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Sperm preparation

Wei Zhou, Geoffry N De Iuliis, Matthew D Dun, Brett Nixon
The testicular spermatozoa of all mammalian species are considered functionally immature owing to their inability to swim in a progressive manner and engage in productive interactions with the cumulus-oocyte complex. The ability to express these key functional attributes develops progressively during the cells' descent through the epididymis, a highly specialized ductal system that forms an integral part of the male reproductive tract. The functional maturation of the spermatozoon is achieved via continuous interactions with the epididymal luminal microenvironment and remarkably, occurs in the complete absence of de novo gene transcription or protein translation...
2018: Frontiers in Endocrinology
Katarzyna Brach, Akiko Hatakeyama, Claude Nogues, Joanna Olesiak-Banska, Malcolm Buckle, Katarzyna Matczyszyn
The anisotropic shape of DNA molecules allows them to form lyotropic liquid crystals (LCs) at high concentrations. This liquid crystalline arrangement is also found in vivo (e.g., in bacteriophage capsids, bacteria or human sperm nuclei). However, the role of DNA liquid crystalline organization in living organisms still remains an open question. Here we show that in vitro, the DNA spatial structure is significantly changed in mesophases compared to non-organized DNA molecules. DNA LCs were prepared from pBluescript SK (pBSK) plasmid DNA and investigated by photochemical analysis of structural transitions (PhAST)...
March 14, 2018: Scientific Reports
Daigo Inoue, Joachim Wittbrodt, Oliver J Gruss
Centrosomes are the main microtubule organizing centers in animal cells. In particular during embryogenesis, they ensure faithful spindle formation and proper cell divisions. As metazoan centrosomes are eliminated during oogenesis, they have to be reassembled upon fertilization. Most metazoans use the sperm centrioles as templates for new centrosome biogenesis while the egg's cytoplasm re-prepares all components for on-going centrosome duplication in rapidly dividing embryonic cells. We discuss our knowledge and the experimental challenges to analyze zygotic centrosome reformation, which requires genetic experiments to enable scrutinizing respective male and female contributions...
March 9, 2018: BioEssays: News and Reviews in Molecular, Cellular and Developmental Biology
Mark D Timken, Sonja B Klein, Martin R Buoncristiani
The efficacy of a DNA differential extraction procedure relies on reducing the amount of non-sperm female DNA carryover into the sperm fraction, while providing a sufficient recovery of male DNA from the sperm cell component. A standard approach to this extraction is to use a mild initial lysis step to digest the female (epithelial cell) component in the mixture, followed by a series of centrifugation and wash steps to further purify the resulting sperm-pellet fraction. This sperm fraction is then digested in the presence of a chemical reducing agent in preparation for DNA extraction...
February 21, 2018: Forensic Science International. Genetics
Jane-Fang Yu, Yu-Hua Lai, Tse-En Wang, Yu-Syuan Wei, Yu-Jia Chang, Sheng-Hsiang Li, Shih-Chien Chin, Radhika Joshi, Hui-Wen Chang, Pei-Shiue Tsai
BACKGROUND: Semen from the chimpanzee species becomes a colloidal solid after ejaculation. The formation of this copulatory plug is believed to prevent additional spermatozoa of subsequent mating events from accessing the ova. However, this naturally preserved strategy hampers the processes for sperm preparation. In this study, we investigated whether collagenase can be used to degelify the semen plug and accelerate the semen liquefaction process in zoo captive chimpanzee species (Pan troglodytes)...
February 27, 2018: BMC Veterinary Research
Eulália M L da Silva, Susannah Rankin
Chromosome structure in both interphase and M-phase cells is strongly influenced by the action of the cohesin and condensin protein complexes. The cohesin complex tethers the identical copies of each chromosome, called sister chromatids, together following DNA replication and promotes normal interphase chromosome structure and gene expression. In contrast, condensin is active largely in M phase and promotes the compaction of individual chromosomes. The Xenopus egg extract system is uniquely suited to analyze the functions of both complexes...
February 23, 2018: Cold Spring Harbor Protocols
Mohamad Irani, Stephen Chow, Derek Keating, Simone Elder, Zev Rosenwaks, Gianpiero Palermo
The objective of this study was to identify sperm score thresholds to achieve satisfactory intrauterine insemination (IUI) success rates according to the response to stimulation with clomiphene citrate (CC). To minimize the confounding effect of female age, we included only CC/IUI cycles of women ≤35 years old. A total of 1,194 CC/IUI cycles were included. Semen volume, concentration, and motility influenced the clinical pregnancy rate (CPR). Normal morphology (≥4%) was associated with a comparable CPR with 3%, 2%, and 1% normal forms (15...
