Cell proliferation and differentiation kinetics during spermatogenesis inHydra carnea

Summary Spermatogenesis inHydra carnea was investigated. The cell proliferation and differentiation kinetics of intermediates in the spermatogenesis pathway were determined, using quantitative determinations of cell abundance, pulse and continuous labelling with³H-thymidine and nuclear DNA measurements. Testes develop in the ectoderm of male hydra as a result of interstitial cell proliferation. Gonial stem cells and proliferating spermatogonia have cell cycles of 28 h and 22 h, respectively. Stem cells undergo four, five or six cell divisions prior to meiosis which includes a premeiotic S+G2 phase of 20 h followed by a long meiotic prophase (22 h).Spermatid differentiation requires 12–29 h. When they first appear, testes contain only proliferating spermatogonia; meiotic and postmeiotic cells appear after 2 and 3 days, respectively and release of mature sperm begins after 4 days. Mature testes produce about 27,000 sperm per day over a period of 4–6 days: about 220 gonial stem cells per testis are required to support this level of sperm differentiation. Further results indicate that somatic (e.g. nematocyte) differentiation does not occur in testes although it continues normally in ectodermal tissue outside testes. Our results support the hypothesis that spermatogenesis is controlled locally in regions of the ectoderm where testes develop.     

Net aerial primary production (NAPP) of the marsh macrophyteScirpus maritimus estimated by a combination of destructive and non-destructive sampling methods

Net aerial primary production (NAPP) of marsh macrophytes is usually estimated either by destructive sampling techniques or by phenometric techniques. Destructive methods, however, are thought to be inaccurate while phenometric techniques are very labour intensive. In this study a new method is presented which allows an accurate and more efficient estimation of NAPP. The method combines destructive sampling to determine end-of-season biomass and phenometric techniques to estimate the mortality of biomass before the end of the season. NAPP is derived through summation of these two estimates. Techniques needed to calculate the precision of the NAPP estimate are provided. The so called hybrid technique was used to estimate NAPP ofScirpus maritimus L. in a brackish marsh along the Westerschelde estuary, the Netherlands. Estimated NAPP was 1372 g m^-2. End-of-season biomass accounted for 1106 g m^-2, while mortality contributed 266 g m^-2. Precision of the end-of-season biomass and the mortality estimates, expressed as coefficient of variation, was 18.2 and 26.0% respectively. The precision of the resultant, NAPP, was higher: 17.2%. These results indicate that NAPP could be estimated with a higher precision than end-of-season biomass. This contradicts the view that the accuracy of NAPP estimates can only be improved at the expense of its precision.     

The response of salt marsh vegetation to tidal reduction caused by the Oosterschelde storm-surge barrier

In 1986 a sluice gate barrier was completed in the mouth of the Oosterschelde estuary. The barrier has been partially or completely closed during 1986 and the first months of 1987. Consequently the high tides were reduced to such a level that the salt marshes were scarcely flooded. Since April 1987 the barrier has been closed on average during two out of 706 high tides a year. Although the barrier allows tidal exchange the tidal flow has been restricted as a result of the reduced width of the mouth of the estuary. This restriction of the tidal flow caused a 26 cm decrease of mean high water in the eastern part of the estuary. As a result the inundation frequencies of the salt marshes decreased.The response of salt-marsh vegetation to this tidal reduction was analyzed using annual records (1982 till 1990) of species composition in 57 permanent plots in two marshes at the southern shore of the estuary. Analysis of the response of individual species to marsh elevation in the pre and the post-barrier situation revealed that most species moved down the marsh elevation gradient. The first axis of an ordination (DCA-1) was significantly negatively related to inundation frequency. Between 1984 and 1990 all plots were displaced towards a significantly higher ordination score, indicating a trend towards a species composition from higher up the marsh. The position of most plots along DCA-1 remained stable until 1985 and started to increase in 1986 or 1987. The vegetation in plots dominated either by Halimione portulacoides or by Spartina anglica, started to change in 1985. This premature change was attributed to frost damage in January 1985. The initially high rate of change along DCA-1 decreased in 1989 and 1990. This would suggest that the vegetation re-equilibrated with the newly established tidal conditions. Further analysis revealed no significant difference in the relation between inundation frequency and sample score along DCA-1 between 1984 and 1990. This corroborates the view that species composition had re-equilibrated with the tidal conditions. Along DCA-2 the samples were displaced towards a significantly higher score as well. The change was attributed to an increase of the perennial Halimione and the annual Suaeda maritima. The annual Atriplex hastata displayed an increase from 1986 till 1988, but strongly declined in 1990. The transient response of this annual was described by DCA-3.During the pre-barrier phase several attempts had been made to predict the vegetation response to tidal reduction. We developed a multivariate model (CCA) with inundation frequency as a constraining factor to describe the relation between inundation frequency and species composition in the pre-barrier phase in 1984. Next we used this model to predict the score of the samples along CCA-1 in the post-barrier phase from the inundation frequencies in 1990. The actual ordination score in 1990 was calculated from the observed species composition. The predicted ordination scores did not differ significantly from the observed ones. In retrospect it is concluded that the response of the vegetation to the reduction of the tides, as far as the fraction of species composition which is related to inundation frequency is concerned, could have been predicted by our model.

