa, thus providing new clues to improve current cryopreservation technologies and treat male factor infertility.
- A Rare Case of a Primary Unilateral Low-Grade Paratesticular Leiomyosarcoma in a 2 Years Old Dog. [Case Reports]
- FVFront Vet Sci 2019; 6:83
- A 2 years old dog was brought to the clinic with complains of testicular enlargement. The tissue was diffusely affected as confirmed by ultrasonographic examination, being the right testicle atrophie…
A 2 years old dog was brought to the clinic with complains of testicular enlargement. The tissue was diffusely affected as confirmed by ultrasonographic examination, being the right testicle atrophied and the right epididymis enlarged, with loss of echotexture and presence of several anechogenic areas. The situation required the excision of the referred testicle and epididymis. Final diagnose made by histopathological analysis was primary unilateral low-grade paratesticular leiomyosarcoma. Scarce bibliography is found on this matter, with several cases reported on human, and none in dog. This case report is therefore an important milestone on the area of small animal oncology directly related to the reproductive tissue.
- Effect of Different Extenders and Dilution Ratios on the Spermatological and Fertilization Parameters of Cyprinus carpio During Cryopreservation. [Journal Article]
- BBBiopreserv Biobank 2019; 17(2):157-162
- Production of quality seeds is a critical need, and cryopreservation serves as an important biotechnological tool in achieving that goal. In the present study, the effects of different extenders and …
Production of quality seeds is a critical need, and cryopreservation serves as an important biotechnological tool in achieving that goal. In the present study, the effects of different extenders and dilution ratios on the spermatological and fertilization parameters of cryopreserved milt were evaluated in common carp brooders. Milt was collected from Cyprinus carpio brooders and diluted and cryopreserved at three dilution ratios such as 1:40, 1:80, and 1:120, using Freshwater Fish Saline (FWFS) and Modified Fish Ringer as extenders and dimethyl sulfoxide as cryoprotectant (90:10). Motility duration, fertilization, and hatching rates were studied after different treatments. Rapid freezing of straws was done for 20 minutes at a freezing rate of -78°C/min. It can be concluded that C. carpio milt can be cryopreserved with FWFS at a dilution ratio of 1:40 to obtain the highest post-thaw motility duration (60.33 ± 1.52 seconds), fertilization rate (87.6% ± 1.52%), and hatching rate (57.1% ± 4.16%). The difference in the values between extenders, dilution ratios, and cryopreserved and fresh milt were statistically significant (p < 0.05) when analyzed using ANOVA.
- Depletion of thiols leads to redox deregulation, production of 4-hydroxinonenal and sperm senescence: a possible role for GSH regulation in spermatozoa. [Journal Article]
- BRBiol Reprod 2018 Nov 10
- Although redox regulation and oxidative stress are important concepts in spermatology, the molecular mechanisms behind these processes are poorly understood. Recent research has revealed that redox h…
Although redox regulation and oxidative stress are important concepts in spermatology, the molecular mechanisms behind these processes are poorly understood. Recent research has revealed that redox homeostasis is extremely important for the maintenance of sperm functionality in the stallion due to the high mitochondrial activity in the spermatozoa of this species. We hypothesized that thiols and particularly glutathione (GSH) are essential for the regulation of stallion sperm functionality. To test this hypothesis, we initially investigated the relationship between sperm function and GSH content, revealing highly significant correlations between GSH, sperm viability, motility and velocity parameters (p < 0.001). Furthermore, the deleterious effects of GSH depletion using menadione and 1,3 Dimethoxy 1,4, naphtoquinone (DMNQ) were able to be prevented by the addition of cysteine, but no other antioxidant. Pre-incubation with cysteine prevented menadione and DMNQ induced damage to sperm membranes after 1 h (viable sperm in control: 80%; menadione treated: 56%; both p < 0.001; pre-incubation with cysteine and treatment with menadione: 80%, controls 62%, DMNQ 44% and pre-incubation with cysteine and treatment with DMNQ 55% p < 0.05) and after 3 h of incubation (control: 78%; menadione: 30%; both p < 0.001, and pre-incubation with cysteine and treatment with menadione: 83%, controls 52.6%, DMNQ 45% and pre-incubation with cysteine and treatment with DMNQ 50% p < 0.05). Similar patterns were also observed for motility and sperm velocity parameters, and pre-incubation with cysteine ameliorated both the menadione- and DMNQ-induced increase in 4-hydroxynonenal (p < 0.001). As cysteine is a pre cursor of GSH, we hypothesized that stallion spermatozoa are able to synthesize this tri-peptide using exogenous cysteine. To test this hypothesis we investigated the presence of two enzymes required to synthesize GSH (Glutathione Synthetase and glutamate-cysteine ligase) and using western blotting and