scholarly journals Dynamics of sperm subpopulations based on motility and plasma membrane status in thawed ram spermatozoa incubated under conditions that support in vitro capacitation and fertilisation

2014 ◽  
Vol 26 (5) ◽  
pp. 725 ◽  
Author(s):  
Olga García-Álvarez ◽  
Alejandro Maroto-Morales ◽  
Manuel Ramón ◽  
Enrique del Olmo ◽  
Pilar Jiménez-Rabadán ◽  
...  

The present study evaluated modifications occurring in thawed ram spermatozoa during incubation in different media that supported in vitro capacitation and fertilisation, and examines how these changes relate to IVF. Thawed sperm samples were incubated under capacitating (Cap) and non-capacitating (non-Cap) conditions for 0, 1 and 2 h and used in an IVF test. During incubation, changes related to membrane status and the motility pattern of spermatozoa were assessed, the latter being used to characterise sperm subpopulations. A significantly greater increase (P ≤ 0.05) in the percentage of spermatozoa with higher membrane fluidity was observed in samples incubated with Cap medium from the beginning of incubation. In addition, changes over time in the distribution of the motile subpopulation were particularly evident when spermatozoa were incubated with Cap medium, with a noted increase in spermatozoa classified as ‘hyperactivated like’, with major changes occurring after 1 h incubation. Both characteristics (i.e. membrane fluidity and the percentage of the hyperactivated-like subpopulation) were significantly related with in vitro fertility, and only sperm samples incubated with the Cap medium were capable of fertilising oocytes. These results support the idea that changes in sperm membrane fluidity and motility pattern (i.e. an increase in hyperactivated spermatozoa) are needed for fertilisation to take place.

1990 ◽  
Vol 10 (3) ◽  
pp. 263-270 ◽  
Author(s):  
J. Pascal Zimmer ◽  
Hans A. Lehr ◽  
Christoph Hübner ◽  
Stephan G. Lindner ◽  
Ralf Ramsperger ◽  
...  

Although most non-human primates, except the chimpanzee and the gibbon in vivo are not infectible by HIV-1, lymphocytes of several of these species can be infected by HIV-1 in vitro.In order to investigate whether the in vitro infectibility of primate lymphocytes might be attributed to plasma membrane adaptation processes or to serum factors, we compared HIV-1 infectibility of cultivated peripheral blood lymphocytes of macaques and of baboons on day one and on day ten of cultivation. These data were correlated to plasma membrane lipid composition and membrane fluidity.We found a correlation between increased HIV-1 in vitro infectibility and changes in plasma membrane lipid composition resulting in decreased membrane fluidity of cultured primate lymphocytes.


Zygote ◽  
2015 ◽  
Vol 24 (3) ◽  
pp. 364-370 ◽  
Author(s):  
Seunghoon Lee ◽  
Yuuki Hiradate ◽  
Yumi Hoshino ◽  
Yeoung-gyu Ko ◽  
Kentaro Tanemura ◽  
...  

SummaryMany studies of the main gap junction protein, Cx43, have been conducted in porcine oocyte research, but they have been limited to investigations of cumulus–oocyte complexes (COCs). In this study, we verified Cx43 not in COCs, but in porcine oocytes during maturation, and conducted a quantitative time course analysis. The location and dynamics of Cx43 were examined by immunocytochemistry and western blotting, respectively. COCs were cultured in NCSU23 medium and processed for immunocytochemistry and western blotting at 0, 14, 28, and 42 h after denuding. A Cx43 signal was detected on oolemmas, transzonal projections and the surface of zona pellucidae. Western blotting showed that Cx43 band density increased from 0 to 14 h, and gradually decreased thereafter. Our results clarified that Cx43 is localized in the ooplasmic membrane through zona pellucidae and its level changes over time during culture in porcine oocytes.


Author(s):  
Paula Piccolo Maitan ◽  
Elizabeth G. Bromfield ◽  
Romy Hoogendijk ◽  
Miguel Ricardo Leung ◽  
Tzviya Zeev-Ben-Mordehai ◽  
...  

