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Sperm preparation

Z G Cai, J H An, Y L Liu, S M Yie, Y Zhang, F P Li, J S Chen, X Wang, J M Morrell, R Hou
The reproductive capacity of captive giant pandas is poor and sperm cryopreservation is necessary for the reproduction and conservation of this species. Cryopreservation, however, leads to a significant decrease in sperm quality, including sperm motility, acrosome integrity and DNA integrity. In the present study, a method was developed based on colloid single layer centrifugation that could significantly improve frozen-thawed sperm quality. Two colloids were compared for post-thaw giant panda sperm preparation; the sperm samples had greater total motility (Colloid 1: 44...
May 15, 2018: Animal Reproduction Science
Justin Sparks, Johannes C Walter
Frog egg extracts represent a powerful approach with which to dissect molecular mechanisms of vertebrate DNA replication and repair. In the classical approach, sperm chromatin is added to a crude egg lysate to form replication-competent nuclei. We subsequently described a procedure that bypasses the requirement for nuclear assembly in DNA replication. In this method, DNA is first added to a high-speed supernatant (HSS) of egg lysate, which mimics the G1 phase of the cell cycle in that it supports replication licensing...
May 16, 2018: Cold Spring Harbor Protocols
Béatrice Horard, Laure Sapey-Triomphe, Emilie Bonnefoy, Benjamin Loppin
BACKGROUND: Anti-Silencing Factor 1 (ASF1) is a conserved H3-H4 histone chaperone involved in both Replication-Coupled and Replication-Independent (RI) nucleosome assembly pathways. At DNA replication forks, ASF1 plays an important role in regulating the supply of H3.1/2 and H4 to the CAF-1 chromatin assembly complex. ASF1 also provides H3.3-H4 dimers to HIRA and DAXX chaperones for RI nucleosome assembly. The early Drosophila embryo is an attractive system to study chromatin assembly in a developmental context...
May 11, 2018: Epigenetics & Chromatin
Karina Radefeld, Sophie Papp, Vitezslav Havlicek, Jane M Morrell, Gottfried Brem, Urban Besenfelder
Conventionally inseminated spermatozoa suffer a dramatic reduction in numbers during their long journey until fertilization. In addition sperm survival seems to be strongly affected by the reconstitution of the female reproductive tract in the post partum period. The purpose of this study was to develop a novel AI technique for cattle that allows the deposition of spermatozoa directly into the ampulla in the immediate vicinity of the fertilization site. This new reproductive biotechnique was investigated with focus on semen origin, sperm dosage, semen preparation and time of insemination...
April 27, 2018: Theriogenology
Akram Hosseini, Mohammad Ali Khalili, Ali Reza Talebi, Azam Agha-Rahimi, Saeed Ghasemi-Esmailabad, Bryan Woodward, Nahid Yari
Cryopreservation and subsequent survival of semen samples with low numbers of human spermatozoa in assisted reproductive technology (ART) facilities can be challenging. The aim of this study was to compare the quality of warmed human spermatozoa following vitrification using direct submerging (DS) in liquid nitrogen (LN2 ) or with LN2 vapour (V). Normozoospermic ejaculates were prepared by the swim-up technique and the motile sperm fraction was divided into three groups: (i) fresh (control); (ii) DS methods in LN2 (DS Group); or (iii) cryopreserved in V (Group V)...
May 11, 2018: Human Fertility: Journal of the British Fertility Society
W-J Zhu
The aim of this study was to evaluate the production of artefacts during preparation and observation of human sperm for ultrastructural morphology and analyse the possible reasons of causing these artefacts. Under the scanning electron microscopy, damaged sperm heads (crack or/and rupture), necks (head-neck or head-midpiece separation) and midpiece (disassembled and denuded axoneme ultrastructures, bent midpiece) were analysed to be the consequence of exogenous effects. Thirty infertile men with teratozoospermia revealed more spermatozoa with damage to head, neck and midpiece than did thirty fertile males (p < ...
May 7, 2018: Andrologia
G Luke Machen, Erin T Bird, Monica L Brown, Dale A Ingalsbe, Milaida M East, Michelle Reyes, Thomas J Kuehl
Semen from asymptomatic men who are being evaluated as male partners in interfile couples have been reported to contain a variety of bacteria. Longitudinal studies of the variation of these bacteria over time and their resistance patterns have not been commonly reported. At our institution, residues from semen samples are routinely evaluated for bacteria, including antibiotic sensitivity profiles. We set out to profile the changes in semen bacteria and antibiotic resistance at our institution over time. A total of 72 semen isolates were examined for type of bacteria and sensitivity to a panel of antibiotics...
April 2018: Proceedings of the Baylor University Medical Center
A Taylor, D Mills, T Wang, N Ntalamagka, S F Cummins, A Elizur
Pheromones are considered to play an important role in broadcast spawning in aquatic animals, facilitating synchronous release of gametes. In oysters, the sperm has been implicated as a carrier for the spawn-inducing pheromone (SIP). In hatchery conditions, male pearl oysters (Pinctata maxima) can be stimulated to spawn through a variety of approaches (e.g. rapid temperature change), while females can only be induced to spawn through exposure to conspecific sperm, thus limiting development of targeted pairing, required for genetic research and management...
