Rhythmic contractions of the ampullar epidermis during metamorphosis of the ascidian Molgula occidentalis

Summary The ampullae of Molgula occidentalis are hollow, tubular extensions of the epidermis. They are ensheathed by a secreted tunic. When they grow out shortly after settlement, the ampullae spread the tunic over the substratum to form a firm attachment for the sessile juvenile. A simple squamous epithelium forms the thin ampullar walls. A glandular, simple columnar epithelium forms the distal tip of each ampulla. The glandular cells probably secrete the adhesive that attaches the tunic to the substratum.Repetitive, peristaltic contractions pass from the base to the distal end of each ampulla. Microsurgery, time-lapse cinemicrography and TEM have been used to analyze this phenomenon. The contractions are mediated by a layer of 4–8 nm microfilaments in the base of the ampullar epithelium.Each juvenile has 7–9 ampullae which contract at different frequencies. Isolated ampullae continue to contract normally for several days. Thus each ampulla has an intrinsic rhythm. Microsurgical experiments suggest that there is no specific region within an ampulla with unique pacemaker properties. It is proposed that communication via gap junctions allows the coordination of ampullar cells into a well organized peristaltic wave.     

THE MECHANISM OF THE NEPHRIDIAL APPARATUS OF PARAMECIUM MULTIMICRONUCLEATUM : II. The Filling of the Vesicle by Action of the Ampullae

Our recent analysis of the nephridial apparatus of Paramecium multimicronucleatum by high-speed cinematography (300 fps at X 250) indicates that before the water expulsion vesicle ("contractile vacuole") is completely voided of fluid during expulsion, the ampullae surrounding and confluent with the vesicle swell with fluid entering from their respective nephridial tubules. Once the membranes of the excretory pore at the base of the excretory canal (leading from the vesicle proper to the outside) have constricted and resealed the excretory pore, the up till then constricted injection tubules of the ampullae which conduct fluid to the vesicle open as waves of contraction along the coacervate gel around the ampulla and proceed along each ampulla from distal to proximal end. The coacervate gel around any one ampulla does not necessarily contract in phase with that of any other ampulla. Each ampulla acts independently. The fluid from the ampullae is thus pumped sequentially, but not in predetermined order, into the water expulsion vesicle, refilling and distending it. Our previous studies (Organ et al., 1968a) suggest that an actomyosinoid ATP-using mechanism may be functional in the ampullary contractions.     

Temporal-spatial co-localisation of tissue transglutaminase (Tgase2) and matrix metalloproteinase-9 (MMP-9) with SBA-positive micro-fibres in the embryonic kidney cortex

Growth of the kidney is a complex process piloted by the collecting duct (CD) ampullae. The dichotomous arborisation and consecutive elongation of this tubular element determines the exact site and time for the induction of nephrons in the overlaying mesenchymal cap condensates. The mechanism by which the CD ampullae find the correct orientation is currently unknown. Recently, we have demonstrated micro-fibres that originate from the basal aspect of the CD ampullae and extend through the mesenchyme to the organ capsule. The micro-fibres are assumed to be involved in the growth and arborisation process of the CD ampulla. Therefore, we have investigated the specific distribution of the micro-fibres during branching morphogenesis. We have also analysed whether the micro-fibres co-localise with extracellular matrix (ECM)-modulating enzymes and whether the co-localisation pattern changes during CD ampulla arborisation. Micro-fibres were detected in all stages of CD ampulla arborisation. Tissue transglutaminase (Tgase2) co-localised with soybean agglutinin (SBA)-positive micro-fibres, whose presence depended upon the degree of CD branching. Matrix metalloproteinase-9 (MMP-9) also co-localised with micro-fibres, but its expression pattern was different from that for Tgase2. Western blotting experiments demonstrated that Tgase2 and MMP-9 co-migrated with SBA-labelled proteins. Thus, the micro-fibres are developmentally modulated by enzymes of the ECM in embryonic kidney cortex. These findings illustrate the importance of micro-fibres in directing CD ampulla growth.     