Classical in vitro fertilization (IVF) is still poorly successful in horses. This lack of success is thought to be due primarily to inadequate capacitation of stallion spermatozoa under in vitro conditions. In species in which IVF is successful, bicarbonate, calcium, and albumin are considered the key components that enable a gradual reorganization of the sperm plasma membrane that allows the spermatozoa to undergo an acrosome reaction and fertilize the oocyte. The aim of this work was to comprehensively examine contributors to stallion sperm capacitation by investigating bicarbonate-induced membrane remodelling steps, and elucidating the contribution of cAMP signalling to these events. In the presence of capacitating media containing bicarbonate, a significant increase in plasma membrane fluidity was readily detected using merocyanine 540 staining in the majority of viable spermatozoa within 15 min of bicarbonate exposure. Specific inhibition of soluble adenylyl cyclase (sAC) in the presence of bicarbonate by LRE1 significantly reduced the number of viable sperm with high membrane fluidity. This suggests a vital role for sAC-mediated cAMP production in the regulation of membrane fluidity. Cryo-electron tomography of viable cells with high membrane fluidity revealed a range of membrane remodelling intermediates, including destabilized membranes and zones with close apposition of the plasma membrane and the outer acrosomal membrane. However, lipidomic analysis of equivalent viable spermatozoa with high membrane fluidity demonstrated that this phenomenon was neither accompanied by a gross change in the phospholipid composition of stallion sperm membranes nor detectable sterol efflux (p > 0.05). After an early increase in membrane fluidity, a significant and cAMP-dependent increase in viable sperm with phosphatidylserine (PS), but not phosphatidylethanolamine (PE) exposure was noted. While the events observed partly resemble findings from the in vitro capacitation of sperm from other mammalian species, the lack of cholesterol removal appears to be an equine-specific phenomenon. This research will assist in the development of a defined medium for the capacitation of stallion sperm and will facilitate progress toward a functional IVF protocol for horse gametes.


2019 ◽  
Vol 121 (6) ◽  
pp. 601-614 ◽  
Author(s):  
Carlos A. Montoya ◽  
Sharon J. Henare ◽  
Peter Zhu ◽  
Shane M. Rutherfurd ◽  
Paul J. Moughan

AbstractThe effect of kiwi fruit at two dietary levels on the adaptation of intestinal fermentation over time in the growing pig was studied. A semi-synthetic fibre-free diet and two semi-synthetic diets containing kiwi fruit as a model fibre source (133 or 266 g/kg (DM basis); 28 or 48 g fibre/kg) were formulated and the diets contained titanium dioxide as an indigestible marker. A total of fourteen ileal cannulated pigs (41 kg body weight) were fed the fibre-free diet for 7 d followed by either the low or high kiwi fruit-containing diets (n 7/diet) for a further 44 d. Ileal digesta and faeces were collected at five times throughout the study. Ileal digesta were fermented (in vitro) with a standard pooled human faecal inoculum, while fresh pig faeces were used as inocula to ferment in vitro a standard purified fibre. Observations were normalised for diet DM intake using the marker. The 16S ribosomal RNA gene copy number of ileal and total faecal bacteria were high for the high-kiwi fruit level diet (P<0·05). The ileal bacteria tended to decrease over time (P<0·1), while the faecal bacteria increased (P<0·05), at the same rate for both diets. The amounts of crude protein and insoluble dietary fibre entering the hindgut changed over time similarly for both diets, whereas for starch it changed only for the low kiwi fruit-containing diet (P<0·05). Changes over time were also observed for the predicted hindgut valeric acid production and butyric acid absorption (P<0·05). In conclusion, adaptational changes over time of some characteristics of intestinal fermentation depended on the dietary level of kiwi fruit.


Cancers ◽  
2021 ◽  
Vol 13 (15) ◽  
pp. 3852
Author(s):  
Federica Foglietta ◽  
Vanessa Pinnelli ◽  
Francesca Giuntini ◽  
Nadia Barbero ◽  
Patrizia Panzanelli ◽  
...  

