267 DIFFERENCES IN BULL SPERM NUCLEAR SHAPE BETWEEN FLOW-SORTED AND NON-SORTED SPERM USING FOURIER HARMONIC ANALYSIS

2009 ◽  
Vol 21 (1) ◽  
pp. 231
Author(s):  
J. R. Schindler ◽  
R. L. Monson ◽  
K. L. Willenburg ◽  
J. J. Parrish
Keyword(s):  
Longitudinal Axis ◽  
Sperm Nucleus ◽  
Sperm Head ◽  
Nuclear Shape ◽  
Harmonic Amplitude ◽  
Head Shape ◽  
Hoechst 33342 ◽  
Fourier Harmonic ◽  
Frozen Semen ◽  
Sorting Process

The objective of the study was to evaluate sperm nuclear shape in sex-sorted and non-sorted semen using Fourier Harmonic Amplitudes (FHA). Frozen semen was obtained from a single commercially available source. Mature bulls (n = 15) from the same breed with an average age of 4.42 ± 2.08 years were collected and the semen was either frozen or X-chromosome sorted using a flow cytometer and then frozen. Frozen straws were transported to the lab and analyzed for FHA. Briefly, straws were thawed and cells were incubated with 1.6 μm Hoechst 33342. Cells were then washed, fixed, dried to a slide and analyzed for nuclear head shape. Harmonic amplitudes 0 to 5 (HA0–HA5), derived from FHA, were previously shown to be an accurate, objective, and repeatable measure of sperm nuclear shape. HA0 describes the overall nuclear size of the sperm, whereas HA1 describes the anterior head, HA2 the length of the sperm along the longitudinal axis, and HA3 to 5 the distal, post-nuclear curvature of the sperm head. Each unit of semen was evaluated for motility and FHA. There was a significant decrease in motility in the sorted group (77 ± 1% v. 54 ± 3%; P < 0.0001). Multivariate ANOVA showed that there were differences between the sorted and non-sorted groups in HA1 to 4 (P < 0.02). Harmonic amplitude means ± SD (microns) for sorted and non-sorted treatments are as follows: HA1 (0.117 ± 0.003 v. 0.109 ± 0.003), HA2 (1.087 ± 0.005 v. 1.063 ± 0.005), HA3 (0.139 ± 0.003 v. 0.130 ± 0.003), and HA4 (0.201 ± 0.004 v. 0.191 ± 0.004), respectively. The nuclear shape of X-sorted sperm is longer and more pinched in both the anterior and posterior head. Interestingly there was no difference in HA0 (P = 0.119) indicating that the overall size of the sperm head is not affected by the sorting process. The differences in harmonic amplitudes may be due to the size and a restricted location of the X v. Y chromosome in the sperm nucleus.

226 DAY LENGTH DOES NOT AFFECT LIVE SPERM NUCLEAR SHAPE IN THE STALLION

2008 ◽  
Vol 20 (1) ◽  
pp. 192
Author(s):  
J. J. Parrish ◽  
C. Mueller ◽  
E. Ludwig ◽  
J. L. Susko-Parrish
Keyword(s):  
Sperm Head ◽  
Nuclear Shape ◽  
Day Length ◽  
Fourier Harmonic ◽  
Glass Slides ◽  
Large Numbers ◽  
Alternative Approach ◽  
Fertility Data ◽  
Sperm Nuclei ◽  
Live Sperm

