Cellular membranes in horses are complex structures that function as selective barriers, enclosing the cell contents and regulating the passage of substances in and out of the cell. These membranes are primarily composed of a phospholipid bilayer interspersed with proteins, cholesterol, and carbohydrates, which contribute to their fluidity and functionality. Cellular membranes play a role in various physiological processes, including signal transduction, cell communication, and maintaining homeostasis. This topic encompasses peer-reviewed research studies and scholarly articles that explore the composition, function, and dynamics of cellular membranes in equine cells, as well as their involvement in health and disease.
Gibson JS, Cossins AR, Ellory JC.Oxygen is essential for all higher forms of animal life. It is required for oxidative phosphorylation, which forms the bulk of the energy supply of most animals. In many vertebrates, transport of O(2) from respiratory to other tissues, and of CO(2) in the opposite direction, involves red cells. These are highly specialised, adapted for their respiratory function. Intracellular haemoglobin, carbonic anhydrase and the membrane anion exchanger (AE1) increase the effective O(2)- and CO(2)-carrying capacity of red cells by approximately 100-fold. O(2) also has a pathological role. It is a very reac...
Olivry T, Borrillo AK, Xu L, Dunston SM, Slovis NM, Affolter VK, Demanuelle TC, Chan LS.Bullous pemphigoid (BP) is an autoimmune subepithelial blistering dermatosis of humans, dogs, cats and pigs. It is characterized by skin-fixed and circulating IgG autoantibodies that target one or both BP antigens. An immunological homologue of BP in humans was diagnosed in two horses with cutaneous and mucosal ulcerations as well as microscopic subepithelial vesiculation. Immunological investigations revealed similar findings for both the horses. Direct immunofluorescence demonstrated the presence of IgG deposited linearly at the dermoepidermal junction in mucosal and skin biopsy specimens. I...
Minelli A, Allegrucci C, Mezzasoma I, Ronquist G, Lluis C, Franco R.Membrane vesicles of horse seminal plasma present at their surface a highly specific serine-type protease, dipeptidyl peptidase IV/CD26, a surface antigen known to characterize human prostasomes. Horse sperm cells expressed at their surface A(1) adenosine receptors (A(1)AR) and ecto-adenosine deaminase (ecto-ADA), both detected by immunoblot analysis, whereas CD26 was visualized at the equatorial segment by immunofluorescence microscopy. In addition to CD26, horse membrane vesicles showed ecto-ADA. The fusion process between horse sperm cells and vesicles was evidenced by confocal microscopy, ...
Minelli A, Moroni M, Martínez E, Mezzasoma I, Ronquist G.Equine seminal plasma was shown to contain membrane vesicles that are similar to the well characterized prostasomes in human seminal plasma. Determination of nucleoside and nucleotide concentrations of these particles have shown that ATP, ADP and adenosine are the main components of the nucleotidic pool. 5' nucleotidase, endopeptidase and dipeptidyl peptidase i.v. activities have been found on the surface of the particles. The interaction between these prostasome-like vesicles and spermatozoa was demonstrated by electron micrograph scans which revealed the steps of a fusion-like process leadin...
Pamplona R, Requena JR, Portero-Otín M, Prat J, Thorpe SR, Bellmunt MJ.The non-enzymatic modification of aminophospholipids with lipoperoxidation-derived aldehydes and glycoxidation-derived products have been reported previously. However, it remains unknown whether intracellular membranes are damaged by these glycoxidation-derived products. To investigate this issue, we tested whether aminophospholipids from mitochondrial membranes are damaged by glycoxidative stress the mitochondrion being identified as the major site of reactive-species production in the cell. We have used a selected-ion-monitoring/gas-chromatography/mass-spectrometry assay for carboxymethyleth...
Minelli A, Moroni M, Trinari D, Mezzasoma I.Ectoenzymic activities capable of hydrolyzing ATP sequentially to adenosine are present on equine epidydimal spermatozoa membranes. Kinetic parameters for ATPase, ADPase and 5'-nucleotidase were obtained by analysis of progress reactions curve when ATP, ADP and AMP were supplied as initial substrates. These values are not different from those found when the substrates were supplied from the preceding reactions. Feed-forward inhibition on 5'-nucleotidase by ATP/ADP was taken into account to fit simulated data to the experimental results. None of the substrates supplied by the preceding reaction...
