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Topic:Spermatogenesis

Spermatogenesis in horses is the process by which spermatozoa are produced in the testes. This complex sequence of events involves the proliferation and differentiation of germ cells, supported by Sertoli cells and influenced by hormonal regulation. Key stages include spermatogonial mitosis, meiosis, and spermiogenesis, resulting in the formation of mature sperm cells. Hormones such as testosterone and follicle-stimulating hormone (FSH) are integral to regulating this process. This topic encompasses research studies and scholarly articles that explore the mechanisms, hormonal influences, and factors affecting spermatogenesis in horses, as well as its implications for equine reproduction and breeding management.
A 5-color flow cytometry panel to assess plasma membrane integrity, acrosomal status, membrane lipid organization and mitochondrial activity of boar and stallion spermatozoa following liquid semen storage.
Animal reproduction science    September 23, 2022   Volume 247 107076 doi: 10.1016/j.anireprosci.2022.107076
Quirino M, Jakop U, Mellagi APG, Bortolozzo FP, Jung M, Schulze M.For a more practically applicable analysis of different sperm characteristics, this study aimed to develop a 5-color flow cytometry (FC) panel to concurrently analyze four sperm parameters in liquid boar and stallion semen, using also a DNA-marker for selecting sperm cell events. From each of thirty extended boar semen doses and twelve stallion semen doses, six aliquots were taken. For evaluating mitochondrial activity (A), degree of lipid disorder of plasma membrane (B), integrity of plasma membrane (C), acrosomal status (D) and marking DNA (E), five aliquots were individually stained with Rh...
Dataset of the sperm proteome of stallions with different motility.
Data in brief    September 5, 2022   Volume 45 108578 doi: 10.1016/j.dib.2022.108578
Gaitskell-Phillips G, Martín-Cano FE, Ortiz-Rodríguez JM, Silva-Rodríguez A, da Silva-Álvarez E, Gil MC, Ortega-Ferrusola C, Peña FJ.This paper provides a detailed set of data on how the stallion sperm proteome differs among stallions with different sperm motilities, although within normal ranges. Findings distinguish proteins that may help to identify stallions of superior sperm motility. Sperm proteins were analyzed using a UHPLC/MS/MS system comprising of an Agilent 1290 infinity series UHPLC coupled to an Agilent 6550 Q-TOF mass spectrometer (Agilent Technologies, Santa Clara, CA, USA). These data can be used to disclose potential targets to identify good sperm samples and to study specific pathways involved in the regu...
Seasonal changes in the expression of molecular markers of stallion germ cells.
Journal of equine veterinary science    August 24, 2022   Volume 118 104109 doi: 10.1016/j.jevs.2022.104109
Shakeel M, Jung H, Yoon D, Yoon M.The economic impacts of infertility and subfertility of stallions greatly influence the horse breeding industry. Self-renewal and differentiation of spermatogonial stem cells are the initial processes to maintain an adequate sperm population. Thus, understanding these processes may provide useful information to reveal the causes and remedies of subfertile and infertile stallions. Stallions are seasonal breeders. About 50% of the sperm population is reduced during the non-breeding season (NBS) in stallions. The seasonal regulation of spermatogenesis renders stallions as ideal models to understa...
Effects of Exercise on Donkey Semen Parameters and Some Physiological Indices.
Journal of equine veterinary science    July 16, 2022   Volume 117 104079 doi: 10.1016/j.jevs.2022.104079
Yang F, Wu S, Yang W, Yu J, Liu B, Zeng S.To study the effects of exercise on donkey semen parameters, Dezhou donkey (Equus asinus) jackasses (n = 18) were assigned to three groups: control (CN, n = 6), 1-hour exercise (1 h EX, n = 6) and 2-hour exercise (2 h EX, n = 6) groups. They were exercised with a walking machine for 3 days/week for 8 weeks. Semen parameters and reproductive hormones were evaluated weekly. The heart rate, rectal temperature, blood haematology indexes, and the body condition score (BCS) were also measured. Results showed that the ejaculation volume increased while the sperm concentration decreased after ...
Multispecies comparative analysis reveals transcriptional specificity during Mongolian horse testicular development.
