A microtubule-based, dynein-dependent force induces local cell protrusions: Implications for neurite initiation

A key event in neurite initiation is the accumulation of microtubule bundles at the neuron periphery. We hypothesized that such bundled microtubules may generate a force at the plasma membrane that facilitates neurite initiation. To test this idea we observed the behavior of microtubule bundles that were induced by the microtubule-associated protein MAP2c. Endogenous MAP2c contributes to neurite initiation in primary neurons, and exogeneous MAP2c is sufficient to induce neurites in Neuro-2a cells. We performed nocodazol washout experiments in primary neurons, Neuro-2a cells and COS-7 cells to investigate the underlying mechanism. During nocodazol washout, small microtubule bundles formed rapidly in the cytoplasm and immediately began to move toward the cell periphery in a unidirectional manner. In neurons and Neuro-2a cells, neurite-like processes extended within minutes and concurrently accumulated bundles of repolymerized microtubules. Speckle microscopy in COS-7 cells indicated that bundle movement was due to transport, not treadmilling. At the periphery bundles remained under a unidirectional force and induced local cell protrusions that were further enhanced by suppression of Rho kinase activity. Surprisingly, this bundle motility was independent of classical actin- or microtubule-based tracks. It was, however, reversed by function-blocking antibodies against dynein. Suppression of dynein expression in primary neurons by RNA interference severely inhibited the generation of new neurites, but not the elongation of existing neurites formed prior to dynein knockdown. Together, these cell biological data suggest that neuronal microtubule-associated proteins induce microtubule bundles that are pushed outward by dynein and locally override inward contraction to initiate neurite-like cell protrusions. A similar force-generating mechanism might participate in spontaneous initiation of neurites in developing neurons. Electronic Supplementry Materials: Supplementary Materials are available in the online version of this article at     

Multi drug resistance-dependent “vacuum cleaner” functionality potentially driven by the interactions between endocytosis, drug size and Pgp-like transporters surface density

In cells, multi drug resistance (MDR) is associated with Pgp-like transporters expression extruding drugs from cellular membranes. MDR is efficiently generated with a relatively small fraction of membrane transporters. As the insertion of drugs into cellular membranes is widespread, there are no reasons why a drug should incorporate the membrane in the vicinity of a transporter. As a result a further elusive hypothesis is usually invoked: these transporters act like “vacuum cleaners” of drugs embedded in the membrane. Nonetheless, how these transporters attract drugs remains obscure. To clarify the “vacuum cleaner” notion, we suggest that during its residency time in cellular membranes, the lateral movement of drugs from their point of insertion to transporters is governed by Brownian’s diffusion, which allows the drugs/transporters interaction. Taking into account the functionality of Pgp-like transporters, namely the extrusion of drugs from the plasma membrane inner leaflet, we characterize how the state of drug resistance is triggered involving: membrane endocytosis, drug physico-chemical properties and the surface density of Pgp-like transporters. In addition, the theory developed provides for the first time a theoretical proof of Lipinski’s second rule with regard to drugs’ size (or MW) selectivity on their permeation across cellular membranes. Electronic Supplementary Material Supplementary material is available in the online version of this article at ) and is accessible for authorized users.     

The apoptotic microtubule network preserves plasma membrane integrity during the execution phase of apoptosis

It has recently been shown that the microtubule cytoskeleton is reformed during the execution phase of apoptosis. We demonstrate that this microtubule reformation occurs in many cell types and under different apoptotic stimuli. We confirm that the apoptotic microtubule network possesses a novel organization, whose nucleation appears independent of conventional γ-tubulin ring complex containing structures. Our analysis suggests that microtubules are closely associated with the plasma membrane, forming a cortical ring or cellular “cocoon”. Concomitantly other components of the cytoskeleton, such as actin and cytokeratins disassemble. We found that colchicine-mediated disruption of apoptotic microtubule network results in enhanced plasma membrane permeability and secondary necrosis, suggesting that the reformation of a microtubule cytoskeleton plays an important role in preserving plasma membrane integrity during apoptosis. Significantly, cells induced to enter apoptosis in the presence of the pan-caspase inhibitor z-VAD, nevertheless form microtubule-like structures suggesting that microtubule formation is not dependent on caspase activation. In contrast we found that treatment with EGTA-AM, an intracellular calcium chelator, prevents apoptotic microtubule network formation, suggesting that intracellular calcium may play an essential role in the microtubule reformation. We propose that apoptotic microtubule network is required to maintain plasma membrane integrity during the execution phase of apoptosis. Electronic Supplementary Material Supplementary material is available in the online version of this article at .     

