On the role of cell surface associated,mucin-like glycoproteins in the pennate diatom Craspedostauros australis (Bacillariophyceae)

Diatoms are single-celled microalgae with silica-based cell walls (frustules) that are abundantly present in aquatic habitats, and form the basis of the food chain in many ecosystems. Many benthic diatoms have the remarkable ability to glide on all natural or man-made underwater surfaces using a carbohydrate- and protein-based adhesive to generate traction. Previously, three glycoproteins, termed FACs (F rustule A ssociated C omponents), have been identified from the common fouling diatom Craspedostauros australis and were implicated in surface adhesion through inhibition studies with a glycan-specific antibody. The polypeptide sequences of FACs remained unknown, and it was unresolved whether the FAC glycoproteins are indeed involved in adhesion, or whether this is achieved by different components sharing the same glycan epitope with FACs. Here we have determined the polypeptide sequences of FACs using peptide mapping by LC–MS/MS. Unexpectedly, FACs share the same polypeptide backbone (termed CaFAP1), which has a domain structure of alternating Cys-rich and Pro-Thr/Ser-rich regions reminiscent of the gel-forming mucins. By developing a genetic transformation system for C. australis, we were able to directly investigate the function of CaFAP1-based glycoproteins in vivo. GFP-tagging of CaFAP1 revealed that it constitutes a coat around all parts of the frustule and is not an integral component of the adhesive. CaFAP1-GFP producing transformants exhibited the same properties as wild type cells regarding surface adhesion and motility speed. Our results demonstrate that FAC glycoproteins are not involved in adhesion and motility, but might rather act as a lubricant to prevent fouling of the diatom surface.     

Das Belichtungspotential der isolierten Retina des Einsiedlerkrebses (Eupagurus bernhardus L.) in Abhängigkeit von den extrazellulären Ionenkonzentrationen

