• Title/Summary/Keyword: egg membrane

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Cell Signaling Mechanisms of Sperm Motility in Aquatic Species

  • Kho, Kang-Hee;Morisawa, Masaaki;Cho, Kap-Seong
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.665-671
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    • 2005
  • Initiation and activation of sperm motility are prerequisite processes for the contact and fusion of male and female gametes at fertilization. The phenomena are under the regulation of cAMP and $Ca^{2+}$ in vertebrates and invertebrates. Mammalian sperm requires $Ca^{2+}$ and cAMP for the activation of sperm motility. Cell signaling for the initiation and activation of sperm motility in the ascidians and salmonid fishes has drawn much attention. In the ascidians, the sperm-activating and attracting factors from unfertilized egg require extracellular $Ca^{2+}$ for activating sperm motility and eliciting chemotactic behavior toward the egg. On the other hand, the cAMP-dependent phosphorylation of protein is essential for the initiation of sperm motility in salmonid fishes. A decrease of the environmental $K^+$ concentration surrounding the spawned sperm causes $K^+$ efflux and $Ca^{2+}$ influx through the specific $K^+$ channel and dihydropyridine-sensitive L-/T-type $Ca^{2+}$ channel, respectively, thereby leading to the membrane hyperpolarization. The membrane hyperpolarization induces synthesis of cAMP, which triggers further cell signaling processes, such as cAMP-dependent protein phosphorylation, to initiate sperm motility in salmonid fishes. This article reviews the studies on the physiological mechanisms of sperm motility and its cell signaling in aquatic species.

Comparison various level ascorbic acid and lycopene additions in semen diluent enhanced sperm quality of Sapudi ram

  • Bintara Sigit;Dyah Maharani;Luis Tavares;Pradita Iustitia Sitaresmi
    • Journal of Animal Science and Technology
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    • v.66 no.5
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    • pp.891-904
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    • 2024
  • The primary cause of sperm quality decline during the freeze-thaw pathway is the peroxidation hazard caused by reactive oxygen species produced by the biological molecules of sperm. Ascorbic acid (Vitamin C) and lycopene are two potent antioxidants that operate to prevent oxidation processes. This study aimed to analyse the effects of ascorbic acid and lycopene on the motility, viability, abnormality and plasma membrane integrity of post-thawed Sapudi rams. Sperm samples were obtained and pooled from six sexually mature Sapudi rams, separated into ten equal proportions and diluted with Tris-egg yolk-glycerol (TEY) extender. Semen was supplemented with 0 (C0; L0), 1 (C1; L1), 2 (C2; L2), 3 (C3; L3) and 4 (C4; L4) mg/100 mL (1%-4%) diluent each of ascorbic acid and lycopene, respectively. Total sperm motility, viability, abnormalities and semen membrane plasma (%) were analysed after thawing. C3 and L3 extenders resulted in higher total motility (p < 0.05) compared to the other extenders, with all treatments higher than that of the control. The extender C3 (p < 0.05) exhibited the highest semen quality. Finally, the current findings show that C3 and L3 can increase the quality of post-thawed Sapudi ram spermatozoa.

Control of $Ca^{2+}$- Influx by $Ca^{2+}$/Calmodulin Dependent Protein Kinase II in the Activation of Mouse Eggs

  • Yoon, Sook-Young;Kang, Da-Won;Bae, In-Ha
    • Development and Reproduction
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    • v.15 no.1
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    • pp.31-39
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    • 2011
  • Change in intracellular $Ca^{2+}$-concentration ($[Ca^{2+}]_i$) is an essential event for egg activation and further development. $Ca^{2+}$ ion is originated from intracellular $Ca^{2+}$-store via inositol 1,4,5-triphosphate receptor and/or $Ca^{2+}$ influx via $Ca^{2+}$ channel. This study was performed to investigate whether changes in $Ca^{2+}$/calmodulin dependent protein kinase II (CaM KII) activity affect $Ca^{2+}$ influx during artificial egg activation with ethanol using $Ca^{2+}$ monitoring system and whole-cell patch clamp technique. Under $Ca^{2+}$ ion-omitted condition, $Ca^{2+}$-oscillation was stopped within 30 min post microinjection of porcine sperm factor, and ethanol-induced $Ca^{2+}$ increase was reduced. To investigate the role of CaM KII known as an integrator of $Ca^{2+}$- oscillation during mammalian egg fertilization, CaM KII activity was tested with a specific inhibitor KN-93. In the eggs treated with KN-93, ethanol failed to induce egg activation. In addition, KN-93 inhibited inward $Ca^{2+}$ current ($I_{Ca}$) in a time-dependent manner in whole-cell configuration. Immunostaining data showed that the voltage-dependent $Ca^{2+}$ channels were distributed along the plasma membrane of mouse egg and 2-cell embryo. From these results, we suggest that $Ca^{2+}$ influx during fertilization might be controlled by CaM KII activity.

