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S Caveney

Publications and source records attributed to S Caveney.

33 records · Page 2Linked to original sources

Active muscle migration during insect metamorphosis.

The R1 abdominal retractor muscles of the insect Tenebrio molitor change position during the course of metamorphosis. These muscles detach from the epidermal tendon cells at their anterior ends, and migrate in a posterior direction, parallel to the body axis, to form completely new attachments shortly before adult emergence. Movement is preceded by the loss of sarcomere structure, and the muscles migrate in a partially dedifferentiated condition, closely accompanied by satellite cells and haemocytes. Movement appears to result from the extension of muscle processes towards the epidermis posterior to the larval attachment sites, which contact reciprocal processes extended from the epidermis. Contacts at the new posterior sites are then reinforced, and relinquished at the anterior. This cycle is subsequently repeated. It is envisaged that migration ceases when the muscles encounter a contour in the epidermal gradient known to specify the position of the adult muscle attachment sites. This positional information may be encoded in the epidermal basal lamina. The muscles then redifferentiate, with concurrent differentiation of new epidermal tendon cells. Development of adult muscle attachments appears to require reciprocal morphogenetic interactions between muscle and epidermis.

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Control of gap junction formation in early mouse embryos.

Intercellular communication via gap junctions begins in the eight-cell stage in early mouse embryos. We have studied the timing of this event in relation to compaction, and have begun to explore some of the possible control mechanisms underlying it. Gap junction formation was inferred by measuring ionic coupling as well as by observing the intercellular transfer of fluorescent dye. Embryos were obtained early on Day 3 of pregnancy by flushing the oviducts of HA/ICR mice that had been mated with CB6F1/J males. Gap junctions were detected only in those embryos which had achieved the fully compacted state. Inhibition of protein synthesis by cycloheximide treatment beginning as early as the late four-cell stage failed to block compaction or the acquisition of gap junctions, demonstrating that the necessary proteinaceous components are present in advance of these events. In order to test the possibility that gap junctions could be induced to form prematurely, fully compacted, communication-competent eight-cell embryos were aggregated with two- or four-cell embryos. Even after 10 hr of aggregation, no interembryonic gap junctions could be detected. Fully compacted eight-cell embryos when aggregated with each other, however, became ionically coupled within 3-5 hr. The number of interembryonic junctional channels was judged to be effectively small, since the aggregated embryos exhibited obvious ionic coupling but very weak dye coupling. In contrast to gap junction formation within embryos, junction formation between embryos was blocked by cycloheximide. These results demonstrate that gap junction formation in early mouse embryos is under precise temporal control, involving the assembly or mobilisation of preexisting components. This stockpile of components is either unavailable or insufficient to allow the formation of additional gap junctions between aggregated communication-competent embryos without new protein synthesis.

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Drugs that block calmoduLin activity inhibit cell-to-cell coupling in the epidermis of Tenebrio molitor.

In many cell systems, the permeability of membrane junctions is modulated by the cytoplasmic level of free Ca++. To examine whether the calcium-dependent regulatory protein calmodulin is involved in this process, the ability of anticalmodulin drugs to influence the cell-to-cell passage of injected current and an organic tracer was tested using standard intracellular glass microelectrode techniques. Several antipsychotics and local anesthetics were found to block junctional communication in the epidermis of the beetle Tenebrio molitor. Treatment of the epidermis with chlorpromazine (0.25 mM) raised intercellular resistance two- to threefold within 20 to 25 min; cell-to-cell passage of electrical current was abolished within 41 +/- 5 min. Loss of electrotonic coupling was accompanied by a block in the cell-to-cell movement of the organic tracer carboxyfluorescein. The reaction is fully reversible, with normal electrotonic coupling being restored within 2 to 4 hr. Other antipsychotics and local anesthetics had similar effects on cell coupling. The order of potency found was: trifluoperazine greater than thioridazine greater than D-butaclamol greater than chlorprothixine = chlorpromazine greater than L-butaclamol greater than dibucaine greater than tetracaine. The relative uncoupling potencies of these drugs correlate well with their known ability to inhibit calmodulin-dependent phosphodiesterase activity. Other anesthetic compounds, procaine and pentobarbital, did not block cell-to-cell communication. Altering the extracellular Ca++ concentration did not affect the rate of uncoupling by antipsychotics, while chelation of extracellular Ca++ with EGTA raised electrotonic coupling. The effect of three metabolic inhibitors on coupling was also examined. Iodoacetate uncoupled the epidermal cells while DNP and cyanide did not. These results are discussed in terms of possible mechanisms by which calmodulin may control junctional communication in this tissue.