Morphological characterization of single fan-shaped body neurons in <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

The fan-shaped body is the largest substructure of the central complex in Drosophila melanogaster. Two groups of large-field neurons that innervate the fan-shaped body, viz., F1 and F5 neurons, have recently been found to be involved in visual pattern memory for “contour orientation” and “elevation” in a rut-dependent manner. The F5 neurons have been found to be responsible for the parameter “elevation” in a for-dependent manner. We have shown here that the F1 neuron also affects visual memory for “contour orientation” in a for-dependent way. With the help of Gal4/UAS and FLP-out techniques, we have characterized the morphological features of these two groups of neurons at single neuron resolution. We have observed that F1 or F5 neurons are groups of isomorphic individual neurons. Single F1 neurons have three main arborization regions: one in the first layer of the fan-shaped body, one in the ventral body, and another in the inferior medial protocerebrum. Single F5 neurons have two arborization regions: one in the fifth layer of the fan-shaped body and the other in the superior medial protocerebrum. The polarity of the F1 and F5 neurons has been studied with the Syt-GFP marker. Our results indicate the existence of presynaptic sites of both F1 and F5 neurons located in the fan-shaped body and postsynaptic sites outside of the fan-shaped body. This work was supported by the “973 Program” (2005CB522804 and 2009CB918702), the National Natural Sciences Foundation of China (30621004, 30625022, and 30770682), and the Knowledge Innovation Program of the Chinese Academy of Sciences (KSCX2-YW-R-28).     

Slow modulation of synaptic transmission by brain-derived neurotrophic factor leads to the central sensitization associated with neuropathic pain

Chronic constriction injury (CCI) of the rat sciatic nerve increases the dorsal horn excitability. This “central sensitization” leads to behavioral manifestations analogous to those related to human neuropathic pain. We found, using whole-cell recording from acutely isolated spinal cord slices, that 7-to 10-day-long CCI increases excitatory synaptic drive to putative excitatory “delay”-firing neurons in the substantia gelatinosa but attenuates that to putative inhibitory “tonic”-firing neurons. A defined-medium organotypic culture (DMOTC) system was used to investigate the long-term actions of brain-derived neurotrophic factor (BDNF) as a possible instigator of these changes. When all five neuronal types found in the substantia gelatinosa were considered, BDNF and CCI produced similar patterns, or “footprints,” of changes across the whole population. This pattern was not seen with another putative “pain mediator,” interleukin 1β. Thus, BDNF decreased synaptic drive to “tonic” neurons and increased synaptic drive to “delay” neurons. Actions of BDNF on “delay” neurons were presynaptic and involved increased mEPSC frequency and amplitude without changes in the function of postsynaptic AMPA receptors. By contrast, BDNF exerted both pre-and post-synaptic actions on “ tonic” cells to reduce the mEPSC frequency and amplitude. These differential actions of BDNF on excitatory and inhibitory neurons contributed to a global increase in the dorsal horn network excitability as assessed by the amplitude of depolarization-induced increases in the intracellular [Ca²⁺]. Experiments with the BDNF-binding protein TrkB-d5 provided additional evidence for BDNF as a harbinger of neuropathic pain. Thus, the cellular processes altered by BDNF likely contribute to “central sensitization” and hence to the onset of neuropathic pain. Neirofiziologiya/Neurophysiology, Vol. 39, Nos. 4/5, pp. 315–326, July–October, 2007.     