Sonodynamic Therapy (SDT) is a new anticancer strategy based on ultrasound (US) technique and is derived from photodynamic therapy (PDT); SDT is still, however, far from clinical application. In order to move this therapy forward from bench to bedside, investigations have been focused on treatment selectivity between cancer cells and normal cells. As a result, the effects of the porphyrin activation by SDT on cancer (HT-29) and normal (HDF 106-05) cells were studied in a co-culture evaluating cell cytotoxicity, reactive oxygen species (ROS) production, mitochondrial function and plasma membrane fluidity according to the bilayer sonophore (BLS) theory. While PDT induced similar effects on both HT-29 and HDF 106-05 cells in co-culture, SDT elicited significant cytotoxicity, ROS production and mitochondrial impairment on HT-29 cells only, whereas HDF 106-05 cells were unaffected. Notably, HT-29 and HDF 106-05 showed different cell membrane fluidity during US exposure. In conclusion, our data demonstrate a marked difference between cancer cells and normal cells in co-culture in term of responsiveness to SDT, suggesting that this different behavior can be ascribed to diversity in plasma membrane properties, such as membrane fluidity, according to the BLS theory.


Author(s):  
N. Seki ◽  
Y. Toyama ◽  
T. Nagano

It is believed that i ntramembra.nous sterols play an essential role in membrane stability and permeability. To investigate the distribution changes of sterols in sperm membrane during epididymal maturation and capacitation, filipin has been used as a cytochemical probe for the detection for membrane sterols. Using this technique in combination with freeze fracturing, we examined the boar spermatozoa under various physiological conditions.The spermatozoa were collected from: 1) caput, corpus and cauda epididymides, 2) sperm rich fraction of ejaculates, and 3)the uterus 2hr after natural coition. They were fixed with 2.5% glutaraldehyde in 0.05M cacodylate buffer (pH 7.4), and treated with the filipin solution (final concentration : 0.02.0.05%) for 24hr at 4°C with constant agitation. After the filipin treatment, replicas were made by conventional freeze-fracture technique. The density of filipin-sterol complexes (FSCs) was determined in the E face of the plasma membrane of head regions.


VASA ◽  
2015 ◽  
Vol 44 (5) ◽  
pp. 355-362 ◽  
Author(s):  
Marie Urban ◽  
Alban Fouasson-Chailloux ◽  
Isabelle Signolet ◽  
Christophe Colas Ribas ◽  
Mathieu Feuilloy ◽  
...  

Abstract. Summary: Background: We aimed at estimating the agreement between the Medicap® (photo-optical) and Radiometer® (electro-chemical) sensors during exercise transcutaneous oxygen pressure (tcpO2) tests. Our hypothesis was that although absolute starting values (tcpO2rest: mean over 2 minutes) might be different, tcpO2-changes over time and the minimal value of the decrease from rest of oxygen pressure (DROPmin) results at exercise shall be concordant between the two systems. Patients and methods: Forty seven patients with arterial claudication (65 + / - 7 years) performed a treadmill test with 5 probes each of the electro-chemical and photo-optical devices simultaneously, one of each system on the chest, on each buttock and on each calf. Results: Seventeen Medicap® probes disconnected during the tests. tcpO2rest and DROPmin values were higher with Medicap® than with Radiometer®, by 13.7 + / - 17.1 mm Hg and 3.4 + / - 11.7 mm Hg, respectively. Despite the differences in absolute starting values, changes over time were similar between the two systems. The concordance between the two systems was approximately 70 % for classification of test results from DROPmin. Conclusions: Photo-optical sensors are promising alternatives to electro-chemical sensors for exercise oximetry, provided that miniaturisation and weight reduction of the new sensors are possible.


2007 ◽  
Author(s):  
Miranda Olff ◽  
Mirjam Nijdam ◽  
Kristin Samuelson ◽  
Julia Golier ◽  
Mariel Meewisse ◽  
...  

2010 ◽  
Author(s):  
Rebecca D. Stinson ◽  
Zachary Sussman ◽  
Megan Foley Nicpon ◽  
Allison L. Allmon ◽  
Courtney Cornick ◽  
...  

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