Purification and Characterization of a Sperm Motility Inhibiting Factor from Caprine Epididymal Plasma

Das, Sujoy; Saha, Sudipta; Majumder, Gopal C.; Dungdung, Sandhya Rekha

doi:10.1371/journal.pone.0012039

Cited by 21 publications

(37 citation statements)

References 34 publications

(53 reference statements)

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“…Hoskins and associates [18,32], in a novel in vitro model, triggered forward motility in the immature (immotile) sperm derived from bovine caput epididymis by incubation in presence of epididymal or seminal plasma and theophylline. Subsequently, from caprine epididymal plasma, pursuing to identify the extracellular proteins and their role in biochemical regulation of sperm motility, Jaiswal et al [33] and Das et al [34] have purified and characterized a motility initiating protein (MIP) and a motility inhibitory factor (MIF), respectively.…”

Section: Motility Initiating Protein and Motility Inhibitory Factormentioning

confidence: 99%

Sperm Motility Regulatory Proteins: A Tool to Enhance Sperm Quality

Dungdung¹,

Bhoumik²,

Saha³

et al. 2016

Insights From Animal Reproduction

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Sperm forward motility is an essential parameter in mammalian fertilization. Studies from our laboratory have identified and characterized a few unique sperm motility regulatory proteins/glycoproteins from the male reproductive fluids and mammalian blood serum. The purified sperm motility-initiating protein MIP from caprine epididymal plasma as well as the forward motility-stimulating factor FMSF and motility-stimulating protein MSP from buffalo and goat serum, respectively, have high efficacy to initiate or increase motility in nonmotile or less motile sperm. "ntibody of sperm motility inhibitory factor MIF-II has the high potential to enhance sperm vertical velocity and forward motility by increasing intracellular cyclic adenosine monophosphate c"MP level. The appearance and disappearance of D-galactose-specific lectin and its receptor along the epididymis has been reported to be involved in motility regulation in spermatozoa. " novel synthetic cryopreservation method and role of lipid to protect membrane damage during cryopreservation have been demonstrated. Motility-promoting proteins may be extremely useful for improving cattle breeding and breeding of endangered species, thereby helping in enhanced production of animal products as well as in the conservation of animals. Isolated proteins and developed cryopreservation technology may also be beneficial in human infertility clinics to increase the chance of fertilization.Keywords: Spermatozoa, Epididymis, Motility regulatory proteins, Cryopreservation, Reproduction © 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. . IntroductionLivestock is a very important subsector of Indian agricultural production system. The overall contribution of the livestock is almost . %. India ranks first in milk production in the world . million tons , which is mostly contributed by cattle and buffalo. The bull concentrates in itself a high economic value and thus need to be maintained on proper nutrition and management to obtain optimum performance in terms of semen production [ ]. The demand for the best males has increased considerably due to a shortage in the number of proven bulls having better semen characteristics for sustaining a successful breeding program [ ].Reproductive techniques facilitate the breeding of farm animals, which ultimately improve milk production and growth in dairy industry. "rtificial insemination "I is a first-generation reproductive biotechnology that profoundly contributed to genetic improvement, particularly in dairy cattle. Such impact would not have been possible without successfully freezing bull semen. Quality control of frozen sperm is of utmost importance for the sperm to be used in "I [ ]. Sperm cryopreservation allows prolonged preservation of semen and a wider use of a male gamete [ ]. Thi...

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Section: Motility Initiating Protein and Motility Inhibitory Factormentioning

confidence: 99%

Sperm Motility Regulatory Proteins: A Tool to Enhance Sperm Quality

Dungdung¹,

Bhoumik²,

Saha³

et al. 2016

Insights From Animal Reproduction

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“…Immobilin was reported as high molecular weight protein and decrease sperm motility by increasing viscoelastic drag. But detailed mechanism of action of these proteins still remains unclear (Das et al, ; Dungdung & Majumder, ; Usselman & Cone, ). Hence, QF is different from other reported sperm motility inhibitory molecules from cauda region.…”

Section: Discussionmentioning

confidence: 99%

“…Esposito et al () showed that soluble adenylyl cyclase (sAC) deficient mice were not able to produce cAMP, thus infertile due to severe sperm‐motility defect. Das et al () reported MIF‐II reduced intracellular cAMP level of spermatozoa thus inhibited motility but detailed mechanism of action was not reported. However, in QF treated mature cauda spermatozoa no significant changes in intracellular cAMP concentration were observed (Figure d), which revealed the sperm motility inhibitory activity of QF was not mediated by modulating intracellular Ca ++ as well as cAMP concentration.…”

Section: Discussionmentioning

confidence: 99%

A novel epididymal quiescence factor inhibits sperm motility by modulating NOS activity and intracellular NO‐cGMP pathway