Salamon Z, Tollin G.Cyclic voltammetry has been used to study the effects of interactions between horse cytochrome c and solid-supported planar lipid membranes, comprised of either egg phosphatidylcholine (PC) or PC plus 20 mol.% cardiolipin (CL), on the redox potential and the electrochemical electron transfer rate between the protein and a semiconductor electrode. Experiments were performed over a wide range of cytochrome c concentrations (0-440 microM) at low (20 mM) and medium (160 mM) ionic strengths. Three types of electrochemical behavior were observed, which varied as a function of the experimental condit...
The Journal of physiologyNovember 15, 1996
Volume 497 ( Pt 1), Issue Pt 1 19-29 doi: 10.1113/jphysiol.1996.sp021746
Ko WH, Chan HC, Wong PY.1. Anion secretion induced by capacitative Ca2+ entry through apical and basolateral membranes of cultured equine sweat gland epithelium was studied using the short-circuit current (Isc) technique. 2. Thapsigargin induced an increase in Isc that could be inhibited when external Ca2+ was chelated by EGTA. 3. The inhibition of the thapsigargin-induced Isc could be reversed by re-addition of Ca2+ to apical or basolateral solutions. The magnitude of the reactivated Isc depended predominantly on basolateral Ca2+ concentration. 4. The magnitude of the reactivated Isc upon basolateral Ca2+ addition i...
Research in microbiologyNovember 1, 1996
Volume 147, Issue 9 687-696 doi: 10.1016/s0923-2508(97)85116-9
Douet JP, Castroviejo M, Dodin A, Bébéar C.The haemolytic action of 125I-labelled thermostable direct haemolysin from Vibrio parahaemolyticus was studied on human and equine erythrocytes. In the first step, the haemolysin bound to the membranes of both erythrocyte species. This binding seemed temperature-independent. Then, for human erythrocytes, haemolysin produced cell disruption, and haemoglobin was released. Following this step, haemolysin was also released in a temperature-dependent manner. In contrast, equine erythrocytes were not disrupted, and no release of haemolysin occurred. The receptors of labelled haemolysin were analysed...
Aurich JE, Kühne A, Hoppe H, Aurich C.Effects of seminal plasma on post-thaw motility and membrane integrity of cryopreserved horse spermatozoa were investigated. Carboxyfluorescein diacetate staining was used for the assessment of sperm membrane integrity. Adding 30% of seminal plasma from stallions with high post-thaw sperm motility to ejaculates from stallions with low post-thaw sperm motility increased progressive motility from 24.0 +/- 1.6 to 34.5 +/- 1.9% (P < 0.05) and membrane integrity from 27.0 +/- 2.1 to 34.3 +/- 2.3% membrane-intact spermatozoa (P < 0.05). Conversely, the addition of seminal plasma from stallions...
Salamon Z, Tollin G.Complex formation between horse heart cytochrome c (cyt c) and bovine cytochrome c oxidase (cco) incorporated into a supported planar egg phosphatidylcholine membrane containing varying amounts of cardiolipin (CL) (0-20 mol%) has been studied under low (10 mM) and medium (160 mM) ionic strength conditions by surface plasmon resonance (SPR) spectroscopy. Both specific and nonspecific modes of cyt c binding are observed. The dissociation constant of the specific interaction between cyt c and cco increases from approximately 6.5 microM at low ionic strength to 18 microM at medium ionic strength, ...
Atwal OS, McDonell W, Staempfli H, Singh B, Minhas KJ.The pulmonary intravascular macrophages (PIMs) of horse contain a unique electron-dense globular surface-coat which is arranged in a linear fashion in conformity with the contours of the cell membrane. The coat is sensitive to heparin treatment and to the digestive effect of lipolytic lipase, suggesting that the coat is predominantly composed of lipoproteins. During the present study, ultrastructural features of PIMs were analysed after exposing horses to halothane inhalation which was chosen as the model agent of lipid-soluble anaesthetic. The surface-coat showed acute sensitivity to halothan...
