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> Anatomy > Cell > Spermatid

Spermatid

Spermatids are the haploid male gametocytes produced during spermatogenesis.
They undergo a complex series of morphological changes, including the formation of the acrosome, the development of the flagellum, and the condensation of the nucleus, to become mature spermatozoa.
The study of spermatid biology is crucial for understanding male fertility and reproductive health.
PubCompare.ai can help optimize your spermatid research by locating the best protocols from literature, preprits, and patents, enhancing reproducibility and accuracy to ensure reliable experimental results.
Explore the power of PubCompare.ai and take your spermatid research to new heights.

Most cited protocols related to «Spermatid»

1
Real-time RT-PCR and Microarray Analysis of Testis Transcripts
For real-time reverse transcription-polymerase chain reaction (RT-PCR), total testis RNA was extracted using Trizol and then DNaseI-treated (Invitrogen). Reverse transcription of polyadenylated RNA was performed with Superscript Reverse Transcriptase II, according to the manufacturer's guidelines (Invitrogen). Real-time PCR was performed using Absolute qPCR SYBR Green ROX mix (ThermoFisher) on an ABI PRISM 7500 machine (Applied Biosystems). PCR reactions were incubated at 95°C for 15 min followed by 40 PCR cycles (5 s at 95°C, 20 s at 60°C, and 45 s at 68°C). Primer sequences are available in Table S3. Samples from four transgenic mice and three nontransgenic siblings (negative controls), all at 2 mo of age, were analyzed. All reactions were carried out in triplicate per assay, and β-actin was included on every plate as a loading control. The difference in PCR cycles with respect to β-actin (ΔCt) for a given experimental sample was subtracted from the mean ΔCt of the reference samples (negative siblings) (ΔΔCt). For the quantification of Sly knock-down, values were further normalized to ΔΔCt values of the spermatid-specific control Acrv1. This was to have a more robust analysis when compared with 2/3MSYq− mice, which have variability in spermatid content.
For microarray analyses, absolute expression values were obtained by single-color hybridizations (Illumina BeadChip, mouse whole-genome array, v2) for three sh367 transgenic individuals and matched littermate controls (negative siblings), and RNA from each individual was hybridized separately. A similar analysis was performed on 2/3MSYq−, 9/10MSYq−, and MSYq− samples and appropriate age/strain-matched controls. In each case, pooled RNA from two or three individuals was used as the sample. Differentially expressed genes were grouped into five categories based on their expression ratios across all genotypes (see Figure S5). Similar microarray analyses were performed on juvenile testes (17 d postpartum) from three sh367 males and three littermate controls (negative siblings). There was no significant change of gene expression between the two groups. Microarray analyses were also performed on purified spermatid fractions from two groups of sh367 transgenic mice, two groups of sh367 negative siblings, and two groups of 2/3MSYq−.
Cocquet J., Ellis P.J., Yamauchi Y., Mahadevaiah S.K., Affara N.A., Ward M.A, & Burgoyne P.S. (2009). The Multicopy Gene Sly Represses the Sex Chromosomes in the Male Mouse Germline after Meiosis. PLoS Biology, 7(11), e1000244.
Publication 2009
Actins Animals, Transgenic Biological Assay Crossbreeding Gene Expression Genes Genome Genotype Lanugo Males Mice, Laboratory Mice, Transgenic Microarray Analysis Oligonucleotide Primers prisma Real-Time Polymerase Chain Reaction Reverse Transcriptase Polymerase Chain Reaction Reverse Transcription RNA, Polyadenylated RNA-Directed DNA Polymerase Sibling Spermatid Strains SYBR Green I Testis trizol
2
Dynamic Gene Regulation in Spermatogenesis