Microtubules and microfilaments coordinate to direct a fountain streaming pattern in elongating conifer pollen tube tips

This study investigates how microtubules and microfilaments control organelle motility within the tips of conifer pollen tubes. Organelles in the 30-m-long clear zone at the tip of Picea abies (L.) Karst. (Pinaceae) pollen tubes move in a fountain pattern. Within the center of the tube, organelles move into the tip along clearly defined paths, move randomly at the apex, and then move away from the tip beneath the plasma membrane. This pattern coincides with microtubule and microfilament organization and is the opposite of the reverse fountain seen in angiosperm pollen tubes. Application of latrunculin B, which disrupts microfilaments, completely stops growth and reduces organelle motility to Brownian motion. The clear zone at the tip remains intact but fills with thin tubules of endoplasmic reticulum. Applications of amiprophosmethyl, propyzamide or oryzalin, which all disrupt microtubules, stop growth, alter organelle motility within the tip, and alter the organization of actin microfilaments. Amiprophosmethyl inhibits organelle streaming and collapses the clear zone of vesicles at the extreme tip together with the disruption of microfilaments leading into the tip, leaving the plasma membrane intact. Propyzamide and oryzalin cause the accumulation of membrane tubules or vacuoles in the tip that reverse direction and stream in a reverse fountain. The microtubule disruption caused by propyzamide and oryzalin also reorganizes microfilaments from a fibrillar network into pronounced bundles in the tip cytoplasm. We conclude that microtubules control the positioning of organelles into and within the tip and influence the direction of streaming by mediating microfilament organization.Electronic Supplementary Material Supplementary material is available in the online version of this article at Abbreviations APM Amiprophosmethyl - FITC Fluorescein isothiocyanate - LATB Latrunculin B     

Primary culture and characteristic morphologies of neurons from the cerebral ganglion of the mud crab, <Emphasis Type="Italic">Scylla paramamosain</Emphasis>

Crustacean neurons, obtained from the cerebral ganglion of the mud crab Scylla paramamosain, were successfully cultured in vitro. They maintained typical morphological characteristics and showed better outgrowth in modified Medium 199 (M199) medium than that in Liebowitz’s L-15 medium. Fetal bovine serum (FBS), muscle extracts, and hemolymph of the mud crab S. paramamosain were added as supplements. Only 20% FBS could promote neuron outgrowth, while muscle extracts and hemolymph of S. paramamosain did not improve neuron outgrowth. For cell dissociation, both collagenase type I and trypsin worked well as determined by initial cell viability and following cell outgrowth potential. More than six kinds of cells with different morphological characteristics were identified in the neuron outgrowth. They were “small cells”, “veilers”, “branchers”, “multipolar cells”, “super-large cell”, and “bipolar cells”. Among all of the cells, bipolar cells were identified for the first time in crustacean neurons culture and they could live longer than other cells. The neurons could grow for more than a week before retraction and eventual degradation.     

The Jackprot Simulation Couples Mutation Rate with Natural Selection to Illustrate How Protein Evolution Is Not Random

Protein evolution is not a random process. Views which attribute randomness to molecular change, deleterious nature to single-gene mutations, insufficient geological time, or population size for molecular improvements to occur, or invoke “design creationism” to account for complexity in molecular structures and biological processes, are unfounded. Scientific evidence suggests that natural selection tinkers with molecular improvements by retaining adaptive peptide sequence. We used slot-machine probabilities and ion channels to show biological directionality on molecular change. Because ion channels reside in the lipid bilayer of cell membranes, their residue location must be in balance with the membrane’s hydrophobic/philic nature; a selective “pore” for ion passage is located within the hydrophobic region. We contrasted the random generation of DNA sequence for KcsA, a bacterial two-transmembrane-domain (2TM) potassium channel, from Streptomyces lividans, with an under-selection scenario, the “jackprot,” which predicted much faster evolution than by chance. We wrote a computer program in JAVA APPLET version 1.0 and designed an online interface, The Jackprot Simulation , to model a numerical interaction between mutation rate and natural selection during a scenario of polypeptide evolution. Winning the “jackprot,” or highest-fitness complete-peptide sequence, required cumulative smaller “wins” (rewarded by selection) at the first, second, and third positions in each of the 161 KcsA codons (“jackdons” that led to “jackacids” that led to the “jackprot”). The “jackprot” is a didactic tool to demonstrate how mutation rate coupled with natural selection suffices to explain the evolution of specialized proteins, such as the complex six-transmembrane (6TM) domain potassium, sodium, or calcium channels. Ancestral DNA sequences coding for 2TM-like proteins underwent nucleotide “edition” and gene duplications to generate the 6TMs. Ion channels are essential to the physiology of neurons, ganglia, and brains, and were crucial to the evolutionary advent of consciousness. The Jackprot Simulation illustrates in a computer model that evolution is not and cannot be a random process as conceived by design creationists.     

