Fertilization environment of the non-copulating marine sculpin, Hemilepidotus gilberti

To clarify the extracellular environment for external fertilization in the non-copulating marine sculpin Hemilepidotus gilberti, sperm motility was measured in NaCl, KCl, mannitol solutions, seawater, and ovarian fluid. Spermatozoa of H. gilberti actively moved in seminal plasma the moment they were removed from the genital papilla. Spermatozoa showed higher motility in NaCl solution at osmolalities between 300–400 mOsmol kg^-1. In KCl and in mannitol solutions, spermatozoa actively moved at osmolalities between 500 and 800 mOsmol kg^-1, and at osmolality 300 mOsmol kg ^-1, respectively. The ovarian fluid was a transparent and viscous gelatinous material, rich in sodium with an osmolality of 340 mOsmol kg^-1. Sperm motility in the ovarian fluid lasted more than 90 min, which was six times longer than in seawater. This sperm motility under conditions isotonic to body fluid is similar to that of copulating marine sculpins rather than to other non-copulating marine fishes. In addition, eggs of H. gilberti could be fertilized in the ovarian fluid. This suggests that external fertilization takes place under physiological conditions similar to the internal conditions of the ovary provided by the ovarian fluid, which isolates the eggs from sea water for several hours after spawning. This manner of fertilization is thought to be one of the evolutionary pre-adaptations allowing copulation among marine sculpins.     

The Proteome of Pig Spermatozoa Is Remodeled During Ejaculation

1. Download : Download high-res image (100KB)
2. Download : Download full-size image

Highlights

• •Proteome of mature boar spermatozoa from cauda epididymal and ejaculated sources were analyzed by iTRAQ-based LC-MS/MS.
• •1,723 sperm proteins identified (974 of Sus scrofa taxonomy).
• •1,602 sperm proteins quantified (960 of Sus scrofa taxonomy).
• •32 Sus scrofa sperm proteins were differentially expressed among sperm sources.
• •The proteome of boar spermatozoa is remodelled during ejaculation.

     

Osmotic regulation in aqueous humor and blood in the euryhaline fish,Eugerres plumieri

• 1.1. The estuarine fish Eugerres plumieri was acclimated to sea-water concentrations ranging from 6 to 85% sea-water.
• 2.2. Serum and aqueous humor osmolalities were moderately well regulated over the range of concentrations studied.
• 3.3. Serum osmolality and aqueous humor osmolalities conformed to the following relations: serum osmolality = (319 ± 3) + (0.56 ± 0.03) (% sea-water); aqueous humor osmolality = (314 ± 4) + (0.35 ± 0.04) (% sea-water).
• 4.4. Aqueous humor osmolality was more strictly regulated than that of serum, indicating that the retina and nervous system of the fish, which are encased in inextensible structures, are well protected from variations in sea-water concentration in order to minimize osmotically induced changes in cell volume.

     

The chemical composition of the follicular fluid of the teleost Clinus dorsalis (Perciformes: Clinidae)

• 1.1. The osmolarity and pH of the follicular fluid was determined and analyses of total glucose, total lipids, total proteins, amino acids, urea, sodium and potassium carried out.
• 2.2. The mean osmolarity of the follicular fluid was found to be 325 mOsm/kg and the mean pH was 7.9.
• 3.3. The embryotrophe was rich in lipids (1092.39 mg/100 ml) and amino acids with the amino acid concentration exceeding normal values for human plasma.

     

Integrated Analyses of Phenotype and Quantitative Proteome of CMTM4 Deficient Mice Reveal Its Association with Male Fertility

1. Download : Download high-res image (158KB)
2. Download : Download full-size image

Highlights

• •CMTM4 is associated with human spermatogenesis and sperm quality.
• •Cmtm4 knockout mouse were generated by CRISPR/Cas9 technology for male fertility research.
• •CMTM4 is required for male fertility but not female fertility.
• •Phenotype and quantitative proteomics of Cmtm4 KO mice reveal an association of CMTM4 with histone-to-protamine exchange, sperm motility and induction of the acrosome reaction.

