A portable sperm cell purification instrument based on continuous flow acoustophoretic separation of sperm cells for on-site forensic sample pretreatment

Lab on a Chip ◽  
2021 ◽  
Author(s):  
Kai Sun ◽  
Han Wang ◽  
Lei Wang ◽  
Ying Lu ◽  
Ran Liu ◽  
...  

Obtaining purified male sperm specimen from the original forensic sample mixtures is a critical procedure in identifying criminal suspect in the forensic analysis of sexual assult crimes. Differential extraction (DE)...

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Kazuki Motomura ◽  
Hidenori Takeuchi ◽  
Michitaka Notaguchi ◽  
Haruna Tsuchi ◽  
Atsushi Takeda ◽  
...  

AbstractDuring the double fertilization process, pollen tubes deliver two sperm cells to an ovule containing the female gametes. In the pollen tube, the vegetative nucleus and sperm cells move together to the apical region where the vegetative nucleus is thought to play a crucial role in controlling the direction and growth of the pollen tube. Here, we report the generation of pollen tubes in Arabidopsis thaliana whose vegetative nucleus and sperm cells are isolated and sealed by callose plugs in the basal region due to apical transport defects induced by mutations in the WPP domain-interacting tail-anchored proteins (WITs) and sperm cell-specific expression of a dominant mutant of the CALLOSE SYNTHASE 3 protein. Through pollen-tube guidance assays, we show that the physiologically anuclear mutant pollen tubes maintain the ability to grow and enter ovules. Our findings provide insight into the sperm cell delivery mechanism and illustrate the independence of the tip-localized vegetative nucleus from directional growth control of the pollen tube.


2008 ◽  
Vol 57 (11) ◽  
pp. 1405-1410 ◽  
Author(s):  
Tidhar Zan Bar ◽  
Ronen Yehuda ◽  
Tomer Hacham ◽  
Sigal Krupnik ◽  
Benjamin Bartoov

Campylobacter fetus subsp. fetus infection can occur in female sheep, causing infertility or abortion. Despite extensive research on the effect of these bacteria on female fertility, little research has been done on the influence of C. fetus subsp. fetus on the male factor. Our objective was to examine the influence of C. fetus subsp. fetus on ram sperm. Motility index, percentage of live spermatozoa, mean αt value (an indication of the chromatin stability of the sperm cell) and percentage of sperm cells expressing the FAS receptor were measured in sperm incubated in the presence or absence of C. fetus subsp. fetus. The motility index and viability of sperm incubated with the bacteria were lower than those of untreated sperm samples after 5 h. In bacteria-incubated sperm cells, the percentage expressing FAS receptor was already significantly elevated at 15 min. Bacteria-incubated sperm showed a greater prevalence of morphological damage. The bacteria were attached to tail and acrosome regions, and the sperm damage was concentrated in both the motility and chromatin regions. Bacteria-infected sperm cells showed a decrease in motility, increase in early acrosome reaction and chromatin damage. Similar effects were induced by incubation of the sperm with supernatants from C. fetus subsp. fetus cultures. Thus this study demonstrates that C. fetus subsp. fetus has a detrimental effect on the quality of ram sperm.