Reproduction in domestic animals = Zuchthygiene    July 13, 2022   Volume 57, Issue 11 1295-1306 doi: 10.1111/rda.14203
Zhang FL, Zhang XY, Zhao JX, Zhu KX, Liu SQ, Zhang T, Sun YJ, Wang JJ, Shen W.Mongolian horses have been bred and used for labor and transport for centuries. Nevertheless, traits of testicular development in Mongolian horses have rarely been studied; particularly, studies regarding the transcriptional regulation characteristics of testicular development are lacking. In this paper, transcription specificity during testicular development in Mongolian horses is highlighted via a multispecies comparative analysis and weighted gene co-expression network analysis (WGCNA). Interestingly, the results showed that most genes were up-regulated in the testes after sexual maturity, ...
Coenzyme Q10 and melatonin protect cryopreserved equine sperm against lipoperoxidation.
Animal reproduction science    June 18, 2022   Volume 243 107027 doi: 10.1016/j.anireprosci.2022.107027
Nogueira BG, Pereira RR, Bitencourt JL, Milan B, Reis WVA, Junior MV, Acácio BR, Sampaio BFB, I L Souza M.The objective of the experiment was to evaluate the effect of the addition of different concentrations of the antioxidants Coenzyme Q10 (CoQ10) and melatonin to equine semen freezing diluent, alone or in combination, during the cryopreservation process. Twenty ejaculates (n = 5 stallions) were divided in groups: Control (C, without the addition of antioxidants), melatonin 0.75 mM (MEL1), melatonin 1.5 mM (MEL2), CoQ10 40 μg/mL (Q1), CoQ10 200 μg/mL (Q2), and CoQ10 40 μg/mL+ 0.75 mM melatonin (Q1 +MEL1). Q1 and Q2 groups demonstrated intact plasma membrane and high mitochondrial me...
Effects of an anti-gonadoliberin releasing hormone vaccine on testicular, epididymal and spermatogenic development in the horse.
Reproduction in domestic animals = Zuchthygiene    May 12, 2022   Volume 57, Issue 8 919-927 doi: 10.1111/rda.14141
Botha AE, Schulman ML, Birrell J, du Plessis L, Laver PN, Soley J, Colenbrander B, Bertschinger HJ.The effects of the GnRH vaccine Improvac on testicular and epididymal morphometrics, histology and spermatogenesis were measured in 19 young (15-20 months) colts randomly assigned to one control (saline, castration at 57 days, n = 6) or either of two GnRH vaccine-treatment groups, T-57 (castration at 57 days, n = 7) or T-100 (castration at 100 days, n = 6), respectively. All were immunized on Day 0 with a single booster on Day 28. Excised testes and epididymides were weighed and processed for histology to measure tubule, epithelial and muscle dimensions, the ratio of interstitial tis...
Evaluation of testicular echotexture with Ecotext as a diagnostic method of testicular dysfunction in stallions.
Theriogenology    March 16, 2022   Volume 185 50-60 doi: 10.1016/j.theriogenology.2022.03.004
Da Silva-Álvarez E, Gaitskell-Phillips G, Ortiz-Rodríguez JM, Serres C, García-Rodríguez B, Gutiérrez-Cepeda L, Martín-Cano FE, Echegaray A....This study aimed to assess if Ecotext, a new software for evaluation of testicular echotexture, is a good method for diagnosis of stallions with testicular dysfunction (TD). Relationships between Ecotext parameters and sperm motility and production, testicular volume, and testicular blood flow were also studied. Ecotext provides a total of six echotexture parameters: Ecotext 1 (black pixels), 2 (white pixels) and 3 (grey pixels), and another 3 parameters related to hypoechogenic areas: Ecotext tubular density (ETD), Ecotext tubular diameter (ETd), and Ecotext tubular area (ETA). Stallions (n ...
Deep-horn Artificial Insemination With Frozen Thawed Semen After re-extension in Autologous Seminal Plasma May Improve Pregnancy Rates in Jennies.
Journal of equine veterinary science    March 10, 2022   Volume 112 103932 doi: 10.1016/j.jevs.2022.103932
Fanelli D, Tesi M, Monaco D, Diaz-Jimenez M, Camillo F, Rota A, Panzani D.In the literature, the very low pregnancy rates after artificial insemination (AI) with frozen semen in donkeys were improved in one study after re-extension of the frozen-thawed semen in autologous seminal plasma. The aims of our study were (1) to describe in vitro post-thaw parameters of donkey jackass semen after re-extension in seminal plasma (SP) or in INRA96 and (2) to compare pregnancy rates in jennies bred with frozen-thawed semen using two different AI protocols. Semen collected from two Amiata donkey stallions, known to be fertile, was frozen in INRA96 supplemented with 2% egg yolk, ...
