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PubMed1994

Antigen processing in earthworms.

The administration of protein antigens into earthworms Lumbricus terrestris and Eisenia foetida induces the formation of antigen-binding protein (ABP) with the maximum response occurring between days 4 and 8. High proteolytic activities observed both in coelomocytes and in coelomic fluids cause rapid antigen degradation; the majority of antigen is digested during the first 24 h. To analyze the role of proteolytic processing of antigen in ABP response in vitro, the intact antigen (ARS-HSA) as well as its proteolytic fragments were added to tissue explant cultures and ABP formation followed by indirect ELISA using mAb to ABP. The response to small fragments was comparable to that induced by intact ARS-HSA. Furthermore, the response to intact antigen was almost completely blocked by the non-toxic serine protease inhibitor Pefabloc, while the response to small fragments was only slightly reduced. The kinetics of response to intact antigen significantly differed from that induced by small (< 3 kDa) fragments. We suggest that proteolytic processing is involved in the stimulation of ABP formation.

Animals↗
PubMed1991

Analysis of the acid and alkaline dissociation of earthworm hemoglobin, Lumbricus terrestris, by front-face fluorescence spectroscopy.

The steady-state fluorescence properties of the multisubunit hemoglobin isolated from the earthworm, Lumbricus terrestris, were studied by front-face fluorometry. Acid and alkaline dissociation of this high-molecular-weight hemoglobin were examined over the pH range 3.7-12.5 using different liganded states (oxy, CO, met). The relative intensity of the emission maximum at 320 nm (exc. 280 nm) is ligand-dependent increasing as follows: oxy less than deoxy less than CO less than met at pH 7.0. The intensity of the emission maximum of oxyhemoglobin at the alkaline acid end point, pH 10.5 (333 nm), is significantly greater than that observed at the acid end point, pH 4.18 (320 nm), suggesting different subunit dissociation. The spectra of oxyhemoglobin at pH 4.18 and the spectrum of carbonmonoxy hemoglobin at pH 7.0 in the presence of 1 M magnesium chloride were almost identical, indicating similar subunit dissociation. Difference spectrum (pH 9.0-7.2) of fluorescence emission (exc. 305) resulted in a maximum at 341 nm, indicative of tyrosinate formation. This suggests that tyrosine(s) may also be located at the subunit interface(s) of this hemoglobin. These studies indicate that several aromatic amino acid residues are associated with the critical sites of subunit interactions within this molecule. Analysis of the fluorescence spectra also suggests that the formation of different subunit species resulting from acid and alkaline dissociation cannot be ruled out.

Animals↗
PubMed1993

Zinc as modulator of oxygenation function and stabilizer of quaternary structure in earthworm hemoglobin.

Blood of the earthworm Pheretima hilgendorfi contains 10.7 mM Ca, 2.0 mM Mg, 75.5 mM Na, 5.9 mM K, 0.9 mM Zn and 0.3 mM Mn. Some of these cations cannot be removed completely from the blood by dialysis, and 36 atoms of Ca, 1-3 atoms of Mg and 1-2 atoms of Zn per 164 atoms of Fe were retained in purified preparation of the hemoglobin (Hb). At physiological pH, oxygen affinity and the Hill coefficient at half saturation (n1/2 value) of the Hb increased in the presence of 100 mM of Ca, Mg or Na. These effects were in the order of Ca > Mg > Na. At physiological concentration, however, the effect of each of these three cations on oxygenation was rather small. On the other hand, Zn gave a remarkable effect at less than 1 mM. This suggests a possibility that Zn acts as a primal modulator for the oxygenation function of the Hb in vivo. Oxygenation data at various pH values in the presence of each cation strongly suggest that Zn binds to a site different from those for the other three cations. Zn at a concentration of only 1 mM protected considerably the dissociation of the whole molecule to smaller components at alkaline pH and Zn thus contributes to the stabilization of the quaternary structure of the Hb.

Animals↗
PubMed1995

A novel gut tetradecapeptide isolated from the earthworm, Eisenia foetida.