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Changing muscle patterns in a segmental epidermal field.

The spatial rearrangements that take place during metamorphosis in the abdominal sternites and associated retractor muscles of the beetle Tenebrio molitor are described. This paper provides the descriptive background needed to consider whether a morphogenetic gradient is involved in specifying the position at which adult muscle attachments develop. (Experimental work in support of this gradient hypothesis is published in a companion paper.) The ventral abdominal retractor muscles are extensively remodelled at metamorphosis so that the adult muscles differ considerably in appearance from the larval muscles from which they are derived. In particular, there is a change in both the absolute and relative positions of the sites of muscle attachment. Rearrangement of muscles takes place during both the prepupal period and the pupal stage. It is achieved by means of two separate and temporally distinct mechanisms. Epidermal remodelling in the pre-pupal period results in the movement of attached retractor muscles (epidermokinetic muscle movement). In the pupal stage, however, the muscles move over the basal surface of the epidermis (myokinetic muscle movement). Myokinetic movements may be brought about by extension of myoblast processes from the metamorphosing muscles. These findings are considered in terms of Poyarkoff's theory that the pupa serves as an integumental mould, approximating the shape of the adult, within which certain adult muscles develop.

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A gradient of morphogenetic information involved in muscle patterning.

The results of grafts performed in the larva of the beetle Tenebrio molitor reveal that the position at which certain adult muscle attachments form is specified by a regulative, rather than a mosaic, mechanism. The results of grafts involving the rotation of squares of integument through 180 degrees or 90 degrees, or the antero-posterior transposition of two adjacent rectangles of integument, show that the site of muscle attachments in the antero-posterior axis of an abdominal sternite is specified by an epidermal segmental gradient of positional information. This gradient is presumed to be identical to the gradient which specifies cuticular patterns in this insect. There is a good correlation between the effect of grafts on adult muscle morphology and cuticular patterns.

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Intercellular communication in insect development is hormonally controlled.

Cellular coupling in the insect epidermis changes in a characteristic way during metamorphosis. In vitro, beta-ecdysone mimics the initial phase of these changes by increasing electrical coupling. Both adenosine 3',5'-monophosphate (cyclic AMP) and Ca2+ reverse natural and beta-ecdysone-stimulated changes, which suggests that ecdysone could work on communication through changes in cyclic AMP and Ca2+ levels. The transient changes in intercellular communication before metamorphosis may reflect the timing of the signals that trigger proliferation and the generation of new spatial patterns in the epidermis.

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Intercellular communication in a positional field. Ultrastructural correlates and tracer analysis of communication between insect epidermal cells.

The junctional membrane in the epidermal cells of the larval beetle (Tenebrio molitor L.) is comprised of macular gap junctions embedded in septate junctions. Ultrastructural and morphometric analysis of the distribution of gap junctions within the segmental epidermis suggests that this junction alone could account for the high electrotonic coupling recorded for the epidermal sheet. Analysis of the lanthanum-impregnated septate junction makes it doubtful that this junction serves as a communicating channel between beetle cells. A new model for the septate junction is presented in which pleated septa, less than 30 A thick, connect adjacent plasma membranes; the septa themselves are interconnected by two interseptal platforms that are coplanar with the plasma membranes. Iontophoretic injection of organic tracers into single epidermal cells suggests that only molecules of less than MW 1000 can transfer between cells through low-resistance junctions.

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