Small granulated cell types in rat superior cervical and coeliac-mesenteric ganglia

Summary In the rat superior cervical and coeliac-mesenteric ganglia we have observed three types of small granulated (SG) cell: Type I cells are characterised by membrane-bounded cytoplasmic granules with a core of variable, moderate to low electron-density, whose limiting membranes are rounded in profile ranging from 50–150 nm in diameter. Type II SG cells contain numerous highly electron-dense, polymorphic cytoplasmic granules ranging from 100–300 nm in diameter. The haloes of Type II cell granules are variable in shape, and the core is often eccentrically located or fragmented. Type III SG cells contain membrane-bounded granules with a core of variable moderate to low electron-density. In profile these granules appear oblong or circular with average dimensions of 170 × 50 nm. All three SG cell types receive cholinergic-type pre-ganglionic terminals whose afferent nature is confirmed by their degeneration following pre-ganglionic neurectomy. Only Type I cells have been observed to donate efferent synapses to dendrites of principal ganglionic neurones and are thus interneuronal.This work was in part supported by a grant from the Medical Research Council. We wish to thank Mr. T.T. Lee for valuable technical assistance and Mr. P.F. Hire and Mr. K. Twohigg for illustrative help     

Deoxyribonucleic acid synthesis in isolated polytene nuclei of Drosophila hydei

DNA synthesis has been studied in polytene nuclei isolated from larval salivary glands of Drosophila hydei. The incubation conditions employed promote maximum incorporation of TTP-H³ and retention of normal polytene chromosome morphology. The chromosome structure is sensitive to the Mg²⁺ concentration; a normal banding pattern is observed between 4 and 10 mM Mg²⁺. At the optimum pH of 7.8, incorporation continues for over an hour. All four deoxyribonucleoside triphosphates are required for maximum incorporation. The reaction is stimulated by 0.6 mmATP and strongly inhibited at higher ATP concentrations. Competition experiments demonstrate that either TDP or TTP is the effective labeled precursor. The labeled product is sensitive to DNase and has a density identical to that of nuclear DNA. Autoradiographs prepared from spread chromosomes demonstrate that discontinuous and continuous labeling patterns observed in vivo are also produced with isolated nuclei in the absence of cytoplasmic factors. Incubation of the isolated nuclei results in a low level of uniform incorporation that is superimposed on the normal autoradiographic pattern obtained after in vivo labeling. This background incorporation can be greatly increased by prior irradiation of the glands. The presence of exogenous DNA during nuclear incubation stimulates total incorporation. These observations demonstrate that the isolated nuclei possess a reserve synthetic capacity. About 20% of the isolated nuclei are inactive in DNA synthesis.This investigation was supported by PHS Research Grant No. 5 R01 GM 16298 from the National Institute of General Medical Sciences.     

Endogenously expressed apolipoprotein E has different effects on cell lipid metabolism as compared to exogenous apolipoprotein E carried on triglyceride-rich particles

Apolipoprotein E (apoE) on model triglyceride-rich particles (TGRP) increases triglyceride (TG) utilization and cholesteryl ester (CE) hydrolysis, independent of its effect on enhancing particle uptake. We questioned whether, under physiological concentrations, endogenously expressed apoE has similar effects on cellular lipid metabolism as compared to exogenous apoE. J774 macrophages, which do not express apoE, were engineered to express endogenous apoE by transfection of human apoE3 cDNA expression constructs (E(+)) or control vectors (E(-)) into the cells. To compare the effects of exogenous apoE and endogenous apoE on TGRP uptake, cells were incubated with or without apoE associated with (3)H-cholesteryl ether-labeled TGRP. Exogenous apoE enhanced TGRP uptake in both E(-) and E(+) cells. E(-) cells displayed significantly higher TGRP uptake than E(+) cells. Sodium chlorate, which inhibits cell proteoglycan synthesis, markedly diminished differences in TGRP uptake between E(-) and E(+) cells, suggesting that endogenous apoE-proteoglycan interaction contributes to differences in uptake between the two cell types. Particle uptake by the LDL receptor, by the LDL receptor related protein, or by scavenger receptors were similar between E(-) and E(+) cells indicating that endogenous apoE expression does not have a general effect on endocytic pathways. Exogenous apoE carried on TGRP stimulated TG utilization and CE hydrolysis in both cell types. However, TG utilization and CE hydrolysis were not affected by endogenous apoE expression. In conclusion, macrophage expression of apoE has very different effects on TGRP metabolism than exogenously supplied apoE. The fluorescence microscopy results in this study showing that exogenous apoE and endogenous apoE were confined in separate cellular compartments support the hypothesis that these differences resulted from distinct intracellular trafficking pathways followed by exogenous apoE bound to TGRP as compared to endogenous cell-expressed apoE.     

