Atomic Force Microscope Analysis of Chromatin Volumes in Human Sperm with Head-Shape Abnormalities1

Lee, J D; Allen, Michael J.; Balhorn, Rod

doi:10.1095/biolreprod56.1.42

Cited by 28 publications

(26 citation statements)

References 34 publications

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“…After rinsing in distilled water, the amembraneous sperm heads were then resuspended in deionized water. This treatment of the samples does not alter the morphology of the sperm head [36]. Each of the sperm head morphology classes reported previously by Moruzzi et al [37] can still be identified and distinguished [36].…”

Section: Sample Preparationmentioning

confidence: 63%

Raman spectroscopy of DNA packaging in individual human sperm cells distinguishes normal from abnormal cells

Huser

Orme

Hollars

et al. 2009

Journal of Biophotonics

110

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Healthy human males produce sperm cells of which about 25–40% have abnormal head shapes. Increases in the percentage of sperm exhibiting aberrant sperm head morphologies have been correlated with male infertility, and biochemical studies of pooled sperm have suggested that sperm with abnormal shape may contain DNA that has not been properly repackaged by protamine during spermatid development. We have used micro‐Raman spectroscopy to obtain Raman spectra from individual human sperm cells and examined how differences in the Raman spectra of sperm chromatin correlate with cell shape. We show that Raman spectra of individual sperm cells contain vibrational marker modes that can be used to assess the efficiency of DNA‐packaging for each cell. Raman spectra obtained from sperm cells with normal shape provide evidence that DNA in these sperm is very efficiently packaged. We find, however, that the relative protein content per cell and DNA packaging efficiencies are distributed over a relatively wide range for sperm cells with both normal and abnormal shape. These findings indicate that single cell Raman spectroscopy should be a valuable tool in assessing the quality of sperm cells for in‐vitro fertilization. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

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Section: Sample Preparationmentioning

confidence: 63%

Raman spectroscopy of DNA packaging in individual human sperm cells distinguishes normal from abnormal cells

Huser

Orme

Hollars

et al. 2009

Journal of Biophotonics

110

View full text Add to dashboard Cite

show abstract

“…For example, mice with reduced levels of protamines due to haploinsufficiency had defective chromatin condensation accompanied by narrowed sperm heads with a reduced curvature, 45 and mice lacking a novel histone-like protein involved in chromatin compaction showed abnormal nuclear morphology and "halos" within densely stained sperm nucleus. 46 While abnormalities in sperm chromatin packaging are more likely to manifest as altered quality, DNA damage and infertility in ejaculated sperm, rather than a difference in head shape, 47 relationships between sperm head morphology and DNA integrity in human sperm have been demonstrated. 48 Thus it is reasonable to assume that processes governing sperm DNA chromatin could impact on sperm head shape.…”

Section: E979623-4mentioning

confidence: 99%

Mechanisms of spermiogenesis and spermiation and how they are disturbed

2014

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Haploid round spermatids undergo a remarkable transformation during spermiogenesis. The nucleus polarizes to one side of the cell as the nucleus condenses and elongates, and the microtubule-based manchette sculpts the nucleus into its species-specific head shape. The assembly of the central component of the sperm flagellum, known as the axoneme, begins early in spermiogenesis, and is followed by the assembly of secondary structures needed for normal flagella. The final remodelling of the mature elongated spermatid occurs during spermiation, when the spermatids line up along the luminal edge, shed their residual cytoplasm and are ultimately released into the lumen. Defects in spermiogenesis and spermiation are manifested as low sperm number, abnormal sperm morphology and poor motility and are commonly observed during reproductive toxicant administration, as well as in genetically modified mouse models of male infertility. This chapter summarizes the major physiological processes and the most commonly observed defects in spermiogenesis and spermiation, to aid in the diagnosis of the potential mechanisms that could be perturbed by experimental manipulation such as reproductive toxicant administration. Signature LesionThere are a number of signature lesions that could indicate a disturbance in spermiogenesis or spermiation. These would include misshapen heads and/or tails of elongating/elongated spermatids. Multinucleated round spermatids may reflect disturbances in spermiogenesis, but could also reflect altered Sertoli cell function. Disruption of spermiation could present itself as spermatid retention at the lumen of post stage VIII tubules, failure of elongated spermatids to ascend to the lumen of stage VII/ VIII tubules, or phagocytosis of mature spermatids at the base of tubules in stages VII through to approximately XII (in mice) or XIV (in rats). The changes might occur in isolation or in combination with one another.

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“…AFM requires simple sample preparation protocols such as usage of physiological saline and air drying of smeared spermatozoa as compared to the use of artificial preservatives and elaborate sample preparation protocols in EM (17). AFM has been successfully used to view the surface topography of sperm heads of numerous mammals (15)(16)(17)(18)(19)(20)(21). Despite the vast development achieved technologically in the process of visualization and image capture, and intensive investigation in the field of molecular reproduction of crustaceans in the past century, some of the intrinsic structures, such as architectural arrangement of testicular lobules, remain elusive and are only hypothetical.…”

Section: Introductionmentioning

confidence: 99%

Probing the architecture of testis and morphology of male germinal cells in the mud crab with the atomic force microscopy

Anbarasu¹,

Kirubagaran²,

Dharani³

et al. 2013

Turk J Biol

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The architecture of testicular lobules, morphology of spermatozoa, and spermatophore in the mud crab, Scylla serrata, are described for the first time employing atomic force microscopy (AFM). A fast and simple squash preparation protocol was followed using crustacean physiological saline without distorting the natural structure. Observations provided new insight into the arrangement of lobules and morphology of testicular assembly. It consists of a close network of lobules of different sizes in a radial manner on radially branching basal tubules. Budding of new lobules is another distinguishable feature noted in the network of testicular organization. A thin membranous envelope of the lobule with undulating processes offered indirect evidence on the possible storage of sperm cells within the lumen of the lobule. Spermatozoa of S. serrata were of different shapes and sizes, and, depending on the maturational status, they displayed changes in the acrosome vesicle with a significant amount of cytoplasm. Vertical constrictions found in the testicular lobules suggest propulsive extrusion of spermatophoric masses into the central lumen. These observations necessitate further study using AFM and involving molecular probes to elucidate the polymorphic nature of male germinal cells, mechanisms of spermatophore formation, and the phylogenetic relationship among crustaceans in general.

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Atomic Force Microscope Analysis of Chromatin Volumes in Human Sperm with Head-Shape Abnormalities1

Cited by 28 publications

References 34 publications

Raman spectroscopy of DNA packaging in individual human sperm cells distinguishes normal from abnormal cells

Raman spectroscopy of DNA packaging in individual human sperm cells distinguishes normal from abnormal cells

Mechanisms of spermiogenesis and spermiation and how they are disturbed

Probing the architecture of testis and morphology of male germinal cells in the mud crab with the atomic force microscopy

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