Hashimoto Y, Reese S, Bragulla H, Budras KD.Carbohydrate moieties of cell membranes in the epidermis of equine skin and hoof were investigated using lectin-histochemistry. The loss of desmosomes during keratinization leads to a reduced lectin affinity. Lectin binding capacity of the intercellular matrix in the stratum corneum of the epidermis is weak compared to that of the intercellular kitt of the hoof. Qualitative differences in lectin binding of the intercellular kitt of different hoof segments correlate with the quality of the horn in the respective areas.
Grosenbaugh DA, Hood DM, Amoss MS, Williams JD.Epidermal growth factor (EGF) receptors were detected in plasma membrane preparations of equine hoof wall laminar tissue at concentrations comparable to that of equine liver. Scatchard analysis of the equilibrium binding data suggested the presence of two classes of EGF binding sites in most of the controls (plasma membranes from clinically normal horses); a high-affinity class and a more numerous low-affinity class. The dissociation constant of the low-affinity class of EGF-specific receptors (KD = 1 x 10(-9)M) is in reasonable agreement with other values established for the EGF receptor. The...
Fayrer-Hosken RA, Caudle AB, Shur BD.beta 1, 4-Galactosyltransferase (GalTase) is localized to the plasma membrane of mouse sperm, in which it mediates the binding of sperm to glycoconjugate residues in the egg zona pellucida. In this study, the presence of subcellular distribution of sperm GalTase were determined in two other mammalian species that yield sufficient sperm for subcellular fractionation. Equine and bovine semen were collected, and the plasma membranes (PM), outer acrosomal membranes (OAM), and inner acrosomal membranes (IAM) were sequentially removed. The purities of the isolated membrane preparations were determin...
Satoh K, Nakao T, Nagai F, Kano I, Nakagawa A, Ushiyama K, Urayama O, Hara Y, Nakao M.Monoclonal antibodies against horse kidney outer medulla (Na+ + K+)-ATPase were prepared. One of these antibodies (M45-80), was identified as an IgM, recognized the alpha subunit of the enzyme. M45-80 had the following effects on horse kidney (Na+ + K+)-ATPase: (1) it inhibited the enzyme activity by 50% in 140 mM Na+ and by 80% in 8.3 mM Na+; (2) it increased the Na+ concentration necessary for half-maximal activation (K0.5 for Na+) from 12.0 to 57.6 mM, but did not affect K0.5 for K+; (3) it slightly increased the K+-dependent p-nitrophenylphosphatase (K-pNPPase) activity; (4) it inhibited p...
Muccioli G, Bellussi G, Ghé C, Pagnini G, Di Carlo R.The binding of 125I-labeled ovine prolactin (125I-oPRL) to membranes from different brain regions of pigeon, rabbit, rat, pig, calf, horse, and ewe was studied. The hypothalamus from rabbit, pig, horse, and pigeon showed a low but specific binding for 125I-oPRL clearly different from the other brain regions examined (cortex and cerebellum), whereas in the brain from rat, calf, and ewe the binding was very small and quite uniform in the various regions. Also the membranes from choroid plexus of rabbit, pig, calf, and horse showed an evident specific binding for prolactin. The binding of 125I-oP...
Yoshihara E, Nakae T.In order to develop the cytotoxic liposome, the cytolytic effect of polycationic liposome was examined. Upon incubation of the stearylamine-containing liposome (stearylamine-liposome) with rabbit erythrocyte, a significant extent of hemolysis was observed. Hemolytic activity of the liposome depends on the amount of stearylamine in the liposome membrane. The plots of the initial rate of hemolysis versus the concentration of stearylamine-liposome showed a sigmoidal curve, suggesting that stearylamine-liposomes act cooperatively on the erythrocyte membrane. Hemolytic activity of stearylamine-lipo...
Brayton CF.Dimethyl sulfoxide (DMSO) is a very simple compound that has stimulated much controversy in the scientific and popular literature. Fig. 1 It is an aprotic solvent. Therapeutic and toxic agents that are not soluble in water are often soluble in DMSO. DMSO has a very strong affinity for water; on exposure to air, pure DMSO is rapidly diluted. DMSO's physiologic and pharmacologic properties and effects are incompletely understood. Properties that are considered to be particularly important to its therapeutic and toxic effects include: its own rapid penetration and enhanced penetration of other su...