Ultrastructure of the ampullae of Lorenzini of <Emphasis Type="Italic">Aptychotrema rostrata</Emphasis> (Rhinobatidae)

Small epidermal pores of the electrosensory ampullae of Lorenzini located both ventrally and dorsally on the disk of Aptychotrema rostrata (Shaw and Nodder, 1794) open to jelly-filled canals, the distal end of which widens forming an ampulla that contains 6 ± 0.7 alveolar bulbs (n = 13). The sensory epithelium is restricted to the alveolar bulbs and consists of receptor cells and supportive cells. The receptor cells are ellipsoid and their apical surfaces are exposed to the alveolar lumen with each bearing a single central kinocilium. Presynaptic bodies occur in the basal region of the receptor cell immediately proximal to the synaptic terminals. The supportive cells that surround receptor cells vary in shape. Microvilli originate from their apical surface and extend into the alveolar lumen. Tight junctions and desmosomes connect the supportive cells with adjacent supportive and receptor cells in the apical region. The canal wall consists of two cell layers, of which the luminal cells are squamous and interconnect via desmosomes and tight junctions, whereas the cells of the deeper layer are heavily interdigitated, presumably mechanically strengthening the canal wall. Columnar epithelial cells form folds that separate adjacent alveoli. The same cells separate the ampulla and canal wall. An afferent sensory nerve composed of up to nine myelinated nerve axons is surrounded by several layers of collagen fibers and extends from the ampulla. Each single afferent neuron can make contacts with multiple receptor cells. The ultrastructural characteristics of the ampullae of Lorenzini in Aptychotrema rostrata are very similar to those of other elasmobranch species that use electroreception for foraging.     

Morphology and function of Malpighian tubules and associated structures in the cockroach, Periplaneta americana.

This paper describes the different regions of the Malpighian tubules and the associated structures (ampulla, midgut, ileum) in the cockroach, Periplaneta americana. There are about 150 tubules in each insect. Each tubule consists of at least three parts. The short distal region is thinner than the other parts and is highly contractile. The middle region comprises most of the tubule length and is composed of primary and stellate cells. Primary cells contain numerous refractile mineral concretions, while stellate cells have smaller nuclei, fewer organelles, simpler brush border, and numerous multivesicular bodies. Symbiont protozoa are sometimes present within the lumen of the middle region near where it opens into the proximal region of the tubule. The latter is a short region that drains the tubular fluid into one of the six ampullae. These are contractile diverticula of the intestine located at the midgut-hindgut junction. The ampulla is highly contractile, and consists of a layer of epithelial cells surrounding a cavity that opens into the gut via a narrow slit lined by cells of unusual morphology. The proximal region of the tubule and the ampulla resemble the midgut in that they have similar micromal origin and reabsorptive function for the proximal region of the tubule and for the ampulla. A number of inclusions found within the tubule cells are described, including peroxisomes and modified mitochondria. Current theories of fluid transport are evaluated with regard to physiological and morphological characteristics of Malpighian tubules. The possible role of long narrow channels such as those between microvilli and within basal folds is considered, as is the mechanism by which these structures are formed and maintained. Also discussed is the role of peroxisomes and symbionts in the excretory process.     

Electron microscopic study of cell surface rings during cell division and morphogenesis of Arthrobacter crystallopoietes.

The whole cell ultrastructure during cell division and morphogenesis of Arthrobacter crystallopoietes was monitored using electron microscopic techniques. Glucose-grown spherical cells were inoculated into succinate-based medium. In this medium, the organism undergoes a morphogenetic cycle consisting of elongation of spheres to rods, exponential growth as rods, and fragmentation of rods to spherical cells. Raised bands or rings that encircled the cells were evident on the cell surface of both sphere- and rod-shaped cells. Many rod-shaped cells possessed two or more rings arranged adjacent to each other in a parallel orientation. At each cell division a new ring was formed on both siblings. However, as predicted by the proposed model of unidirectional cell growth and by maintaining a ring from the previous generation, unequal numbers of rings were observed on sibling cells. Only one ring was visible on most of the spherical inoculum cells, but in some cases a second ring perpendicular to the other ring was observed. Parallel rings were found on spherical cells resulting from fragmentation or reductive cell division of rods during the stationary growth phase. Thus, these spheres could be distinguished from inoculum spheres containing a single ring or perpendicular orientation of rings. The number of rings per cell and arrangement of rings on the cell surface of sibling cells after cell division, but before cell separation, are discussed with respect to cell age, cell division, and sphere-rod-sphere morphogenesis of A. crystallopoietes.     