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Publication 2018
Cloning Vectors Gene Expression Gene Expression Regulation Genes Germ Cells Phase Transition Sequence Analysis Spermatid Spermatocytes Spermatogenesis Spermatogonia Transcription Factor Transcription Initiation Site
3
Isolation of Enriched Spermatid Fractions
To obtain cell fractions enriched in spermatids, an adapted protocol from the trypsin method described by Meistrich [53] was used. Testes from a group of four to five adult mice from the same genotype (i.e., three groups of sh367 transgenic mice, three groups of sh367-negative siblings, and two groups of 2/3MSYq−) were used for the study. Testes were dissected and chopped in 20 ml of DMEM (GIBCO) and treated with 2.5 mg/ml trypsin (GIBCO) and 50 µg/ml DNase I (Sigma) for 30 min at 31°C with stirring. After adding fetal calf serum (GIBCO) (final concentration of 8%), the cells were passed through a 100-µm filter. Cells were then centrifuged at room temperature (500 g, 15 min), resuspended in DMEM 0.5% bovine serum albumin (Sigma) with 50 µg/ml DNase I, and cooled on ice. Cells were counted and checked for clumps before proceeding with the elutriation. Cell integrity was checked using Trypan blue. Fractions enriched in different testis cell types were separated with a JE-6B elutriator (Beckman) with conditions described before [54] (link). Collected fractions were washed in PBS, and cell pellets were frozen down at −80°C. Fraction content was assessed based on cell morphology after DAPI staining. Fractions #6 contained >90% round spermatids and were used for RNA and protein analyses.
Cocquet J., Ellis P.J., Yamauchi Y., Mahadevaiah S.K., Affara N.A., Ward M.A, & Burgoyne P.S. (2009). The Multicopy Gene Sly Represses the Sex Chromosomes in the Male Mouse Germline after Meiosis. PLoS Biology, 7(11), e1000244.
Publication 2009
Adult Cells DAPI Deoxyribonucleases Fetal Bovine Serum Freezing Genotype Mice, Transgenic Mus Pellets, Drug Proteins PRSS2 protein, human Serum Albumin, Bovine Sibling Spermatid Testis Trypan Blue Trypsin
4
Live Imaging of Spermatocyte Cytokinesis
To follow cytokinesis in living primary spermatocytes, we modified our previous protocol (Savoian et al., 2000 (link)). We observed our living cells in “open” chambers. These were constructed by adhering clean No. 1 1/2 thickness glass coverslips to the underside of aluminum slides in which the center-most portion had been removed, leaving a depression into which the cells could be placed. Testes were isolated from adult flies and their cells spread under Voltalef 10s oil (Elf Atochem) onto the attached coverslips for subsequent filming. Time-lapse imaging was performed on a microscope (model Axiovert 200; Carl Zeiss MicroImaging, Inc.) outfitted with excitation, emission, and neutral density filterwheels (Prior Scientific) and a PIFOC z-axis focus drive (Physik Instruments). Cells were imaged with a 100× (NA 1.4) lens and long working distance DIC condenser (NA 0.55). Specimens were illuminated with heat and UV filtered, shuttered light using the appropriate filter wheel combinations through an EGFP/DsRed filter cube (Chroma Technology Corp.). At each 1 min time interval, six, near-simultaneous fluorescence (EGFP) and transmitted light (DIC) optical sections (1 μm step size) were captured with a camera (model Coolsnap HQ; Roper Scientific) using a 2 × 2 bin. Image acquisition was controlled through the Metamorph software package (Universal Imaging Corp.) running on a PC. To determine fluorescence intensity distributions, maximum intensity projections were analyzed using the linescan function along the length of the spindle as shown in the figures with a line width of 30 pixels. The values were exported into Excel (Microsoft) for plotting before being incorporated into the figures.