Cavatorta P, Crippa PR, Ito AS, Casali E, Ferrari MB, Masotti L.In the present paper we report a comparative study of physical properties and biochemical composition of isolated melanosomal membranes extracted from bovine eyes and from an equine spleen melanoma. Some biophysical characteristics of such membranes were obtained by steady-state and time resolved fluorescence spectroscopy using DPH as fluorescent probe. By these methods we have measured both static fluorescence polarization and fluorescence lifetimes and from the experimental data we have calculated the rotational correlation times by Perrin's equation. Since dynamic and static parameters, suc...
Montgomery I, Jenkinson DM, Elder HY.Sweating in the horse had little effect on the ultrastructure of the glandular duct, other than on the lumen which enlarged. The fundus secretory cells, which in the resting gland were packed with vesicles, gradually lost them as sweating progressed until, after 4 hours of activity, few remained. Sweat appeared to be largely the product of secretion (a) by fluid transport, probably involving a region of complex cellular interdigitations adjacent to the basement membrane and (b) by exocytosis of vesicles, although a secondary mechanism of vesicle loss by micro-apocrine secretion may occur. Howe...
Murayama JI, Takeshita K, Tomita M, Hamada A.Crude glycophorin fraction was prepared from horse erythrocyte membranes by extraction with lithium diiodosalicylate and partition in aqueous phenol. Two glycophorins, designated glycophorins HA and HB, were isolated by two different techniques: preparative gel electrophoresis in the presence of sodium dodecyl sulfate and ion-exchange chromatography in the presence of the nonionic detergent Ammonyx LO. Each glycophorin formed at least two bands on gel electrophoresis, which corresponded to a dimeric form and a monomeric form. Glycophorin HA, the major component, had a blocked amino-terminus an...
Slauson DO, Hahn FF.Advances in the understanding of human respiratory disease can come from careful clinical studies of the diseases as they occur in man, but such studies are naturally limited in terms of experimental manipulation. In the last 2 decades, an increasingly complex plethora of experimental respiratory disease models has been developed and utilized by investigators, but relatively less attention has been paid to the naturally occurring pulmonary diseases of animals as potential models. This paper is aimed at presenting selected examples of spontaneous pulmonary disease in animals that may serve as e...
Swaney LM, Breese SS.Haemophilus equigenitalis, a proposed new species of Haemophilus and the causative agent of contagious equine metritis, a venereal disease of the horse, had ultrastructural characteristics of gram-negative bacteria. The organism additionally had a small, threadlike capsule that was removed by heating in phosphate-buffered saline solution. Heating also detached the outer membrane from the cytoplasmic membrane. The capsule could only be demonstrated when bacterial were stained with ruthenium red during the preparation of ultrathin sections. The gross morphology of newly isolated organisms (rodli...
Gürtler LG, Yeboa DA, Cleve H.The membrane components of equine, porcine and ovine erythrocytes were separated by sodium dodecylsulfate polyacrylamide gel electrophoresis and subsequently incubated with the radioiodinated lectins from lentils (LCH), castorbeans (RCA), Phaseolus beans (L-PHA), gorse seeds (UEH-F) and from vineyard snails (HPA). The following individual glycoproteins could be labeled: gp 26, 33, 100 and 320 in horse erythrocytes, gp 24, 46, 75, 130 and 210 in swine and gp 24, 57, 100 and 210 in sheep erythrocytes.
Apukhovska LI, Vendt VP, Ivashkevych SP.Sterols of the whole erythrocytes, hemoglobin, membranes and their protein fractions were studied. Erythrocytes and their components were established to contain, besides cholesterol, other substances of the sterol nature. Cholesterol, 7-dehydrocholesterol and other substances of the cholesterol series are practically completely linked with the lipoproteid fractions of the erythrocyte membrane whereas all sterols found in the native erythrocytes in negligible amounts are bound with hemoglobin.
Kuwano A, Ueno T, Katayama Y, Nishiyama T, Arai K.Between the laminar epidermis and the laminar dermis of laminar region (LR) in equine foot, it can be observed the basement membrane zone (BMZ), which is composed of a basement membrane and its accompaniments like the hemidesmosome and anchoring fibril. Alteration in the BMZ in equine laminitis is possibly related with not only development but also recovery outcome and recurrence of this disease. However, there is little known about the structure of the BMZ during the recovery phase of this disease. To assess the condition of the BMZ of LR affected by chronic laminitis, the tissue was examined...