Golgi membranes are absorbed into and reemerge from the ER during mitosis

Quantitative imaging and photobleaching were used to measure ER/Golgi recycling of GFP-tagged Golgi proteins in interphase cells and to monitor the dissolution and reformation of the Golgi during mitosis. In interphase, recycling occurred every 1.5 hr, and blocking ER egress trapped cycling Golgi enzymes in the ER with loss of Golgi structure. In mitosis, when ER export stops, Golgi proteins redistributed into the ER as shown by quantitative imaging in vivo and immuno-EM. Comparison of the mobilities of Golgi proteins and lipids ruled out the persistence of a separate mitotic Golgi vesicle population and supported the idea that all Golgi components are absorbed into the ER. Moreover, reassembly of the Golgi complex after mitosis failed to occur when ER export was blocked. These results demonstrate that in mitosis the Golgi disperses and reforms through the intermediary of the ER, exploiting constitutive recycling pathways. They thus define a novel paradigm for Golgi genesis and inheritance.     

The spatial configuration of taxonomic biodiversity along a tropical elevational gradient: α‐, β‐, and γ‐partitions

Biodiversity at larger spatial scales (γ) can be driven by within‐site partitions (α), with little variation in composition among locations, or can be driven by among‐site partitions (β) that signal the importance of spatial heterogeneity. For tropical elevational gradients, we determined the (a) extent to which variation in γ is driven by α‐ or β‐partitions; (b) elevational form of the relationship for each partition; and (c) extent to which elevational gradients are molded by zonation in vegetation or by gradual variation in climatic or abiotic characteristics. We sampled terrestrial gastropods along two transects in the Luquillo Mountains. One passed through multiple vegetation zones (tabonuco, palo colorado, and elfin forests), and one passed through only palm forest. We quantified variation in hierarchical partitions (α, β, and γ) of species richness, evenness, diversity, and dominance, as well as in the content and quality of litter. Total gastropod abundance linearly decreased with increasing elevation along both transects, but was consistently higher in palm than in other forest types. The gradual linear decline in γ‐richness was a consequence of opposing patterns with regard to α‐richness (monotonic decrease) and β‐richness (monotonic increase). For evenness, diversity, and dominance, α‐partitions and γ‐partitions evinced mid‐elevational peaks. The spatial organization of gastropod biodiversity did not mirror the zonation of vegetation. Rather, it was molded by: (a) elevational variation in productivity or nutrient characteristics, (b) the interspersion of palm forest within other forest types, and (c) the cloud condensation point acting as a transition between low and high elevation faunas. Abstract in Spanish is available with online material.     

Abundant and diverse fungal microbiota in the murine intestine

Enteric microbiota play a variety of roles in intestinal health and disease. While bacteria in the intestine have been broadly characterized, little is known about the abundance or diversity of enteric fungi. This study utilized a culture-independent method termed oligonucleotide fingerprinting of rRNA genes (OFRG) to describe the compositions of fungal and bacterial rRNA genes from small and large intestines (tissue and luminal contents) of restricted-flora and specific-pathogen-free mice. OFRG analysis identified rRNA genes from all four major fungal phyla: Ascomycota, Basidiomycota, Chytridiomycota, and Zygomycota. The largest assemblages of fungal rRNA sequences were related to the genera Acremonium, Monilinia, Fusarium, Cryptococcus/Filobasidium, Scleroderma, Catenomyces, Spizellomyces, Neocallimastix, Powellomyces, Entophlyctis, Mortierella, and Smittium and the order Mucorales. The majority of bacterial rRNA gene clones were affiliated with the taxa Bacteroidetes, Firmicutes, Acinetobacter, and Lactobacillus. Sequence-selective PCR analyses also detected several of these bacterial and fungal rRNA genes in the mouse chow. Fluorescence in situ hybridization analysis with a fungal small-subunit rRNA probe revealed morphologically diverse microorganisms resident in the mucus biofilm adjacent to the cecal and proximal colonic epithelium. Hybridizing organisms comprised about 2% of the DAPI (4',6-diamidino-2-phenylindole, dihydrochloride)-positive organisms in the mucus biofilm, but their abundance in fecal material may be much lower. These data indicate that diverse fungal taxa are present in the intestinal microbial community. Their abundance suggests that they may play significant roles in enteric microbial functions.     