Author(s):  
Akila CR ◽  
Dinesh Babu J ◽  
Sravan Kumar P ◽  
Vinaya B

Mycotoxins represent poisonous materials produced via certain types of growths. Among different mycotoxins, aflatoxins are viewed as plainly hazardous, taking into account that they are portrayed as cancer-causing for creatures and individuals. The admission of aflatoxins by means of feeds or nourishments should cause pernicious results on creatures' or people's wellbeing. Exploration on creatures has indicated that the overall casing circumstance just as a portion of the blood boundaries, particularly those of the liver may be adversely influenced with aflatoxin the executives. As to conceptive device, in spite of the fact that not remarkably examined, a few specialists bolster the helpless impacts of aflatoxins both on women or on grown-up guys. More precisely, in male, the scale and weight of the genital organs, the spermatogenesis, the number, the motility and the morphology of sperm cells just as hormones' fixations can be influenced after presentation of the creatures to aflatoxins, making barrenness inconveniences additional normal. Most examination allude to lab and significantly less to viable creatures, while least complex two investigations look for counsel from the doable issues of barrenness on men in view of aflatoxins. Since imitation is made one out of the greatest fundamental areas of creature cultivation, exceptional intrigue should be paid to supplements with the goal that the chance of the aflatoxin admission by creatures could be evacuated, the creature wellbeing extraordinarily in regards to the conceptive gadget may be covered and financial misfortunes could be enhanced.


1992 ◽  
Vol 102 (3) ◽  
pp. 487-494 ◽  
Author(s):  
R.K. Naz ◽  
K. Ahmad ◽  
P. Kaplan

The presence and role of c-ras proteins were investigated in mature human sperm cells. The v-H-ras monoclonal antibody (mAb) against the c-ras protein, p21, reacted specifically with the acrosomal region of methanol-fixed as well as unfixed-live capacitated and non-capacitated human sperm cell in the indirect immunofluorescence technique. The v-H-ras mAb predominantly recognized c-ras protein of 21 kDa on the Western blot of lithium diiodosalicylate (LIS)-solubilized human sperm preparation. The incubation of sperm cells with v-H-ras mAb affected the sperm cell function in the human sperm penetration assay. The antibody significantly reduced the acrosome reaction and release of acrosin activity from the sperm cells. There was no effect of the mAb on percentage motility, although the mAb significantly affected various motility characteristics such as linearity, amplitude of lateral head displacement (ALH) and beat frequency, the motility parameters involved in the hyperactivation phenomenon of sperm cells leading to capacitation and acrosome reaction. These results suggest that the c-ras or c-ras-like proteins are present in mature sperm cell and may have a role in capacitation and/or acrosome reaction of human sperm cell.


1998 ◽  
Vol 76 (10) ◽  
pp. 1822-1836 ◽  
Author(s):  
Martin N DiLauro ◽  
Wayne Kaboord ◽  
Rosemary A Walsh ◽  
William F Krise ◽  
Michael A Hendrix

Atlantic sturgeon (Acipenser oxyrhynchus) and lake sturgeon (Acipenser fulvescens) sperm-cell morphologies were examined using scanning electron microscopy. Major differences were found in four of nine metrics, all in the head region of the cell. Atlantic sturgeon sperm cells were much shorter than those of lake sturgeon. Anterior head width exceeded posterior head width, in contrast to the arrangement in lake sturgeon sperm cells. Lake sturgeon sperm cells are nearer in size to those of other sturgeons than are Atlantic sturgeon sperm cells. Comparisons were made with sperm-cell structures known from other sturgeon species, including the Russian sturgeon (Acipenser gueldenstaedti colchicus), stellate sturgeon (Acipenser stellatus), Chinese sturgeon (Acipenser sinensis), and white sturgeon (Acipenser transmontanus). Variation in cell morphology may indicate evolutionary relationships. In addition, the fine structure of Atlantic sturgeon sperm cells was examined using transmission electron microscopy and selected metrics are described. The cell possesses a distinct acrosome, a midpiece, and a single flagellum. A comparison is made with ultrastructural details of the sperm cells of stellate and white sturgeons. Similarities among these species include radial symmetry about the longitudinal axis, an elongate shape, a distinct acrosome, and the presence of endonuclear canals. Noteworthy differences include a smaller total length and width than stellate and white sturgeon sperm cells. The main sperm-cell body is approximately 4 µm long and the flagellum about 37 µm long, resulting in a total cell length of about 41 µm. Also, the Atlantic sturgeon sperm cell possesses only two membraned endonuclear canals, in contrast to the arrangement in white and stellate sturgeons, where three such organelles are found. A structural connection of unknown function between the nuclear fossa and proximal centriole is also present in the Atlantic sturgeon sperm cell. Sperm-cell nuclei of white and stellate sturgeons are elongate trapezoids, with the anterior end narrower, whereas in Atlantic sturgeon the anterior portion of the trapezoid is wider than the posterior. Structural similarities between species may indicate a commonality of ancestral and evolutionary relationships that may have taxonomic implications. Ultrastructure suggests a closer evolutionary relationship between the white and stellate sturgeon than between either of these species and the Atlantic sturgeon. The present findings may be used by biologists studying the reproductive physiology, forensics, taxonomy, and genetics of sturgeons.