Fourier harmonic analysis (FHA) of sperm nuclei is a precise and objective method to evaluate shape of the sperm head, with the calculated harmonic amplitudes highly related to male fertility. The FHA approach has been developed for use in the bull and the boar but has not yet been applied to the stallion. Direct utilization of the previous fluorescent approaches to identify and image live sperm nuclei in the bull cannot be used in the stallion due to the increased thickness of the post-nuclear region and thin anterior region of the sperm head. An alternative approach was developed in which live and motile sperm were isolated after filtration of an ejaculate through a Sephadex G-15 column. The resulting live sperm were sonicated briefly to separate tails and heads. The heads were isolated on a 45–90 discontinous Percoll gradient, fixed with paraformaldehyde (0.2%), centrifuged onto glass slides, and dried. The slides were then stained with eosin (1%), cleared with water, and dried again; Permount was added, followed by a coverslip. Slides were imaged with phase contrast microscopy; digital images were acquired and evaluated with custom software to identify perimeter coordinates of sperm nuclei. The perimeter coordinates were next converted to Fourier harmonic amplitudes 0–5 (HA0–HA5) using trigonomic regression at 1 degree equally spaced angles. Fertility of bulls were previously reported to be most related to changes in HA0 and HA2 but no information is available on stallions. As fertility data on stallions is limited, to evaluate FHA in the equine, the day length (period of light) was increased in January from the ambient 9–10 h to 16. Semen samples from 5 light horse stallions were collected at weekly intervals for 8 weeks following the increase in light. It was hypothesized that fertility would increase for each stallion over the course of the experiment, as testosterone increases and spermatogenesis improves with increasing day length, as previously shown. Each week semen samples were evaluated for FHA live sperm nuclei, as described above. All parameter means and variations were recorded on 100 randomly selected sperm nuclei per semen sample evaluated. There was no difference relative to week 0 in the least squares mean or SD of HA0, HA1, HA2, HA3, HA4, or HA5 over the 8 weeks (P > 0.05). However stallions were consistently different for all HAs (P < 0.05). The overall mean HA0–HA5 � SEM were 1.973 � 0.38, 0.087 � 0.002, 0.721 � 0.021, 0.051 � 0.004, 0.208 � 0.014, and 0.031 � 0.002, respectively. Even though libido increased during the experiment, confirming the effect of light on the stallions, no affect on sperm nuclear shape or its variation was detected using FHA. Based on work in other species involving numbers of sperm inseminated, if large numbers of sperm from these stallions were inseminated in mares, we would predict no change in fertility due to season. Further research is needed to confirm this prediction.

PSXII-17 Fertility prediction model for Nilli-Ravi buffalo bulls through Fourier harmonic analysis of sperm

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 424-424
Author(s):  
John J Parrish ◽  
Javeria Arshad ◽  
M A Awan ◽  
S Akhter
Keyword(s):  
Harmonic Analysis ◽  
Nuclear Shape ◽  
Low Fertility ◽  
Harmonic Amplitude ◽  
High Fertility ◽  
Fourier Harmonic ◽  
The Mean ◽  
Sperm Nuclei ◽  
Live Sperm

Abstract A model to predict Nili-ravi buffalo bull fertility was developed based on Fourier harmonic analysis of sperm. Seventeen bulls with 3032 AI records were categorizes based on fertility rate (FR) as low (36.5±0.2, n = 6: SD&lt; ˗1 from mean FR), medium (39.9±0.2, n = 3; SD +1 to -1 from mean FR) and high fertility (41.4±0.1, n = 8; SD &gt; +1 from mean FR). Cryopreserved semen samples from these bulls were investigated for Fourier harmonic analysis of sperm nuclear shape. Hoechst-33342 and YOYO-1 fluorescent stains were used to identify live and dead sperm. Digital images were analyzed to get sperm nuclear perimeter points at different phase angles to generate Fourier functions. Mean harmonic amplitude (HA) 0 was different (P &lt; 0.05) for 1700 live vs. 1294 dead sperm from the 17 bulls, thus live sperm were used for remaining analyses. The mean, variance, skewness and kurtosis values of 100 live sperm nuclei/bull were compared for HA0-5 between high (n = 6) and low (n = 6) fertility groups, considering equal number of bulls in each category. The mean HA2 (0.739±0.01 vs 0.686±0.00) and 4 (0.105±0.001 vs 0.007±0.001) were higher in high vs low fertility group respectively (P &lt; 0.05). Sperm nuclear perimeter among high fertility group sperm was more elongated. There was also an increased skewness of HA0 as fertility increased (P &lt; 0.05). Discriminant analysis defined a fertility model by using mean HA4, skewness HA0 and variance HA2, that resulted in 91.7% bulls into their correct fertility group upon cross-validation (canonical correlation=0.928; P &lt; 0.05). Higher values of mean HA4, skewness HA0 and variance HA2 increase the chance of bulls being placed in the high fertility group. In conclusion, sperm nuclear shape in Nili-ravi buffalo bull is related to in vivo fertility. A fertility model using reported discriminant measures could be used to objectively identify Nili-ravi buffalo bulls of varying fertility.