     

Effect of salinity on the osmotic,chloride, total protein and calcium concentrations in the hemolymph of the prawn Peneaus monodon (fabricius)

• 1.1. Osmolality and chloride concentrations in the hemolymph of Penaeus monodon became stable 1 day after molting in 32 ppt, while total protein and calcium concentrations remained stable throughout the molting cycle. When intermolt (≥ 36 hr postmolt) animals were transferred from control (32 ppt) to experimental (8–40 ppt) salinities, osmolality, chloride and total protein, but not calcium, concentrations in the hemolymph achieved steady state values 24–48 hr after transfer.
• 2.2. The hemolymph osmolality was a linear function (slope = 0.28) of medium osmolality at salinities between 8 and 40 ppt. It was isosmotic to seawater at 698 mOsm (10 g prawns) and 752 mOsm (30 g), and was hyperosmotic to the medium below isosmotic concentrations, and hypoosmotic to those above.
• 3.3. Hemolymph chloride concentration was isoionic to seawater at 334 mM, and was hyperregulated below isoionic concentrations, and hyporegulated to those above.
• 4.4. P. monodon maintained its hemolymph calcium concentration between 6.4 and 10 mM when medium salinities increased from 8 to 40 ppt.
• 5.5. Total protein concentration in the hemolymph was independent of medium salinity (8–40 ppt) and hemolymph osmolality (540–850 mOsm).

     

A halogenated amino acid-containing sperm activating peptide and its related peptides isolated from the egg jelly of sea urchins,Tripneustes gratilla,Pseudoboletia maculata,Strongylocentrotus nudus,Echinometra mathaei and Heterocentrotus mammillatus

• 1.1. Twenty-eight peptides were isolated from the egg jelly of sea urchins, Tripneustes gratilla, Pseudoboletia maculata, Strongylocentrotus nudus, Echinometra mathaei (type A and B) and Heterocentrotus mammillatus and their amino acid sequences were determined.
• 2.2. Two of the peptides obtained from T. gratilla egg jelly possessed a bromophenylalanine (Br-Phe) residue in their sequences (Gly-(Br-Phe)-Asn-Leu-Asn-Gly-Gly-Gly-Val-Gly and Gly-(Br-Phe)-Asp-Leu-Asn-Gly-Gly-Gly-Val-Gly).
• 3.3. All of the peptides elevated cyclic GMP concentrations in the spermatozoa of the respective sea urchin and caused a shift in the apparent mol. wt of a major sperm protein of the respective sea urchin.
• 4.4. They stimulated respiration rates of the spermatozoa of Hemicentrotus pulcherrimus as well as their own species.
• 5.5. One-half maximal concentrations of the peptides for respiratory stimulation of H. pulcherrimus spermatozoa were between 10⁻¹¹ M and 10⁻⁹ M except a methionine-containing peptide which was about 10⁻⁷ M.

     

Anuran amphibia which are not acclimable to high salt,tolerate high plasma urea

• 1.1. The capacity of five anuran Amphibians (Bufo viridis B. regularis, Rana ridibunda, Hyla arborea and Pelobates syriacus) to acclimate to NaCl and urea solutions was investigated.
• 2.2. All species could be acclimated to relatively high concentrations of urea solutions, while only Bufo viridis and Hyla arborea could be acclimated to 500 mOsm/kg or higher NaCl solutions.
• 3.3. The plasma urea concentration in B. viridis and H. arborea was elevated to levels over 140 mmol/1.
• 4.4. The sum of plasma sodium and chloride concentrations did not increase over 400 mmol/l in any species.
• 5.5. Urine osmolality, which was normally low, increased, but never exceeded the plasma osmolality.
• 6.6. In the urea acclimation conditions, urine electrolytes diminished, similarly in all species in this study.
• 7.7. It is concluded that anuran Amphibians can tolerate high plasma urea concentrations, but only those species which can elevate it, either through retention or net synthesis, can be acclimated to high salt solutions.

     

A comparative study of the basic nuclear proteins from sperm of bivalve molluscs

• 1.1. Basic nuclear proteins from spermatozoa of the three mollusc species belonging to the class Bivalvia have been analyzed using one- and two-dimensional electrophoresis.
• 2.2. Four nuclear basic proteins have been purified and their amino acid compositions determined.
• 3.3. In these spermatozoa histone-type proteins coexist with protamine-like proteins.
• 4.4. The protamine-like proteins that have been studied show different electrophoretic behavior but in general are similar, with a high content of lysine, arginine, alanine and serine.
• 5.5. Interspecific variability has been found for the H1-like histone.