Filho JS, Corcini CD, Santos FCC, Anciuti AN, Gatti NLS, Anastacio E, Mielke R, Nogueira CEW, Curcio BR, Varela AS. BACKGROUND: Supplementation of sperm diluents to reduce the damage caused by the freeze-thaw cycle is broadly used in equine semen cryopreservation. Objective: The present study aimed at determining the most appropriate quercetin supplementation in equine freezing extender. Methods: Quercetin at four different concentrations (0.25, 0.5, 0.75 or 1 mM) was added in the sperm freezing diluent before the freeze-thaw cycle. The spermatozoa population was analyzed by flow cytometry and a statistical analysis was conducted to detect significant differences between control and treated samples. R...
Oldenhof H, Heutelbeck A, Blässe AK, Bollwein H, Martinsson G, Wolkers WF, Sieme H.The aim of this study was to evaluate inter-individual variability in osmotic properties of stallion spermatozoa and its correlation with cryosurvival. In addition, temperature dependency of hypo-osmotic tolerance and membrane fluidity were studied. Stallion sperm membranes exhibited good resistance towards hypotonic stress in the 15-30 °C temperature range, whereas membrane stability was found to be decreased at 4 and 37 °C. Bull spermatozoa showed greater hypo-osmotic tolerance compared with stallion spermatozoa, especially at temperatures above 30 °C, which coincided with decreased membr...
Olivry T, Borrillo AK, Xu L, Dunston SM, Slovis NM, Affolter VK, Demanuelle TC, Chan LS.Bullous pemphigoid (BP) is an autoimmune subepithelial blistering dermatosis of humans, dogs, cats and pigs. It is characterized by skin-fixed and circulating IgG autoantibodies that target one or both BP antigens. An immunological homologue of BP in humans was diagnosed in two horses with cutaneous and mucosal ulcerations as well as microscopic subepithelial vesiculation. Immunological investigations revealed similar findings for both the horses. Direct immunofluorescence demonstrated the presence of IgG deposited linearly at the dermoepidermal junction in mucosal and skin biopsy specimens. I...
Greube A, Müller K, Töpfer-Petersen E, Herrmann A, Müller P.Seminal plasma of mammalians contains, among others, proteins that are characterized by the fibronectin (Fn) type II module. Our knowledge about the structure and the physiological function of seminal Fn type II proteins mainly originates from studies on PDC-109, the bovine representative of this protein family. The present work focuses on the equine protein SP-1/2 (also named HSP-1/2) with particular emphasis on its interaction with lipid membranes by employing the intrinsic protein fluorescence and a number of spin-labeled and fluorescent lipid analogues. The results indicate that the intera...
Contreras-Mendez LA, Medrano A.There are many protocols for horse sperm cryopreservation, but results are inconsistent; sperm survival after freeze-thawing is usually poor; in consequence, fertility is low. The objective of this work was to see whether slow cooling before freezing to minus 3 °C instead of +5 °C, the traditional target temperature, could improve horse sperm cryosurvival, capability to carry out capacitation and the acrosome reaction induced by progesterone. Spermatozoa from five stallions were packaged in straws and slowly cooled to +5 °C. Half of the straws were frozen directly and the other half was ...
Asami K.Horse erythrocytes, murine lymphoblasts (L5178Y) and lipid vesicles that were treated with dipicrylamine (DPA) as a lipophilic ion were studied by dielectric spectroscopy over a frequency range of 10 Hz to 10 MHz. The DPA-treated cells and lipid vesicles showed low-frequency (LF) dielectric dispersion around 1-10 kHz in addition to β-dispersion due to the Maxwell-Wagner effect. The LF dispersion corresponds to that found in previous electrorotation (ROT) studies on DPA-treated cells, being due to the translocation of mobile ions in the plasma membranes. Analysis of the LF dispersion based on ...