Changes in Activity of Glyoxylate Cycle Enzymes During Myxospore Development in Myxococcus xanthus

Activities of the glyoxylate cycle enzymes isocitrate lyase (EC 4.1.3.1) and malate synthase (EC 4.1.3.2) were assayed in extracts prepared at different stages of myxospore formation in liquid cultures of Myxococcus xanthus. Activities of both enzymes attained peak values during conversion of rods to spheres. Isocitrate lyase activity decreased after reaching its peak value. Malate synthase activity also declined but at a much slower rate. The loss of isocitrate lyase activity could be prevented by the addition of chloramphenicol to cultures early in myxospore formation (during the initial rise in enzyme activity), but not by such addition at later stages of myxospore formation. The increase in glyoxylate cycle enzymes was not observed in a mutant unable to form myxospores in liquid culture under conditions suitable for morphological conversion of the wild type, or in wild-type cells incubated in the absence of an inducer for myxospore formation. It is concluded that the changes in the glyoxylate cycle enzymes represent regulatory phenomena associated with the development of the myxospore.     

Niche Overlap and Network Specialization of Flower-Visiting Bees in an Agricultural System

Different resource use strategies manifest as differences in the realized niches of species. Niche segregation may involve several dimensions of the niche, such as diet, space, and time. We measured the level of redundancy and complementarity of a bee–plant interaction network in an agricultural system. Because flower resource diversity is high and resource abundance associated with flowering phenology varies throughout the year, we hypothesized that trophic overlap in the community would be low (i.e., high niche complementarity). In contrast, we expected a combination of physiological constraints and exploitation competition to create high temporal overlap, leading to high redundancy in the time of use of floral resources. Dietary overlap was low (NO_ih?=?0.18): niches of 88% of species pairs had less than 30% overlap. In contrast, temporal overlap was intermediate (NO_ih?=?0.49): niches of 65% of species pairs had 30% to 60% overlap. Network analysis showed that bees separated their dietary niches and had intermediate complementary specialization (H₂′?=?0.46). In terms of their temporal niches (H₂′?=?0.12), bees were generalists, with high temporal redundancy. Temperature was not a key factor in the determination of niche overlap, suggesting that environmental factors do not likely have a primary role in determining high redundancy in the temporal use of floral resources. Rather, temporal overlap is likely associated with the timing of nectar production by flowers. Our results suggest that bees partition a wide variety of available floral resources, resulting in low dietary overlap and intermediate temporal overlap.     

A comprehensive framework for the evaluation of metacommunity structure

The metacommunity framework is a powerful platform for evaluating patterns of species distribution in geographic or environmental space. Idealized patterns (checkerboard, Clementsian, evenly spaced, Gleasonian and nested distributions) give the framework shape. Each pattern represents an area in a multidimensional continuum of metacommunity structures; however, the current approach to analysis of spatial structure of metacommunities is incomplete. To address this, we describe additional non‐random structures and illustrate how they may be discerned via objective criteria. First, we distinguish three distinct forms of species loss in nested structures, which should improve identification of structuring mechanisms for nested patterns. Second, we define six quasi‐structures that are consistent with the conceptual underpinnings of Clementsian, Gleasonian, evenly spaced and nested distributions. Finally, we demonstrate how combinations of structures at smaller spatial extents may aggregate to form Clementsian structure at larger extents. These refinements should facilitate the identification of best‐fit patterns, associated structuring mechanisms, and informative scales of analysis and interpretation. This conceptual and analytical framework may be applied to network properties within communities (i.e. structure of interspecific interactions) and has broad application in ecology and biogeography.