February 20, 2018: Gynecological Endocrinology
Floyd D Wilson, Deirdre I Johnson, Danny L Magee, Frederic J Hoerr
Testicular histomorphometric methods were developed for the investigation of fertility and hatchability problems in commercial broiler-breeder roosters. Sertoli cell (SC) evaluations were emphasized in view of their role in influencing embryonic testicular organogenesis and spermatogenesis in adults. H&E stained sections prepared from formalized tissues were examined. SC evaluations were mostly made from computer generated images using image analysis software, but were also compared to direct counting with ocular grid reticle...
February 14, 2018: Poultry Science
J W Lee, J H Cha, S H Shin, H J Cha, J H Kim, C K Park, K A Pak, J S Yoon, S Y Park
This study was to investigate whether the sexual abstinence period (SAP) recommended by the World Health Organization (WHO) affects clinical outcomes. We compared the rate of clinical outcomes between 2-7 and ≥8 days of SAP in first fresh embryo transfer after intracytoplasmic sperm injection (ICSI) in groups of young maternal age (YMA: <38 years) and old maternal age (OMA: ≥38 years). We conducted a retrospective study of 449 first ICSI cycles with a normal ovarian response. SAP was identified before collecting the semen samples...
February 20, 2018: Andrologia
Toru Hyakutake, Kotaro Mori, Koichi Sato
Mammalian spermatozoa in organisms with internal fertilization are required to swim in the cervical and oviductal mucus, whose rheological properties differ substantially from those of water. Moreover, on the way to the oviduct, a change in sperm motility called hyperactivation may occur. In the present study, we focused on the motion characteristics of hyperactivated bovine sperm and investigated the effect of the surrounding fluid on motility. We prepared two kinds of polyacrylamide with high-viscosity non-Newtonian fluid properties, similar to the actual cervical and oviductal mucus...
February 11, 2018: Journal of Biomechanics
Nisar A Wani, Seungbum Hong
Experiments were conducted to investigate the development of in vitro matured camel oocytes after their intra-cytoplasmic sperm injection (ICSI) with epididymal sperm collected from slaughtered male camels. Ovaries and testes were collected from a local slaughterhouse in normal saline solution (NSS) at 37 °C and on ice (0-1 °C), respectively. Cumulus-oocyte complexes (COCs) aspirated from the follicles were randomly distributed to 4-well culture plates (20-25 COCs/well) containing 500 μL of maturation medium and cultured at 38...
February 8, 2018: Theriogenology
I Ahmed, S Abdelateef, M Laqqan, H Amor, M A Abdel-Lah, M E Hammadeh
The purpose of this study was to determine influence of extended incubation time on sperm chromatin condensation and DNA strand breaks and their effect on fertilisation rate. Forty couples undergoing ICSI therapy were included. Semen was prepared by PureSperm gradient centrifugation and divided into two parts. The first part (G1) was used immediately for ICSI, whereas the second part (G2) was kept in the incubator at 37°C, 5% and 90% Humidity for 5 hr, and thereafter, the capacitated spermatozoa were used for ICSI...
February 14, 2018: Andrologia
Mario Binelli, Angela Maria Gonella-Diaza, Fernando Silveira Mesquita, Claudia Maria Bertan Membrive
In cattle, the oviduct is a tubular organ that connects the ovary and the uterus. The oviduct lumen stages a dynamic set of cellular and molecular interactions to fulfill the noble role of generating a new individual. Specific anatomical niches along the oviduct lumen provide the appropriate microenvironment for final sperm capacitation, oocyte capture and fertilization, and early embryo development and transport. To accomplish such complex tasks, the oviduct undergoes spatially and temporally-regulated morphological, biochemical, and physiological changes that are associated with endocrine events of the estrous cycle...
February 2, 2018: Biology
U Punjabi, D De Neubourg, H Van Mulders, W Cassauwers, K Peeters
To validate semen preparation via density gradient centrifugation, we took into account the input via the semen sample, the action generated by technical and equipment characteristics and the output measured by the level of performance. Longer periods of abstinence reduced % yield, but increased viscosity and incomplete samples collected had no effect. Under controlled technical and equipment characteristics, precision and reproducibility were validated for density gradient. Additionally, as a good laboratory practice, internal and external quality control measures were implemented to guarantee the level of performance...