Structure and Role of the Renette Cell and Caudal Glands in the Nematode Sphaerolaimus gracilis (Monhysterida)

Ultrastructure of the renette cell and caudal glands was studied in the free-living aquatic nematode Sphaerolaimus gracilis. The renette cell occurred posterior to the esophageal-intestinal junction and opened through an ampulla to a ventral pore behind the nerve ring. The caudal gland system of the tail consisted of two gland cells opening through separate pores and 2 to 3 other gland cells of a different type opening through a common pore. The renette cell and the two caudal gland cells were similar and both contained secretory granules, 0.5-1.5 μm in diameter. The material released attached the nematode to the substrate. The renette ampulla was surrounded by a specialized cell, the ampulla cell, which had characteristics of myoepithelium. A plug or valve structure connected to the ampulla cell may regulate the output of the secretory material. The ampulla cell is able to contract and thus is probably under direct neuronal control. Other cells in the renette ampulla region of body cavity were termed supporting cells. Living, cold-relaxed nematodes were attached to sediment particles in the renette pore region and at the tail tip. Release from sediment particles was mechanical at the renette cell discharge site but appeared to be chemical at the caudal gland. In behavioral experiments, nematodes in a water current had the ability to release a thread from the caudal glands while maintaining contact with a sediment particle attached to the tail end. If the thread was strong enough, it also could be used to change location. Nematodes anchored by the thread from the caudal glands to a sediment particle could float in water currents until they attached themselves to another sediment particle with the help of secretions from the renette cells.     

A morphological analysis of the ampullae of Lorenzini in selected skates (Pisces,Rajoidei)

A comparison of the ampullae of Lorenzini among 40 species of skates (Rajoidei) demonstrates a close relationship between inferred electroreceptive capabilities and feeding mechanisms. Three general lines of morphological modifications are noted. (1) Whereas the majority of ampullary pores are located on the ventral surface of the dorsoventrally flattened body, the relative proportion of ventral pores is significantly lower on species inhabiting aphotic waters. (2) The ventral pores on more piscivorous species are distributed over a larger portion of body surface than they are on those species that feed primarily on invertebrates. Ventral pores in this latter group are more noticeably concentrated around the mouth and their densities on the adult are inversely related to the overall mobility of preferred prey species. (3) The size of each ampulla and the number of alveoli associated with it are directly related to the habitat depth occupied by each species. Shallow-water species have smaller ampullae with fewer alveoli than deeper-dwelling (> 1,000 m) species. The general distribution of ampullary pores on deep dwelling rajoids appears to compensate for reduced visual input, whereas their relative densities are a measure of the system's resolution and reflect major differences in feeding strategies. The increased ampullary size and complexity observed in deep-sea rajoids provides mechanisms to increase both the sensitivity and signal-to-noise ratios.     

Ecological Consequences of Altering the Timing Mechanism for Metamorphosis in Anural Ascidians

SYNOPSIS. In the present study the timing of metamorphosis inan anural ascidian, Molgula pacifica, was compared to metamorphosisin a urodele species Boltenia villosa. Metamorphosis in M. pacificawas triggered at a fixed time in development (32–36 hoursafter fertilization), just prior to hatching. In contrast, metamorphosiswas triggered in B. villosa after the hatched larvae respondedto substrate cues. The timing of metamorphosisin B. villosawas often delayed for up to four days, whereas delays in M.pacifica were not observed. An antibody, termed Epi-3, was foundto cross-react exclusively with epidermal cells in both species.The binding of FITC-labelled Epi-3 was very low prior to metamorphosisand then it increased dramatically after metamorphosis was triggered.The cytoplasm of ampulla tip cells and the tunic immediatelysurrounding each ampulla showed the highest levels of Epi-3fluorescence. The histological and ultrastructural featuresof the ampulla cells suggest that Epi-3 antibody recognizesgranules localized in the apical cytoplasm. How the evolution of an internal "clock" mechanism responsiblefor initiating metamorphosis may be beneficial to anural speciesis discussed. One possibility is that the anural type of timingmechanism reduces mortality rates during this critical phaseof its life cycle.     