April 28, 2018: Marine Biotechnology
Beata Irena Cejko, Sławomir Krejszeff, Daniel Żarski, Sylwia Judycka, Katarzyna Targońska, Dariusz Kucharczyk
The effect of carp pituitary homogenate (CPH, n = 7) at a dose of 2.0 mg kg-1 and (D-Arg6 , Pro9 NET)-sGnRH + domperidone (Ovaprim, n = 7) at a dose of 0.5 ml kg-1 in northern pike (Esox lucius) sperm maturation under controlled conditions was examined. On the control group, 0.9% NaCl at a dose of 1.0 ml kg-1 (n = 7) was used. Sperm was collected 48 h following injection. Sperm quantity (total sperm volume, total sperm production and sperm concentration), sperm motility using the Computer-assisted sperm analysis (CASA) system and sperm and seminal plasma quality (sperm pH, seminal plasma osmotic pressure and seminal plasma pH) were determined for each male separately in each group...
April 19, 2018: Animal Reproduction Science
M A Awan, S Akhter, A U Husna, M S Ansari, B A Rakha, A Azam, S Qadeer
The free radical scavenging activity (RSA) of Nigella sativa extract and its efficiency for cryopreservation of buffalo spermatozoa was investigated. In experiment 1, Nigella sativa extract was prepared and evaluated for RSA using 2,2-diphenyl-1-picrylhydrazyl assay. The results showed increased pattern of RSA at 1%-5% of Nigella sativa extract. In experiment 2, buffalo semen from three bulls (24 ejaculates) was incubated at 0%, 0.1%, 0.3%, 0.5%, 1%, 1.5%, 2%, 3%, 4%, 5% and 6% extract to assess in vitro tolerability to Nigella sativa in terms of progressive motility (PM)...
April 26, 2018: Andrologia
Marta Gòdia, Fabiana Quoos Mayer, Julieta Nafissi, Anna Castelló, Joan Enric Rodríguez-Gil, Armand Sánchez, Alex Clop
The study of the boar sperm transcriptome by RNA-seq can provide relevant information on sperm quality and fertility and might contribute to animal breeding strategies. However, the analysis of the spermatozoa RNA is challenging as these cells harbor very low amounts of highly fragmented RNA, and the ejaculates also contain other cell types with larger amounts of non-fragmented RNA. Here, we describe a strategy for a successful boar sperm purification, RNA extraction and RNA-seq library preparation. Using these approaches our objectives were: (i) to evaluate the sperm recovery rate (SRR) after boar spermatozoa purification by density centrifugation using the non-porcine-specific commercial reagent BoviPureTM ; (ii) to assess the correlation between SRR and sperm quality characteristics; (iii) to evaluate the relationship between sperm cell RNA load and sperm quality traits and (iv) to compare different library preparation kits for both total RNA-seq (SMARTer Universal Low Input RNA and TruSeq RNA Library Prep kit) and small RNA-seq (NEBNext Small RNA and TailorMix miRNA Sample Prep v2) for high-throughput sequencing...
April 26, 2018: Systems Biology in Reproductive Medicine
Charrid Resgalla, Marcus Vinícius Máximo, Mirella do Nascimento Brasil, Marcos Luiz Pessatti
The aim of this study was to improve the methodological procedure for the evaluation of sea urchin (Lytechinus variegatus) sperm sensitivity in MTT (3-(4,5-dimethylthiazol-2yl)-2,5 diphenyltetrazolium bromide) enzyme reduction assays with the formation of formazan (purple color) in the interior of viable cells. Assays were carried out with the reference toxicants sodium dodecyl sulfate (SDS), copper, zinc, cadmium and ammonium, using a sperm solution previously activated in sea water and a sperm solution prepared in sea water containing 400 μg L-1 verapamil, which enabled activation of the sperm to occur only when exposed to the toxicants...
April 24, 2018: Ecotoxicology
So Maezawa, Kazuteru Hasegawa, Masashi Yukawa, Naoki Kubo, Akihiko Sakashita, Kris G Alavattam, Ho-Su Sin, Andrey V Kartashov, Hiroyuki Sasaki, Artem Barski, Satoshi H Namekawa
Repressive H3K27me3 and active H3K4me2/3 together form bivalent chromatin domains, molecular hallmarks of developmental potential. In the male germline, these domains are thought to persist into sperm to establish totipotency in the next generation. However, it remains unknown how H3K27me3 is established on specific targets in the male germline. Here, we demonstrate that a germline-specific Polycomb protein, SCML2, binds to H3K4me2/3-rich hypomethylated promoters in undifferentiated spermatogonia to facilitate H3K27me3...
