Outer Dendritic Segments Research Articles

Cellular organization and fine structure of type II microtrich sensilla in gammaridean amphipods (Crustacea)

The cellular organization of type II microtrich sensilla was studied in male Anonyx lilljeborgi Boeck, 1871 (Lysianassoidea) by light and transmission electron microscopy. The sensillum consists of two bipolar sensory neurons and three concentric sheath cells. The sensory cell bodies are subepidermal. In each sensillum both dendrites are enclosed by the thecogen cell process. The inner dendritic segments are filled with mitochondria and lucent vesicles and expand in the epidermis into a spindle-shaped swelling. One of the neurons gives rise to two cilia and the second to a single cilium. These three outer dendritic segments lie in the receptorlymph cavity. The dendritic sheath, secreted by the thecogen cell process, completely ensheaths the outer dendritic segments. The trichogen (middle) cell and the tormogen (outer) cell incompletely enclose the thecogen cell, but their processes form autojunctions around the dendritic sheath in the apical epidermis. In premolt, the trichogen cell processes project into the exuvial space. The cytoplasm of the tormogen cell and the bordering epidermal cells contains coarse osmiophilic inclusions. All the cells of the sensillum are joined by desmosomes. The sensilla structurally resemble chemosensory (gustatory) insect sensilla.

A Lobster Phospholipase C-β That Associates with G-Proteins in Response to Odorants

A cDNA clone encoding a protein of 1116 amino acids with significant homology to beta-isoforms of phospholipase C was isolated from lobster olfactory organ cDNA libraries and named lobPLCbeta. This cDNA hybridized predominantly to a 9 kb transcript in RNA from olfactory organ, pereiopod, brain, and eye-eyestalk and to several smaller minor transcripts only in eye-eyestalk. An antiserum raised to the C terminus of lobPLCbeta detected immunoreactivity in a single 130 kDa band in olfactory aesthetasc hairs, olfactory organ, pereiopod, dactyl, and brain. In eye-eyestalk this 130 kDa band was abundant, and minor bands of 100, 79, and 57 kDa also were detected. In cross sections of the aesthetasc hairs, immunoreactivity was detected in the outer dendritic segments of the olfactory receptor neurons, the site of olfactory transduction. A complex odorant caused lobPLCbeta immunoreactivity to increase in membrane fractions and decrease in soluble fractions of homogenates of aesthetasc hairs. The odorant also increased the amount of lobPLCbeta in immunoprecipitates of Galphaq and Gbeta from homogenates of aesthetasc hairs. These results support the conclusion that lobPLCbeta mediates olfactory transduction.

TEM studies on the lattice organs of cirripede cypris larvae (Crustacea, Thecostraca, Cirripedia)

Lattice organs consist of five pairs of sensory organs situated on the dorsal carapace in cypris larvae of the Crustacea Cirripedia. The lattice organs in cypris larvae of Trypetesa lampas (Acrothoracica) and Peltogaster paguri (Rhizocephala) represent the two main types found in cirripedes, but only minor differences exist at the TEM level. Each lattice organ is innervated by two bipolar, primary receptor cells. The inner dendritic segment of each receptor cell carries two outer dendritic segments. The outer dendritic segments contain modified cilia with a short ciliary segment (9×2+0 structure). Two sheath cells envelop the dendrite except for the distal ends of the outer dendritic segments. This distal end enters a cavity in the carapace cuticle and reaches a terminal pore situated at the far end of the cavity. The cuticle above the cavity is modified. In both species the epicuticle is partly perforated by numerous small pores and the underlying exocuticle is much thinner and less electron dense than the regular exocuticle. Lattice organs very probably have a chemosensory function and are homologous with the sensory dorsal organ of other crustacean taxa.

Development of specific responses in antennal taste hairs after ecdysis. An electrophysiological investigation of the cockroach, Periplaneta brunnea

Spike generation in cockroach antennal taste hairs of nymphs appears to follow the same rules as in adults analysed previously. After ecdysis, taste hairs on the proximal segments responded earlier than those on distal ones. Shorter hairs became active before longer ones. The mechanoreceptor responded before the chemoreceptors. During a transient period, 3–5 hours after ecdysis, all four chemoreceptors and also the mechanoreceptor were sensitive to 150 mM KCl. Spikes elicited by 75 mM KCl were generated predominantly in the outer dendritic segments. They did not differ in shape and frequency when 60 mM sucrose was added. The earliest sign of a specific reaction to sucrose was observed approximately 4–5 hours after ecdysis, when the mixture of sucrose and KCl elicited more spikes than KCl alone.