immunocytochemistry approaches we detected both enzymes in stallion spermatozoa. The inhibition of glutamate cysteine ligase reduced the recovery of GSH by addition of cysteine after depletion, suggesting that stallion spermatozoa may use exogenous cysteine to regulate GSH. Other findings supporting this hypothesis were changes in sperm functionality after BSO treatment and changes in GSH and GSSG validated using HPLC-MS, showing that BSO prevented the increase in GSH in presence of cysteine, although important stallion to stallion variability occurred and suggested differences in expression of glutamate cysteine ligase. Mean concentration of GSH in stallion spermatozoa was 8.2 ± 2.1 μM/109 spermatozoa, well above the nanomolar ranges per billion spermatozoa reported for other mammals. These novel findings reveal new avenues for the treatment of stallion infertility and for the development of better stallion sperm preservation technologies based on a more comprehensive understanding of redox regulation in these highly specialized cells.
- Male reproductive health statement (XIIIth international symposium on Spermatology, may 9th-12th 2018, Stockholm, Sweden. [Journal Article]
- BCBasic Clin Androl 2018; 28:13
- On the occasion of the XIIIth International Symposium on Spermatology held from 9 to 13 May 2018 in Stockholm (Sweden), participants (guest speakers and audience) collectively felt the need to make a…
On the occasion of the XIIIth International Symposium on Spermatology held from 9 to 13 May 2018 in Stockholm (Sweden), participants (guest speakers and audience) collectively felt the need to make a public statement on the general issue of male reproductive health. Our intention is to raise awareness of what we believe is a neglected area of research despite alarming situations around the world. The disclosure strategy desired by the co-authors is to bring it to the attention of the greatest number partly by considering co-publication in the various periodicals dealing with Reproductive Biology and Andrology. BaCA's editorial office accepted this mission and found it natural that our periodical, the official journal of the French Andrology Society (SALF), should carry this message.
- Flow cytometry analysis of spermatozoa: Is it time for flow spermetry? [Review]
- RDReprod Domest Anim 2018; 53 Suppl 2:37-45
- Flow cytometry is increasingly used in research and also in clinical andrology. Recent developments in instrumentation, availability of probes and bioinformatics expand the possibilities of flow cyto…
Flow cytometry is increasingly used in research and also in clinical andrology. Recent developments in instrumentation, availability of probes and bioinformatics expand the possibilities of flow cytometry well beyond the classical two parametric analyses in use. In this paper, an overview of recent developments in flow cytometry will be presented under the perspective of the authors; aspects such a multicolor assays and computational cytometry will be discussed as well.
- Ultrastructural, cytochemistry and electron tomography analysis of Caryophyllaeides fennica (Schneider, 1902) (Cestoda: Lytocestidae) reveals novel spermatology characteristics in the Eucestoda. [Journal Article]
- PRParasitol Res 2018; 117(10):3091-3102
- Spermatozoon formation in Caryophyllaeides fennica (Schneider, 1902) is characterised by the following: (1) apical electron-dense material in the zone of differentiation, (2) typical striated roots s…
Spermatozoon formation in Caryophyllaeides fennica (Schneider, 1902) is characterised by the following: (1) apical electron-dense material in the zone of differentiation, (2) typical striated roots situated unconventionally in opposite directions in early stages of spermiogenesis, (3) intercentriolar body composed of three electron-dense and two electron-lucent plates, (4) free flagellum and a flagellar bud that correspond to a greatly reduced flagellum and (5) rotation of free flagellum and a flagellar bud to the median cytoplasmic process at 90°. The development of two flagella of significantly unequal length clearly supports a derived form of spermiogenesis in the Caryophyllidea. New for cestodes is a finding of two additional striated roots situated opposite each other, in conjunction with both the flagellar bud and free flagellum. Mutual position of additional striated roots and typical striated roots is parallel in early stages and perpendicular in advanced stages of spermiogenesis. A complete proximodistal fusion gives rise to a mature spermatozoon consisting of one axoneme, parallel cortical microtubules, a nucleus and a moderately electron-dense cytoplasm with glycogen particles, detected by a technique of Thiéry (J Microsc 6:987-1018, 1967), in the principal regions (II, III, IV). Electron tomography analysis of the free flagellum and one axoneme of a mature spermatozoon of C. fennica provides clear evidence, for the first time, that two tubular structures are present in the central axonemal electron-dense core. Phylogenetically important aspects of spermiogenesis of the Caryophyllidea with one axoneme, and other cestodes with one or two axonemes, are briefly reviewed and discussed.