Molecular phylogenies and mating study data in Polyporus with special emphasis on group “Melanopus” (Basidiomycota)

“Melanopus” is an unranked infrageneric group within Polyporus, a genus comprising wood-decaying homobasidiomycetes. This study compiles new information on the mating systems, phylogeny, and geographic distribution of the group. We report tetrapolar mating systems in “Melanopus,” as well as in the more distantly related Polyporus umbellatus. “Melanopus” is addressed using internal transcribed spacer (ITS) rDNA sequence data, comparing resolving power of several phylogenetic methods and depths of sequence information, including recoded secondary structure data informed by the primary sequence data. The latter, inferred structure of the ITS2 rRNA was insufficient in phylogenetic reconstruction on its own. Intercollection compatibility studies showed Polyporus varius collections from both sides of the Atlantic to be intercompatible, the same was found for Polyporus tubaeformis collections from California and Scotland. Molecular analysis supports P. tubaeformis conspecifity of Scottish and Californian collections with a Norwegian collection. Several other herbarium specimens inaccessible to successful PCR and sequencing were redetermined as P. tubaeformis by morphological characters. This improved on the current knowledge of distribution of the species, tentatively adding Idaho, North Carolina, and Germany’s Mecklenburg-Western Pomerania.Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Pilocarpine-induced epileptiform activity of isolatedCA1 hippocampal neurons

Cellular mechanisms of pilocarpine-induced epileptiform activity in isolatedCA1 andCA3 hippocampal neurons were studied using a current-clamp technique. Isolated unipolar neurons dominate after enzymatic treatment of theCA1 tissue, whereas large multipolar neurons are mainly located in theCA3 area. Measurements of the membrane potential in these two groups of neurons showed that most of them are “silent” cells. Only a small group of neurons from theCA3 area displayed spontaneous electrical activity. Pilocarpine, a well known epileptogenic compound, induced rhythmic waveform changes in the membrane potential in some “silent” unipolar neurons from theCA1 area, whereas in another more numerous group of neurons it induced only steady depolarization of the membrane. Application of tetrodotoxin, the selective blocker of Na⁺ channels, blocked generation of action potentials induced inCA1 neurons by pilocarpine, but exerted no effect on waveform shifts of the membrane potential. It is suggested that the mechanism underlying epileptogenic action of pilocarpine on the brain activity in rats is based on the induction of waveform changes in the membrane potential in unipolar neurons of theCA1 hippocampal area.     

Biosemiosis and the Cellular Basis of Mind

The human brain is a complex organ made up of neurons and several other cell types, and whose role is processing information for use in elicitation of behaviors. To accomplish this, the brain requires large amounts of energy, and this energy is obtained by the oxidation of glucose (Glc). However, the question of how the oxidation of Glc by individual neurons in brain results in their collective ability to rapidly generate feats of cognition that allow them to recognize the nature of the universe in which they live and to communicate this information remains unclear. In this article, insights into this process are provided by first considering the brain’ s homeostatic “operating system” for supply of energy to stimulated neurons, and how this system defines the basic unit of brain “structure”. This is followed by consideration of the brain’s “two-cell” neuronal communication mechanism which defines the basic unit of brain “function”. Finally, an analysis of the nature of frequency-encoded “neuronal languages” that enable ensembles of neurons to translate energy derived from the oxidation of Glc into a collective “mind”, the aggregate of all brain processes including those involving perception, thought, insight, foresight, imagination and behavior.     