Montages were created in Adobe Photoshop®. Each fluorescence image is the maximum intensity projection of the six individual optical sections for that time point. The DIC image is the corresponding, single central section.
Testes squashes to evaluate onion stage spermatids were made using standard protocols and viewed by phase-contrast microscopy.
GFP-tubulin–expressing S2 cells (Goshima and Vale, 2003 (link)) were grown on clean No. 1 1/2 coverslips. These were mounted on 65 μl Gene frames (ABgene) filled with complete Schneider's medium directly before filming. Z-series were acquired every 15–30 s on an Ultraview rs spinning disk confocal system attached to a microscope using a 100× (NA 1.3) lens and a 2 × 2 bin. Montages and videos are the maximum intensity projections of z-series.
Inoue Y.H., Savoian M.S., Suzuki T., Máthé E., Yamamoto M.T, & Glover D.M. (2004). Mutations in orbit/mast reveal that the central spindle is comprised of two microtubule populations, those that initiate cleavage and those that propagate furrow ingression. The Journal of Cell Biology, 166(1), 49-60.
Publication 2004
Adult Allium cepa Aluminum Cells Cucurbita Cytokinesis Diptera Epistropheus Fluorescence Genes Lens, Crystalline Light Microscopy Microscopy, Phase-Contrast Reading Frames Spermatid Spermatocytes Testis Tubulin Vision
5
Ultrastructural Analysis of Testes
Testes were dissected after perfusion fixation with 4% PFA in PBS under anesthesia, and immersed in 4% PFA for 6 h at 4°C. The organs were sliced into 2 mm × 2 mm × 2 mm pieces with safety
razors, immersed in 1% glutaraldehyde in 30 mM HEPES (pH 7.8) overnight at 4°C, and washed three times (5 min each) in 30 mM HEPES. Tissues were postfixed in 1% OsO4 and 0.5%
potassium ferrocyanide in 30 mM HEPES for 1 h at room temperature. After being washed with distilled water, samples were dehydrated with a graded series of ethanol solutions (50, 70, 90%) on
ice, and in 100% ethanol for 10 min at room temperature. Dehydrated samples were incubated twice for 5 min in 100% propylene oxide (PO), and then placed in a mixture of PO and epoxy resin
for 1 h at room temperature. Sample tissues were incubated in a pure epoxy resin mixture twice for 1 h at room temperature, and embedded in epoxy resin for 2 days at 60°C. Eighty nm
ultrathin sections were cut and stained with 2% uranyl acetate solution for 30 min, briefly washed three times with distilled water, stained with a lead staining solution for 2 min, and
washed three times with distilled water. The sample were examined using a JEM-1400 Plus electron microscope (JEOL, Tokyo, Japan) at 80 kV with a CCD Veleta 2K × 2K camera (Olympus). Stages
of the epithelial cycle were identified based on the morphological characteristics of the spermatids, in particular their nucleus and acrosomic system [2 ].
SHIMADA K., KATO H., MIYATA H, & IKAWA M. (2019). Glycerol kinase 2 is essential for proper arrangement of crescent-like mitochondria to form the mitochondrial sheath during mouse spermatogenesis. The Journal of Reproduction and Development, 65(2), 155-162.
Publication 2019
Anesthesia Cell Nucleus Electron Microscopy Epoxy Resins Ethanol Glutaral HEPES Perfusion potassium ferrocyanide propylene oxide Spermatid Testis Tissues uranyl acetate