Voltage Dependent N Type Calcium Channel in Mouse Egg Fertilization

  • Eum, Jin Hee;Park, Miseon;Yoon, Jung Ah;Yoon, Sook Young
    • Development and Reproduction
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    • v.24 no.4
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    • pp.297-306
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    • 2020
  • Repetitive changes in the intracellular calcium concentration ([Ca2+]i) triggers egg activation, including cortical granule exocytosis, resumption of second meiosis, block to polyspermy, and initiating embryonic development. [Ca2+]i oscillations that continue for several hours, are required for the early events of egg activation and possibly connected to further development to the blastocyst stage. The sources of Ca2+ ion elevation during [Ca2+]i oscillations are Ca2+ release from endoplasmic reticulum through inositol 1,4,5 tri-phosphate receptor and Ca2+ ion influx through Ca2+ channel on the plasma membrane. Ca2+ channels have been characterized into voltage-dependent Ca2+ channels (VDCCs), ligand-gated Ca2+ channel, and leak-channel. VDCCs expressed on muscle cell or neuron is specified into L, T, N, P, Q, and R type VDCs by their activation threshold or their sensitivity to peptide toxins isolated from cone snails and spiders. The present study was aimed to investigate the localization pattern of N and P/Q type voltage-dependent calcium channels in mouse eggs and the role in fertilization. [Ca2+]i oscillation was observed in a Ca2+ contained medium with sperm factor or adenophostin A injection but disappeared in Ca2+ free medium. Ca2+ influx was decreased by Lat A. N-VDCC specific inhibitor, ω-Conotoxin CVIIA induced abnormal [Ca2+]i oscillation profiles in SrCl2 treatment. N or P/Q type VDC were distributed on the plasma membrane in cortical cluster form, not in the cytoplasm. Ca2+ influx is essential for [Ca2+]i oscillation during mammalian fertilization. This Ca2+ influx might be controlled through the N or P/Q type VDCCs. Abnormal VDCCs expression of eggs could be tested in fertilization failure or low fertilization eggs in subfertility women.

Green tea extract addition into a Tris-based egg yolk extender improves Bali bull sperm quality

  • Ragil Angga, Prastiya;Tri Wahyu, Suprayogi;Aldea Erian, Debora;Ani, Wijayanti;Anny, Amalia;Deny, Sulistyowati;Aras Prasetiyo, Nugroho
    • Animal Bioscience
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    • v.36 no.2
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    • pp.209-217
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    • 2023
  • Objective: The conservation of Bali bulls, the Indonesian native breed of cattle, is crucial for cattle breeding in Indonesia. To guarantee the spread of Bali bulls through artificial insemination the quality of the frozen semen must be high. To this end, adding an extender material to semen that increases spermatozoa's survival during cryopreservation is important. Green tea extract (GTE) can be used as cryoprotectant because its high antioxidant activity can help avoid reactive oxygen species formation. Methods: Semen of five Bali bulls from the National Artificial Insemination Center at Singosari, Indonesia was collected routinely twice a week. First, fresh semen inspection was performed to determine the feasibility of using Bali bulls as animal samples. The extender used in this study was Tris-based egg yolk. The samples were divided into four treatments: T0, no GTE added to the extender; T1, 0.05 mg GTE plus 100 mL extender; T2, 0.10 mg GTE plus 100 mL extender; and T3, 0.15 mg GTE plus 100 mL extender. The semen freezing process was conducted according to standard procedures and sperm quality parameters, i.e., sperm motility, viability, abnormalities, and membrane integrity observed pre-freezing and post-thawing. Results: There were significant differences in total motility, progressive motility, viability, and integrity membrane of Bali bull sperm at both pre-freezing and post-thawing after adding GTE into the extender. In contrast, there were no differences in abnormalities among treatments. Conclusion: Adding GTE at a 0.15 mg into 100 mL Tris-based egg yolk extender can improve the quality of cryopreserved Bali bull sperm.