Ultrastructure of the proximal region of somatic cilia in Paramecium tetraurelia

The morphology of the transition zone between the terminal plate of the basal body and the 9 + 2 region of the somatic (non-oral) cilium has been examined in Paramecium tetraurelia. Freeze-fracture and thin- section techniques disclosed both membrane specializations and various internal structural linkages. Freeze-fracture material revealed sets of particles interrupting the unit membrane. The more distal of these form plaquelike arrays while the proximal set of particles forms the ciliary "necklace." The plaque regions correspond to anionic sites on the outer membrane surface as revealed by binding of polycationic ferritin. Both the plaque particles and the necklace particles appear to be in contact with outer doublet microtubules via a complex of connecting structures. In the interior of the transition zone an axosomal plate supports an axosome surrounded by a ring of lightly packed material. Only one of the two central tubules of the axoneme reaches and penetrates the axosome. Below the axosomal plate four rings, each approx. 20 nm wide, connect adjacent outer doublets. An intermediate plate lies proximal to these rings, and a terminal plate marks the proximal boundary of this zone. Nine transitional fibers extend from the region of the terminal plate to the plasmalemma. The observations described above have been used to construct a three-dimensional model of the transition region of "wild-type" Paramecium somatic cilia. It is anticipated that this model will be useful in future studies concerning possible function of transition-zone specializations, since Paramecium may be examined in both normal and reversed ciliary beating modes, and since mutants incapable of reverse beating are available.     

Functional anatomy of the valves in the ambulacral system of sea urchins (Echinodermata,Echinoida)

Summary The water vascular system of sea urchins is examined with special reference to the valves positioned between the radial vessel and the ampullae of the tube feet. The lips of the valve protrude into the ampulla. Thus the valve functions mainly like a check valve that allows the unidirectional flow of fluid towards the ampulla. Each ampulla-tube foot compartment acts as a semi-autonomous hydraulic system. The lumina of the ampulla and the tube foot are lined with myoepithelia except for the interconnecting channels that pierce the ambulacral plate. The contraction of the ampulla results in an increasing hydraulic pressure that protrudes the tube foot, provided that the valve is closed. The retraction of the tube foot results in a backflow of fluid independent of the condition of the valve. The lips of the valve are folds of the hydrocoel epithelium. The pore slit lies in the midline. The perradial faces of the lips are covered with the squamous epithelium of the lateral water vessel. The ampullar faces are specialized parts of the ampulla myoepithelium. Turgescent cells which form incompressible cushions take the place of the support cells. The valve myocytes run parallel to the pore slit and form processes that run along the base of the ampulla and the perradial channel up to the podial retractor muscle. The findings lead to the hypothesis of multiple control of the ampulla-tube foot system: (1) The mutual activity of the ampulla and the tube foot is indirectly controlled by the lateral and podial nerves which release transmitter substances that diffuse through the connective tissue up to the muscle layers. (2) A muscle-to-muscle conduction causes the simultaneous contraction of the ampulla or the podial retractor muscles. (3) The valve muscles are directly controlled by the processes of the valve myocytes which make contact with the podial retractor. In extreme conditions a backflow of hydrocoel fluid towards the radial water vessel occurs.     

M ring, S ring and proximal rod of the flagellar basal body of Salmonella typhimurium are composed of subunits of a single protein, FliF.

The flagellar basal body, a major part of the flagellar motor, consists of a rod and four rings. When the fliF gene of Salmonella typhimurium, which was previously shown to code for the component protein of the M ring, was cloned and overexpressed in Escherichia coli, the FliF subunits formed ring structures in the cytoplasmic membrane. Electron microscopic observation of the purified ring structures revealed that each was composed of two adjacent rings and a short appendage extending from the center of the rings. Antibodies raised against the purified FliF protein decorated both the M and S rings of the intact basal body. We conclude that the FliF protein is the subunit protein of the M ring, and of the S ring and of part of the proximal rod of the flagellar basal body.     