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Biosynthesis of the sesquiterpene patchoulol from farnesyl pyrophosphate in leaf extracts of Pogostemon cablin (patchouli): mechanistic considerations

Several mechanistic alternatives have been proposed for the enzyme-catalyzed, electrophilic cyclization of farnesyl pyrophosphate to the tricyclic sesquiterpene alcohol patchoulol, which is the characteristic component of the essential oil of Pogostemon cablin (patchouli). These alternatives include schemes involving deprotonation-reprotonation steps and the intermediacy of the monocyclic and bicyclic olefins germacrene and bulnesene, respectively, and involving a 1,3-hydride shift with only tertiary cationic intermediates and without any deprotonation-reprotonation steps. Analytical studies, based on analyses of P. cablin leaf oil at different stages of plant development, and in vivo time-course investigations, using 14CO2 and [14C]sucrose, gave no indication that germacrene and bulnesene were intermediates in patchoulol biosynthesis. A soluble enzyme system from P. cablin leaves was prepared, which was capable of converting farnesyl pyrophosphate to patchoulol, and isotopic dilution experiments with both labeled and unlabeled olefins were carried out with this system to confirm that sesquiterpene olefins did not participate as fre intermediates in the transformation of the acyclic precursor to patchoulol. Patchoulol derived biosynthetically from [12,13-14C;1-3H]farnesyl pyrophosphate was chemically degraded to establish the overall construction pattern of the product. Similar studies with [12,13-14C;6-3H]farnesyl pyrophosphate as a precursor eliminated deprotonation steps to form bound olefinic intermediates in the biosynthesis of patchoulol, while providing supporting evidence for the hydride shift mechanism.     

Characteristics of rat jejunal transport of tryptophan.

The parameters of rat jejunal transport of tryptophan have been examined. The interactions between tryptophan and lysine or methionine have been reexamined, and some aspects of the trans effects of cellularly accumulates amino acids have been studied. It has been demonstrated that: (1) The influx of tryptophan across the jejunal brush border (Jmc-Trp) can be accounted for by the carrier of alpha-aminomonocarboxylic acids alone. (2) Tryptophan competes with lysine for the carrier of basic amino acids across the brush border membrane without itself being transported by this carrier. (3) Lysine has neither cis nor trans effects on Jmc-Trp, whereas intracellular tryptophan is highly inhibitory to Jmd-Lys. (4) The intracellular concentration of lysine and of tryptophan, [Lys]c and [Trp]c, are unaffected by tryptophan and lysine, respectively, although the transmural fluxes, from the mucosal side to the serosal side, Jms, of lysine, Jms-Lys, and of tryptophan, Jms-Trp, are inhibited by tryptophan and lysine, respectively. The latter effects thus represent inhibitory interactions at the basolateral membrane. (5) Methionine is a potent cis and transinhibitor of Jmc-Trp, but stimulated Jms-Trp and reduces [Trp]c. (6) Methionine causes trans acceleration of the influx of lysine across the brush border membrane, Jmc-Lys, but has no effect on the influx of galactose, Jmc-Gal. (7) Leucine causes trans inhibition of Jmc-Leu. (8) Tryptophan does not cause cis inhibition of Jmc-Gal, but is a strongtransinhibitor of Jmc-Gal. (9) Cellularly accumulated tryptophan appears to accelerate the eventual decline in transepithelial potential difference and short-circuit current. These results are consistent with the conclusions that: (1) Tryptophan is transported across the brush border membrane by the carrier of neutral amino acids alone, but leaves the cell across the basolateral membrane by a mechanism used by lysine also. (2) Leucine, methionine and probably tryptophan have a transeffect on the transport of neutral amino acids across the brush border membrane which may represent a phenomenon which can appropriately be termed decelerating exchange diffusion. (3) Cellularly accumulated tryptophan has a strong and indiscriminate depressive effect on all transport functions of rat jejunal epithelium.