Ghosh

Mukherjee

Bhoumik

et al. 2018

Journal Cellular Physiology

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Mature and potentially motile spermatozoa stored in cauda epididymis in an inactive state for approximately 30 days; however, during ejaculation they regain motility. To understand the actual molecular mechanism of the sperm quiescence during caudal stay, a proteinaceous quiescence factor (QF) has been purified from caprine epididymal plasma to apparent homogeneity. In the present study complete purification, detailed characterization as well as mechanistic pathway of QF has been described. QF is purified to 215-fold with 45% activity recovery. It is a 59 kDa monomeric protein with isoelectric point 5.8 and optimally active at pH 7.5. Circular dichroism spectroscopy and atomic force microscopy study confirm its α-helical secondary structure and globular tertiary conformation. QF is a thermo-stable protein as higher temperature does not alter its helical structure. N-terminal amino acid sequencing and MALDI analysis of QF did not find 100% similarity with any available protein of the database, proved its novelty. QF at 2 μM dose inhibits sperm progressive forward motility within 10 min. This motility inhibitory activity of QF is mediated by reducing NOS enzyme activity and subsequently decreasing the intracellular NO and cGMP concentration. It does not modulate intracellular Ca and cAMP concentration. QF has no adverse effect on DNA integrity and morphology of spermatozoa. Motility inhibitory action of QF is reversible. Thus, the role of QF in maintaining energy saving quiescence state of mature cauda spermatozoa and its reactive nitrogen species reducing activity may lead to a new direction for storage of spermatozoa and idiopathic male infertility.

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“…Equine semen is routinely submitted to centrifugation prior to cryopreservation to reduce the adverse effects of seminal plasma on post-thaw quality/motility (Moore et al, 2005). Likewise, the centrifugation of epididymal sperm can be also recommended for removal of epididymal fluid, once it suppresses the sperm movement (Usselman and Cone, 1983;Acott and Carr, 1984;Jeng et al, 1993;Das et al, 2010). The centrifuged samples, treated or nontreated, showed similarity in motion parameters (TM, VAP, ALH and RAP), confirming the hypothesis that epididymal sperm develops full motility when the epididymal fluid is removed and then exposed to the freezing extender.…”

Section: Discussionmentioning

confidence: 99%

Pentoxifylline effects on capacitation and fertility of stallion epididymal sperm

Guasti

Monteiro

Maziero

et al. 2017

Animal Reproduction Science

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The aims of this study were to determinate whether pentoxifylline (PTX) increases the motion parameters of fresh and frozen-thawed equine epididymal spermatozoa, to evaluate the tyrosine phosphorylation of frozen-thawed epididymal sperm in the presence of PTX and to determine whether the PTX-treatment of stallion epididymal sperm prior to freezing improves the fertility response of mares to a reduced number of spermatozoa per insemination dose. Fifty epididymis were flushed with a skim milk based extender with or without PTX. The pre-treatment with PTX enhanced the sperm motility after being harvested (P<0.05); however the freeze-thaw process did not alter the sperm kinematics between control and treated samples (P>0.05). Plasma membrane integrity did not differ between control and PTX group after recovery and after thawing (P>0.05), as observed in tyrosine phosphorylation, which the PTX treatment did not alter the percentage of tail-associated immunofluorescence of cryopreserved epididymal sperm (P>0.05). For the fertility trial, different insemination groups were tested: 800×10 epididymal sperm (C800); 100×10 epididymal sperm (C100); 100×10 epididymal sperm recovered in an extender containing PTX (PTX100). The conception rates for C800; C100 and PTX100 were 68.7% (11/16); 31.5% (5/16) and 50% (8/16), respectively. The conception rate did not differ among groups (P>0.05), however, a low number of animals was used in this study. A trend toward significance (P=0.07) was observed between C800 and C100 groups. In conclusion, PTX has no deleterious effect on sperm motility, viability and capacitation of cryopreserved stallion epididymal sperm. The conventional artificial insemination with 100×10 sperm recovered with PTX ensures acceptable conception rates and maximize the limited number of doses of cryopreserved stallion epididymal sperm.

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Purification and Characterization of a Sperm Motility Inhibiting Factor from Caprine Epididymal Plasma

Cited by 21 publications

References 34 publications

Sperm Motility Regulatory Proteins: A Tool to Enhance Sperm Quality

Sperm Motility Regulatory Proteins: A Tool to Enhance Sperm Quality

A novel epididymal quiescence factor inhibits sperm motility by modulating NOS activity and intracellular NO‐cGMP pathway

Pentoxifylline effects on capacitation and fertility of stallion epididymal sperm

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