Territorial Pheromones of Female Red-backed Salamanders

Pheromones provide an important source of communication during social interactions of caudate amphibians. To further examine their use in territorial defense, we performed a laboratory experiment to test the hypothesis that non-courting female red-backed salamanders (Plethodon cinereus) deposit pheromones in or on fecal pellets, as males are known to do during territorial advertisement. Four conditions were tested: (1) a burrow marked with a female's own pellet vs. a burrow marked by a conspecific female's pellet, (2) own vs. unmarked burrows, (3) conspecific vs. unmarked burrows, and (4) paired unmarked burrows as a control. Females nose-tapped (for olfactory cues) their own and conspecific pellets about equally. However, they spent significantly more time in both threat and submissive behavior toward the conspecific pellets and spent significantly more time in their own marked burrows. We infer that female P. cinereus do deposit pheromones in or on fecal pellets and that these pellets may be used to advertise territories. The behavioral responses of females toward pellets of other females were more aggressive than those of males (in a previous study) toward pellets of other males.     

Inhibition of TRAP-induced angiogenesis by the tripeptide Phe-Pro-Arg, a thrombin-receptor-derived peptide analogue

Summary We have recently shown that thrombin promotes angiogenesis by a mechanism independent of fibrin formation. In the present paper, we investigated the effect of the thrombin-receptor-activating tetradecapeptide (TRAP_1–14, S₄₂FLLRNPNDKYEPF₅₅) for its effects on angiogenesis in the chick chorioallantoic membrane (CAM) system of angiogenesis. A dose-dependent promotion of angiogenesis is evident with TRAP. In contrast, a thrombin-receptor-derived tripeptide analogue H-Phe-Pro-Arg-OH (FPR), which was designed based on the S₄₂FLLR₄₆ sequence, caused an inhibition of angiogenesis in the CAM, and when it was combined with TRAP it caused a complete reversal of the angiogenesis-promoting effect of TRAP. These results indicate that the proteolytic exposure of the receptor N-terminal tetradecapeptide by thrombin can activate the post-thrombotic events related to angiogenesis. These events can be modulated by constrained peptide analogues such as FPR.     

CD44 standard and variant isoform expression in normal human skin appendages and epidermis

 CD44 isoforms have been implicated in tumor progression and metastasis formation. This study presents a thorough immunohistochemical analysis of CD44 standard and isoform expression in normal human skin appendages and epidermis applying monoclonal antibodies against CD44s, CD44v3, -v4, -v5, -v6, and -v9. An improved immunohistochemical protocol with microwave-based antigen retrieval in paraffin sections and heavy metal amplification of the diaminobenzidine reaction product provided enhanced resolution and sensitivity as compared to studies on frozen sections. The hair follicle, the seborrheic and eccrine sweat glands were strongly positive for all CD44 isoforms studied. In the latter, the clear cells but not the dark (intercalated) cells were positive. The sudoriferous ducts adjacent to the glands were weakly positive for all CD44 isoforms and strongly positive near the skin surface. In the apocrine glands, the basal cells showed only a moderate positivity. The myoepithelial cells expressed only CD44s. In the epidermis, all CD44 isoforms were detectable, with strongest CD44 immunostaining in the lower third of the stratum spinosum and weaker staining in the stratum basale and the upper two-thirds of the stratum granulosum. The stratum granulosum and corneum were unreactive. Thus, a regional and cell type-specific CD44 expression was revealed. Accepted: 10 May 1996     

Localization and Characterization of a Novel 20 kDa Polypeptide in the Chloroplast of the Green Alga Dunaliella salina

Recent work with the green alga Dunaliella salina showed thepresence of a {small tilde}20 kDa chloroplast protein that wasrecognized by polyclonal antibodies raised against the isolatedLHC-II [Webb M.R. and Melis A. (1995) Plant Physiol. 107: 885].In this report, a characterization of the {small tilde}20 kDapolypeptide is presented. It is shown that it is localized inthe chloroplast envelope membrane of D. salina. The abundanceof this protein is constant on a per cell basis and independentof the light regime during cell growth. The {small tilde}20kDa polypeptide is easily degraded to a {small tilde}19 kDaproduct during sample preparation. A limited amino acid sequenceof 21 residues from the free N-terminus of the {small tilde}19kDa product was obtained. On the basis of this partial sequence,it was concluded that the {small tilde}20 kDa polypeptide isnot a degradation product of a known LHC-II but rather a novelprotein. The {small tilde}20kDa polypeptide did not cross-reactwith antibodies raised against the Cbr (carotene biosynthesis-related)gene product and showed a different electrophoretic mobilityfrom the latter. Light-shift experiments suggest that the {smalltilde}20 kDa polypeptide is not an ELIP (early light-inducibleprotein). Possible functions of the {small tilde}20 kDa proteinare discussed. ¹Permanent address: Department of Biochemistry, University oflund, PO Box 124, S-221 00 Lund, Sweden     