Dynamic apical surface rings in superficial layer cells of koi Cyprinus carpio scale epidermis

This study examined the novel ring‐shaped structures found in the apical surface of individual cells of the scale epidermis of koi Cyprinus carpio. These apical rings are highly dynamic structures with lifetimes ranging from a few to several minutes. While several ring forms were observed, the predominant ring morphology is circular or oval. Two distinct ring forms were identified and designated type I and type II. Type I rings have a well‐defined outer border that encircles the surface microridges. Type II rings are smooth‐surfaced, dinner‐plate‐like structures with membranous folds or compressed microridges in the centre. Type II rings appear less frequently than type I rings. Type I rings form spontaneously, arising from swollen or physically interrupted microridges but without initially perturbing the encircled microridges. After persisting for up to several minutes the ring closes in a centripetal movement to form a circular or irregular‐shaped structure, the terminal disc. The terminal disc eventually disappears, leaving behind a submembranous vesicle‐like structure, the terminal body. Type I rings can undergo multiple cycles of formation and closing. Recycling epidermal apical rings form through centrifugal expansion from the terminal disc followed by apparent contraction back to the disc structure, whereupon the cycle may repeat or cease. The findings demonstrate a novel skin surface structure in fishes and are discussed with respect to communication with the external aqueous environment.     

Initial diameter of the polar body contractile ring is minimized by the centralspindlin complex

Polar body formation is an essential step in forming haploid eggs from diploid oocytes. This process involves completion of a highly asymmetric cytokinesis that results in a large egg and two small polar bodies. Unlike mitotic contractile rings, polar body contractile rings assemble over one spindle pole so that the spindle must move through the contractile ring before cytokinesis. During time-lapse imaging of C. elegans meiosis, the contractile ring moved downward along the length of the spindle and completed scission at the midpoint of the spindle, even when spindle length or rate of ring movement was increased. Patches of myosin heavy chain and dynamic furrowing of the plasma membrane over the entire embryo suggested that global cortical contraction forces the meiotic spindle and overlying membrane out through the contractile ring center. Consistent with this model, depletion of myosin phosphatase increased the velocity of ring movement along the length of the spindle. Global dynamic furrowing, which was restricted to anaphase I and II, was dependent on myosin II, the anaphase promoting complex and separase, but did not require cortical contact by the spindle. Large cortical patches of myosin during metaphase I and II indicated that myosin was already in the active form before activation of separase. To identify the signal at the midpoint of the anaphase spindle that induces scission, we depleted two proteins that mark the exact midpoint of the spindle during late anaphase, CYK-4 and ZEN-4. Depletion of either protein resulted in the unexpected phenotype of initial ingression of a polar body ring with twice the diameter of wild type. This phenotype revealed a novel mechanism for minimizing polar body size. Proteins at the spindle midpoint are required for initial ring ingression to occur close to the membrane-proximal spindle pole.     

Spatiotemporal mapping of ex vivo motility in the caecum of the rabbit

We used high definition radial, strain rate and intensity spatiotemporal mapping to quantify contractile movements of the body and associated structures of the rabbit caecum when the terminal ileum was being perfused with saline at a constant rate. This perfusion caused gradual distension of the caecum as a result of relative restriction of outflow from the ampulla caecalis. The body of the caecum exhibited two patterns of motility that appeared autonomous, i.e. occurred independently of any contractile activity at the inlet or outlet. Firstly, the pattern that we termed ladder activity consisted of an orderly sequential contraction of bundles of axially oriented circular muscle between the spiral turns of longitudinal muscle and proceeded either from base to tip or from tip to base at a similar frequency and velocity. Secondly, less-localised, rapidly propagating synchronous contractions of both circular and longitudinal muscle, which were more common when the caecum was distended, that were termed mass peristalsis. Movements of the ileum and sacculus rotundus occurred at the same frequency and were broadly coordinated. Distension of the distal sacculus occurred synchronously with contraction of the ileum and did not propagate in an orderly manner across the structure, i.e. was instantaneous. This pattern was consistent with hydrostatic distension. Contractions propagated through the ampulla caecalis in either an orad or an aborad direction at a similar frequency to, and broadly correlated with, those in the ileum. The frequencies of distension of the sacculus and of contraction in the ileum and ampulla were momentarily augmented during mass peristalsis. The authors conclude that there was some coordination between the contractile activity of the terminal ileum and the caecal ampulla during periods of ongoing inflow from the ileum and between these structures and the caecum during mass peristalsis.     