- Bottlenose dolphin (Tursiops truncatus) sperm revisited: Motility, morphology and ultrastructure of fresh sperm of consecutive ejaculates. [Journal Article]
- ARAnim Reprod Sci 2018 Jun 22
- Computer aided sperm analysis systems allow detailed examination of sperm motility and morphology variables, which are important for the understanding of the spermatology of a species and the develop…
Computer aided sperm analysis systems allow detailed examination of sperm motility and morphology variables, which are important for the understanding of the spermatology of a species and the development of assisted reproductive techniques. Cetacean biology is too complex to study in the wild and data from captive individuals provide an important alternative for the conservation of these charismatic animals. The present study evaluates ejaculate and sperm characteristics, including sperm motility, kinematic variables and quantitative sperm morphology and ultrastructure, of consecutive ejaculates from Atlantic bottlenose dolphins (Tursiops truncatus). Sperm concentrations and total and progressive motilities were greater in the second than the first ejaculate, with all ejaculates being of very high quality (6.9-1127 × 106/ml sperm concentration, 75% to 91% total motility and 89% to 96% normal sperm). Most sperm in an ejaculate (≥84%) were highly (VCL>150 μm/s) and progressively motile with very few abnormal sperm. The sperm have small heads, a short but very bulky midpiece and a long tail. Detailed sperm morphometrics using CASA indicated there were similarities from one ejaculate to the next. The large mitochondria with extensive cristae mitochondriales are tightly packed in the midpiece resulting in a large midpiece volume. All the semen and sperm characteristics indicate high quality sperm and support the assumption that a multimale mating system is present in T. truncatus.
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- Progesterone stimulates the long-distance migration of capacitated ram spermatozoa through viscous media under geotactic condition. [Journal Article]
- TTheriogenology 2018 Sep 15; 118:7-15
- Forward progressive motility of spermatozoa is an essential prerequisite for reproductive success, and sperm navigation is assisted by guidance mechanisms that may depend on micro-environmental facto…
Forward progressive motility of spermatozoa is an essential prerequisite for reproductive success, and sperm navigation is assisted by guidance mechanisms that may depend on micro-environmental factors. In the present study, we performed an integrated analysis of long-distance ram sperm migration in vitro that combined two environmental factors (10 μM progesterone and a geotactic effect) and the physiological status of the cells (capacitation treatment). A penetration assay was used in which spermatozoa had to travel 20 mm in a viscous medium (two media of differing viscosity: acrylamide and hyaluronic acid) through a tube device. The number of migrating spermatozoa, the physiology of the cells (motility analyzed using a CASA system; acrosomal status, viability and active mitochondria evaluated by flow cytometry; DNA fragmentation index calculated by quantitative PCR) and the morphometry of sperm heads (performed using an image analysis system) were evaluated after long-distance sperm migration. Ram sperm capacitation significantly stimulates cell migration through viscous media under geotactic conditions, and this effect is enhanced by progesterone induction. The rheological characteristics of viscous media have a marked impact on ram sperm migration, and acrylamide more favorably facilitates navigation over a large distance. The migrating spermatozoa are morphologically better adapted (high ellipticity) for displacement in viscous media and exhibit remarkably depleted mitochondrial membrane potential.