35 The effect of different cooled storage protocols on frozen-thawed equine semen.
Reproduction, fertility, and development    March 2, 2022   Volume 34, Issue 2 252-253 doi: 10.1071/RDv34n2Ab35
van Heule M, Verstraete MH, Blockx Z, De Blende P, Dini P, Daels P.No abstract available
Is stallion epididymal fluid phosphoproteome affected by the equine reproductive season?
Polish journal of veterinary sciences    February 19, 2022   Volume 24, Issue 4 487-495 doi: 10.24425/pjvs.2021.139973
Dyrda K, Orzołek A, Ner-Kluza J, Wysocki P.Phosphorylation and dephosphorylation of proteins are considered to be the most important processes in sperm maturation during epididymal transit. The main aim of this study was to isolate and identify phosphoproteins from the epididymal milieu obtained from reproductively mature stallions during and out of the breeding season. With the use of 1D-PAGE and nanoLC-MS/MS, we identified phosphoproteins that fulfil various functions: regulatory, transport, motility, ubiquitination, chaperone, antioxidant, apoptotic and enzymatic. Moreover, we characterized tyrosine, serine and threonine phosphoryla...
Use of biochemical and protein profiles of seminal plasma to prediction of semen quality and fertility in stallions.
Polish journal of veterinary sciences    February 19, 2022   Volume 24, Issue 4 505-514 doi: 10.24425/pjvs.2021.139975
Stelletta C, Alberti S, Cil B, Tekin K, Tirpan MB, Arganaraz M, Akcay E, Daskin A.The identification of various substances in seminal plasma has opened the way to study their functionality. It was aimed to identify the electrophoretic protein profile (EPP) and biochemical parameters (BP) of seminal plasma (SP) as predictors of semen quality and fertility in stallion. Forty-six ejaculates from 7 fertile stallions, aged between 6-26 years, were collected from May to July and 117 mares were used to obtain fertility data. For each ejaculate, volume, sperm motility, concentration were determined and seminal plasma samples were collected to perform one- -dimensional electrophores...
What is the relevance of seminal plasma from a functional and preservation perspective?
Animal reproduction science    February 17, 2022   Volume 246 106946 doi: 10.1016/j.anireprosci.2022.106946
Andrade AFC, Knox RV, Torres MA, Pavaneli APP.When preserving sperm in the liquid or cryopreserved state, seminal plasma (SP) components within ejaculates can alter fertilizing capacity of these gametes. Depending on the species or how semen is collected, volume and concentration of SP components varies considerably. The SP contains substances essential for maintenance of sperm viability and fertility; however, these components can be deleterious depending on quantity, or duration of time before there is removal of SP from sperm in semen processing. Substances that impair (e.g., BSP - bull; HSP-1 - stallion; Major seminal plasma protein P...
Use of Equine Sperm Cryopreservation Techniques as a Conservation Method of Donkey Germplasm.
Journal of equine veterinary science    January 29, 2022   Volume 111 103888 doi: 10.1016/j.jevs.2022.103888
Monteiro RA, Cunha RM, Guerra MMP, de Almeida VM, Peña-Alfaro CE, Silva SV.The aim of this study was to test equine semen cryopreservation techniques for the conservation of donkey germplasm. Ejaculates of three male donkeys were used (n = 18; six ejaculates per donkey; six repetitions), collected by the artificial vagina method. To remove the seminal plasma, the ejaculates were split and submitted to filtration or centrifugation methods. To assess the freezing method, each fraction was submitted to the automated system or the conventional system, and groups were formed: automated centrifuge, automated filtrate, conventional centrifuge and conventional filtrate. Af...
Proteomic analysis of spermatozoa reveals caseins play a pivotal role in preventing short-term periods of subfertility in stallions†.
