scholarly journals Comparison of electronic volume and forward scatter principles of cell selection using flow cytometry for the evaluation of acrosome and plasma membrane integrity of bull spermatozoa

2014 ◽  
Vol 85 (8) ◽  
pp. 719-728 ◽  
Author(s):  
Fride Berg Standerholen ◽  
Frøydis Deinboll Myromslien ◽  
Elisabeth Kommisrud ◽  
Erik Ropstad ◽  
Karin Elisabeth Waterhouse
Keyword(s):  
Flow Cytometry ◽  
Plasma Membrane ◽  
Membrane Integrity ◽  
Cell Selection ◽  
Forward Scatter ◽  
Plasma Membrane Integrity

scholarly journals Simultaneous evaluation of plasma membrane integrity, acrosomal integrity, and mitochondrial membrane potential in bovine spermatozoa by flow cytometry

Zygote ◽  
2015 ◽  
Vol 24 (4) ◽  
pp. 529-536 ◽  
Author(s):  
Chihiro Kanno ◽  
Sung-Sik Kang ◽  
Yasuyuki Kitade ◽  
Yojiro Yanagawa ◽  
Yoshiyuki Takahashi ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Plasma Membrane ◽  
Membrane Potential ◽  
Fluorescence Microscopy ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Membrane Integrity ◽  
Plasma Membrane Integrity ◽  
Bovine Spermatozoa ◽  
Acrosomal Integrity

SummaryThe present study aimed to develop an objective evaluation procedure to estimate the plasma membrane integrity, acrosomal integrity, and mitochondrial membrane potential of bull spermatozoa simultaneously by flow cytometry. Firstly, we used frozen–thawed semen mixed with 0, 25, 50, 75 or 100% dead spermatozoa. Semen was stained using three staining solutions: SYBR-14, propidium iodide (PI), and phycoerythrin-conjugated peanut agglutinin (PE–PNA), for the evaluation of plasma membrane integrity and acrosomal integrity. Then, characteristics evaluated by flow cytometry and by fluorescence microscopy were compared. Characteristics of spermatozoa (viability and acrosomal integrity) evaluated by flow cytometry and by fluorescence microscopy were found to be similar. Secondly, we attempted to evaluate the plasma membrane integrity, acrosomal integrity, and also mitochondrial membrane potential of spermatozoa by flow cytometry using conventional staining with three dyes (SYBR-14, PI, and PE–PNA) combined with MitoTracker Deep Red (MTDR) staining (quadruple staining). The spermatozoon characteristics evaluated by flow cytometry using quadruple staining were then compared with those of staining using SYBR-14, PI, and PE–PNA and staining using SYBR-14 and MTDR. There were no significant differences in all characteristics (viability, acrosomal integrity, and mitochondrial membrane potential) evaluated by quadruple staining and the other procedures. In conclusion, quadruple staining using SYBR-14, PI, PE–PNA, and MTDR for flow cytometry can be used to evaluate the plasma membrane integrity, acrosomal integrity, and mitochondrial membrane potential of bovine spermatozoa simultaneously.

scholarly journals Effect of Seminal Plasma Protein Fractions on Stallion Sperm Cryopreservation

2020 ◽  
Vol 21 (17) ◽  
pp. 6415
Author(s):  
Filipa Bubenickova ◽  
Pavla Postlerova ◽  
Ondrej Simonik ◽  
Jitka Sirohi ◽  
Jiri Sichtar
Keyword(s):  
Flow Cytometry ◽  
Plasma Membrane ◽  
Plasma Protein ◽  
Seminal Plasma ◽  
Protein Fraction ◽  
Membrane Integrity ◽  
Protein Fractions ◽  
Heparin Binding ◽  
Plasma Membrane Integrity ◽  
Acrosomal Membrane

