CGRP immunoreactivity and NADPH-diaphorase in afferent nerves of the rat penis

Calcitonin gene-related peptide (CGRP)-immunoreactive afferent nerve fibers are abundant in the rat penis. In addition, NADPH-diaphorase, which stains for nitric oxide synthase, has been localized within both autonomic and sensory dorsal root ganglia (DRG) and may be part of an important biochemical pathway involved in penile tumescence. The purpose of this study was: 1) to examine the circuitry of afferent nerves that are CGRP immunoreactive from the L6 DRG, 2) to examine the possibility that there are NADPH-diaphorase-positive afferent fibers from the L6 DRG to the rat penis, and 3) to examine the localization and colocalization of CGRP and NADPH-diaphorase within L6 DRG afferent perikarya. Calcitonin gene-related peptide immunostaining in the penis was eliminated following a bilateral transection of the pudendal nerves, but was unchanged following a bilateral transection of the pelvic splanchnic or hypogastric nerves. The NADPH-diaphorase staining was not altered by any of the nerve transections. Injection of the retrograde axonal tracer fluorogold (FG) into the dorsum penis labeled perikarya in the L6 DRG. Although the majority of FG-labeled perikarya contained neither CGRP nor NADPH-diaphorase, small subpopulations of perikarya contained either CGRP immunoreactivity, NADPH-diaphorase, or both. A unilateral pudendal nerve transection virtually eliminated (>99%) FG labeling in the ipsilateral L6 DRG. These data suggest that NADPH-diaphorase and CGRP are present, either together or separately, within a subpopulation of penile afferent perikarya. In addition, CGRP-immunoreactive afferent nerve fibers reach the penis primarily via the pudendal nerves. Finally, NADPH-diaphorase-positive penile afferents may be another important source of nitric oxide (NO) for penile tumescence.     

Huge increase in bacterivores on freshly killed barley roots

Abstract Adding fresh roots to intact soil cores resulted in marked increases in microbial and microfaunal activity at the resource islands. Microbial activity increased in two phases following root addition. Respiratory activity and concentration of respiratory enzyme (dehydrogenase) in soil adhering to the roots was very high during the first three weeks resulting in anaerobic conditions in the soil. After a period of low respiratory activity and enzyme content, these quantities increased from 6 to 20 weeks, but not enough to maintain anaerobic conditions. Numbers of protozoa peaked earlier than the nematodes. Based on yield coefficients of microbes and bacterivores, the increase in bacterivores was in accordance with root-induced respiration activity. In soil adhering to roots, numbers of bacterial grazers (protozoa and nematodes) were up to 80 and 30 times higher, respectively, than in the surrounding soil. This effect is up to 20 times higher than observed around live root systems, which may suggest that the rhizosphere effect on microbivores could for the major part result from the decomposition of dead segments of the root system.     

Synthesis of the biologically active β-subunit of human nerve growth factor in Escherichia coli

The gene(NGFB) encoding the β subunit of mature human nerve growth factor (hNGFB) was subcloned into the pJLA503 expression vector under the control of bacteriophage promoters p_R and p_L, and expressed in Escherichia coli. The recombinant protein represented approximately 3% of the total cellular protein. Biologically active hNGFB was solubilized (0.2% total NGFB) and purified by cation-exchange chromatography and it yielded two bands on polyacrylamide-gel electrophoresis under nonreducing conditions, corresponding to the monomeric (14 kDa) and homodimeric (26.5 kDa) forms of the molecule. Both hNGFB forms were immunopositive on Western blots with rabbit anti-NGFB antibodies; however, following additional purification, only the species corresponding to the hNGFB homodimer was biologically active on cultured chicken dorsal root ganglion neurons. These results demonstrate the feasibility of synthesizing the biologically active form of hNGFB in E. coli.     

Structure of the Bombyx mori fibroin light-chain-encoding gene: upstream sequence elements common to the light and heavy chain.