Functional morphology of a double-walled multiporous olfactory sensillum: the sensillum coeloconicum of Bombyx mori (Insecta, Lepidoptera)

The fine structure of coeloconic sensilla of Bombyx mori was studied in cryofixed specimens. These sensilla belong to the category of double-walled wall-pore sensilla. The pegs are ∼10 μm long, located in pits on the dorsal side of the antennal branches, and longitudinally grooved in their distal half (grooved surface ∼30 μm 2). The central lumen contains the outer dendritic segments of usually five receptor cells, and is surrounded by up to 15 partially fused cuticular fingers. The peripheral lumina of these cuticular fingers are filled with material resembling wax-canal filaments. Radial spoke channels (∼600 per peg), each 10–20 nm wide, connect the central lumen with the longitudinal groove channels. Groove and spoke channels are assumed to mediate the transport of odorant molecules from the outer epicuticular surface layers to the sensory dendrites. Thus the double-walled wall-pore sensilla represent a bauplan essentially different from single-walled wall-pore sensilla; the reason, however, why the two types are found together throughout the insect orders remains enigmatic. Other peculiar features of the coeloconic sensilla of the silkmoth are invaginations of the outer dendritic segments and direct contacts between the receptor cell somata. The latter may be the structural correlate to electrophysiological observations indicative of peripheral interaction between the receptor neurons. All three auxiliary cells have elaborately folded apical plasma membranes studded with portasomes and associated with an abundance of mitochondria; basally they often contact tracheal branches. As compared to the auxiliary cells of the single-walled olfactory sensilla of the same species, all the mentioned features are much more prominent and hint to a higher ion pumping activity at the border to the sensillum-lymph cavities.

Fine structure and physiology of the infrared receptor of beetles of the genus Melanophila (Coleoptera: Buprestidae)

Buprestid beetles of the genus Melanophila (Coleoptera: Buprestidae) possess paired thoracic pit organs. Each pit houses about 70 tightly packed infrared sensilla that enable the beetles to detect forest fires at long range. The cuticular apparatus of a single infrared sensillum consists of an endocuticular sphere with an average diameter of 15 μm. Each sphere is innervated from below by the dendritic outer segment of a single sensory neuron that shows the same ultrastructure as a typical hair mechanoreceptor (e.g., sensillum trichodeum). Several transitional stages between cuticular mechanoreceptors and infrared sensilla can be found in the neighbourhood of the infrared sensillum field (intermediate sensilla and so-called suppressed systems). First electrophysiological recordings show that the receptors at the bottom of the pit are true infrared receptors. The sensilla respond in a fast phasic manner to infrared radiation and even an exposition time of 2 ms is sufficient to release a single spike. All morphological and physiological findings suggest that the infrared sensilla have evolved from ordinary cuticular mechanoreceptors and that mechanical events are still part of the transduction process. A model about the possible function of the infrared receptors of Melanophila is presented.

Fine Structure of the Putative Pheromone-Sensitive Sensilla on the Antennae of Andraca bipunctata (Lepidoptera: Bombycidae)

The antennae of Andraca bipunctata Walker are sexually dimorphic; those of males are bipectinate with 43 pairs of branches, and those of females are filiform without branches. In both sexes, most sensilla are on the frontal (windward, ventral) side, but the types of sensilla differ in the 2 sexes. The main type in males is sensillum trichodeum (s. trichodeum), and in females are s. basiconicum and the short s. chaeticum. Each putative pheromone-sensitive sensillum, trichodeum, is innervated by 2 bipolar neurons containing many mitochondria, multivesicular bodies, rough endoplasmic reticulum, microtubles, and lamellar bodies. The dendrite is subdivided into inner and outer dendritic segments between the ciliary region, which is characterized by proximal and distal bodies. The outer dendritic segment, which contains microtubules, penetrates the cuticular canal and extends into the hair shaft. Outside the neurons, thecogen, trichogen, and tormogen cells surround the neural bodies, forming the inner, intermediate, and outer sheath below the hair shaft. This s. trichodeum has multipores and each pore has 2-3 pore tubules within the pore canal. These pore tubules extend into the hair lumen and probably help the pheromone molecules contact the dendritic membrane receptor, because most pheromones are water insoluble.

Structure of the aesthetasc (olfactory) sensilla of the blue crab, Callinectes sapidus : transformations as a function of salinity