Influence of aqueous extract of Annona muricata leaves in Tris-egg yolk extender on storage of spermatozoa from West African Dwarf goat (Capra hircus)

  • Mohamadou Adamou;Dongmo Nguedia Arius Baulland;Ngo Bahebeck Pierrette;Tchoffo Herve;Chongsi Margaret Mary Momo;Noudjio Kezeter Claude;Nnanga, Germaine Estelle Mvondo;Ngwemetah Nathalie;Adamu Yusufa Njeiri;Ngoula Ferdinand
    • Journal of Animal Reproduction and Biotechnology
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    • v.39 no.3
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    • pp.179-193
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    • 2024
  • Background: Because oxidative stress can induce decreased quality of caprine semen during the storage, there has been limitation for the use of stored semen in the assisted reproductive technologies. The present study, therefore, assesses the potential of Annona muricata (A. muricata) to reduce semen storage associated-damages. Methods: Semen was collected by electro-ejaculation from ten bucks, and extended with Tris-egg yolk (TEY) supplemented with A. muricata leaf aqueous extract (SAE) at 20 (SAE20), 40 (SAE40), and 80 (SAE80) ㎍/mL. Sperm variables including motility, motion characteristics, viability, membrane functionality, and DNA integrity were assessed at different storage periods (6, 24, 48, and 72 hr). In addition, oxidative stress indicators in the extender supplemted with SAE were also assessed for each group. Results: By adding SAE at 80 ㎍/mL in TEY, the storage of goat buck semen was improved, resulting in reduced loss of sperm motility, viability, DNA fragmentation, and membrane integrity during chilled storage at 4℃ for up to 72 hr. In addition, enrichment of TEY extender with SAE significantly (p < 0.05) reduced malondialdehyde, an indicator of oxidative stress, compared to the negative control. Conclusions: Supplementation of SAE in TEY extender can reduce buck spermatozoa liquid storage associated damages due to oxidative stress.

Ultrastructure of the Fertilized Egg Envelope from Hyphessobrycon serpae, Characidae, Teleost (경골어류 카라신과 Hyphessobrycon serpae의 수정란 난막 미세구조)

  • Kim, Dong-Heui;Deung, Young-Kun;Lee, Kyu-Jae
    • Applied Microscopy
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    • v.35 no.2
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    • pp.89-96
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    • 2005
  • The ultrastructures of the fertilized egg envelope from Hyphessobrycon serpae belonging to Characidae was studied using scanning and transmission electron microscopes to get systematic fundamental data for classification of species and to confirm whether micropyle is a common trait of Characidae or not. The fertilized egg was of colorless, transparent, spherical, adhesive and demersal type. There were not oil droplets in vitelline membrane and attached structures in the outside of fertilize egg envelope. The egg envelope had a single micropyle resembling the pathway of sperm in the area of the animal pole. The micropyle was surrounded by 13 to 15 protruded lines of the egg envelope in a radiated form. The outer surface of fertilized egg envelope was covered by reticular adhesive fibrous structures and irregularly arranged by pore canals. The fertilized egg envelope consisted of three distinct layers an outer adhesive fibrous layer with high electron density, a middle layer with pore canals, and an inner layer consisting of 6 to 7 lamellae alternating layers with interlamellae of lower electron density. These ultrastructural characters of fertilized egg envelope form Hyphessobrycon serpae can be utilized in taxonomy of teleost, and as fundamental data for study on early development of fertilized egg. It seems that the morphology of micropyle is a common trait of Characidae

The Oogenesis of Chinese minnow, Leuciscinae, Teleostei (경골어류 황어아과 버들치의 난자형성과정)