Corticotropin-releasing factor (CRF) expression in postnatal and adult rat sacral parasympathetic nucleus (SPN)

The neural control of micturition undergoes marked changes during the early postnatal development. During the first few postnatal weeks, the spinal micturition reflex is gradually replaced by a spinobulbospinal reflex pathway that is responsible for micturition in adult animals. Upregulation of brainstem regulation of spinal micturition pathways may contribute to development of mature voiding patterns. We examined the expression of corticotropin-releasing factor (CRF), present in descending projections from Barrington's nucleus to the sacral parasympathetic nucleus (SPN), in postnatal (P0–P36) and adult Wistar rats (P60–90). CRF-immunoreactivity (IR) was present predominantly in the SPN region, although some staining was also observed in the dorsal horn and dorsal commissure in L5–S1 spinal segments. CRF-IR in spinal cord regions was age dependent (R ²=0.87–0.98). The majority of the CRF-IR in the lumbosacral spinal cord was eliminated by complete spinalization (2–3 weeks). Double-label immunohistochemistry was combined with quantitative confocal laser scanning microscopy to quantify the number and percentage of colocalization between CRF-immunoreactive varicosities and preganglionic somas or proximal neurites in the SPN in postnatal and adult rats. Results demonstrate an age-dependent upregulation of CRF-IR in the SPN region and specifically in association with preganglionic parasympathetic neurons identified with neuronal nitric oxide synthase (nNOS)-IR. CRF-immunoreactive varicosities on or within a 1 μm perimeter of nNOS-immunoreactive somas or proximal neurites also increased with postnatal age. The upregulation of CRF-IR in bulbospinal projections to the SPN may contribute to mature voiding reflexes. This work was supported in part through NIH grants DK051369, DK060481, DK065989, NS040796.     

Terahertz vibrational properties of water nanoclusters relevant to biology

Water nanoclusters are shown from first-principles calculations to possess unique terahertz-frequency vibrational modes in the 1–6 THz range, corresponding to O–O–O “bending,” “squashing,” and “twisting” “surface” distortions of the clusters. The cluster molecular-orbital LUMOs are huge Rydberg-like “S,” “P,” “D,” and “F” orbitals that accept an extra electron via optical excitation, ionization, or electron donation from interacting biomolecules. Dynamic Jahn–Teller coupling of these “hydrated-electron” orbitals to the THz vibrations promotes such water clusters as vibronically active “structured water” essential to biomolecular function such as protein folding. In biological microtubules, confined water-cluster THz vibrations may induce their “quantum coherence” communicated by Jahn–Teller phonons via coupling of the THz electromagnetic field to the water clusters’ large electric dipole moments.     

“Not <Emphasis Type="Italic">Here</Emphasis>”: Making the Spaces and Subjects of “Global Health” in Botswana

This essay argues that what makes “global health” “global” has more to do with configurations of space and time, and the claims to expertise and moral stances these configurations make possible, than with the geographical distribution of medical experts or the universal, if also uneven, distribution of threats to health. Drawing on a study of public–private partnerships supporting Botswana’s HIV/AIDS treatment program, this essay demonstrates ethnographically the processes by which “global health” and its quintessential spaces, namely “resource-limited” or “resource-poor settings,” are constituted, reinforced, and contested in the context of medical education and medical practice in Botswana’s largest hospital. Using Silverstein’s work on orders of indexicality, I argue that the terms of “global health” are best understood as chronotopic, and demonstrate how actors orient themselves and others spatio-temporally, morally, and professionally by using or refuting those terms. I conclude by arguing that taking “global health” on its own terms obscures the powerful forces by which it becomes intelligible. At stake are the frames within which medical anthropologists understand their objects of study, as well as the potential for the spaces of “global health” intervention to expand ever outward as American medical personnel attempt to calibrate their experiences to their expectations.     