The aesthetasc sensilla of the euryhaline blue crab, Callinectes sapidus, are innervated by the dendrites of from 40 to 160 bipolar chemosensory neurons. Each dendrite forms two cilia within the basal portion of the sensillum, and these subsequently branch yielding approximately 10 outer dendritic segments per neuron. Auxiliary cells surround the inner dendritic segments and also ensheathe the outer dendritic segments up to the terminus of the ”constricted region” (a zone in which there is a slight narrowing of the aesthetasc). Crystal violet staining suggesting access of odor stimuli is limited to that portion of the sensillum distal to the constricted region. In freshwater-acclimated blue crabs the length and level of branching in the dendrites extending beyond the constricted region is significantly reduced relative to that of seawater-acclimated animals (mean lengths: 150 µm versus 517 µm, respectively). After transfer of freshwater-acclimated crabs to seawater there is a rapid increase in length of the outer dendritic segments, reaching 60% of that for seawater-acclimated crabs by 48 h. A similar time course for regrowth is seen for seawater-acclimated crabs in which the outer dendritic segments have been osmotically ablated. Conversely, with rapid transfer of seawater-acclimated animals to lower salinities, there is a correspondingly rapid reduction in length of the outer dendritic segments. The reduced length of the outer dendritic segments in freshwater-acclimated animals may reflect the effective distance over which an appropriate osmotic/ionic microenvironment for neural function can be maintained within the aesthetasc.

STRUCTURE AND DISTRIBUTION PATTERNS OF AESTHETASCS AND MALE-SPECIFIC SENSILLA IN LOPHOGASTER TYPICUS (MYSIDACEA)

The aesthetasc sensilla of the mysid Lophogaster typicus are distributed on the lateral flagellum of the antennules (first antennae). They essentially conform to the typical aesthetasc structure. Compared to aesthetascs described in other crustaceans, there are divergences in this structure: there is only 1 outer dendritic segment per sensory cell, and the outer part of the hair is flattened in Lophogaster. In males, the lateral flagella also carry a varying number of slender sensilla, located between the aesthetascs. The male-specific sensilla are innervated by monoand biciliary sensory neurons. The phylogenetic implications of the morphology and distribution patterns of the aesthetascs and male-specific sensilla are considered.

Responses and spike generation in the largest antennal taste hairs of Periplaneta brunnea Burm.

The largest taste hair type of the 20 distal antennal segments is investigated electrophysiologically. Two chemoreceptors respond to potassium chloride, tripotassium citrate but not to choline salts, another one to sucrose, but not to D-glucose and only slightly to D-fructose. No response was obtained with pure water. Spikes with a leading negative phase indicate that the primary process controls an apical spike generator localized in the membranes of the outer dendritic segment. The succeeding basal spike consists of two positive phases, that are interpreted as the result of two further spike generation zones, one at the dendritic inner segments and the other at the origin of the axon. Spike activity in the mechanoreceptor also starts with negative going spikes, that are followed by a series of solely positively going ones. TTX applied to the hair tip inhibits the apical spike generator of the chemo- and mechanoreceptor cells.

Correlation between dendrite diameter and action potential amplitude in sex pheromone specific receptor neurons in male Ostrinia nubilalis (Lepidoptera: Pyralidae)

Outer dendritic segments of olfactory receptor neurons tuned to sex pheromone components were measured morphometrically on the antenna of male European corn borers. Ostrinia nubilalis, to determine if a correlation exists between the diameter of the outer dendritic segment and the spike amplitude. The olfactory sensilla investigated each contained three receptor cells. Two cells were each specific for one of the two pheromone components, (Z)-11-tetradecenyl acetate (Z11-14:OAc) and ( E)-11-tetradecenyl acetate (E11-14:OAc). Two strains of cornborers (Z and E) differ as to which of the two pheromone components is the main one. In both strains a large difference could be observed between the spike amplitudes elicited in the receptor cells by the two pheromone components, the main component always eliciting the large spike. In F1-hybrids (EZ) of these two strains, producing both pheromone components in similar quantities, the spike amplitudes were equal in the two pheromone-specific receptor cells. The third cell responded specifically to a behavioural antagonist. ( Z)-9-tetradecenyl acetate (Z9-14:OAc) in both the parental and hybrid strains, and always showed the smallest spike amplitude. In a morphometric study, the outer dendritic segments were shown to differ more in diameter between the largest and second largest cell in the two parental strains than in the hybrid strain, while the smallest diameter cell did not differ between the different strains. These results imply that receptor cells with larger diameter produce spikes with greater amplitude. The data also show that all three types of receptor neurons display outer dendritic segments with strong variation in the diameter along the length of the segment, and with a pronounced taper towards the tip.

The tuft organs of the human body louse, pediculus humanus corporis—cryofixation study of a thermo-/ hygrosensitive sensillum

The tuft organs of the human body louse, Pediculus humanus corporis, display a complex cuticular apparatus without wall pores, consisting of the cuticular cone, cuticular pocket, and the sensillar peg with 5–6 tuft processes. The cuticular wall of the sensillar peg exhibits several layers of differing electron density tightly surrounding the dendrites. The sensillar peg is innervated by the unbranched dcndrites of two receptor cells which do not reach into the tuft processes. A third receptor cell forms a lamellar outer dendritic segment which wraps around the two other dendrites and terminates before reaching the base of the sensory peg. This type of receptor cell is named stratoterminal while for those which end within the cuticular apparatus the term conoterminal is proposed. Five auxiliary cells are associated with each tuft organ. The pore organs display a porous cuticular plate innervated by the branching dendrites of four receptor cells. A fifth receptor cell terminates more proximal and resembles the stratoterminal cell of the tuft organ. From the fine structure it is evident that the lull organs represent thermo-hygrosensitive sensilla whereas the pore organs may have an olfactory function.