A mutation study of sperm head shape and motility in the mouse: lessons for the clinic

Andrology ◽  
2014 ◽  
Vol 3 (2) ◽  
pp. 174-202 ◽  
Author(s):  
P. de Boer ◽  
M. de Vries ◽  
L. Ramos
Keyword(s):  
Sperm Head ◽  
Head Shape

scholarly journals Distribution of DNA damage in the human sperm nucleus: implications of the architecture of the sperm head

2020 ◽  
Vol 22 (4) ◽  
pp. 401
Author(s):  
MaríaPaz Herráez ◽  
Silvia González-Rojo ◽  
Cristina Fernández-Díez ◽  
Marta Lombó
Keyword(s):  
Dna Damage ◽  
Human Sperm ◽  
Sperm Nucleus ◽  
Sperm Head

scholarly journals Core Histones Are Constituents of the Perinuclear Theca of Murid Spermatozoa: An Assessment of Their Synthesis and Assembly during Spermiogenesis and Function after Gametic Fusion

2021 ◽  
Vol 22 (15) ◽  
pp. 8119
Author(s):  
Lauren E. Hamilton ◽  
Morgan Lion ◽  
Luis Aguila ◽  
João Suzuki ◽  
Genevieve Acteau ◽  
...  
Keyword(s):  
Chromatin Remodeling ◽  
De Novo ◽  
Sperm Nucleus ◽  
Sperm Head ◽  
Cell Fractionation ◽  
Cellular Processes ◽  
Functional Potential ◽  
Core Histones ◽  
Perinuclear Theca ◽  
And Function

The perinuclear theca (PT) of the eutherian sperm head is a cytoskeletal-like structure that houses proteins involved in important cellular processes during spermiogenesis and fertilization. Building upon our novel discovery of non-nuclear histones in the bovine PT, we sought to investigate whether this PT localization was a conserved feature of eutherian sperm. Employing cell fractionation, immunodetection, mass spectrometry, qPCR, and intracytoplasmic sperm injections (ICSI), we examined the localization, developmental origin, and functional potential of histones from the murid PT. Immunodetection localized histones to the post-acrosomal sheath (PAS) and the perforatorium (PERF) of the PT but showed an absence in the sperm nucleus. MS/MS analysis of selectively extracted PT histones indicated that predominately core histones (i.e., H3, H3.3, H2B, H2A, H2AX, and H4) populate the murid PT. These core histones appear to be de novo-synthesized in round spermatids and assembled via the manchette during spermatid elongation. Mouse ICSI results suggest that early embryonic development is delayed in the absence of PT-derived core histones. Here, we provide evidence that core histones are de novo-synthesized prior to PT assembly and deposited in PT sub-compartments for subsequent involvement in chromatin remodeling of the male pronucleus post-fertilization.

scholarly journals Sexual selection on protamine and transition nuclear protein expression in mouse species

2014 ◽  
Vol 281 (1783) ◽  
pp. 20133359 ◽  
Author(s):  
Lena Lüke ◽  
Polly Campbell ◽  
María Varea Sánchez ◽  
Michael W. Nachman ◽  
Eduardo R. S. Roldan
Keyword(s):  
Sexual Selection ◽  
Sperm Competition ◽  
Nuclear Protein ◽  
Sperm Head ◽  
Nuclear Proteins ◽  
Sperm Chromatin ◽  
Head Shape ◽  
Mouse Species ◽  
The Relationship ◽  
Protamine 2

Post-copulatory sexual selection in the form of sperm competition is known to influence the evolution of male reproductive proteins in mammals. The relationship between sperm competition and regulatory evolution, however, remains to be explored. Protamines and transition nuclear proteins are involved in the condensation of sperm chromatin and are expected to affect the shape of the sperm head. A hydrodynamically efficient head allows for fast swimming velocity and, therefore, more competitive sperm. Previous comparative studies in rodents have documented a significant association between the level of sperm competition (as measured by relative testes mass) and DNA sequence evolution in both the coding and promoter sequences of protamine 2. Here, we investigate the influence of sexual selection on protamine and transition nuclear protein mRNA expression in the testes of eight mouse species that differ widely in levels of sperm competition. We also examined the relationship between relative gene expression levels and sperm head shape, assessed using geometric morphometrics. We found that species with higher levels of sperm competition express less protamine 2 in relation to protamine 1 and transition nuclear proteins. Moreover, there was a significant association between relative protamine 2 expression and sperm head shape. Reduction in the relative abundance of protamine 2 may increase the competitive ability of sperm in mice, possibly by affecting sperm head shape. Changes in gene regulatory sequences thus seem to be the basis of the evolutionary response to sexual selection in these proteins.