Drosophila RhoGAP68F is a putative GTPase activating protein for RhoA participating in gastrulation

The Rho family small GTPases Rho, Rac, and Cdc42 regulate cell shape and motility through the actin cytoskeleton. These proteins cycle between a GTP-bound “on” state and a GDP-bound “off” state and are negatively regulated by GTPase-activating proteins (GAPs), which accelerate the small GTPase’s intrinsic hydrolysis of bound GTP to GDP. Drosophila RhoGAP68F is similar to the mammalian protein p50RhoGAP/Cdc42GAP, which exhibits strong GAP activity toward Cdc42. We find that, despite the strong similarities between RhoGAP68F and p50RhoGAP/Cdc42GAP, RhoGAP68F is most effective as a GAP for RhoA. These in vitro data are supported by the in vivo analysis of mutants in RhoGAP68F. We demonstrate through the characterization of two alleles of the RhoGAP68F gene that RhoGAP68F participates in gastrulation of the embryo, a morphogenetic event driven by cell constriction that involves RhoA signaling. We propose that RhoGAP68F functions as a regulator of RhoA signaling during gastrulation.     

A model for neuron firing with exponential decay of potential resulting in diffusion equations for probability density

A neuron is assumed to receive synaptic input of both excitatory and inhibitory natures from a large number of neighboring neurons; it is also assumed that a large number of such impulses are required to raise the neuron’s transmembrane potential to its threshold potential, at which it “fires” or “spikes”. The model is similar to one of Gerstein and Mandelbrot, except that in the absence of input an exponential decay of potential toward a resting level is introduced. Computational methods of determining the spike timeinterval distribution are discussed, along with the inverse problem of estimating the parameters of the system from observed spike time-interval data.     

How Willing Are You to Accept Sexual Requests from Slightly Unattractive to Exceptionally Attractive Imagined Requestors?

In their classic study of differences in mating strategies, Clark and Hatfield (1989, Journal of Psychology and Human Sexuality, 2, 39–54) found that men and women demonstrated a striking difference in interest in casual sex. The current study examined the role of an imagined requestor’s physical attractiveness (slightly unattractive, moderately attractive, and exceptionally attractive) on men’s and women’s willingness to accept three different requests (go out, come to apartment, go to bed) as reflected in answers to a questionnaire. We tested two hypotheses with a sample of 427 men and 443 women from three countries. Hypothesis 1 states that men, relative to women, will demonstrate a greater willingness to accept the “come to apartment” and “go to bed” requests but not the “go out” request for all three levels of requestor attractiveness. This hypothesis reflects Clark and Hatfield’s main findings. Hypothesis 2 states that the physical attractiveness of a potential partner will have a greater effect on women’s than on men’s willingness to accept all three requests, and particularly for the explicit request for casual sex. The results partially supported Hypothesis 1 and fully supported Hypothesis 2. The discussion highlights limitations of the current research and presents directions for future research.     

Postsynaptic regulation of the development and long-term plasticity of Aplysia sensorimotor synapses in cell culture