Seminal plasma (SP) is the natural environment for spermatozoa and contains a number of components, especially proteins important for successful sperm maturation and fertilization. Nevertheless, in standard frozen stallion insemination doses production, SP is completely removed and is replaced by a semen extender. In the present study, we analyzed the effects of the selected seminal plasma protein groups that might play an important role in reducing the detrimental effects on spermatozoa during the cryopreservation process. SP proteins were separated according to their ability to bind to heparin into heparin-binding (Hep+) and heparin-non-binding (Hep−) fractions. The addition of three concentrations—125, 250, and 500 µg/mL—of each protein fraction was tested. After thawing, the following parameters were assessed: sperm motility (by CASA), plasma membrane integrity (PI staining), and acrosomal membrane integrity (PNA staining) using flow cytometry, and capacitation status (anti-phosphotyrosine antibody) using imaging-based flow cytometry. Our results showed that SP protein fractions had a significant effect on the kinematic parameters of spermatozoa and on a proportion of their subpopulations. The 125 µg/mL of Hep+ protein fraction resulted in increased linearity (LIN) and straightness (STR), moreover, with the highest values of sperm velocities (VAP, VSL), also this group contained the highest proportion of the fast sperm subpopulation. In contrast, the highest percentage of slow subpopulation was in the groups with 500 µg/mL of Hep+ fraction and 250 µg/mL of Hep− fraction. Interestingly, acrosomal membrane integrity was also highest in the groups with Hep+ fraction in concentrations of 125 µg/mL. Our results showed that the addition of protein fractions did not significantly affect the plasma membrane integrity and capacitation status of stallion spermatozoa. Moreover, our results confirmed that the effect of SP proteins on the sperm functionality is concentration-dependent, as has been reported for other species. Our study significantly contributes to the lack of studies dealing with possible use of specific stallion SP fractions in the complex puzzle of the improvement of cryopreservation protocols. It is clear that improvement in this field still needs more outputs from future studies, which should be focused on the effect of individual SP proteins on other sperm functional parameters with further implication on the success of artificial insemination in in vivo conditions.

Comparision of DNA fragmentantion and plasma membrane integrity between chilled and frozen semen of bulls

2014 ◽  
Author(s):  
Mello Papa Patricia de ◽  
Carlos Ramires Neto ◽  
Priscilla Nascimento Guasti ◽  
Rosiara Rosaria Dias Maziero ◽  
Yame F R Sancler-Silva ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Integrity ◽  
Plasma Membrane Integrity ◽  
Frozen Semen

scholarly journals Plasma membrane integrity in health and disease: significance and therapeutic potential

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Catarina Dias ◽  
Jesper Nylandsted
Keyword(s):  
Plasma Membrane ◽  
Therapeutic Potential ◽  
Calcium Influx ◽  
Membrane Integrity ◽  
Cell Types ◽  
Physical Parameters ◽  
Membrane Repair ◽  
Plasma Membrane Integrity ◽  
Repair Mechanisms ◽  
And Function

AbstractMaintenance of plasma membrane integrity is essential for normal cell viability and function. Thus, robust membrane repair mechanisms have evolved to counteract the eminent threat of a torn plasma membrane. Different repair mechanisms and the bio-physical parameters required for efficient repair are now emerging from different research groups. However, less is known about when these mechanisms come into play. This review focuses on the existence of membrane disruptions and repair mechanisms in both physiological and pathological conditions, and across multiple cell types, albeit to different degrees. Fundamentally, irrespective of the source of membrane disruption, aberrant calcium influx is the common stimulus that activates the membrane repair response. Inadequate repair responses can tip the balance between physiology and pathology, highlighting the significance of plasma membrane integrity. For example, an over-activated repair response can promote cancer invasion, while the inability to efficiently repair membrane can drive neurodegeneration and muscular dystrophies. The interdisciplinary view explored here emphasises the widespread potential of targeting plasma membrane repair mechanisms for therapeutic purposes.

scholarly journals Plasma membrane integrity: implications for health and disease

