In-vitro effect of γ-amino butyric acid and progesterone on sperm motility and acrosome reaction in patients with varicocele before and after varicocelectomy

At the avian body temperature of 40 °C, intact fowl spermatozoa require Ca2+for the initiation of motility and a combination of both Ca2+and homogenized inner perivitelline layer (IPVL) together to induce the acrosome reaction. Within the range of 1–100 μmol/l, neither PD 150606 (a Ca2+-dependent calpain inhibitor) nor Y-27632 (an inhibitor of Ca2+-dependent Rho-kinase) were able to inhibit the acrosome reaction induced by the presence of Ca2+and IPVL. However, PD 150606, although not Y-27632, was able to inhibit sperm motility initiated by Ca2+, as well as motility initiated by calyculin A – a specific inhibitor of protein phosphatases, which also initiates sperm motility at 40 °C. The addition of PD 150606 did not reduce the ATP concentrations of intact spermatozoa, nor the motility of demembranated spermatozoa. Immunoblot analysis of sperm extract using a polyclonal antibody against calpain 12 revealed a cross-reacting protein of approximately 80 kDa. These results suggest that Rho-kinase is not involved in the regulation of the acrosome reaction or of motility in fowl spermatozoa. In contrast, calpain appears to be involved in the regulation of flagellar movement, but not izn that of the acrosome reaction. Furthermore, it seems that endogenous calpain is present in the cytoplasmic matrix and/or the plasma membrane, but not retained in the axoneme and/or accessory cytoskeletal components.

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Acrosomal integrity and capacitation are not influenced by sperm cryopreservation in the giant panda

Reproduction ◽

10.1530/rep.1.00034 ◽

2004 ◽

Vol 127 (5) ◽

pp. 547-556 ◽

Cited By ~ 26

Author(s):

R E Spindler ◽

Y Huang ◽

J G Howard ◽

P Wang ◽

H Zhang ◽

...

Keyword(s):

Sperm Motility ◽

Giant Panda ◽

Calcium Ionophore ◽

Sperm Cryopreservation ◽

Management Tools ◽

Giant Pandas ◽

Before And After ◽

Cryopreservation Protocol ◽

Acrosomal Integrity

Sperm cryopreservation and artificial insemination are important management tools for giant panda breeding and the preservation of extant genetic diversity. This study examined the influence of freeze–thawing on sperm function, specifically capacitation. Sperm from nine giant pandas were assessed before and after rapid (− 40 and − 100 °C/min) cryopreservation by incubation in HEPES-buffered Ham’s F10 medium with and without the capacitation accelerators, 3-isobutyl-1-methylxanthine (IBMX) and dibutyryl cyclic AMP (dbcAMP). At 0, 3 and 6 h of exposure, aliquots were assessed for sperm motility traits and capacitation, defined as the proportion of sperm with intact acrosomes following exposure to solubilised zonae pellucidae (ursid or felid) or calcium ionophore subtracted from the proportion of sperm with intact acrosomes before exposure. Although mean±s.e.m. sperm motility post-thaw (56.1 ± 3.9% at 0 h) was less (P < 0.05) than pre-freeze (71.7 ± 6.0%), there was no difference (P > 0.05) in the proportion of acrosome-intact sperm (fresh, 93.0 ± 1.7% versus cryopreserved–thawed, 81.7 ± 4.7% at 0 h). Incidence of capacitation was greater (P < 0.05) in fresh sperm incubated with capacitation accelerators IBMX and dbcAMP (9 h: 50.9 ± 1.1) compared with fresh sperm incubated without accelerators (9 h: 41.2 ± 1.1%). Frozen–thawed sperm preincubated without accelerators underwent capacitation (49.6 ± 1.1%) to a greater extent (P < 0.05) compared with these fresh counterparts. Thawed samples with (9 h: 45.9 ± 1.4%) and without accelerators (9 h: 41.2 ± 1.1%) did not differ (P > 0.05) during the 9-h incubation. We conclude that giant panda spermatozoa (1) undergo capacitation in vitro with or without chemical accelerators and (2) withstand a rapid cryopreservation protocol, including retaining normal acrosomal integrity and functional capacitation ability.