The monosynaptic component of the neuronal circuit that mediates the withdrawal reflex of Aplysia californica can be reconstituted in dissociated cell culture. Study of these in vitro monosynaptic connections has yielded insights into the basic cellular mechanisms of synaptogenesis and long-term synaptic plasticity. One such insight has been that the development of the presynaptic sensory neurons is strongly regulated by the postsynaptic motor neuron. Sensory neurons which have been cocultured with a target motor neuron have more elaborate structures—characterized by neurites with more branches and varicosities—than do sensory neurons grown alone in culture or sensory neurons that have been cocultured with an inappropriate target cell. Another way in which the motor neuron regulates the development of sensory neurons is apparent when sensorimotor cocultures with two presynaptic cells are examined. In such cocultures the outgrowth from the different presynaptic cells is obviously segregated on the processes of the postsynaptic cell. By contrast, when two sensory neurons are placed into cell culture without a motor neuron, thier processes readily grow together. In addition to regulating the in vitro development of sensory neurons, the motor neuron also regulates learning-related changes in the structure of sensory neurons. Application of the endogenous facilitatory trasmitter serotonin (5-HT) causes long-term facilitation of in vitro sensorimotor synapses due in part to growth of new presynatpic varicosities. But 5-HT applied to sensory neurons alone in cultuer does not produce structural changes in these cells. More recently it has been found that sensorimotor synapses in cell culture can exhibit long-term potentiation (LTP). Like LTP of some hippocampal synapses, LTP of in vitro Aplysia syanpses is regulated by the voltage of the postsynaptic cell. Pairing high-frequency stimulation of sensory neurons with strong hyperpolarization of the motor neuron blocks the induction of LTP. Moreover, LTP of sensorimotor synapses can be induced in Hebbian fashion by pairing weak presynaptic stimulation with strong postsynaptic depolarization. These findings implicate a Habbian mechanism in classical conditioning in Aplysia. They also indicate that Hebbian LTP is a phylogenetically ancient form of synaptic plasticity. 1994 John Wiley & Sons, Inc.     

Scanning electron microscopy ofMycoplasma pneumoniae on the membrane of individual ciliated tracheal cells

Summary Cell monolayer cultures were prepared from hamster tracheal explants by a collagenase exposure and subsequent incubation in Waymouth’s MAB 87/3 medium. The epithelial outgrowth occurred on glass cover slips. Cilia on the monolayers continued to beat normally after the “parent” explant was removed. Monolayer cultures infected withMycoplasma pneumoniae had significant amounts of attachment. A morphological analysis of the attachment was conducted with scanning electron microscopy. Clusters, cocci, and filaments ofM. pneumoniae all attached to the epithelial cells, but the filaments were especially common. Mycoplasmas were seen in association with both ciliated and nonciliated cell membranes. On ciliated cells, mycoplasmas were on the ciliary strands and on the cell membrane. When located immediately adjacent to or in between cilia, mycoplasmas were oriented vertically with the constricted attachment tip oriented down toward the host cell membrane. When located more than a micron away from the ciliary fibers, mycoplasmas lay horizontally along the epithelial cell membrane. The photographic data suggest that clusters or “sperules” of mycoplasmas may liberate individual mycoplasmas that attach to the cell membrane. It appears that the receptor sites forM. pneumoniae are rather uniformly distributed along the ciliated cell membrane, and are not restricted to the interciliary areas. Electron microscopy was done with the cooperation of Dr. R. Macleod and the staff of the Center for Electron Microscopy at the University of Illinois. Critical editorial review was provided by C. Dayton. This investigation was supported in part by grants to M. G. G. from the National Institute of Allergy and Infectious Diseases (AI 12559) and the National Heart, Lung, and Blood Institute (HL 23806), Bethesda, Maryland.     

QTL analysis of fruit components in the progeny of a Rennell Island Tall coconut (Cocos nucifera L.) individual

We investigated the genetic factors controlling fruit components in coconut by performing QTL analyses for fruit component weights and ratios in a segregating progeny of a Rennell Island Tall genotype. The underlying linkage map of this population was already established in a previous study, as well as QTL analyses for fruit production, which were used to complement our results. The addition of 53 new markers (mainly SSRs) led to minor amendments in the map. A total of 52 putative QTLs were identified for the 11 traits under study. Thirty-four of them were grouped in six small clusters, which probably correspond to single pleiotropic genes. Some additional QTLs located apart from these clusters also had relatively large effects on the individual traits. The QTLs for fruit component weight, endosperm humidity and fruit production were found at different locations in the genome, suggesting that efficient marker-assisted selection for yield can be achieved by selecting QTLs for the individual components. The detected QTLs descend from a genotype belonging to the “Pacific” coconut group. Based on the known molecular and phenotypic differences between “Pacific” and “Indo-Atlantic” coconuts, we suggest that a large fraction of coconut genetic diversity is still to be investigated by studying populations derived from crosses between these groups. Electronic Supplementary Material Supplementary material is available for this article at and is accessible for authorized users.