Most recents protocols related to «Spermatid»

1
Sperm DNA Methylation Analysis in Valproic Acid-Treated Mice
Sperm DNA was extracted as described above from VPA-treated mice, VPA_F1, and control mice (n = 3 each). Subsequently, DNA methylation analysis was performed using WGBS, and DNA bisulfite conversion was performed using an EZ DNA Methylation-Gold Kit (Zymo Research, Orange, CA, USA). Further, DNA libraries were constructed using the Accel-NGS Methyl-Seq DNA Library Kit (Integrated DNA Technologies, Coralville, IA, USA) or Abclonal Scale Methyl-DNA Lib Prep Kit for Illumina (Abclonal, Tokyo, Japan) according to the manufacturer’s instructions. The DNA libraries were used for 150 bp paired-end sequencing on a Novaseq 6000 platform (Illumina, San Diego, CA, USA). Adapter trimming was performed using the Trim Galore 0.5.0 program (RRID: SCR_011847) and Trimmomatic program [28 (link)]. The resulting reads were quality checked using FastQC version 0.11.7 [29 ]. The reads were aligned to the mouse genome mm10 using Bismark version 0.22.3 [30 (link)] with default parameters. Gene annotation and methylation level calculations were performed using the methylKit package [31 (link)] in R (https://www.r-project.org/). Promoters were defined as regions 2 kb upstream of the transcription start sites. The DMCs were identified as cytosines in the promoter regions whose methylation rate showed a P-value < 0.05, as determined by t-test, and the mean methylation rate difference was ≥ 20%. Similarly, regions that contained at least 10 CpGs within 300 bp and whose average difference in methylation rates was ≥ 20% were extracted as DMRs. The genomic regions rich in acetylated histones K9 and K27 located in the vicinity of the DMRs (± 1 kb) were identified using the Peak Browser of ChIP-Atlas [32 (link)], where the threshold for significance was set to 50 (q-value < 1E-05) and cell type as mouse male germ line (testis, male germ cells, spermatogonia, spermatogenic cells, round spermatids, and spermatids). Pathway analysis using IPA software version 68752261 (QIAGEN, Valencia, CA, USA) was performed for genes containing DMC in the promoter region.
Sakai K., Hara K, & Tanemura K. (2023). Testicular histone hyperacetylation in mice by valproic acid administration affects the next generation by changes in sperm DNA methylation. PLOS ONE, 18(3), e0282898.
Publication 2023
Cells cytidylyl-3'-5'-guanosine Cytosine DNA Chips DNA Library DNA Methylation Gene Annotation Genes Genome Germ Cells Germ Line Gold Histones hydrogen sulfite Males Methylation Mus Sperm Spermatid Spermatogenesis Spermatogonia Testis Transcription Initiation Site
2
Ultrastructural Analysis of Mammalian Gametes
Mice eyecups, mice semen cells collected from shredded unilateral caudal epididymis, mice testicular tissues, and human ejaculated sperm were subjected to TEM assay, respectively. Tissues were fixed in 2.5% glutaradehyde at 4°C overnight immediately after enucleation. Eyecups were dissected as described for TEM, and only the remaining posterior eyecups were used for the following experiments. Samples were then post-fixed in 1% osmic acid at 4°C for an hour, stained in aqueous 3% uranyl acetate for 2 hr, dehydrated with ascending concentrations of acetone, embedded in epoxy resin, cut into ultra-thin slides, and stained in 0.3% lead citrate. Ultrastructure of mice retina and human spermatozoa was visualized using a JEM-1010 electron microscope (JEOL, Tokyo, Japan). Ultrastructure of mice spermatids cells were observed with a Philips CM100 electron microscope (Philips, Amsterdam, North-Holland). Ultrastructure of testicular tissues was visualized using a FEI Tecnai G2 Spirit Bio TWIN electron microscope (Thermo, Waltham, MA, USA).
For SEM assay, mice sperms collected from shredded unilateral caudal epididymis and human ejaculated sperm were used. Human and mice spermatozoa were immersed in 2.5% glutaraldehyde at 4°C overnight, fixed in 1% osmic acid supplemented with 1.5% K3[Fe(CN)3] at 4°C for an hour, steeped in 1% thiocarbohydrazide for an hour, post-fixed in 1% osmic acid at 4°C for an hour, and soaked in 2% uranyl acetate solution at 4°C overnight. The treated samples were then progressively dehydrated with an ethanol and isoamyl acetate gradient on ice, and dried with a CO2 critical-point dryer (Eiko HCP-2, Hitachi Ltd., Tokyo, Japan). The specimens were subsequently mounted on aluminum stubs, sputter coated by an ionic sprayer meter (Eiko E-1020, Hitachi Ltd.), and visualized using a FEI Nova NanoSEM 450 SEM (Thermo, Waltham, MA, USA).