Biology of reproduction    January 14, 2022   Volume 106, Issue 4 741-755 doi: 10.1093/biolre/ioab225
Griffin RA, Swegen A, Baker MA, Ogle RA, Smith N, Aitken RJ, Skerrett-Byrne DA, Fair S, Gibb Z.Stallions experience transient fluctuations in fertility throughout the breeding season. Considering pregnancy diagnoses cannot be ascertained until ~14 days postbreeding, the timely detection of decreases in stallion fertility would enhance industry economic and welfare outcomes. Therefore, this study aimed to identify the proteomic signatures reflective of short-term fertility fluctuations and to determine the biological mechanisms governing such differences. Using liquid chromatography-mass spectrometry (LC-MS/MS), we compared the proteomic profile of semen samples collected from commercia...
An Exploration of Current and Perspective Semen Analysis and Sperm Selection for Livestock Artificial Insemination.
Animals : an open access journal from MDPI    December 15, 2021   Volume 11, Issue 12 doi: 10.3390/ani11123563
Zuidema D, Kerns K, Sutovsky P.Artificial insemination of livestock has been a staple technology for producers worldwide for over sixty years. This reproductive technology has allowed for the rapid improvement of livestock genetics, most notably in dairy cattle and pigs. This field has experienced continuous improvements over the last six decades. Though much work has been carried out to improve the efficiency of AI, there are still many areas which continue to experience improvement, including semen analysis procedures, sperm selection techniques, sperm sexing technologies, and semen storage methods. Additionally, the use ...
Purification, molecular characterization and ligand binding properties of the major donkey seminal plasma protein DSP-1.
International journal of biological macromolecules    December 1, 2021   Volume 194 213-222 doi: 10.1016/j.ijbiomac.2021.11.177
Alim S, Cheppali SK, Laitaoja M, Talluri TR, Jänis J, Swamy MJ.Fibronectin type-II (FnII) family proteins are the major proteins in many mammalian species including bull, horse and pig. In the present study, a major FnII protein has been identified and isolated from donkey (Equus hemionus) seminal plasma, which we refer to as Donkey Seminal Plasma protein-1 (DSP-1). The amino acid sequence determined by mass spectrometry and computational modeling studies revealed that DSP-1 is homologous to other mammalian seminal plasma proteins, including bovine PDC-109 (also known as BSP-A1/A2) and equine HSP-1/2. High-resolution LC-MS analysis indicated that the prot...
Molecular Cytogenetic and Y Copy Number Analysis of a Reciprocal ECAY-ECA13 Translocation in a Stallion with Complete Meiotic Arrest.
Genes    November 26, 2021   Volume 12, Issue 12 doi: 10.3390/genes12121892
Castaneda C, Ruiz AJ, Tibary A, Raudsepp T.We present a detailed molecular cytogenetic analysis of a reciprocal translocation between horse (ECA) chromosomes Y and 13 in a Friesian stallion with complete meiotic arrest and azoospermia. We use dual-color fluorescence in situ hybridization with select ECAY and ECA13 markers and show that the translocation breakpoint in ECAY is in the multicopy region and in ECA13, at the centromere. One resulting derivative chromosome, Y;13p, comprises of ECAY heterochromatin (ETSTY7 array), a small single copy and partial Y multicopy region, and ECA13p. Another derivative chromosome 13q;Y comprises of E...
Characteristics of diluted-stored and post-thawed semen of Hutsul stallions.
Acta veterinaria Hungarica    November 25, 2021   doi: 10.1556/004.2021.00050
Bugno-Poniewierska M, Kij-Mitka B, Podstawski Z, Długosz B, Lasek O, Mrowiec P, Tischner M.The use of frozen semen lowers the risk of disease transmission, eliminates geographical limitations and supports the implementation of genetic resource protection programs. However, due to the very rare use of frozen semen from Hutsul stallions, their genetic material is not secured in sperm banks, and very little information is available about their semen, including its suitability for cryopreservation, and sperm survival rates after thawing. The aim of this study was to analyse basic parameters such as sperm motility, vitality and morphology in diluted-stored and post-thawed Hutsul semen, u...
An Update on Semen Physiology, Technologies, and Selection Techniques for the Advancement of In Vitro Equine Embryo Production: Section II.
Animals : an open access journal from MDPI    November 20, 2021   Volume 11, Issue 11 doi: 10.3390/ani11113319
Orsolini MF, Meyers SA, Dini P.As the use of assisted reproductive technologies (ART) and in vitro embryo production (IVP) expand in the equine industry, it has become necessary to further our understanding of available semen selection techniques. This segment of our two-section review will focus on the selection of spermatozoa based on quality and sex for equine intracytoplasmic sperm injection (ICSI), as well as current and future developments in sperm sorting technologies. Ultimately, novel methods of semen selection will be assessed based on their efficacy in other species and their relevance and future application towa...