     

Biochemical aspects of Balanus hameri haemolymph,together with some comparative haemolymph data for Balanus balanus and Lepas anatifera

• 1.1. The concentration of protein in the haemolymph of Balanus hameri ranged from 2.0 to 17.3 mg/ml, and the lipid from 1.4 to 7.7 mg/ml; the haemolymph protein and lipid levels increased significantly prior to cross-fertilization.
• 2.2. The protein and lipid concentrations in Balanus balanus haemolymph were 8.1 and 1.7 mg/ml respectively.
• 3.3. The lipid concentration of Lepas anatifera haemolymph was 1.2 mg/ml.
• 4.4. The neutral lipid and phospholipid components of B. hameri and L. anatifera haemolymph were the same, with the major components of the phospholipid fraction being phosphatidyl ethanolamine and phosphatidyl choline.
• 5.5. The osmolarity (970.4 mOsm), chloride ion concentration (501.3 m-eq/l) and pH (7.29) of B. hameri haemolymph were also determined.

     

Identification and characterization of vitellin in a hermophrodite shrimp,Pandalus kessleri

• 1.1. A female specific protein (FSP, vitellogenin) in hemolymph and its related ovarian protein (vitellin) of Pandalus kessleri were studied by means of electrophoretical and immunological procedures.
• 2.2. The vitellin was purified from vitellogenic ovaries using hydroxylapatite, DEAE cellulose and Sepharose 6B columns, consecutively.
• 3.3. The vitellin had a molecular weight of approximately 560 kD and was composed of two subunits, 81 and 110 kD, respectively.
• 4.4. The vitellogenin concentrations in the hemolymph increased as vitellogenesis in the ovarian oocytes advanced and dropped markedly after the release of mature eggs.

     

Induction of locomotor behavior in the giant African snail,Achatina fulica

• 1.1. The locomotor-inducting factor of the giant African snail, Achatina fulica, was examined.
• 2.2. Snails showed nocturnal circadian behavior in relative humidity at least over 50%. Although the rhythmicity was independent of light and darkness, it was disturbed easily by hydration, and hydrated snails continued to locomote throughout the day. For induction of locomotor behavior, relative humidity over 50% was the fundamental factor and water is shown to be the limiting factor for the endogeneous circadian oscillator.
• 3.3. The integument of snails showed a higher water permeability. Through the integument, hemolymph osmolality changed easily according to hydration and dehydration from about 120 to 400 mOsm/kg H₂O. Circadian behavior was induced in snails in which hemolymph osmolality ranged from about 130 to 230 mOsm/kg H₂O.
• 4.4. By hydration, hemolymph osmolality in quiescent and estivated snails which have higher osmolality decreased gradually and then they began to locomote according to the degree of dilution, and vice versa. The induction of behavior in these snails was controlled by low hemolymph osmolality.
• 5.5. Together with the endogeneous rhythmicity, water environment was shown to be the key factor for the induction of locomotor behavior.
• 6.6. Based on these results, the mechanisms of the induction of locomotor behavior in terrestrial pulmonates are proposed.

     

Hyperosmotic properties of the fluids of the perivisceral coelom and water vascular system of starfish kept under stable conditions

• 1.1. Osmotic measurements were made on the perivisceral coelomic and water vascular fluids of 4 species of northwest Pacific starfish and their stable sea-water media.
• 2.2. Mean levels of both fluids were hyperosmotic in every species, often at statistically significant levels.
• 3.3. For all species combined, mean hyperosmolality (mosmol/kg ± SE) of perivisceral coelomic fluid was 1.49 ± 0.17, and water vascular fluid 6.07 ± 0.74.
• 4.4. The hyperosmotic nature of these fluids contributes to water balance, working in conjunction with madreporitic inflow and other factors.

     

Influence of sex hormones on the chick gastrointestinal transit and emptying

• 1.1. Gastrointestinal (GI) transit and emptying of male and female 15-day-old chickens treated with testosterone, estradiol and progesterone was measured by means of ¹⁴C-polyethylene glycol-4000.
• 2.2. All the administered sex hormones increased GI motility at the shortest time (0.5 and 1 hr) after the marker administration, but decreased GI motility at the longest times (2 and 4 hr). This motor pattern agrees with the known anabolic role of sex hormones.
• 3.3. We conclude that testosterone and estradiol increased GI motility and intestinal inhibitory reflexes. Thus, chicks' and mammals' GI motility were modified by testosterone and estradiol in a similar form.
• 4.4. The effect of progesterone on the chick GI motility was contrary to that observed in mammals. This may happen because of increased inhibitory GI motor reflexes or direct inhibition of visceral smooth muscle activity.
• 5.5. No statistical differences were observed between the sexes, which could be explained by the sexual immaturity of chicks.
• 6.6. Chicks constitute good biological material to study the influence of sex hormones on avian GI motility.