A novel bioactive tetradecapeptide, GFKDGAADRISHGFamide, was isolated from the gut of the oligochaete annelid, Eisenia foetida, using the isolated anterior gut (crop-gizzard) as a bioassay system. A highly homologous peptide, GFRDGSADRISHGFamide, was also purified from the whole body of another species of earthworm, Pheretima vittata. These peptides were termed Eisenia tetradecapeptide (ETP) and Pheretima tetradecapeptide (PTP), respectively. Both the peptides showed a potent excitatory action on spontaneous contractions of the anterior gut with a threshold as low as 10(-10)-10(-9) M. These peptides were significantly homologous to molluscan tetradecapeptides and, to a lesser extent, to arthropodan tridecapeptides that have been reported to date. All these peptides seem to be evolutionally related to each other.

Amino Acid Sequence↗
PubMed1992

Comparative toxicity of polychlorinated biphenyls to earthworms Eisenia foetida and Lumbricus terrestris.

Effects of polychlorinated biphenyls (PCB; Aroclor 1254) in the manure worm, Eisenia foetida, on survival (LC50/LD50), and ability of coelomic leukocytes (also called coelomocytes) to form secretory rosettes (SR) and erythrocyte rosettes (ER) with, and to phagocytose antigenic rabbit red blood cells were determined and compared with those published for the earthworm, Lumbricus terrestris. Using a 5-day filter paper contact exposure protocol, LC50 and LD50 were 30.4 microg cm(-2) and 4500 microg g(-1) dry mass, respectively. Nominal PCB exposure concentrations of 5.0 and 10.0 microg cm(-2) resulted in tissue levels of 1400 and 2900 microg g(-1) dry mass. These body burdens resulted in significant reduction in SR formation by 18 and 52%, respectively. ER formation and phagocytosis were reduced 52 and 61%, respectively, only at the higher tissue concentration. Compared to L. terrestris, E. foetida: (1) accumulated considerably more PCB at each exposure concentration; (2) showed lower LC50, but higher LD50; and (3) exhibited effects on coelmocytes only at tissue PCB concentrations that caused some mortality. In terms of lethality and immunomodulation of SR, ER and phagocytosis, E. foetida appears to be more resistant to PCB than L. terrestris.

Journal Article↗
PubMed1988

Immunohistochemical demonstration of S-100 protein in the brain neurosecretory cells of invertebrates (insects and earthworms).

This immunohistochemical study demonstrated the presence of S-100 protein (-like substance) in the nervous systems of some invertebrate animals. The immunoreactivity of S-100 protein was detected in some neurosecretory cells, but not in glial cells in the brains (cerebral ganglia) of silkworms, cockroaches and earthworms. The immunoreactive nerve fibers extended to the neurohemal organs. These results suggested the possibility that S-100 protein (-like substance) may act as a neurohormone or carrier protein in invertebrate animals.

Animals↗
PubMed1994

Calcium wave propagation in the giant axon of the earthworm.

We examined a spatio-temporal pattern of intracellular free Ca2+ concentration in the giant axons of the earthworm, Eisenia foetida, with a fluorescent imaging technique using conforcal laser-scanning microscope and calcium indicator. 'Calcium Green 1'. Electrical tetanic stimulation applied to the nerve cord induced calcium waves along the giant axon. The calcium waves propagated both anteriorly and posteriorly with various speeds, and sometimes were split into several waves with different velocities. The results suggest that some types of calcium-releasing mechanisms may be associated with the calcium wave propagation.

Animals↗
PubMed1985

Chemo-attractant for the garter snake: characterization of vomeronasally-mediated response-eliciting components of earthworm wash--III.

Heating a 0.1 N HCl solution of lyophilized garter snake, Thamnophis sirtalis, chemoattractant obtained from earthworm, Lumbricus terrestris, surface washings, in a boiling water bath, and separation by gel chromatography, gave two fractions, F2, of high molecular weight and containing all the chemoattractant activity, and F4, of low molecular weight. The activity is lost between 15 and 30 min of heating. A similarly treated 0.1 N NaOH solution of lyophilized washings gave no F2 fraction and an F4 fraction in increasing amount. All the chemoattractant activity is lost between 0 and 15 min. Separations were monitored by assays for protein and carbohydrate.

Animals↗
PubMedDate not supplied

Distributions of polymorphisms among pathways of carbohydrate metabolism in the earthworm Eisenia fetida (Oligochaeta).