  • Kim, Dong-Heui;Chang, Byung-Soo;Jung, Han-Suk;Teng, Yung-Chien;Kim, Seok;Lee, Kyu-Jae
    • Applied Microscopy
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    • v.39 no.3
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    • pp.237-243
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    • 2009
  • Chinese minnow, Rhynchocypris oxycephalus is a teleost belonging to Leuciscinae, Cyprinidae. The oogenesis and ultrastructure of egg envelope in Chinese minnow were investigated by light and electron microscopes. The ovary was of white yellowish and ellipsoidal shape with the major axis 30 mm and the minor axis 7mm. Cytoplasm of oogonia was basophilic and many nucleoli were located at inside of nuclear membrane. In primary oocytes, yolk vesicles were distributed only in the marginal area and egg envelope was not formed on the outside of an egg. In secondary oocytes, the egg envelope was formed and yolk vesicles in the cytoplasm were increased than the earlier stage. The basophilic substance of cytoplasm was changed to acidic. In case of matured egg, thickness of egg envelope and size of egg were increased. The yolk vesicles were changed to yolk mass in accordance with development. The outer surface of egg envelope was covered by microvilli-structures, and had a micropyle on the area of animal pole. Egg envelope consisted with 2 layers, an adhesive outer layer with microvilli-structures and fibrillar inner layer. In conclusion, the oogenesis of Chinese minnow was characterized by the increase in cell size, the formation and accumulation of yolk, and the decrease of basophilic substance in the cytoplasm. The oogenesis of Chinese minnow seems to share common patterns in Cyprinidae, but these ultrastructural unique characters of egg envelope can be utilized in taxonomy of teleost.

Ultrafiltration and Separation Process Optimization of Hen Egg White Lysozyme as Natural Antimicrobial Enzyme (천연 항균 효소제 난백 lysozyme의 한외여과 조건 최적화)

  • Lee, Eun-Young;Woo, Gun-Jo
    • Korean Journal of Food Science and Technology
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    • v.30 no.2
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    • pp.397-406
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    • 1998
  • Hen egg white lysozyme (HEWL) is very valuable as a natural preservative in food processing due to its selective bactericidal activity. HEWL which traditionally isolated by crystallization or freeze drying was simply separated from 13 different hen egg white (HEW) proteins by a single-step ultrafiltration. Freeze dried HEW (0.25%, w/v) dissolved in a citrate-phosphate buffer (pH 4.6) was ultrafiltered with a PM30 membrane under various operating conditions, by changing concentration, temperature, transmembrane pressure $({\triangle}P_T)$, and stirring speed. Optimum separation conditions were decided when maximal flux was obtained. Under the optimum separation conditions, the effect of membrane material and fouling on flux as time passed as well as lysozyme concentration, protein concentration, specific activity (SA) in the permeate were measured. Best separation conditions of HEWL with PM30 membrane were sample concentration 0.25%, temperature $35^{\circ}C$, ${\Delta}P_T\;30\;psi$, and stirring speed 300 rpm. During the first 12 min, the flux of YM30 was higher, but at the steady-state it was lower than that of PM30. The SA of the PM30 permeate was over 2 times higher in spite of the lysozyme and protein concentration being lower than that of YM30 permeate. The flux of 5 times used PM30 decreased 30% compared to a new PM30, but both had the same tendency in flux decrease when time passed. Both of them reached a steady-state after 35 min and remained at 70% of the initial flux. In the PM30 permeate, the lysozyme concentration and SA were 110 units/mL and 2,821 units/mg protein, respectively. Therefore, PM30 membrane separation was very effective for separation of antimicrobial lysozyme.

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Adhesive Membrane of Oocyte in Korean Cobitid Species (Pisces, Cobitidae) (한국산 기름종개속 어류의 난모세포의 부착막)

  • 김익수;박종영
    • The Korean Journal of Zoology
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    • v.38 no.2
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    • pp.212-219
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    • 1995
  • Through the histolosical observation of ovary in the nine species of the genus Cobitis, we found that specific adhesive membranes were attached to the outer zone radials during the vitellosenesis. These adhesive membranes could be classified into four forms as follows: 1) granular form of Colitis lutheri, C. striate, and C. sinensis, 2) villous form of C. longicorpus, C. konensis koreensis, C. k. pumilus, and C. granoei, 3) saw-like form of C. rotundicaudata, and 4) filamentous form of C. chou. The various forms of adhesive membrane of egg in the cobitid fishes are showed a species specificitv with reference to their habitats and spawning property.

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