February 2, 2018: Andrologia
Xiaoli Chen, Mingyuan Liang, Dong Wang
The preparation of spermatogonial stem cell (SSC) transplant recipients laid the technical foundation for SSC transplant technology and the understanding of spermatogenesis mechanisms. Busulfan is commonly used to prepare recipients for mouse SSC transplantation; however, its safety and efficiency have been questioned. This review summarizes the relationship between SSCs and Sertoli cells (SCs), and the mechanism of busulfan toxicity against sperm cells. We concluded that the proliferation, differentiation, and apoptosis of SSCs are regulated by SCs...
January 19, 2018: Cytotechnology
Gideon O Alade, Jones O Moody, Olanrewaju R Awotona, Saburi A Adesanya, Daowan Lai, Peter Proksch
AIM: To isolate the spermicidal constituents of Sacoglottis gabonensis. MATERIALS AND METHODS: The ethanolic extract with partitioned fractions of Sacoglottis gabonensis stem bark were subjected to sperm immobilization assay. The most active EtOAc fraction was further purifi ed by column and Semi-Preparative High Performance Liquid Chromatography to give compounds which were characterized by spectroscopic methods (UV, LC/MS, and NMR). The compound(s) was also tested for sperm immobilization activity...
December 20, 2017: Folia Medica
Şule Ayla, Gülden Tunalı, Bülent E Bilgiç, Kenan Sofuoğlu, A Arman Özdemir, Gamze Tanrıverdi, Semra Özdemir, B Cem Soner, Bahar Öztürk, Serçin Karahüseyinoğlu, Esra Güler Aslan, Ismail Seçkin
PURPOSE: Sperm processing (e.g., centrifugation) used in preparation for assisted reproduction can result in excessive generation of reactive oxygen species (ROS) and potential sperm damage. The use of antioxidants during sperm processing has been shown to prevent iatrogenic sperm damage, including DNA damage. In this study, we evaluated the effect of caffeic acid phenethyl ester (CAPE) on oxidative stress mediated sperm dysfunction and DNA damage. METHODS: Semen samples were obtained to liquefy at room temperature...
January 8, 2018: Acta Histochemica
Kiran Kumar Soni, Li Tao Zhang, Bo Ram Choi, Keshab Kumar Karna, Jae Hyung You, Yu Seob Shin, Sung Won Lee, Chul Young Kim, Chen Zhao, Han-Jung Chae, Hye Kyung Kim, Jong Kwan Park
CONTEXT: MOTILIPERM was prepared as a mixture of extracts of three medicinal herbs [roots of Morinda officinalis How (Rubiaceae), outer scales of Allium cepa L. (Liliaceae) and seeds of Cuscuta chinensis Lamark (Convolvulaceae)]. OBJECTIVE: To investigate the role of reactive oxygen species (ROS)-based endoplasmic reticulum (ER) stress in a rat model of varicocele and the therapeutic efficacy of MOTILIPERM in this model. MATERIALS AND METHODS: Sixty male rats were divided into five experimental groups: a normal control group (CTR + vehicle), a control group administered MOTILIPERM 200 mg/kg (CTR + M 200), a varicocele-induced control group (VC + vehicle) and two varicocele-induced groups administered MOTILIPERM 100 (VC + M 100) or 200 (VC + M 200) mg/kg for 4 weeks...
December 2018: Pharmaceutical Biology
M K Panner Selvam, R Henkel, R Sharma, A Agarwal
Oxidation-reduction potential describes the balance between the oxidants and antioxidants in fluids including semen. Various artificial culture media are used in andrology and IVF laboratories for sperm preparation and to support the development of fertilized oocytes under in vitro conditions. The composition and conditions of these media are vital for optimal functioning of the gametes. Currently, there are no data on the status of redox potential of sperm processing and assisted reproduction media. The purpose of this study was to compare the oxidation-reduction potential values of the different media and to calibrate the oxidation-reduction potential values of the sperm wash medium using oxidative stress inducer cumene hydroperoxide and antioxidant ascorbic acid...
January 3, 2018: Andrology
Monika A Ward, Ryuzo Yanagimachi
Intracytoplasmic sperm injection (ICSI) is the injection of a single spermatozoon directly into the cytoplasm of an oocyte using an injection pipette. Spermatozoa that are used for ICSI do not need to be motile or alive. Indeed, for mouse ICSI, the sperm tail is typically broken off before injection into the oocyte, resulting essentially in a nuclear transfer. The protocol describes creation of the injection pipettes, preparation of oocytes and spermatozoa, and steps of the ICSI procedure. The technique can be used to create both hybrid and inbred mice, as well as transgenic animals...
January 2, 2018: Cold Spring Harbor Protocols
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