Summary 1.The action potential of the isolated retina of the hermit crab Eupagurus bernhardus L. resulting from exposure to light has been measured with external electrodes under constant stimulus conditions.Three measurements of the retinal action potentials (RAP's) were taken to observe the changes of the RAP's quantitatively: a) The amplitude h _max of the maximum; b) the amplitude h _e of the plateau, measured at the end of the stimulus, for taking the shape quotient h _max /h _e ; c) the peak-amplitude-time t _max. 2.The RAP's of the retina in a standard physiological saline are compared with those of the retina in salines of different ionic composition while osmotic pressure and p _h were kept constant. 3.Increasing K⁺-concentration reduces the amplitude h _max of the RAP's gradually, which is zero at 500 mM K⁺/l. The peak-time t _max decreases with increasing K⁺-concentration up to 50 mM K⁺/l, whereas at higher concentrations it increases. In contrast to this fact h _max/h _e increases up to 50 mM K⁺/l and decreases above this concentration (Fig. 3). 4.Increasing Ca⁺⁺-concentration reduces h _max (zero above 350 mM Ca⁺⁺/l) and t _max. h _max/h _e rises up to a Ca⁺⁺-concentration of about 30 mM Ca⁺⁺/l; whereas at a higher Ca⁺⁺-concentration it decreases again (Fig. 6). When the Ca⁺⁺-concentration is very low the fall of the RAP is much slowed down and the plateau h _e extremly rises. In a Ca⁺⁺-free saline which contained 1 mM/l Ethylendiamintetraacetic acid (EDTA) the retina lost its irritability reversibly (Fig. 8). 5.The amplitude of the RAP's is augmented with increasing Mg⁺⁺-concentration up to 10–30 mM Mg⁺⁺/l and decreases above this concentration (Fig. 11). When the saline contains virtually no other cations but Mg⁺⁺ (367 mM/l) the amplitude of the RAP is small (20%) but not zero. 6.In a buffered isotonic NaCl-solution as well as in a saline in which all the Cl^--ions are substituted by SO₄ ^---ions the amplitude of the RAP's is higher but the shape of the RAP's is changed in the same way as in other solutions with very low concentrations of Ca⁺⁺. 7.All the changes of the RAP's described so far are reversible. 8.Even when the retina is kept in a salt solution containing sodium in a very low concentration (ca. 3–5 mM/l, the sodium substituted by choline⁺-ions) for 5 hours the amplitude h _max of the RAP does not change significantly but the shape: the peaktime t _max is longer, h _max/ h _e is much greater. Afterwards when the retina is brought into standard saline again, the amplitude h _max increases, t _max remains almost unchanged and h _max/h _e , decreases strongly (Fig. 17). 9.Substitution of all the Na⁺-, Ca⁺⁺- and Mg⁺⁺-ions by choline⁺-ions results in a decrease of the amplitude h _max, a lengthening of t _max and a small increase of h _max/h _e . 10.Substitution of all the NaCl by glucose decreases the amplitude h _max, lengthens t _max very much and decreases the value of h _max/h _e but little. Afterwards, when the retina is brought into standard saline again the effect of the glucose solution on the amplitude h _max is only little reversible: h _max increases very little, t _max decreases strongly and h _max/h _e increases (Fig. 21). 11.With increasing external K⁺-concentration the resting potential decreases. The changes of the resting potential cause the changes in the shape of the RAP's. 12.The presence of a small concentration of Ca⁺⁺-ions outside of the cell membrane is obviously necessary for the ability of the cell membrane of the photoreceptor to increase its ionic permeability consequent to stimulation by light. Above a Ca⁺⁺-concentration of about 1 mM/l the raise of permeability of the cell membrane during illumination is smaller with increasing Ca⁺⁺-concentration. The velocity of the changes in permeability is augmented, especially of those changes concerned with the fall of the RAP. The effect of Mg⁺⁺-ions is somewhat similar to that of Ca⁺⁺-ions, but much weaker. 13.The changes of the RAP are mainly determined by the low Ca⁺⁺-content when the retina stays in the NaCl- or sulfate saline. 14.Choline⁺-ions probably greatly increase the raise of permeability of the cell membrane for the divalent cations Ca⁺⁺ and Mg⁺⁺ during excitation. 15.It is suggested that under normal conditions in Eupagurus the amplitude of the RAP is determined primarily by the Na⁺-concentration gradient over the receptor cell membrane. But also the divalent cations Ca⁺⁺ and Mg⁺⁺ contribute to the amplitude of the RAP, especially after the retina has been treated with choline chloride.

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Herrn Professor Dr. C. Kosswig zum 60. Geburtstag überreicht.Der Deutschen Forschungsgemeinschaft danke ich für großzügige Unterstützung, Herrn Peter Thomsen und meiner Frau für technische Hilfe.     

Autosomal control of lampbrush-loop formation during spermatogenesis in Drosophila hydei by a gene also affecting somatic sex determination

Summary The Y chromosome of Drosophila hydei carries information that is necessary for the development of the spermatozoa. In primary spermatocytes Y chromosomal genes become active: five of the male fertility factors form giant lampbrush loops. Our prior work indicated interactions between the Y chromosomal genes and autosomal loci. It is of interest to identify loci regulating the activity of the Y chromosomal genes. We, therefore, screened a total of about 14,000 chromosomes (X, 2, 3 and 4) for mutations that interfere with the expression of the lampbrush loops. Two mutations with substantial effects on the loop morphology were recovered. One of them, a recessive male sterile mutation (ms (3) 5) on chromosome 3, is described in this paper. Its homozygous state results in a complete absence of all Y chromosomal lampbrush loops at 26° C; at 18° C the loops are formed. Temperature shifts with homozygous males indicate that the function early during the spermatogonial stage is crucial for the development of lampbrush loops in the primary spermatocyte. Meiosis is entirely absent in the male, but normal in females. Females homozygous for ms (3) 5 display a maternal effect, which reduces the viability and fertility of homozygous daughters and produces sons with signs of intersexuality. Linkage studies indicated that the effect on the male germ line and the maternal effects cannot be separated and may hence be induced by a single gene.