Gallardo Bolaños JM, Balao da Silva C, Martín Muñoz P, Plaza Dávila M, Ezquerra J, Aparicio IM, Tapia JA, Ortega Ferrusola C, Peña FJ.To investigate the mechanisms inducing sperm death after ejaculation, stallion ejaculates were incubated in BWW media during 6 h at 37°C. At the beginning of the incubation period and after 1, 2, 4 and 6 h sperm motility and kinematics (CASA), mitochondrial membrane potential and membrane permeability and integrity were evaluated (flow cytometry). Also, at the same time intervals, active caspase 3, hydrogen peroxide, superoxide anion (flow cytometry) and Akt phosphorylation (flow cytometry) were evaluated. Major decreases in sperm function occurred after 6 h of incubation, although after 1 h ...
Okamura M, Yokoyama N, Takabatake N, Okubo K, Ikehara Y, Igarashi I.In the present study, we investigated the effects of protease pretreatments of host erythrocytes (RBC) on the in vitro growth of bovine Babesia parasites (Babesia bovis and B. bigemina) and equine Babesia parasites (B. equi and B. caballi). The selected proteases, trypsin and chymotrypsin, clearly modified several membrane proteins of both bovine and equine RBC, as demonstrated by SDS-PAGE analysis; however, the protease treatments also modified the sialic acid content exclusively in bovine RBC, as demonstrated by lectin blot analysis. An in vitro growth assay using the protease-treated RBC sh...
Research in microbiologyNovember 1, 1996
Volume 147, Issue 9 687-696 doi: 10.1016/s0923-2508(97)85116-9
Douet JP, Castroviejo M, Dodin A, Bébéar C.The haemolytic action of 125I-labelled thermostable direct haemolysin from Vibrio parahaemolyticus was studied on human and equine erythrocytes. In the first step, the haemolysin bound to the membranes of both erythrocyte species. This binding seemed temperature-independent. Then, for human erythrocytes, haemolysin produced cell disruption, and haemoglobin was released. Following this step, haemolysin was also released in a temperature-dependent manner. In contrast, equine erythrocytes were not disrupted, and no release of haemolysin occurred. The receptors of labelled haemolysin were analysed...
Murayama JI, Takeshita K, Tomita M, Hamada A.Crude glycophorin fraction was prepared from horse erythrocyte membranes by extraction with lithium diiodosalicylate and partition in aqueous phenol. Two glycophorins, designated glycophorins HA and HB, were isolated by two different techniques: preparative gel electrophoresis in the presence of sodium dodecyl sulfate and ion-exchange chromatography in the presence of the nonionic detergent Ammonyx LO. Each glycophorin formed at least two bands on gel electrophoresis, which corresponded to a dimeric form and a monomeric form. Glycophorin HA, the major component, had a blocked amino-terminus an...
Gürtler LG, Yeboa DA, Cleve H.The membrane components of equine, porcine and ovine erythrocytes were separated by sodium dodecylsulfate polyacrylamide gel electrophoresis and subsequently incubated with the radioiodinated lectins from lentils (LCH), castorbeans (RCA), Phaseolus beans (L-PHA), gorse seeds (UEH-F) and from vineyard snails (HPA). The following individual glycoproteins could be labeled: gp 26, 33, 100 and 320 in horse erythrocytes, gp 24, 46, 75, 130 and 210 in swine and gp 24, 57, 100 and 210 in sheep erythrocytes.
Grosenbaugh DA, Hood DM, Amoss MS, Williams JD.Epidermal growth factor (EGF) receptors were detected in plasma membrane preparations of equine hoof wall laminar tissue at concentrations comparable to that of equine liver. Scatchard analysis of the equilibrium binding data suggested the presence of two classes of EGF binding sites in most of the controls (plasma membranes from clinically normal horses); a high-affinity class and a more numerous low-affinity class. The dissociation constant of the low-affinity class of EGF-specific receptors (KD = 1 x 10(-9)M) is in reasonable agreement with other values established for the EGF receptor. The...
Silva DM, Holden SA, Lyons A, Souza JC, Fair S.The aim of the present study was to assess the effect of the addition of docosahexaenoic acid (DHA) on the in vitro quality of cooled and frozen-thawed stallion semen. In Experiment 1, semen from 10 stallions was collected (three ejaculates per stallion). Semen was diluted to 100×106 spermatozoa mL-1 with 0.02mM vitamin E (VE) and 0, 1, 10 or 20ng mL-1 DHA and frozen. Semen was thawed and total motility (TM), rapid progressive motility (PM), acrosome integrity, membrane fluidity and morphology were assessed. In Experiment 2, semen from three stallions was collected (three ejaculates per stall...