2007 ◽  
Vol 19 (1) ◽  
pp. 315
Author(s):  
W. B. Feitosa ◽  
M. P. Milazzotto ◽  
E. A. L. Martins ◽  
A. M. Rocha ◽  
J. L. Avanzo ◽  
...  

Mammalian sperm cells have a spontaneous ability to take up exogenous DNA in a process regulated by specific protein. However, little is known about the effect of exogenous DNA on sperm cell association. The aim of this work was to evaluate the effect of size, concentration, and structure of exogenous DNA sequence in apoptosis and oncosis induction and the association with spermatozoa. To evaluate the effect of size and concentration of exogenous DNA, sequences of 2.2, 5.5, and 8.5 kb were used at the concentrations of 500 ng (Experiment 1) or 5 × 1011 molecules (Experiment 2). To assess the effect of structure (Experiment 3), 3 sequences of approximately 500 bp were used, containing 65.7% of AT (AT sequence), 46.4% of AT (IN sequence), and 38.6% of AT (GC sequence). To evaluate apoptosis and oncosis in all treatments, sperm cells were incubated with exogenous DNA for 1 h, stained with 2 µL of Yo-Pro (100 µM) for 20 min, and 10 µL of propidium iodide (6 µM) for 10 min, and then analyzed by flow cytometry. To evaluate the transfection rates, sperm cells, after incubation, were separated from non-associated exogenous DNA by 2.5% gel electrophoresis. DNA (genomic and exogenous) was extracted and association rates obtained by real-time PCR. Data were submitted to ANOVA and compared by Tukey's test (P ≤ 0.05). The number of viable, apoptotic, oncosis, or in initial apoptosis/late oncosis spermatozoa incubated with different concentrations or sequences of exogenous DNA did not differ among groups. In Experiment 1, a higher DNA association with sperm cells was observed in the 5.5 kb sequence in comparison with the 2.2 and 8.5 kb. In Experiment 2, the 8.5 kb sequence had the lower association with the spermatozoa in comparison with the 2.2 kb and 5.5 kb. In Experiment 3, there was a higher interaction of the AT sequence when compared to IN and GC sequences. Reference data suggest that big sequences of exogenous DNA have advantages in the interaction with spermatozoa. The present work is the first one to show that the advantage of the big sequences on interaction became a disadvantage in relation to the ability of these cells to internalize this DNA. Table 1. Quantification of different exogenous DNA associated to sperm cells Financial support was provided by FAPESP 03/10234-7 and 03/07456-8.


Reproduction ◽  
2005 ◽  
Vol 130 (5) ◽  
pp. 615-626 ◽  
Author(s):  
Anke Kurz ◽  
Dagmar Viertel ◽  
Andreas Herrmann ◽  
Karin Müller

