Parameters that specify the timing of cytokinesis.

One model for the timing of cytokinesis is based on findings that p34(cdc2) can phosphorylate myosin regulatory light chain (LC20) on inhibitory sites (serines 1 and 2) in vitro (Satterwhite, L.L., M.H. Lohka, K.L. Wilson, T.Y. Scherson, L.J. Cisek, J.L. Corden, and T.D. Pollard. 1992. J. Cell Biol. 118:595-605), and this inhibition is proposed to delay cytokinesis until p34(cdc2) activity falls at anaphase. We have characterized previously several kinase activities associated with the isolated cortical cytoskeleton of dividing sea urchin embryos (Walker, G.R., C.B. Shuster, and D.R. Burgess. 1997. J. Cell Sci. 110:1373-1386). Among these kinases and substrates is p34(cdc2) and LC20. In comparison with whole cell activity, cortical H1 kinase activity is delayed, with maximum levels in cortices prepared from late anaphase/telophase embryos. To determine whether cortical-associated p34(cdc2) influences cortical myosin II activity during cytokinesis, we labeled eggs in vivo with [(32)P]orthophosphate, prepared cortices, and mapped LC20 phosphorylation through the first cell division. We found no evidence of serine 1,2 phosphorylation at any time during mitosis on LC20 from cortically associated myosin. Instead, we observed a sharp rise in serine 19 phosphorylation during anaphase and telophase, consistent with an activating phosphorylation by myosin light chain kinase. However, serine 1,2 phosphorylation was detected on light chains from detergent-soluble myosin II. Furthermore, cells arrested in mitosis by microinjection of nondegradable cyclin B could be induced to form cleavage furrows if the spindle poles were physically placed in close proximity to the cortex. These results suggest that factors independent of myosin II inactivation, such as the delivery of the cleavage stimulus to the cortex, determine the timing of cytokinesis.     

Root system architecture and gravitropism in the oil palm.

The oil palm (Elaeis guineensis Jacq.) has a root system consisting of primary (or order 1) roots, which are either orthogravitropic (R1 VD, with positive gravitropism) or diagravitropic (R1 H). Their statenchyma have very similar characteristics (mainly vacuolated, large cells). However, their statoliths sediment along the longitudinal wall in R1 H and along the distal wall in R1 VD (furthest cell wall from the apical meristem, opposite the proximal wall). Order 2 roots may have vertical upward (R2 VU) or downward growth (R2 VD) or even horizontal growth (R2 H). In all cases, the statoliths are located near the lower wall of the statocyte (distal in R2 VD, proximal in R2 VU and longitudinal in R2 H). Order 3 roots are usually agravitropic. When they grow upwards, R3 VU, their amyloplasts are located near the proximal wall. Likewise, the growth direction of R4 varies, but they have little or no statolith sedimentation. Roots with marked gravitropism (positive or negative) have amyloplasts that can sediment along different walls. But, irrespective of amyloplast position in the statocytes, the direction of root growth may be stable. The relation between the different reactions of roots and different sensitivity to auxin or to a curvature-halting signal is discussed.     

A recessive mutation leading to vertebral ankylosis in zebrafish is associated with amino acid alterations in the homologue of the human membrane-associated guanylate kinase DLG3.

We describe the characterization of the zebrafish homologue of the human gene DLG3. The zebrafish dlg3 gene encodes a membrane-associated guanylate kinase containing a single PDZ domain. This gene was cloned using a gene-trap construct inserted in the gene's first intron. The insertion co-segregates with a viable mutation called humpback (hmp), which leads to formation of ankylotic vertebrae in adult fishes. Insertion and mutation have both been mapped to chromosome 12, in a segment which is syntenic with region p12 to q12 of human chromosome 17. The hmp mutant phenotype, however, appears to be due to two point mutations in the guanylate kinase domain rather than to the transgene insertion itself. The results of this study are discussed in the light of the possible function of the guanylate kinase domain.     

PHYLOGENETIC ANALYSIS OF THE MALE ONTOGENETIC PROGRAM IN AQUATIC AND TERRESTRIAL MONOCOTYLEDONS

The male program of ontogeny in flowering plants encompasses the events from meiosis of microsporocytes to fertilization. Three main sequences are discussed; the deposition of cell walls, changes in cytoplasmic organelles, and the program of nuclear divisions leading to the formation of two sperm cells and a vegetative cell in each pollen grain. Variations in these ontogenetic sequences are particularly apparent in the monocotyledons, which exhibit diversity in pollen morphology, wall structure, and mode of pollination. The male program of development has been compared in selected terrestrial monocotyledons belonging to the Liliaceae and Gramineae and aquatic members of the Cymodoceaceae, Najadaceae, and Zannichelliaceae. A total of 26 characters from the male program are discussed and then used to construct a cladogram derived only from developmental data for the five species. The polarity of only a few of the character transformations has been determined directly by observation of developmental sequences; most have been interpreted by outgroup analysis.     

The development of anoxia following occlusion

Following arteriolar occlusion, tissue oxygen concentration decreases and anoxic tissue eventually develops. Although anoxia first appears in the region most distal to the capillary at the venous end, it eventually spreads throughout the entire region of supply. In this paper the changing oxygen concentration, from the time of occlusion until the tissue is entirely anoxic, is examined mathematically. The equations governing oxygen transport to tissue are solved by iterating a nonlinear integral equation. This solution is valid until anoxia first appears. After anoxia develops it is necessary to solve a moving boundary problem. This is done using the method of matched asymptotic expansions, and accurate solutions are obtained for a wide range of physiological conditions.     

Metabolism of carbon tetrachloride to phosgene

Aerobic incubation of hepatic microsomal fractions in the presence of carbon tetrachloride, NADPH and cysteine resulted in the formation of phosgene which was identified by gas chromatography/mass spectrometry as the adduct, 2-oxothiazolidine-4-carboxylic acid, formed by its reaction with cysteine. [¹³C]-Carbon tetrachloride was metabolized to 2-[¹³C]-oxothiazolidine-4-carboxylic acid the , when carbon tetrachloride was incubated in the presence of [¹⁸O]-O₂, 2- [¹⁸O]-oxothiazolidine-4-carboxylic acid was formed. The reaction was inhibited by carbon monoxide showing the involvement of the cytochrome P-450-dependent mixed function oxidase system. The metabolism of carbon tetrachloride to phosgene may play a role in the production of hepatotoxicity by this compound.