Montgomery I, Jenkinson DM, Elder HY.Sweating in the horse had little effect on the ultrastructure of the glandular duct, other than on the lumen which enlarged. The fundus secretory cells, which in the resting gland were packed with vesicles, gradually lost them as sweating progressed until, after 4 hours of activity, few remained. Sweat appeared to be largely the product of secretion (a) by fluid transport, probably involving a region of complex cellular interdigitations adjacent to the basement membrane and (b) by exocytosis of vesicles, although a secondary mechanism of vesicle loss by micro-apocrine secretion may occur. Howe...
Satoh K, Nakao T, Nagai F, Kano I, Nakagawa A, Ushiyama K, Urayama O, Hara Y, Nakao M.Monoclonal antibodies against horse kidney outer medulla (Na+ + K+)-ATPase were prepared. One of these antibodies (M45-80), was identified as an IgM, recognized the alpha subunit of the enzyme. M45-80 had the following effects on horse kidney (Na+ + K+)-ATPase: (1) it inhibited the enzyme activity by 50% in 140 mM Na+ and by 80% in 8.3 mM Na+; (2) it increased the Na+ concentration necessary for half-maximal activation (K0.5 for Na+) from 12.0 to 57.6 mM, but did not affect K0.5 for K+; (3) it slightly increased the K+-dependent p-nitrophenylphosphatase (K-pNPPase) activity; (4) it inhibited p...
Aurich C, Seeber P, Müller-Schlösser F.To maintain the fertility of stallion spermatozoa during cooled storage, extender media are added to semen. In this study, three semen extenders were compared: EquiPro which contains defined caseinates and whey proteins instead of dried skim milk. The extender is provided in dry form and dissolved in distilled water prior to use. EquiPro TM has the same composition as EquiPro but is provided in a sterilized ready-to-use liquid form. AndroMed-E contains soybean lecithin as protein source. Semen was collected from seven stallions. Ejaculates were divided into three aliquots, diluted with the dif...
Pessoa GA, Martini AP, Trentin JM, Minela T, Fiorenza MF, Rubin MIB.The aim of this study was to compare the sperm separation technique using filtration through glass wool compared with just diluted cooled semen. Eighteen ejaculates were collected from 6 pony stallions of the Brazilian pony breed. Evaluations were done on pH, osmolarity, total motility, membrane functionality (HOST), membrane integrity (CFDA/PI), morphology and mitochondrial viability (MTT) in fresh, 24 and 48 h of cooled semen at 5°C. After dilution, the half of the extended semen was cooled (control group). The other half was cooled after filtration trough glass wool (filtered group). Reta...
Contreras MJ, Arias ME, Silva M, Cabrera P, Felmer R.Cryopreservation of stallion semen is less efficient than other species such as bovine. This is mainly because of the greater susceptibility of stallion sperm to the freezing damage that generates oxidative stress and plasma membrane injury, resulting in DNA fragmentation and cell death. These data suggest the need to develop new strategies of sperm cryopreservation that can improve the efficiency of this technique in stallions by reducing or preventing membrane damage and cell death. The present study aimed to evaluate the effect of adding membrane stabilizers to the freezing medium and asses...
Portier K, Guichardant M, Debouzy JC, Crouzier D, Geraud I, Kirschvink N, Lekeux P, Fellmann N, Coudert J.Whether direct exposure to different concentrations (0%, 13%, 100%) of oxygen may affect horse erythrocyte membrane fluidity (EMF) and fatty acid (FA) composition was studied during 1 (T60) and 2h (T120) exposure. EMF was investigated at the head group level and hydrophobic core thanks to phosphorus nucleus 31 ((31)P) nuclear magnetic resonance ((31)P NMR) and electronic paramagnetic resonance (EPR) using two spin probes: 5-nitroxydestearic acid and 16-doxylstearic acid. Lipid structure of the membranes was studied by gas liquid chromatography. 4-Hydroxy-2E-nonenal was also analyzed as a marke...