BMC Biology ◽  
2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Dustin A. Ammendolia ◽  
William M. Bement ◽  
John H. Brumell
Keyword(s):  
Plasma Membrane ◽  
Membrane Damage ◽  
Membrane Integrity ◽  
Whole Body ◽  
Plasma Membrane Integrity ◽  
Plasma Membrane Damage ◽  
Cellular Environment ◽  
Health And Disease ◽  
Repair Pathways ◽  
The Impact

AbstractPlasma membrane integrity is essential for cellular homeostasis. In vivo, cells experience plasma membrane damage from a multitude of stressors in the extra- and intra-cellular environment. To avoid lethal consequences, cells are equipped with repair pathways to restore membrane integrity. Here, we assess plasma membrane damage and repair from a whole-body perspective. We highlight the role of tissue-specific stressors in health and disease and examine membrane repair pathways across diverse cell types. Furthermore, we outline the impact of genetic and environmental factors on plasma membrane integrity and how these contribute to disease pathogenesis in different tissues.

Assessment of the Functional Integrity of Hepatocytes: A Brief Review

1993 ◽  
Vol 21 (3) ◽  
pp. 324-329
Author(s):  
Jeffrey R. Fry ◽  
Alison H. Hammond
Keyword(s):  
Plasma Membrane ◽  
Trypan Blue ◽  
Membrane Integrity ◽  
Blue Dye ◽  
Plasma Membrane Integrity ◽  
Functional Integrity ◽  
Cellular Integrity

A variety of approaches to assessment of cellular integrity exist, based on tests of integrity of the plasma membrane, tests of metabolic competence, and asessment of morphology. By definition, such approaches address different aspects of cellular integrity and hence are not interchangeable indices of cellular integrity. Accordingly, it would be most appropriate to characterise hepatocyte preparations on the basis of more than just trypan blue dye exclusion (a test of plasma membrane integrity) as is customary. A scheme for the choice of the most appropriate mix of tests of cellular integrity is presented.

Toxic and osmotic effects of glycerol on human granulocytes

1984 ◽  
Vol 247 (5) ◽  
pp. C382-C389 ◽  
Author(s):  
W. J. Armitage ◽  
P. Mazur
Keyword(s):  
Plasma Membrane ◽  
Osmotic Stress ◽  
Cell Volume ◽  
Membrane Integrity ◽  
Fluorescein Diacetate ◽  
Major Fraction ◽  
Plasma Membrane Integrity ◽  
Human Granulocytes ◽  
Osmotic Effects

Human granulocytes are damaged by exposure to concentrations of glycerol as low as 0.5 M. We therefore investigated the addition of glycerol to granulocytes and its subsequent dilution under various conditions to try to distinguish between toxic and harmful osmotic effects of glycerol. The lesion caused by glycerol at 0 degree C was expressed as a loss of plasma membrane integrity (as visualized by fluorescein diacetate) only after incubation (greater than or equal to 1 h) at 37 degrees C. This damage was not ameliorated when osmotic stress was lessened by reducing the rates of addition and dilution of glycerol to keep the computed cell volume within 80-170% of isotonic cell volume. However, when osmotic stress was reduced further by increasing the temperature of addition and dilution of glycerol from 0 degree C to 22 degrees C, the tolerance of the cells to 1 M glycerol increased somewhat. Reducing exposure to glycerol to 3 min or less at 0 degree C greatly increased survival, but this time was too short to allow glycerol to equilibrate intracellularly. Finally, the presence of extra impermeant solute (NaCl or sucrose) in the medium to reduce the equilibrium cell volume to 60% of isotonic cell volume enabled granulocytes to survive 30-min exposure to 1 M glycerol at 0 degree C, but cells had to remain shrunken during the 37 degrees C incubation to prevent the loss of membrane integrity. Suspensions that contained damaged granulocytes formed aggregates when incubated at 37 degrees C, and these aggregates were responsible for a major fraction of the observed loss in viability.

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