     

Integration of Two In-depth Quantitative Proteomics Approaches Determines the Kallikrein-related Peptidase 7 (KLK7) Degradome in Ovarian Cancer Cell Secretome

1. Download : Download high-res image (59KB)
2. Download : Download full-size image

Highlights

• •Kallikrein-related peptidase 7 is over expressed in ovarian cancer.
• •Quantitative PROTOMAP and TAILS approaches identified putative substrates of KLK7.
• •Pro-MMP10 is activated by KLK7.
• •KLK7 cleaves thrombospondin 1 and IGFBP6 in vitro.

     

Proteome-wide Tyrosine Phosphorylation Analysis Reveals Dysregulated Signaling Pathways in Ovarian Tumors

1. Download : Download high-res image (94KB)
2. Download : Download full-size image

Highlights

• •Multiplexed PTM assays on HuProt array were developed using ovarian tumor lysates.
• •Proteome-wide Tyr phosphorylation with 102 ovarian tumors were performed and analyzed.
• •19 kinases were predicted to have elevated activities in ovarian tumor.
• •Elevated activities of PTK2 and PTK2B were confirmed in ovarian cancer cell lines.

     

Identification and partial characterization of plasma membrane polypeptides of Crithidia guilhermei,crithidia deanei and Crithidia oncopelti

• 1.1. Components of the cell surface of Crithidia guilhermei, Crithidia deanei and Crithidia oncopelti were radioiodinated by the iodogen technique. The distribution of proteins in the detergent-poor (DPP) and detergent-enriched phase (DRP) were studied using a phase separation technique in Triton X-114 and one- and two-dimensional polyacrylamide gel electrophoresis in sodium dodecyl sulphate (1D and 2D SDS-PAGE).
• 2.2. Significant differences were noted in the proteins present in the DRP when the three species were compared.
• 3.3. Two major bands with mol. wt 28,000 and 56,000 and motility in the pH gradient of 7.4 and 6.3, respectively, were observed in C. guilhermei, but not discernible in C. deanei and C. oncopelti.
• 4.4. One polypeptide with mol. wt 50,000 and p1 4.9 was identified in the DRP of C. deanei.
• 5.5. A broad band with mol. wt 68,000–140,000 and pI 4.7–5.5 was clearly observed in the DRP of C. deanei and one or two polypeptides only present in the DPP were observed in the three Crithidia species analyzed.
• 6.6. Our observations show that C. guilhermei has characteristic surface polypeptides not found in C. deanei and C. oncopelti.
• 7.7. Our results, in association with those reported by others, show that the phase separation using Triton X-114 offers a simple approach to the separation and further analysis of a select group of proteins from the bulk of the cellular proteins.

     

Identification of TEX101-associated Proteins Through Proteomic Measurement of Human Spermatozoa Homozygous for the Missense Variant rs35033974,

1. Download : Download high-res image (74KB)
2. Download : Download full-size image

Highlights

• •Missense variant rs35033974 resulted in significantly reduced levels of human TEX101 protein in seminal plasma and spermatozoa.
• •Differential proteomics revealed TEX101-associated testis-specific proteins, including LY6K, which were down-regulated in rs35033974^hh spermatozoa.
• •Deep proteome of human spermatozoa, including some “missing” proteins, was identified.

     

A Robust and Versatile Automated Glycoanalytical Technology for Serum Antibodies and Acute Phase Proteins: Ovarian Cancer Case Study

1. Download : Download high-res image (109KB)
2. Download : Download full-size image

Highlights

• •Quantitative high-throughput glycoanalytical technology as a diagnostic tool for ovarian cancer detection.
• •Multiplexed approach harnessing N-glycan data for six glycoproteins from a single biological sample.
• •Detailed characterization of human serum N-glycans from antibodies IgG, IgM and IgA and acute phase proteins transferrin, haptoglobin and alpha-1-antitrypsin.
• •Structural differences in antibody and acute phase protein glycosylation for mechanistic insights.