Ertmer F, Oldenhof H, Schütze S, Rohn K, Wolkers WF, Sieme H.If the physiological balance between production and scavenging of reactive oxygen species (ROS) is shifted towards production of ROS this may result in accumulation of cell damage over time. In this study stallion spermatozoa were incubated with xanthine and xanthine oxidase (X-XO) to artificially generate defined levels of superoxide and hydrogen peroxide resulting in sub-lethal oxidative damage. The effects of X-XO treatment on various sperm characteristics were studied. Special emphasis was placed on sperm osmotic tolerance pre-freeze and its correlation with cryosurvival, given that cryopr...
Tkachev AV, Tkacheva OL.The article presents the results of the studies of cytotoxic effect of zearalenone and T-2 toxin on sperm of horses and bulls during incubation and after thawing according to the technology of sperm obtaining and cryopreservation in Kharkov. We first have shown in vitro toxic effects of different concentrations of zearalenone and T-2 toxin (from 0.5 to 0.01 mM) on the membrane stability, as well as quantitative and qualitative indicators of semen in stallions and bulls before and after freezing and thawing. It has been found that the biological activity of the native sperm in 1 h after additio...
Apukhovska LI, Vendt VP, Ivashkevych SP.Sterols of the whole erythrocytes, hemoglobin, membranes and their protein fractions were studied. Erythrocytes and their components were established to contain, besides cholesterol, other substances of the sterol nature. Cholesterol, 7-dehydrocholesterol and other substances of the cholesterol series are practically completely linked with the lipoproteid fractions of the erythrocyte membrane whereas all sterols found in the native erythrocytes in negligible amounts are bound with hemoglobin.
Hashimoto Y, Reese S, Bragulla H, Budras KD.Carbohydrate moieties of cell membranes in the epidermis of equine skin and hoof were investigated using lectin-histochemistry. The loss of desmosomes during keratinization leads to a reduced lectin affinity. Lectin binding capacity of the intercellular matrix in the stratum corneum of the epidermis is weak compared to that of the intercellular kitt of the hoof. Qualitative differences in lectin binding of the intercellular kitt of different hoof segments correlate with the quality of the horn in the respective areas.
Içli S, Soleimani M, Oldenhof H, Sieme H, Wriggers P, Wolkers WF.Cryopreservation can be used to store equine oocytes for extended periods so that they can be used in artificial reproduction technologies at a desired time point. It requires use of cryoprotective agents (CPAs) to protect the oocytes against freezing injury. The intracellular introduction of CPAs, however, may cause irreversible osmotic damage. The response of cells exposed to CPA solutions is governed by the permeability of the cellular membrane towards water and the CPAs. In this study, a mathematical mass transport model describing the permeation of water and CPAs across an oocyte membrane...
Schara M, Nemec M, Falnoga I, Kobal AB, Kveder M, Svetek J.The action of mercuric chloride and methyl mercuric chloride on the membrane lateral domain organization of bovine, equine, and canine erythrocytes was studied. Electron paramagnetic resonance (EPR) spectra of spin-labeled erythrocytes were analyzed with respect to their lateral domain structure. Continuous alteration of the membrane domain populations revealed that mercuric compounds affect the membrane via the evolution of toxic events in the cells.
Minelli A, Moroni M, Trinari D, Mezzasoma I.Ectoenzymic activities capable of hydrolyzing ATP sequentially to adenosine are present on equine epidydimal spermatozoa membranes. Kinetic parameters for ATPase, ADPase and 5'-nucleotidase were obtained by analysis of progress reactions curve when ATP, ADP and AMP were supplied as initial substrates. These values are not different from those found when the substrates were supplied from the preceding reactions. Feed-forward inhibition on 5'-nucleotidase by ATP/ADP was taken into account to fit simulated data to the experimental results. None of the substrates supplied by the preceding reaction...
Cunha Dos Santos FC, Morrell JM, Nunes MM, Nogueira CE, Curcio BR, Malschitzky E.Epididymal sperm cryopreservation represents the ultimate option to preserve spermatozoa of valuable stallions. Objective: The study aims to evalute whether single layer centrifugation (SLC) prior to cryopreservation or after post-thawing improves the quality of stallion epididymal sperm. Methods: Epididymal sperms of stallions were harvested (N=20). Sperm samples were subjected to treatments: conventional centrifugation, SLC prior to cryopreservation (SLC-PC) or SLC post-thaw (SLC+). All samples were cryopreserved, thawed and evaluated. SLC+ were thawed, single layer cenrifuged and resuspende...