Two overlapping genomic clones containing the fibroin light-chain (Fib-L)-encoding gene (Fib-L) were obtained from the cosmid library of the silkworm, Bombyx mori J-139, by hybridization with the Fib-L cDNA clone. Sequencing of the 14.6-kb region revealed that Fib-L was 13472 bp long containing seven exons, and that the gene contained a large first intron which occupied about 60% of the gene. Comparison of restriction patterns of the J-139 Fib-L with those of eight other B. mori breeds producing normal-level fibroin demonstrated that considerable restriction-fragment length polymorphisms were present in regions containing the first intron and the 3′-flanking sequence. However, sizes of the Fib-L mRNA and the Fib-L polypeptide were very similar among the nine breeds tested, suggesting that the exon sequences and the splice signals were all well conserved. 5′-Flanking regions of Fib-L and the fibroin heavy-chain (Fib-H)-encoding gene (Fib-H) compared in this study contained three 18-30-bp sequences of high similarity and many 8-10-bp common elements, six of which coincided with the binding sites of homeodomain proteins. Gel retardation assays with the nuclear extracts of the posterior and middle silk glands suggested that protein factors present in the posterior silk-gland nuclei could bind to a set of those common upstream elements.     

Ecophysiology of Aufwuchs-eating cichlids in Lake Tanganyika: niche separation by trophic specialization

Synopsis The Aufwuchs-eating cichlids of Lake Tanganyika show clear trophic differences that are correlated to their morphology, physiology and foraging behaviour. The species are grouped into three categories of relative intestinal length according to their feeding habits. A correlation between the intestinal length and the diet could be demonstrated, ranging from around 2.5 for species ingesting more animal food, to 7.8 for detritivorous and microalgivorous species. The relative intestinal length of domesticTropheus moorii, raised in aquaria was significantly lower than that of wild individuals by a factor of 1.7, demonstrating a wide range of phenotypic adaptability. The activities of trypsin and amylase were at an equal level in four Aufwuchseating species, but the activity of laminarinase of a detritivorous-microalgivorous species (Petrochromis orthognathus) was 2.6 times higher than that of an algivorous species (Tropheus moorii). The laminarinase seems to be an excellent marker enzyme for detritivorous or microalgivorous feeding.     

The problem of the acts duration measurement in relation to the “focal-animal” technique of behavioral recording

The focal-animal technique assumes the continuous recording of the behavior of an individual during a certain time interval. The length of this interval (sampling unit) can be problematic when estimating the duration of behavioral acts. Two acts from the behavioral repertoire of the ant Leptothorax fuenteiwere focused on in this work at different ranges of temporal scales. Analyzing these acts we show the possibility of existence of a sampling artifact, in such a way that the estimates of the durations of the acts would be forced to depend upon the length of the sampling unit that is being used.     

Genetic and physical mapping of the patatin genes in potato and tomato

Summary Genes for the major storage protein of potato, patatin, have been mapped genetically and physically in both the potato and tomato genomes. In potato, all patatin genes detected by the cDNA clone pGM01 map to a single locus at the end of the long arm of chromosome 8. By means of pulsed field gel electrophoresis (PFGE) it was possible further to delimit this locus, containing 10–15 copies of the gene, to a maximum size of 1.4 million base pairs. Hybridizations with class-specific clones suggest that the locus is at least partially divided into domains containing the two major types of patatin genes, class I and II. In tomato, patatin-homologous sequences were found to reside at the orthologous locus at the end of chromosome 8. The approximately three copies in tomato were localized by PFGE to a single fragment of 300 kilobases. Whereas the class II-specific 5 promoter sequences reside in tomato at the same locus as the coding sequences, the single class I-specific copy of the 5 promoter sequences was localized on chromosome 3 with no coding sequence attached to it. A clone from this chromosome 3 locus of tomato was isolated and by restriction fragment length polymorphism mapping it could be further shown that a similar class I-specific sequence also exists on chromosome 3 of potato. As in tomato, this copy on chromosome 3 is not linked to a coding sequence for patatin. The results are discussed with respect to genome evolution and PFGE analysis of complex gene families.     