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92 EVALUATION OF BOAR SPERM FUNCTIONALITY AFTER A CUSHIONED CENTRIFUGATION TECHNIQUE

Reproduction Fertility and Development ◽

10.1071/rdv18n2ab92 ◽

2006 ◽

Vol 18 (2) ◽

pp. 154

Author(s):

J. Gadea ◽

S. Martínez-Miró ◽

G. Decuadro-Hansen ◽

C. Matás

Keyword(s):

Seminal Plasma ◽

Acrosome Reaction ◽

Sperm Viability ◽

Lipid Packing ◽

Lipid Disorder ◽

Merocyanine 540 ◽

Centrifuge Tube ◽

Before And After ◽

Centrifugation Technique

Separation of sperm from seminal plasma is required in most semen freezing procedures. Semen is typically subjected to centrifugation to concentrate sperm into a pellet and allow removal of the seminal plasma prior to dilution in freezing extender. Centrifugation is a relatively effective method to recover sperm, however, the process also causes considerable sperm damage. The use of a dense, inert, and isotonic solution as a cushion in the bottom of the centrifuge tube allows a greater centrifugation speed to be applied and results in greater sperm recovery. The aim of the present work was to evaluate the effects of this cushioned centrifugation technique on in vitro sperm viability and functionality. Sperm-rich fractions from 16 fertile boars were diluted and cooled to 15�C; then subsamples were centrifuged by one of two different techniques. A standard method (SM), 800 g for 10 min in 50-mL tubes (Westendorf et al. 1975 Dtsch. Tier�rztl. Wschr. 82, 261-267) and a cushioned method (CM), 1000 g for 20 min using 45 mL of diluted semen on 5 mL of an isotonic iodixanol solution (60% w/v gradient) were performed. Sperm samples were stained with merocyanine 540 (M540) and Yo-Pro 1 (Harrison et al. 1996 Mol. Rep. Dev. 45, 378-391) to detect changes in lipid packing disorder of the plasma membrane. Another set of sperm samples was incubated in the presence of (0.7 �M) 22,72-dichlorodihydrofluorescein diacetate (Gadea et al. 2005 J. Androl. 26, 396-404) to estimate production of reactive oxygen species (ROS). A final set of sperm samples was stained with peanut aggultinin-fluorscein isothiocyanate (PNA-FITC) and propidium iodide to evaluate the acrosome reaction. All of these parameters were evaluated by flow cytometry before and after centrifugation. ANOVA analysis revealed that centrifugation altered lipid packing disorder and viability. Raw semen (RS) had a larger number of viable low lipid disorder sperm than centrifuged semen (RS = 86.9a vs. SM = 81.64b vs. CM = 80.6b, P < 0.01) and a decreased number of dead sperm cells (RS = 9.5a vs. SM = 15.0b vs. CM = 16.3b, P < 0.01). However, the cushioned and standard centrifugation methods yielded similar results for all the parameters measured. No significant differences were found for generation of ROS or in the number of sperm exhibiting the acrosome reaction. In conclusion, compared to the standard centrifugation method, this simple cushioned modification is a more efficient means of processing boar semen for freezing because significantly less sample losses are detected; also, it provides similar levels of sperm viability and functionality, and consequently a higher number of doses per ejaculation can be produced.

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Giant panda (Ailuropoda melanoleuca) spermatozoon decondensation in vitro is not compromised by cryopreservation

Reproduction Fertility and Development ◽

10.1071/rd06030 ◽

2006 ◽

Vol 18 (7) ◽

pp. 767 ◽

Cited By ~ 6

Author(s):

Rebecca E. Spindler ◽

Huang Yan ◽

JoGayle Howard ◽

Wang PengYan ◽

Zhang Hemin ◽

...

Keyword(s):

Giant Panda ◽

Acrosome Reaction ◽

Ailuropoda Melanoleuca ◽

Genetic Management ◽

Giant Pandas ◽

Before And After ◽

In Captivity ◽

Oocyte Cytoplasm ◽

Natural Breeding

Natural breeding of giant pandas in captivity is compromised, making artificial insemination and spermatozoa cryopreservation essential for genetic management. This study examined the influence of freeze–thawing on traditional parameters such as motility and spermatozoon functionality, specifically decondensation in vitro. Giant panda spermatozoa were assessed before and after rapid cryopreservation (4°C to –130°C over 2 min) in liquid nitrogen vapour. Spermatozoa pre-incubated in medium for 6 h were co-incubated with cat zonae (2 zonae μL–1) for 30 min to effect capacitation and an acrosome reaction. Spermatozoa were then mixed with mature cat oocyte cytoplasm (2 cytoplasm μL–1) for 4 h and evaluated for decondensation. Frozen spermatozoa were less motile (P < 0.05) than fresh counterparts immediately post-thawing, but not after 6 h incubation. There were more (P < 0.05) spermatozoa with completely diffused chromatin post-thaw (10.4 ± 1.3%; mean ± s.e.m.) compared to fresh counterparts (5.1 ± 1.0%). However, there was no overall difference (P > 0.05) in the incidence of decondensation between fresh (4 h, 69.8 ± 5.9%) and thawed (4 h, 71.5 ± 4.9%) spermatozoa after exposure to cat oocyte cytoplasm. It is concluded that the ‘rapid’ method now used to cryopreserve giant panda spermatozoa has little impact on spermatozoon decondensation.