An Update on Semen Physiology, Technologies, and Selection Techniques for the Advancement of In Vitro Equine Embryo Production: Section I.
Animals : an open access journal from MDPI    November 13, 2021   Volume 11, Issue 11 doi: 10.3390/ani11113248
Orsolini MF, Meyers SA, Dini P.As the use of assisted reproductive technologies (ART) and in vitro embryo production (IVP) expand in the equine industry, it has become necessary to further our understanding of semen physiology as it applies to overall fertility. This segment of our two-section review will focus on normal sperm parameters, beginning with development and extending through the basic morphology of mature spermatozoa, as well as common issues with male factor infertility in IVP. Ultimately, the relevance of sperm parameters to overall male factor fertility in equine IVP will be assessed.
Effect of Mare Age on Transcript Abundance of Connexins-37 and -43, Zona Pellucida Proteins, and Sperm Binding.
Journal of equine veterinary science    October 21, 2021   Volume 108 103796 doi: 10.1016/j.jevs.2021.103796
Bresnahan DR, Lupole RE, Stilz CR, Carnevale EM.Zona pellucida (ZP) proteins are important for fertilization and sperm binding and are closely associated with cumulus cells. Communication between cumulus and oocytes is facilitated by intracellular membrane channels composed of connexins. The extent aging impacts potential differences in fertilization and reductions in fertility is not well understood. This study characterized age-related differences in transcript abundance of ZP proteins and connexins in cells from ovarian follicles. Additionally, differences in sperm binding to oocytes from old and young mares was evaluated. For experiment...
A Novel QTL and a Candidate Gene Are Associated with the Progressive Motility of Franches-Montagnes Stallion Spermatozoa after Thaw.
Genes    September 25, 2021   Volume 12, Issue 10 1501 doi: 10.3390/genes12101501
Gmel AI, Burger D, Neuditschko M.The use of frozen-thawed semen is an important reproduction tool to preserve the biodiversity of small, native horse breeds such as the Franches-Montagnes (FM). However, not all stallions produce cryotolerant semen with a progressive motility after thaw ≥ 35%. To improve our understanding of the genetic background of male fertility traits in both fresh and frozen-thawed semen, we performed genome-wide association studies (GWAS) on gel-free volume, sperm cell concentration, total sperm count, and progressive motility in fresh and frozen-thawed semen from 109 FM stallions using 335,494 genome-...
Evaluation of Stallion Testicular Cell Types by Flow Cytometry.
Journal of equine veterinary science    September 24, 2021   Volume 107 103778 doi: 10.1016/j.jevs.2021.103778
Serafini R, Varner DD, Love CC.Flow cytometry procedures can be used for evaluation of both spermatogenic efficiency and diagnose disorders of stallion spermatogenesis. Aims of this study were to compare two testicular sample acquisition techniques (needle aspirate-N and tissue wedge-T) and results when using flow cytometry and histology procedures. Testicular cell types were stained with acridine orange, and nine regions (R2 to R10) were identified and enumerated following acquisition by either N or T. Testes were also grouped and analyzed by size and sexual maturity (Small [immature] compared with Large [mature]) and used...
Extracellular Reactive Oxygen Species (ROS) Production in Fresh Donkey Sperm Exposed to Reductive Stress, Oxidative Stress and NETosis.
Antioxidants (Basel, Switzerland)    August 27, 2021   Volume 10, Issue 9 1367 doi: 10.3390/antiox10091367
Yánez-Ortiz I, Catalán J, Mateo-Otero Y, Dordas-Perpinyà M, Gacem S, Yeste N, Bassols A, Yeste M, Miró J.Jenny shows a large endometrial reaction after semen influx to the uterus with a large amount of polymorphonuclear neutrophils (PMN) migrating into the uterine lumen. PMN act as a sperm selection mechanism through phagocytosis and NETosis (DNA extrudes and, together with proteins, trap spermatozoa). While a reduced percentage of spermatozoa are phagocytosed by PMN, most are found to be attached to neutrophil extracellular traps (NETs). This selection process together with sperm metabolism produces a large amount of reactive oxygen species (ROS) that influence the reproductive success. The pres...