Gene expression in nematode-infected plant roots

Summary A major pathogen of potato plants (Solanum tuberosum) is the potato cyst nematode (Globodera spp.), which induces localized redifferentiation of a limited number of host cells to form a specialized feeding-site termed the syncytium. A novel strategy utilizing the polymerase chain reaction (PCR) was employed to construct a cDNA library from dissected potato roots highly enriched in syncytial material. The library was differentially screened with cDNA probes derived from the infected root tissue from a compatible interaction and from healthy root tissue. Characterization of one gene identified by the library screen indicated an expression pattern that correlated with events in the immediate vicinity of the pathogen after syncytial establishment. The strategy for library construction and screening could be applicable to the study of gene expression in any plant-pathogen interaction in which the limited supply of cells at the interface of the two organisms precludes a more traditional approach.     

Root hydrotropism of an agravitropic pea mutant, ageotropum

We have partially characterized root hydrotropism of an agravitropic pea mutant, ageotropum (from Pisum sativum L. cv. Weibull's Weitor), without interference of gravitropism. Lowering the atmospheric air humidity inhibited root elongation and caused root curvature toward the moisture-saturated substrate in ageotropum pea. Removal of root tips approximately 1.5 mm in length blocked the hydrotropic response. A computer-assisted image analysis showed that the hydrotropic curvature in the roots of ageotropum pea was chiefly due to a greater inhibition of elongation on the humid side than the dry side of the roots. Similarly, gravitropic curvature of Alaska pea roots resulted from inhibition of elongation on the lower side of the horizontally placed roots, while the upper side of the roots maintained a normal growth rate. Gravitropic bending of Alaska pea roots was apparent 30 min after stimulation, whereas differential growth as well as curvature in positive root hydrotropism of ageotropum pea became visible 4–5 h after the continuous hydrostimulation. Application of 2,3,5-triiodobenzoic acid or ethyleneglycol-bis-( β -aminoethylether)-N,N,N',N'-tetraacetic acid was inhibitory to both root hydrotropism of ageotropum pea and root gravitropism of Alaska pea. Some mutual response mechanism for both hydrotropism and gravitropism may exist in roots, although the stimulusperception mechanisms differ from one another.     

Ammonium and nitrate uptake by barley genotypes in diurnally fluctuating root temperatures simulating till and no-till conditions

The morphological development and N uptake patterns of spring barley (Hordeum vulgare L.) genotypes of Northern European (Nordic) and Pacific Northwest US (PNW) origin were compared under two diurnally fluctuating root temperature regimes in solution culture. The two regimes, 15/5°C and 9/5°C day maximum/night minimum temperatures, simulated soil temperature differences between tilled vs. heavy-residue, no-till conditions, respectively, observed during early spring in eastern Washington. Previous field experiments indicated that some of the Nordic genotypes accumulated more N and dry matter than the PNW cultivars during early spring under no-till conditions. The objective of this experiment was to determined whether these differences 1) are dependent on the temperature of the rooting environment, and 2) are correlated with genotypic differences in NH₄ ⁺ and NO₃ ^– uptake. Overall, shoot N and dry matter accumulation was reduced by 40% due to lower root temperatures during illumination. Leaf emergence was slowed by 14 to 22%, and tiller production was also inhibited. All genotypes absorbed more ammonium than nitrate from equimolar solutions, and the proportion of total N absorbed as NH₄ ⁺ was slightly higher in the 9/5°C than the 15/5°C regime. A Finnish genotype, HJA80201, accumulated significantly more shoot N than the PNW cultivars, Clark and Steptoe, and also more than a Swedish cultivar, Pernilla, in the 9/5°C regime. In the 15/5°C regime Steptoe did not differ in shoot N from the Nordic genotypes, while Clark remained significantly lower. These differences were not correlated to relative propensity for N form. Root lengths of the Nordic genotypes were significantly greater than the PNW genotypes grown under the 9/5°C regime, while the root lengths in the warmer root temperture regime were not significantly different among genotypes. Higher root elongation rates under low soil temperature conditions may be an inherent adaptive mechanism of the Nordic genotypes. Overall, the data indicate that lower maximum daytime temperatures of the soil surface layer likely account for a significant portion of the growth reductions and lower N uptake observed in no-till systems.