Acrosome morphogenesis in Lumbricus terrestris

Summary Acrosome morphogenesis commences in the juxtanuclear cytoplasm at the posterior end of spermatids of Lumbricus terrestris. A dense rod-shaped structure and the Golgi apparatus together participate first in forming an acrosome vesicle that contains the acrosome granule, and somewhat later shape the conical base of the acrosome in the cytoplasm beneath the vesicle. Cytoplasmic flow may account for the migration of the immature acrosome to the apical surface of the nucleus of the spermatid. Manchette microtubules play a key role in the final modelling of the acrosome. Sheathed by the manchette the acrosome elongates to 3–4 times its pre-attachment length. The conical base of the acrosome then extends anteriorly to enclose the acrosome vesicle. A dense rod emerging from the rod-shaped granule occupies an indentation of the base of the acrosome vesicle. The mature acrosome of Lumbricus is an extremely complex structure about 5–7 microns long and is bounded by the plasmalemma of the spermatozoon.This study was supported by a research training grant GM-00582-07 from the Public Health Service.     

Parvalbumin-immunoreactive material in the earthworm, Lumbricus terrestris

Parvalbumin-like material was localized using an immunoeytochcmieal method, in neurons of the central nervous system and in cells intermingled in the skin of the earthworm. Lumbricus terreslris L. Parvalbumin-immunoreactive material was found in the cytoplasm of perikarya and neutrites, not in the nucleoplasm. In contrast to vertebrates, Lumbricus musculature did not contain parvalbumin-immunoreactive material.     

Cadmium-induced metallothionein in earthworms (Lumbricus terrestris)

Studies are continuing to compare the common earthworm (Lumbricus terrestris) to rodents by their reactions to toxic levels of metals. A recent study suggests that LD₅₀ of selenium compounds were about ten times less in worms than in rodents, but the toxicity of cadmium was of the same order of magnitude between these diverse species. An interesting finding is that the common earthworm produces a Cd-bound protein, metallothionein, after the injection of cadmium salts, and, similar to rodents, two Cd-bound isoproteins can be isolated by a Sephacryl S-200 column and monitored by both atomic absorption and liquid scintillation spectroscopy.     

The nitrogen excretory metabolism of Lumbricus terrestris

The use of the earthworm as a laboratory animal for studying the effects of starvation on nitrogen metabolism is discussed. Simple techniques and methods are presented allowing in vivo physiological responses to be compared with changes in the enzyme potential of gut tissue.     

Occurrence of ceramide-glycanase in the earthworm, Lumbricus terrestris

We have detected the presence of ceramide-glycanase in the earthworm, Lumbricus terrestris. We have also devised a simple method for the preparation of this enzyme from the earthworm. This enzyme cleaved the linkage between the ceramide and the glycan chain in LacCer, GbOse3Cer, GbOse4Cer, GbOse5Cer, GM3, GM2, GM1 and GD1a. By using tritium-labeled GM2 as substrate, the optimum pH of this enzyme was found to be between pH 4 and 4.5. In the earthworm, the ceramide-glycanase was mainly found in the muscle. The intestine was found to contain a very low level of this enzyme. Because of their easy availability, earthworms should become a convenient source for the preparation of ceramide-glycanase.     

Subunit distribution of calcium-binding sites in Lumbricus terrestris hemoglobin

The giant, 3.6-MDa hexagonal bilayer hemoglobin (Hb) of Lumbricus terrestris consist of twelve 213-kDa globin subassemblies, each comprised of three disulfide-bonded trimers and three monomer globin chains, tethered to a central scaffolding of 36–42 linkers L1–L4 (24–32 kDa). It is known to contain 50–80 Ca and 2–4 Cu and Zn; the latter are thought to be responsible for the superoxide dismutase activity of the Hb. Total reflection X-ray fluorescence spectrometry was used to determine the Ca, Cu, and Zn contents of the Hb dissociated at pH 2.2, the globin dodecamer subassembly, and linker subunits L2 and L4. Although the dissociated Hb retained 20 Ca²⁺ and all the Cu and Zn, the globin subassembly had 0.4 to 3 Ca²⁺, depending on the method of isolation, and only traces of Cu and Zn. The linkers L2 and L4, isolated by reversed-phase high-pressure liquid chromatography at pH 2.2, had 1 Ca per mole and very little Cu and Zn. Electrospray ionization mass spectrometry of linker L3 at pH 2.2 and at neutral pH demonstrated avid binding of 1 Ca²⁺ and additional weaker binding of 7 Ca²⁺ in the presence of added Ca²⁺. Based on these and previous results which document the heterogeneous nature of the Ca²⁺-binding sites in Lumbricus Hb, we propose three classes of Ca²⁺-binding sites with affinities increasing in the following order: (i) a large number of sites (>100) with affinities lower than EDTA associated with linker L3 and dodecamer subassembly, (ii) 30 sites with affinities higher than EDTA occurring within the cysteine-rich domains of linker L3 and dodecamer subassembly, and (iii) 25 very high affinity sites associated with the linker subunits L1, L2, and L4. It is likely that the low-affinity type (i) sites are the ones involved in the effects of 1–100 mM Group IIA cations on Lumbricus Hb structure and function, namely increased stability of its quaternary structure and increased affinity and cooperativity of its oxygen binding.     