Effects of moderate drought on ascorbate peroxidase and glutathione reductase activities in mesophyll and bundle sheath cells of maize

Mesophyll and bundle sheath cells of maize leaves ( Zea mays L.) both contain the enzymes ascorbate peroxidase (AP; EC 1.11.1.11) and glutathione reductase (GR; EC 1.6.4.2) which are involved in hydrogen peroxide detoxification. Since bundle sheath cells of maize are deficient in photosystem II and have high CO₂ levels, oxidative stress may be less severe in these cells than in mesophyll cells. The present study was conducted to determine if AP and GR activity levels preferentially increase in mesophyll cells relative to bundle sheath cells when plants are subjected to moderate drought. Although drought inhibited the growth of greenhouse-grown plants, it did not affect the levels of protein, chlorophyll or AP. GR was unaffected by drought in whole leaf tissue and mesophyll cells, but did increase slightly in bundle sheath cells. This slight increase is of questionable biological importance. AP and GR activity levels were similar in mesophyll cells, bundle sheath cells and in whole leaf tissue. The data suggest that moderate drought has little effect on enzymes of the hydrogen peroxide scavenging system and that mesophyll and bundle sheath cells may be exposed to similar levels of hydrogen peroxide.     

Molecular characterization and expression of a tobacco histone H1 cDNA

We have isolated a 1104 bp tobacco cDNA clone (H1c12) which includes an 846 bp open reading frame. This encodes a polypeptide of 282 amino acid residues and represents the largest plant H1 histone identified so far. The structure of the deduced protein shows the classical tripartite organization of the H1-type linker histones. The expression of the tobacco H1 histone gene(s) corresponding to the H1c12 cDNA clone was examined during different developmental stages. We found that, at the level of steadystate mRNA, expression of gene(s) encoding this H1 histone was rapidly induced in germinating seeds. The H1 gene was expressed in all tissues examined. However, its expression was higher in tissues known to contain meristematic cells. Furthermore, in the leaves of mature plants accumulation of the H1 mRNA exhibits a very characteristic oscillation. This latter finding indicates that, at least in fully developed plants, the expression of this type of H1 histone gene(s) is modulated by a diurnal cycle.     

Distinct response of Medicago suspension cultures and roots to Nod factors and chitin oligomers in the elicitation of defense-related responses

The induction of plant defense-related responses by chitin oligomers and the Rhizobium meliloti lipo-chito-oligosaccharide nodulation signals (Nod factors) in Medicago cell cultures and roots was investigated by following the expression of genes encoding enzymes of the isoflavonoid biosynthetic pathway, such as chalcone synthase, chalcone reductase, isoflavone reductase, as well as genes encoding a pathogenesis-related protein and a peroxidase. In suspension-cultured cells, all genes except the peroxidase gene were induced by both the R. meliloti Nod factor NodRm-IV(C16:2,S) and chitin oligomers with a minimum of three sugar residues. However, activation of these genes was not elicited by the symbiotically inactive, desulfated NodRm-IV(C16:2). Moreover, the cells were more sensitive to the chitin oligosaccharides than to the Nod factor. Analysis of flavonoids in Medicago microcallus cultures revealed differences between cells treated with N -acetyl-chitotetraose and those treated with Nod factor and demonstrated increased production of the phytoalexin medicarpin in the presence of Nod factor. In Medicago roots, none of the tested genes was activated by the N -acetylchitotetraose, whereas the Nod factor at micro-molar concentration enhanced transient expression of the isoflavonoid biosynthetic genes. The differential responses to Nod factors and chitin oligomers suggest that Medicago cells possess distinct perception systems for these related molecules.