Zhu T., Zhang Y., Sheng X., Zhang X., Chen Y., Zhu H., Guo Y., Qi Y., Zhao Y., Zhou Q., Chen X., Guo X, & Zhao C. (2023). Absence of CEP78 causes photoreceptor and sperm flagella impairments in mice and a human individual. eLife, 12, e76157.
Publication 2023
Acetone Aluminum Biological Assay Cells Citrate Desiccation Electron Microscopy Epididymis Epoxy Resins Ethanol Glutaral Homo sapiens Ions isoamyl acetate Mus Osmium Tetroxide Plant Embryos Retina Sperm Spermatid thiocarbohydrazide Tissues Twins uranyl acetate
3
Proteomic Analysis of Cep78 Knockout Mice
Testicular tissues of Cep78+/− and Cep78−/− mice were lysed with Pierce IP lysis buffer (Thermo, Waltham, MA, USA) supplemented with 1% protease inhibitor cocktail 100× (Selleck Chemicals, Houston, TX, USA), respectively. Lysates were revolved for 1 hr at 4°C and centrifuged at 40,000 g for 1 hr. Supernatants were precleared with 20 μL of protein A/G magnetic beads (Millipore, Billerica, MA, USA) at 4°C for 1 hr. IP was performed using the Pierce co-IP kit (Thermo, Waltham, MA, USA). 5 µg of antibody or IgG was subjected for incubation per sample. For in vivo Cep78 IP, the custom-made mouse Cep78 antibody against antigen p457-741 of mouse Cep78 (NP_932136.2) was used. Eluted proteins were then subjected to SDS-PAGE, silver stain, immunoblotting, and MS. For MS, silver-stained gel lanes were carefully cut into small pieces and were then processed for MS analysis. Briefly, after trypsin digestion overnight, peptides were desalted using stage tips, re-suspended in 0.1% formic acid (v/v) and subjected to LC-MS (Guo et al., 2011 (link); Guo et al., 2010 (link); Hu et al., 2013 (link); Fan et al., 2020 (link)).
For MS on elongating spermatids lysates of Cep78+/− and Cep78−/− mice, we isolated spermatid cells with STA-PUT. Lysed spermatid cells were digested overnight at 37°C with trypsin and subjected to the Tandem Mass Tag （TMT） labeling. For MS analyses, each fraction was analyzed using an Orbitrap Fusion Lumos mass spectrometer (Thermo Finnigan, San Jose, CA) coupled with Easy-nLC 1200 (Thermo Finnigan, San Jose, CA) and was separated by a High PH reverse phase separation microcapillary column (ACQUITY BEH C18 Column, 1.7 μm, 300 μm × 150 mm) at a flow rate of 4 μL/min in a 128 min linear gradient (3% buffer B for 14 min, 8% buffer B for 1 min, 29% buffer B for 71 min, 41% buffer B for 12 min, 100% buffer B for 9 min, and 3% buffer B for 21 min; solvent A: 20 mM Ammonium formate, PH = 10; solvent B: 100% ACN, 0.1% FA). Sample were collected at a rate of one tube per minute, a total of 30 components. Easy1200 and fusion Lumos were used for series identification.
Zhu T., Zhang Y., Sheng X., Zhang X., Chen Y., Zhu H., Guo Y., Qi Y., Zhao Y., Zhou Q., Chen X., Guo X, & Zhao C. (2023). Absence of CEP78 causes photoreceptor and sperm flagella impairments in mice and a human individual. eLife, 12, e76157.
Publication 2023
Antigens Buffers Cells Digestion formic acid formic acid, ammonium salt G-substrate GTP-Binding Proteins Immunoglobulins Mus Peptides Proteins SDS-PAGE SERPINA1 protein, human Silver Solvents Spermatid Staphylococcal Protein A Tissues Trypsin
4
Isolation of Spermatid Cells Using STA-PUT
We isolated spermatid cells with STA-PUT method, following Bryant et al., 2013 (link)’s protocol. In brief, after removal of tunica vaginalis, testis tissue of Cep78+/− and Cep78−/− mice (n=6 for Cep78+/− and Cep78−/− mice) was digested with Collagenase IV (Gibco, RI, USA), Trypsin (Gibco, RI, USA), and DNaseI (Bomei, Hefei, China) in shaking water bath at 33°C for 5 min, filtered with 100 μm and 40 μm nylon filters (Corning, NY, USA), re-suspended in 0.5% bovine serum albumin (Sigma-Aldrich, St. Louis, MO, USA) solution, and settled in the BSA gradient in the sedimentation chamber of STA-PUT aparatus for 2 hr and 15 min. Cell fractions were obtained, stained by Hoechst 33342 (Sigma-Aldrich, St. Louis, MO, USA), and examined with a Zeiss Axio Observer A1 inverted microscope (ZEISS, Oberkochen, Germany) to assess the purity of spermatid cells.
Zhu T., Zhang Y., Sheng X., Zhang X., Chen Y., Zhu H., Guo Y., Qi Y., Zhao Y., Zhou Q., Chen X., Guo X, & Zhao C. (2023). Absence of CEP78 causes photoreceptor and sperm flagella impairments in mice and a human individual. eLife, 12, e76157.
Publication 2023
Bath Collagenase HOE 33342 Microscopy Mus Nylons Serum Albumin, Bovine Spermatid Testis Tissues Trypsin
5
Comprehensive Biological Analyses of C. elegans