Die Wasserstoffionenkonzentration im Darm von Lumbricus terrestris L.

Zusammenfassung Die Reaktion des Darminhalts normal gehaltener Regenwürmer (stets Lumbricus terrestris L.) wurde mit drei verschiedenen Methoden geprüft. Indikatorstreifchen in den Darminhalt eingesenkt, Chinhydronelektrode und Glaselektrode (bei der Verdünnung 11) ergeben übereinstimmende Werte. Im Zusammenhang mit der schlechten Pufferung des Darminhalts schwanken die Werte bei verschiedenen Tieren nicht unbeträchtlich. Trotz dieser Schwankungen gliedert sich die Reaktion in aufeinanderfolgenden Darmabschnitten bei jedem Tier gesetzmäßig. Auf den alkalischen Pharynx (6,9–7,8; im Mittel 7,3) folgt der schwach saure bis schwach alkalische Drüsen- und Muskelmagen (6,7–7,6; im Mittel 7,0). Der Anfangsabschnitt des Intestinums (18.–40. Segment) ist bei allen untersuchten Tieren der sauerste Abschnitt (6,47–7,23; im Mittel 6,8); im 40.–60. Segment findet man eine mittlere Reaktion von 7,2, im 60.–80. Segment erreicht der Intestinuminhalt den alkalischsten Punkt (7,65–8,05, im Mittel 7,9), um von hier an wieder abzufallen: 7,1 im 80.–100. Segment (6,82–7,30) und 7,3 im 100. bis zum 120. Segment (7,03–7,37).     

Zur neuronalen Regulation des Wasserhaushalts bei Lumbricus terrestris L.

Zusammenfassung Im Dehydrations-Experiment tolerieren Regenwürmer Flüssigkeitsverluste bis zu 70% ihres Körpergewichts. Hierbei ergeben sich hohe Anforderungen an die Anpassungsfähigkeit von Nephridien, Darm und Integument, die im Dienste des Wasser-und Elektrolythaushalts stehen. Um die Abhängigkeit dieser Organe von zentralnervösen Einflüssen zu studieren, wurden bei Lumbricus terrestris Ober-, Unterschlundganglion sowie ein Bauchmarkabschnitt zwischen dem 10.–15. Segment hinter dem Clitellum entfernt. In diesen Segmenten wurden 30, 60 und 120 min nach Versuchsbeginn der Schleimgehalt von Epidermiszellen, die Kerngröße von Darmepithel- und Chloragogen-Zellen sowie der innere und äußere Tubulusumfang von Nephridien cytometrisch bestimmt. In gleichen Intervallen wurde im Oberschlundganglion der Nucleinsäure- und Proteingehalt Gomoripositiver Neurone mikrospektrographisch gemessen. Aus dem Vergleich von cytometrischen und mikrospektrographischen Daten geht hervor: 1. Der nephridiale Stäbchenkanal steht am stärksten unter neuroendokrinem Einfluß. 2. Darmepithel- und Chloragogen-Zellen hängen sowohl von zentraler, neuroendokriner als auch von segmentaler, nervöser Kontrolle ab. 3. Die Schleimsekretion von Epidermiszellen wird wahrscheinlich über segmentale Nerven gesteuert; sie zeigt jedoch die ausgeprägteste Autonomie. Somit stehen die an der Regulation des Wasser- und Elektrolythaushalts beteiligten Organe von Lumbricus terrestris unter einer doppelten Kontrolle durch peptiderge sekretorische Neurone des Oberschlundganglions und segmentale Neurone des Bauchmarks.

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Neuronal regulatory mechanisms of the water balance in Lumbricus terrestris L.

Summary Earthworms tolerate loss of fluids up to 70% of their total body weight. The adaptive capacity must thus be very high in organs which control the exchange of water and electrolytes in nephridia, the intestine and skin. In order to study the influence of the central nervous system on adaption, the supra- and suboesophageal ganglion and the 10th–15th segments behind the clitellum were removed. In segments deprived of ventral nerve cord the mucous content of epidermal cells, the nuclear area of intestinal epithelial cells and chloragogen cells together with the circumference of nephridial tubules were measured 30, 60 and 120 min after placing Lumbricus terrestris in a standardized dry environment. Protein and nucleic acid contents of Gomori-positive neurons in the supraoesophageal ganglion were measured at these times. Comparison of the results of cytometric and microspectrographic measurements shows that 1) the distal segment of the wide nephridial tube is under neuroendocrine control, 2) intestinal epithelial cells and chloragogen cells are influenced by segmental neurons and central secretory nervous elements, 3) mucous secretion is autonomous but may be controlled by segmental nerve impulses. Therefore, water- and electrolyte exchange in Lumbricus terrestris is regulated by secretory peptidergic neurons of the supraoesophageal ganglion and by nerve cell complexes in the ventral nerve cord.

Geschäftsführender Direktor: Prof. Dr. A. Oksche.