Tunic cells in pyrosomes (Thaliacea, Urochordata): cell morphology, distribution, and motility

Abstract. Cellular components of the tunic were histologically examined in 3 pyrosome species representing all 3 genera of the order: Pyrosoma atlanticum, Pyrosomella verticillata , and Pyrostremma spinosum . Three cell types are distributed in the tunic. Tunic amebocytes, irregularly shaped and motile, often contain granules and/or phagosomes. Spherical tunic cells contain many round vesicles with eosinophilic and acidic materials. Tunic net cells form a cellular network in which their long filopodia connect with one another. The net cells are densely distributed just beneath the tunic surface lining the common cloacal cavity and may produce tension to maintain the colony shape. The presence of net cells suggests a phylogenetic relationship between pyrosomes and some aplousobranch ascidians. Test fibers are multicellular cords that run in the tunic and connect the zooids. In P. atlanticum , they are attached to they are attached to the epidermal cells of the zooids, and transverse cloacal muscles are attached to the other (proximal) side of the epidermal cells. The test fibers may mediate coordination of the zooids and control muscle contraction.     

On the ampullary organs of the South-American paddle-fish Sorubim lima (Siluroidea,Pimelodidae)

Summary Ampullary organs were found in the epidermis of the paddle-fish Sorubim lima; they are distributed all over the skin surface of the fish but are particularly densely grouped in the head region and on the dorsal surface of the paddle. Histological and electron microscopical observations show that their structure is similar to the type of cutaneous ampullary organs characteristic of other Siluroidea. Composed of a relatively large mucus-filled ampulla, the organ possesses a short and narrow canal which leads to the outer epidermal surface. The wall of the ampulla is formed of several layers of flat epidermal cells. In general four sensory cells, each one surrounded by supporting cells, compose the sensory epithelium at the bottom of the ampulla. The inner surface of the sensory cells in contact with the ampullary mucus bears only microvilli. The contact between the nerve endings and the sensory cells show the characteristic structure of an afferent neuro-sensory junction. Two ampullae are innervated in some cases by the same afferent nerve fibre.The author expresses her gratitude to Dr. Szabo for his scientific advice during her stay in Gif sur Yvette     

Relating tree growth to rainfall in Bolivian rain forests: a test for six species using tree ring analysis

Many tropical regions show one distinct dry season. Often, this seasonality induces cambial dormancy of trees, particularly if these belong to deciduous species. This will often lead to the formation of annual rings. The aim of this study was to determine whether tree species in the Bolivian Amazon region form annual rings and to study the influence of the total amount and seasonal distribution of rainfall on diameter growth. Ring widths were measured on stem discs of a total of 154 trees belonging to six rain forest species. By correlating ring width and monthly rainfall data we proved the annual character of the tree rings for four of our study species. For two other species the annual character was proved by counting rings on trees of known age and by radiocarbon dating. The results of the climate–growth analysis show a positive relationship between tree growth and rainfall in certain periods of the year, indicating that rainfall plays a major role in tree growth. Three species showed a strong relationship with rainfall at the beginning of the rainy season, while one species is most sensitive to the rainfall at the end of the previous growing season. These results clearly demonstrate that tree ring analysis can be successfully applied in the tropics and that it is a promising method for various research disciplines.     

Test cell migration and tunic formation during posthatching development of the larva of the ascidian, Ciona intestinalis

Morphological changes in the tunic layers and migration of the test cells during swimming period in the larva of the ascidian, Ciona intestinalis , were observed by light and electron microscopy. The swimming period was divided into three stages. In stage 1, further formation of juvenile tunic layer started only in the larval trunk and neck region. In stage 2, the layer became swollen in the ventral and dorsal sides of the neck region and in stage 3, the swelling expanded backward. Concomitantly with these changes, the outermost larval tunic layer (outer cuticular layer), which had been formed before hatching, also swelled in the neck region in stage 2 and formed two humps in stage 3, although the layer did not change in the tail region during the swimming period. Test cells that were present over the entire larval tunic layer in stage 1 began to move from the surface of the fin toward that of the side of the body in stage 2, and finally gathered to form six bands running radially from the anterior end to the posterior end of the trunk region and aligned along the lateral sides of body in the tail region in stage 3. In electron microscopic observations, pseudopodia protruding from the test cells invaded the larval tunic, following which they extended proximate to the juvenile tunic in the trunk region. In the tail region, which had no juvenile tunic layer as that described, the pseudopodia invaded and remained adjacent to the surface of the epidermis or the sensory cilia protruded from the epidermis. Metamorphosis of the larvae, further tunic formation, degradation of adhesive papilla, attachment of larva to the substratum and tail resorption commenced after these morphological changes occurred. The possible role of the test cells in metamorphosis is discussed.