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Publication 2023
Animals Apoptosis Biopharmaceuticals Fertility Germ Line Human Body Oocytes prisma Sperm Spermatid Stem Cells

Top products related to «Spermatid»

1
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Hoechst 33342 is a fluorescent dye that binds to DNA. It is commonly used in various applications, such as cell staining and flow cytometry, to identify and analyze cell populations.
2
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TRIzol reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components designed for the isolation of total RNA, DNA, and proteins from a variety of biological samples. The reagent maintains the integrity of the RNA while disrupting cells and dissolving cell components.
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More about "Spermatid"

Spermatids are the precursor cells that develop into mature sperm during the process of spermatogenesis.
These haploid male gametocytes undergo a complex series of transformations, including the formation of the acrosome, the development of the flagellum, and the condensation of the nucleus.
The study of spermatid biology is crucial for understanding male fertility and reproductive health.
PubCompare.ai can help optimize your spermatid research by locating the best protocols from literature, preprints, and patents, enhancing reproducibility and accuracy to ensure reliable experimental results.
Hoechst 33342 is a fluorescent dye commonly used to stain and visualize spermatids and other cells.
DNase I is an enzyme that can be used to digest DNA, which is important for isolating and analyzing spermatid RNA and proteins.
TRIzol reagent is a common method for extracting RNA from spermatids and other cell types.
The Agilent 2100 Bioanalyzer and HiSeq 2500 are powerful tools for analyzing the quality and quantity of RNA and DNA extracted from spermatids.
Triton X-100 is a detergent that can be used to lyse spermatid cells and extract their contents.
DMEM is a culture medium that can be used to maintain spermatids and other cell types in vitro.
Bovine serum albumin is a common additive in cell culture media that can help support spermatid growth and viability.
Turbo DNase is an enzyme that can be used to remove any contaminating DNA from spermatid RNA preparations, and Trypsin is an enzyme that can be used to dissociate spermatids and other cells from tissue samples.
By leveraging these tools and techniques, PubCompare.ai can help you take your spermatid research to new hieghts, enhancing your ability to study this crucial aspect of male reproductive biology.