Germ Cell Transplantation in Stallion Testes.
Journal of equine veterinary science    August 21, 2021   Volume 106 103748 doi: 10.1016/j.jevs.2021.103748
Jung H, Yoon M.The production of donor-derived sperm using spermatogonial stem cell transplantation has been studied in various animals including mice, rats, goats, boar, dogs, sheep, and monkeys. However, germ cell transplantation has not been applied in stallions. The objective of this study was to produce donor germ cell-derived sperm using germ cell transplantation in stallions. Donor germ cells were transplanted into the parenchyma of 3 recipient stallions that had been treated with busulfan IV injections of 15 mg/kg body weight. For the preparation of donor single germ cells, tissue (20 g) from each te...
Sexual Differentiation and Primordial Germ Cell Distribution in the Early Horse Fetus.
Animals : an open access journal from MDPI    August 17, 2021   Volume 11, Issue 8 2422 doi: 10.3390/ani11082422
Scarlet D, Handschuh S, Reichart U, Podico G, Ellerbrock RE, Demyda-Peyrás S, Canisso IF, Walter I, Aurich C.It was the aim of this study to characterize the development of the gonads and genital ducts in the equine fetus around the time of sexual differentiation. This included the identification and localization of the primordial germ cell population. Equine fetuses between 45 and 60 days of gestation were evaluated using a combination of micro-computed tomography scanning, immunohistochemistry, and multiplex immunofluorescence. Fetal gonads increased in size 23-fold from 45 to 60 days of gestation, and an even greater increase was observed in the metanephros volume. Signs of mesonephros atrophy wer...
Cooled storage of semen from livestock animals (part I): boar, bull, and stallion.
Animal reproduction science    August 12, 2021   Volume 246 106822 doi: 10.1016/j.anireprosci.2021.106822
Wiebke M, Hensel B, Nitsche-Melkus E, Jung M, Schulze M.This review is part of the Festschrift in honor of Dr. Duane Garner and provides an overview of current techniques for cooled storage of semen from livestock animals. The first part describes the current state of the art of liquid semen preservation in boars, bulls, and stallions, including the diluents, use of additives, processing, temperature, and cooling of semen. The species-specific physiology and varying extents of cold shock sensitivity are taken into consideration. In addition, factors influencing the quality of cooled-stored semen are discussed. Methods, trends, and the most recent a...
The Proteome of Equine Oviductal Fluid Varies Before and After Ovulation: A Comparative Study.
Frontiers in veterinary science    August 5, 2021   Volume 8 694247 doi: 10.3389/fvets.2021.694247
Fernández-Hernández P, Marinaro F, Sánchez-Calabuig MJ, García-Marín LJ, Bragado MJ, González-Fernández L, Macías-García B.Equine fertilization cannot be performed in the laboratory as equine spermatozoa do not cross the oocyte's zona pellucida . Hence, a more profound study of equine oviductal fluid (OF) composition at the pre-ovulatory and post-ovulatory stages could help in understanding what components are required to achieve fertilization in horses. Our work aimed to elucidate the proteomic composition of equine OF at both stages. To do this, OF was obtained postmortem from oviducts of slaughtered mares ipsilateral to a pre-ovulatory follicle ( = 4) or a recent ovulation ( = 4); the samples were kept at -80°...
Andrological and cytogenetic investigations of an infertile Przewalski’s stallion.
Acta veterinaria Hungarica    July 22, 2021   Volume 69, Issue 2 189-193 doi: 10.1556/004.2021.00027
Kútvölgyi G, Brabender K, Andersson M, Javkhlan A, Nagy S, Páble T, Egerszegi I, Hidas A, Soós I, Kovács A.The case of an 8-year-old, sexually active but infertile Przewalski's stallion (Equus ferus przewalskii) was studied. Besides the infertility, the stallion also showed permanent problems with its body condition, being obviously weaker than all the other group members. The horse was kept in a separate place for two years with 12 mares in its harem group (six mares had foals earlier); however, none of the mares covered got pregnant. Andrological and cytogenetic investigations revealed underdeveloped testes, arrested spermatogenesis, azoospermia, and XY/XXY/X0 mosaicism. We classify the case as a...