One of the essential properties of mammalian, including sperm, plasma membranes is a stable transversal lipid asymmetry with the aminophospholipids, phosphatidylserine (PS) and phosphatidylethanolamine (PE), typically in the inner, cytoplasmic leaflet. The maintenance of this nonrandom lipid distribution is important for the homeostasis of the cell. To clarify the relevance of lipid asymmetry to sperm function, we have studied the localization of PS in boar sperm cell membranes. By using labeled annexin V as a marker for PS and propidium iodide (PI) as a stain for nonviable cells in conjunction with different methods (flow cytometry, fluorescence and electron microscopy), we have assessed the surface exposure of PS in viable cells during sperm genesis, that is, before and during capacitation as well as after acrosome reaction. An approach was set up to address also the presence of PS in the outer acrosome membrane. The results show that PS is localized in the cytoplasmic leaflet of the plasma membrane as well as on the outer acrosome membrane. Our results further indicate the cytoplasmic localization of PS in the postacrosomal region. During capacitation and acrosome reaction of spermatozoa, PS does not become exposed on the outer surface of the viable cells. Only in a subpopulation of PI-positive sperm cells does PS became accessible upon capacitation. The stable cytoplasmic localization of PS in the plasma membrane, as well as in the outer acrosome membrane, is assumed to be essential for a proper genesis of sperm cells during capacitation and acrosome reaction.


1967 ◽  
Vol 45 (1) ◽  
pp. 27-30 ◽  
Author(s):  
Liang-Yu Wu

Structural differences in the spermatheca and sperm cells of Ditylenchus and Tylenchus were found to be of significant value in the separation of these two genera. In Ditylenchus, the spermatheca is a long tubular structure located between the uterus and the oviduct. The sperm cell consists of two parts: a dense nucleus and a large translucent vesicle. In Tylenchus, the spermatheca, when present, is in the form of a diverticulum of various shapes, arising at the junction of the uterus and the oviduct. The translucent vesicle is absent within the sperm cell.


PLoS ONE ◽  
2021 ◽  
Vol 16 (7) ◽  
pp. e0253628
Author(s):  
Sarah K. Lamar ◽  
Nicola J. Nelson ◽  
Jennifer A. Moore ◽  
Helen R. Taylor ◽  
Susan N. Keall ◽  
...  

Successful reproduction is critical to the persistence of at-risk species; however, reproductive characteristics are understudied in many wild species. New Zealand’s endemic tuatara (Sphenodon punctatus), the sole surviving member of the reptile order Rhynchocephalia, is restricted to 10% of its historic range. To complement ongoing conservation efforts, we collected and characterized mature sperm from male tuatara for the first time. Semen collected both during mating and from urine after courting contained motile sperm and had the potential for a very high percentage of viable sperm cells (98%). Scanning electron microscopy revealed a filiform sperm cell with distinct divisions: head, midpiece, tail, and reduced end piece. Finally, our initial curvilinear velocity estimates for tuatara sperm are 2–4 times faster than any previously studied reptile. Further work is needed to examine these trends at a larger scale; however, this research provides valuable information regarding reproduction in this basal reptile.


2019 ◽  
Vol 62 (2) ◽  
pp. 557-564
Author(s):  
Mustafa Erkan Özgür ◽  
Zeynep Maraş ◽  
Selim Erdoğan

Abstract. In this study, semen seminal plasma contents and the motility of sperm cells were determined in Capoeta trutta via a computer-assisted sperm analysis system. In addition, we evaluated the relationship between semen seminal plasma ions and the velocities of sperm cells. Although the predominant ions were K (206.84±20.61 mg L−1) and Na (128.06±23.82 mg L−1) in the semen seminal plasma, Ca (14.05±4.13 mg L−1) and Mg (3.35±0.44 mg L−1) were not predominate according to our results. However, partially strong relationships between the curvilinear velocity value (VCL) and K (R2=0.67; p<0.05) were found, while it was moderate with Mg (R2=0.48; p<0.05). There was a weak relationship with Na (R2=0.17; p<0.05) and Ca (R2=0.34; p<0.05). In our results, while the trace metals were determined as Zn > Al > B > Li > Cu in semen seminal plasma, they are not correlated with sperm cell velocities. Finally, we hope that the present information on the motility parameters of Capoeta trutta in this paper will eventually help artificial insemination in reproduction practices.


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