1. Activities and genetic banding patterns of 36 isozymes in carbohydrate metabolism were detected by spectrophotometry and starch-gel electrophoresis, respectively, in the earthworm Eisenia fetida. 2. Polymorphisms were not distributed randomly among metabolic pathways, activity levels, or gene copy numbers. 3. In glycolysis and the Krebs cycle, 16% of the loci were polymorphic and polymorphisms occurred only when multiple copies of the loci existed. 4. In other pathways, 45% of the loci were polymorphic and the distribution of polymorphisms was independent of gene copy number. 5. Polymorphisms may affect metabolic phenotypes and natural selection may have led to conserved biochemical activity in glycolysis and the Krebs cycle.

Animals↗
PubMed1992

Immunological analysis of chemoattractive proteins from earthworm to garter snakes.

1. Two sets of polyclonal antibodies to two highly purified prey-derived snake-attractive proteins, a low molecular weight (3000) protein and a 20,000 mol. wt protein, were generated in rabbits. 2. They are immunospecific for their respective purified immunogens and do not cross-react with each other. 3. Eight prey-derived proteins that elicit attack by garter snakes (Thamnophis sirtalis) from earthworms (Lumbricus terrestris) were analyzed with these antibodies, and can be assigned to three distinct groups on the basis of their antigenic properties. 4. Unfolding or denaturation of the low molecular weight protein did not alter its antigenic activity to its polyclonal antibodies, suggesting the antigenic epitopes contain contiguous amino acid sequences. 5. In contrast, unfolding of the 20,000 mol. wt protein resulted in a loss of its binding with antibodies, suggesting that the epitope of this protein contains noncontiguous amino acid sequences. 6. The snake-attractivity of the 20,000 mol. wt protein could not be neutralized by reacting it with its antiserum, suggesting that the antigenic determinant (the epitope) of the antigen is not an integral part of the attractive domain of the ES20 protein. 7. In contrast, the attractivity of the purified low molecular weight protein could be neutralized by the polyclonal antibodies.

Animals↗
PubMed1993

Mitogenicity of the earthworm's (Eisenia foetida) insulin-like proteins.

1. Biologically active glycolipoprotein complex (G-90), isolated from whole earthworm tissue extract (Eisenia foetida), was separated into seven fractions by gel-filtration. 2. It has been shown by radioimmunoassay that each of the fractions, except the lightest one, is cross-reactive with porcine anti-insulin antibodies. Molecules that possess such activity were detected by immunoblotting. 3. All fractions, except the heaviest and the lightest one, stimulate mammalian normal and transformed cell proliferation in serum-free conditions in vitro. The intensity of stimulation depends on cell type. Stimulation is completely abolished if the medium is supplemented with fetal calf serum.

Animals↗
PubMed1993

Serological evidence and amino acid sequence of ubiquitin-like protein isolated from coelomic fluid and cells of the earthworm Eisenia fetida andrei.

1. A small protein of M(r) 10 kDa has been isolated by reverse-phase chromatography of the basic proteins contained in the coelomic fluid and cell lysate of the earthworm Eisenia fetida andrei. 2. The protein crossreacted in dot-blot with an anti-bovine ubiquitin antiserum. 3. Its N-terminal primary structure was determined by automatic Edman degradation on 26 consecutive amino acids and showed 69% (based on the 26 amino acids) or 82% (based on the first 19 consecutive amino acids) identity with many ubiquitins and similar charge and hydrophobicity profiles and secondary structure conformation.

Amino Acid Sequence↗
PubMed1995

Alterations in polyamine levels of nematode, earthworm, leech and planarian during regeneration, temperature and osmotic stresses.

Free-living nematodes, Caenorhabditis elegans and Dorylaimus fodori, contain putrescine and spermidine. Putrescine, spermidine and spermine occur in the parasitic Nematoda, Ascaris suum, Anisakis simplex and Dirofilaria immitis. Earthworms, Eisenia foetida, Tubifex hattai and Pheretima communissima and the leech, Hirudo nipponia (belonging to Annelida) and the planarian, Dugesia japonica (belonging to Platyhelminthes) contain homospermidine and spermine in addition to putrescine and spermidine. Regenerated heads of E. foetida and D. japonica are rich in putrescine indicating the stimulation of its synthesis during regeneration. Putrescine and spermidine levels temporarily increase after heat shock in C. elegans, E. foetida and D. japonica and cold shock and hypertonic osmotic shock treatments in D. japonica.

Animals↗
PubMedDate not supplied

Neuronal localization of pancreatic polypeptide (PP) and vasoactive intestinal peptide (VIP) immunoreactivity in the earthworm (Lumbricus terrestris).