April 23, 2018: Proceedings of the National Academy of Sciences of the United States of America
Esmat Mangoli, Ali Reza Talebi, Morteza Anvari, Fatemeh Taheri, Mahboobeh Vatanparast, Tahereh Rahiminia, Akram Hosseini
OBJECTIVE: Aim of this study was to evaluate the effects of vitamin C on sperm parameters, sperm chromatin quality and apoptosis resulted of vitrification in neat semen and prepared spermatozoa of normozoospermic samples. MATERIAL AND METHODS: Forty semen samples from normozoospermic men were included in this prospective study. Each sample was divided into five groups. Group I: control or fresh semen, group II: semen prepared by swim-up method and then vitrified, group III: neat semen was vitrified, group IV: vitamin C (600 μM) was added to prepared spermatozoa and then vitrified and group V: vitamin C (600 μM) was added to neat semen and then vitrified...
April 2018: Taiwanese Journal of Obstetrics & Gynecology
Ramin Nasimi Doost Azgomi, Afshar Zomorrodi, Hossein Nazemyieh, Seyed Mohammad Bagher Fazljou, Homayoun Sadeghi Bazargani, Fatemeh Nejatbakhsh, Arezoo Moini Jazani, Yadollah Ahmadi AsrBadr
Introduction: Withania somnifera (WS) also known as ashwagandha is a well-known medicinal plant used in traditional medicine in many countries for infertility treatment. The present study was aimed at systemically reviewing therapeutic effects of WS on the reproductive system. Methods: This systematic review study was designed in 2016. Required data were obtained from PubMed, Scopus, Google Scholar, Cochrane Library, Science Direct, Web of Knowledge, Web of Science, and manual search of articles, grey literature, reference checking, and expert contact...
2018: BioMed Research International
Kutluk Oktay, Brittany E Harvey, Ann H Partridge, Gwendolyn P Quinn, Joyce Reinecke, Hugh S Taylor, W Hamish Wallace, Erica T Wang, Alison W Loren
Purpose To provide current recommendations about fertility preservation for adults and children with cancer. Methods A systematic review of the literature published from January 2013 to March 2017 was completed using PubMed and the Cochrane Library. An Update Panel reviewed the identified publications. Results There were 61 publications identified and reviewed. None of these publications prompted a significant change in the 2013 recommendations. Recommendations Health care providers should initiate the discussion on the possibility of infertility with patients with cancer treated during their reproductive years or with parents/guardians of children as early as possible...
April 5, 2018: Journal of Clinical Oncology: Official Journal of the American Society of Clinical Oncology
K Andraszek, D Banaszewska, B Biesiada-Drzazga
Many of the methods used to stain semen result in very pronounced coloring of the sperm, but unfortunately they do not distinguish their individual structures, which play a key role in the fertilization process. Hence the aim of this study was to identify sperm structures using two staining techniques in the semen of roosters from breeding flocks. The subject of the study was the sperm of roosters from a Ross 308 breeding flock. To capture the differences in the dimensions of sperm subjected to the effect of different chemical substances in dyes, microscope slides were stained by two techniques: with an AgNO3 solution and by a differential method (eosin-nigrosin test)...
April 3, 2018: Poultry Science
Zoeisha S Chinoy, Frédéric Friscourt, Chantelle J Capicciottia, Philip Chiu, Geert-Jan Boons
Complex N-glycans of glycoproteins of the zona pellucida (ZP) of human oocytes have been implicated in the binding of spermatozoa. The termini of these unusual bi-, tri-, and tetra-antennary N-glycans consist of the tetrasaccharide sialyl-Lewisx (SLex), which was previously identified as the minimal epitope for sperm binding. We describe here the chemo-enzymatic synthesis of highly complex tri-antennary N-glycans derived from ZP carrying SLex moieties at the C-2 and C-2' arm and a sialyl-Lewisx-Lewisx (SLex-Lex) residue at the C-6 antenna and two closely related analogs...
March 30, 2018: Chemistry: a European Journal
Selvakumar Narasimman, Dhanasekar Krishnamoorthy, Carmel D Buelah, Munuswamy Natesan
This study focused on the quality of sperm obtained from spermatophores of cadaveric shrimp after long-term storage. Spermatophores were collected using the stripping method, which has resulted in maximum sperm viability when this approach was previously used. Cryoprotectants toxicity assessment of samples was conducted using dimethyl sulfoxide (DMSO), methanol (MeOH), ethylene glycol (EG), glycerol (Gly), dimethyl acetamide (DMA) and propylene glycol (PG) at different concentrations (5%, 10% and 30% v/v), prepared in Ca-F saline...
March 9, 2018: Animal Reproduction Science
Yuksel Oguz, Ismail Guler, Ahmet Erdem, Mehmet Firat Mutlu, Seyhan Gumuslu, Mesut Oktem, Nuray Bozkurt, Mehmet Erdem
PURPOSE: To compare the effect of two different sperm preparation techniques, including swim-up and gradient methods on sperm deoxyribonucleic acid (DNA) fragmentation status of semen samples from unexplained and mild male factor subfertile patients undergoing intrauterine insemination (IUI). DESIGN: A prospective randomized study was conducted in 65 subfertile patients, including 34 unexplained and 31 male factor infertility to compare basal and post-procedure DNA fragmentation rates in swim-up and gradient techniques...
March 23, 2018: Journal of Assisted Reproduction and Genetics
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