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The in vitro effect of benzo[a]pyrene on human sperm hyperactivation and acrosome reaction

Fertility and Sterility ◽

10.1016/j.fertnstert.2009.02.031 ◽

2010 ◽

Vol 94 (2) ◽

pp. 595-598 ◽

Cited By ~ 10

Author(s):

Dyutiman Mukhopadhyay ◽

Parag Nandi ◽

Alex C. Varghese ◽

Rohit Gutgutia ◽

Samir Banerjee ◽

...

Keyword(s):

Acrosome Reaction ◽

Human Sperm ◽

Vitro Effect

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The in vitro effect of emergency contraception doses of levonorgestrel on the acrosome reaction of human spermatozoa

Contraception ◽

10.1016/j.contraception.2005.04.005 ◽

2005 ◽

Vol 72 (3) ◽

pp. 225-228 ◽

Cited By ~ 13

Author(s):

Karen Saboya Brito ◽

Luis Bahamondes ◽

Josiane A.A. Nascimento ◽

Luciana de Santis ◽

María José Munuce

Keyword(s):

Emergency Contraception ◽

Acrosome Reaction ◽

Human Spermatozoa ◽

Vitro Effect

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4-[2-Allylsulfanyl-1-(carboxymethyl-carbamoyl)-ethylcarbamoyl]-2-amino-butyric acid: evaluation as topoisomerase inhibitor using in vitro assay and molecular docking study

Medicinal Chemistry Research ◽

10.1007/s00044-014-1263-y ◽

2014 ◽

Vol 24 (5) ◽

pp. 1893-1900 ◽

Cited By ~ 4

Author(s):

Pachamuthu Pratheebaa ◽

Perumal Perumal ◽

Jayaraman Angayarkanni ◽

Narayanan SundaraBaalaji ◽

Thayumanavan Palvannan

Keyword(s):

Molecular Docking ◽

Butyric Acid ◽

Docking Study ◽

In Vitro Assay ◽

Molecular Docking Study ◽

Vitro Assay ◽

Amino Butyric Acid ◽

Topoisomerase Inhibitor

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STUDY OF THE IN VITRO EFFECT ON GLOMERULAR ALBUMIN PERMSELECTIVITY OF SERUM BEFORE AND AFTER RENAL TRANSPLANTATION IN FOCAL SEGMENTAL GLOMERULOSCLEROSIS1,2

Transplantation Journal ◽

10.1097/00007890-199712270-00014 ◽

1997 ◽

Vol 64 (12) ◽

pp. 1711-1715 ◽

Cited By ~ 20

Author(s):

Yann Godfrin ◽

Jacques Dantal ◽

Sabine Perretto ◽

Dan Hristea ◽

Christophe Legendre ◽

...

Keyword(s):

Renal Transplantation ◽

Before And After ◽

Vitro Effect

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Effect of Natural Fermentation of Sorghum on Resistant Starch Molecular Structure and Fermentation Property

Journal of Chemistry ◽

10.1155/2020/9835214 ◽

2020 ◽

Vol 2020 ◽

pp. 1-11

Author(s):

YunFei Ge ◽

WeiHao Wang ◽

Meng Shen ◽

ZiYue Kang ◽

Juan Wang ◽

...

Keyword(s):

Molecular Structure ◽

Molecular Weight ◽

Butyric Acid ◽

Resistant Starch ◽

Short Chain ◽

In Vitro Fermentation ◽

Percentage Content ◽

Research Results ◽

Before And After

Relevant research results have suggested that fermentation can increase the content of sorghum amylose chains and their retrogradation value. Therefore, this study explored the effect of fermentation pretreatment on the yield, digestibility, molecular structure, and in vitro fermentation property of sorghum-resistant starch by conducting fermentation pretreatment of sorghum and extracting the resistant starch from fermented sorghum with pressure-heat compound enzyme method. The results were as follows. After fermentation pretreatment, the yield of sorghum-resistant starch increased, the digestibility of sorghum-resistant starch reduced, the laminated structure size on the surface of the particles became more uniform, and the stacking mode became more neat and denser. The sorghum-resistant starch prepared before and after fermentation did not produce new chemical groups, and its functional group peak remained unchanged. After fermentation, the weight-average molecular weight of sorghum-resistant starch was elevated, and the percentage content of high- and low-molecular substances increased and decreased, respectively, compared with that of the unfermented sorghum-resistant starch. The percentage content of short-chain branches in the branched chain increased, whereas that of the long-chain branches decreased; the crystallinity of sorghum-resistant starch after fermentation decreased, and the intensity of X-diffraction peak changed slightly before and after fermentation. According to the results of the in vitro fermentation experiments, the fermentation broth of sorghum-resistant starch had the highest content of butyric acid and short-chain fatty acid. Research results reveal that, after fermentation pretreatment, sorghum-resistant starch presented increased yield, more complex molecular structure, heavier molecular weight and more uniform surface morphology, more efficient butyric acid generation, and greater fermentation rate than unfermented sorghum-resistant starch.

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