Nerve fibres and cell bodies displaying vasoactive intestinal polypeptide (VIP) or pancreatic polypeptide (PP) immunoreactivity were demonstrated in ganglia of the earthworm (Lumbricus terrestris). VIP cell bodies were found in the most anterior ganglion of the ventral nerve cord, the subpharyngeal ganglion. Immunoreactive nerves were seen running in the center of the cord until about the 10th segment. PP cell bodies were found in the cerebral ganglion where VIP was lacking, in the subpharyngeal ganglion and in more posteriorly located ganglia of the ventral nerve cord. PP nerve fibres could be followed below the 10th segment of the cord.

Animals↗
PubMed1995

Action potentials pass severed earthworm MGA segments reconnected by laser and electric field pulses.

Active and passive electrical signals transmit across the earthworm (Lumbricus terrestris) medial giant axon (MGA) segment after reconnection induced by laser and electric field pulses. Laser pulses of 308 nm, 15 ns pulse duration, and pulse energy 5-50 microJ in the objective plane induced axon reconnection in 12 of 133 (9%) preparations. Electric field pulses of 200-800 V, 100 microseconds duration, induced axon reconnection in 15 of 188 (8%) preparations. Electrical continuity is possible after the reconnection of severed axons induced by laser and electric field pulses.

Action Potentials↗
PubMed1984

Esterases of earthworms--III. Electrophoresis reveals that Eisenia fetida (Savigny) is two species.

Polyacrylamide gel electrophoresis of esterases in earthworms classified as Eisenia fetida (Savigny) revealed the existence of two species. The classification based upon esterase electromorphs coincided with classification into two colour variants formerly described as subspecies under the names E. fetida typica and E. fetida unicolor (André, 1963). Two main groups of esterases, acetylesterases and carboxylesterases were found. Because of less variability within species and higher stability the acetylesterases are regarded as the most useful in taxonomy. Sterile hybrids, with hybrid esterase patterns were produced at a low rate in the laboratory, but were never found with certainty in the wild populations. Eisenia fetida (Savigny) and Eisenia unicolor (André) are the names proposed for the two species.

Animals↗
PubMed1985

The action of cholinomimetic and cholinolytic agents, hemicholinium-3 and alpha- and beta-bungarotoxin on the body wall muscle of the earthworm, Lumbricus terrestris.

Strips of muscle, approximately 12 segments in length, were prepared from the body wall of the earthworm, Lumbricus terrestris, from which the nerve cord and viscera had been removed. Contractions to electrical stimulation and acetylcholine agonists were recorded using an isometric transducer. A range of nicotinic and muscarinic agonists and antagonists were tested on this preparation and the results indicate that the acetylcholine receptor on this muscle cannot be classified as either nicotinic or muscarinic. Hemicholinium-3 abolished electrically induced muscle twitches at concentrations which had no effect on the acetylcholine response. Alpha-Bungarotoxin blocked the responses to both electrical stimulation and acetylcholine while beta-bungarotoxin blocked the contractions induced by electrical stimulation but potentiated the acetylcholine contraction.

Acetylcholine↗
PubMed1991

Isolation, partial purification, and characterization of the cytochrome P-450-dependent monooxygenase system from the midgut of the earthworm Lumbricus terrestris.

1. Cytochrome P-450 was purified from microsomes of the midgut of the earthworm Lumbricus terrestris up to a maximal specific content of 5.5 nmol P-450/mg protein. 2. At least 3 different cytochromes P-450 with apparent molecular weights of 48,000, 51,000 and 53,000 were identified by SDS-PAGE. 3. Western blot analysis with various polyclonal antibodies did not show structural epitopes common to the cytochromes P-450 of rodents or yeast and L. terrestris. 4. The microsomes contained about 43 pmol P-450/mg protein corresponding to 0.51 nmol P-450/g midgut and 64 pmol P-450/g body weight, respectively, and converted benzyloxyresorufin into resorufin with a Vmax of 2.12 pmol resorufin/min.mg protein and a Km of 770 nM benzyloxyresorufin at 25 degrees C, pH 8.0. 5. The microsomes exhibited a NADPH-cytochrome P-450 reductase activity of 9.4 nmol cytochrome c/min.mg protein. 6. The apparent molecular weight of the threefold-purified reductase was 63,000.

Animals↗