Electron microscopic study of sperm head differentiation in the lizard Agama stellio

1987 ◽  
Vol 65 (12) ◽  
pp. 2959-2968 ◽  
Author(s):  
Hameed Al-Hajj ◽  
Suha Janakat ◽  
Fahmi Mahmoud
Keyword(s):  
Nuclear Envelope ◽  
Microscopic Study ◽  
Electron Microscopic Study ◽  
Longitudinal Axis ◽  
Sperm Head ◽  
Fibrous Layer ◽  
Electron Microscopic ◽  
Acrosomal Vesicle ◽  
Acrosomal Membrane ◽  
Chromatin Material

Early differentiation of the spermatid of Agama stellio is demonstrated by two anterior nuclear depressions, occupied by two proacrosomal vesicles, which fuse to form one vesicle. Later, this vesicle exhibits an acrosomal granule in its midposterior portion. The space between the posterior acrosomal membrane and the nuclear envelope is occupied by a subacrosomal fibrous layer which later exhibits a subacrosomal granule posterior to the acrosomal granule. The acrosomal vesicle and the nuclear depression flatten and later elongate. The acrosomal granule spreads and assumes the inverted V shape of the acrosomal vesicle, and the subacrosomal material assumes a feathery shape capping the nuclear prolongation. The subacrosomal granule on top of this feathery material forms a long, cross-striated subacrosomal rod which extends towards the tip of the acrosome. The chromatin material undergoes condensation into spirally oriented fibers, which eventually become homogeneous and dense. This process is accompanied by a change in the orientation of the manchette microtubules, which initially occur as rings around the nucleus and are eventually found parallel to the longitudinal axis of the nucleus.

Identification of sperm morphometric subpopulations in the canine ejaculate: do they reflect different subpopulations in sperm chromatin integrity?

Zygote ◽  
2007 ◽  
Vol 15 (3) ◽  
pp. 257-266 ◽  
Author(s):  
I. Núñez-Martinez ◽  
J.M. Moran ◽  
F.J. Peña
Keyword(s):  
Semen Quality ◽  
Principal Component ◽  
Sperm Head ◽  
Data Matrix ◽  
Sperm Chromatin ◽  
Head Width ◽  
Dna Integrity ◽  
Linear Regression Models ◽  
Head Shape ◽  
Shape Factors

SummaryA statistical approach using sequentially principal component analysis (PCA) clustering and discriminant analysis was developed to disclose morphometric sperm subpopulations. In addition, we used a similar approach to disclose subpopulations of spermatozoa with different degrees of DNA fragmentation. It is widely accepted that sperm morphology is a strong indicator of semen quality and since the sperm head mainly comprises the sperm DNA, it has been proposed that subtle changes in sperm head morphology may be related to abnormal DNA content. Semen from four mongrel dogs (five replicates per dog) were used to investigate DNA quality by means of the sperm chromatin structure assay (SCSA), and for computerized sperm morphometry (ASMA). Each sperm head was measured for nine primary parameters: head area (A), head perimeter (P), head length (L), head width (W), acrosome area (%), midpiece width (w), midpiece area (a), distance (d) between the major axes of the head and midpiece, angle (θ) of divergence of the midpiece from the head axis; and four parameters of head shape: FUN1 (L/W), FUN2 (4π A/P2), FUN3 ((L – W)/(L + W)) and FUN 4 (π LW/4A). The data matrix consisted of 2361 observations, (morphometric analysis on individual spermatozoa) and 63 815 observations for the DNA integrity. The PCA analysis revealed five variables with Eigen values over 1, representing more than 79% of the cumulative variance. The morphometric data revealed five sperm subpopulations, while the DNA data gave six subpopulations of spermatozoa with different DNA integrity. Significant differences were found in the percentage of spermatozoa falling in each cluster among dogs (p < 0.05). Linear regression models including sperm head shape factors 2, 3 and 4 predicted the amount of denatured DNA within each individual spermatozoon (p < 0.001). We conclude that the ASMA analysis can be considered a powerful tool to improve the spermiogram.

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