The Fine Structure of Trichobothria in Moss Mites with Special Emphasis onAcrogalumna longipluma(Berlese, 1904) (Oribatida, Acari, Arachnida)

AbstractUsing SEM and TEM techniques, the trichobothria ofAcrogalumna longipluma(Galumnidae, Pterogasterina) were studied in relation to a variety of other moss mites. Each sensillum is composed of a mostly solid bothridial seta and a very complex socket, the bothridium. The setal base bends sharply as it passes through the six chambers of the bothridium, which are arranged in an S‐shaped configuration. The proximal end of the seta is an elongated oval and inserted into a thin socket membrane provided with radiating suspension fibres. This peculiar shape of the setal base proper and probably also the existence of connecting pieces are assumed to provide directionality to the sensillum. Two differently shaped tubular bodies are found under the setal base, of which only one is in contact with the seta. The tubular bodies are surrounded by peculiar ‘dense tubes’, i.e. derivatives of the dendritic sheath. The rather thick, outer dendritic segments curve through an extensive receptor lymph cavity and terminate with ciliary regions. The inner dendritic segments are only short. Perikarya and axons as well as the enveloping cells do not show any peculiarities. The trichobothria of moss mites most probably represent vibration receptors reacting to substrate and/or air‐borne stimuli. The variety of shapes and the complexity, which is not found to this extent in any other arthropod group, are discussed in relation to ecophysiological demands.

Ultrastructural localization of 5'AMP odorant receptor sites on the dendrites of olfactory receptor neurons of the spiny lobster

A unique probe--biotinylated adenosine-5'-monophosphate (5'AMP-biotin)--was used in transmission electron microscopic (TEM) studies to localize 5'AMP odorant binding sites on the dendrites of olfactory receptor neurons in the aesthetasc sensilla of the spiny lobster, Panulirus argus. This probe is capable of both binding to and exciting 5'AMP-sensitive olfactory receptor neurons, as revealed through biochemical and electrophysiological assays. TEM studies showed that 5'AMP-biotin binding sites are distributed along the entire dendritic region that is exposed to odorants, including the transitional zone (between the inner and outer dendritic segments, including the ciliary segment) and all of the outer dendritic segment. The density of 5'AMP binding sites per micron2 of membrane is similar along the length of the olfactory dendrite. However, the relative number of 5'AMP-biotin binding sites per micron2 of sensillar area diminishes in the distal 30% of the aesthetasc due to a decrease in the amount of dendritic membrane in that region. The distribution of these 5'AMP binding sites is therefore much more extensive than that of enzymes that inactivate 5'AMP--5'ectonucleotidase/phosphatase--which are restricted to the transitional zone (Gleeson et al., 1991). Taken together, these results suggest that 5'AMP-biotin is labeling 5'AMP-specific olfactory receptor sites that are located along the entire outer dendritic segment and that can be coupled to olfactory transduction. This study represents the first in situ localization of specific olfactory receptor sites using a specific, functionally defined ligand.

Signal transmission in lobster olfactory receptor cells: Functional significance of electrotonic structure analysed by a compartmental model

The electrotonic structure of lobster olfactory receptor cells was evaluated using general purpose simulation software in a compartmental model derived from electron-microscopic reconstruction. The model with non-uniform membrane resistance (R m) was used to (i) simulate current spread and (ii) determine if the electronic structure of the cell improves signal recognition in the soma. The odor-evoked conductance change in dendrites was calculated according to the Michaelis-Menten equation with the assumption that the outer dendritic segments function as independent stimulus detectors. The inflection point of the concentration-response function measured in the soma was shifted to lower concentrations relative to that measured in the ciliary (outer dendritic) arbor. The shift, which was greater for inputs with lower efficacy (represented in the model by smaller Hill coefficients) and for the dynamic phase of the response than for the steady-state phase, effectively increased the selectivity of the somatic response. Randomized input distributed uniformly to progressively more restricted areas of the ciliary arbor showed that stimulation of larger areas (presumably the entire ciliary arbor) decreased the statistical variability of the somatic response.

Ultrastructure and distribution patterns of sensilla on the palps of caddisflies (Trichoptera)

Sensilla on the palps of 23 species of Trichoptera representing 16 families, were examined by scanning and transmission electron microscopy. Most of the sensilla found can be assigned to well-known categories, such as sensilla chaetica (mechanosensory and bimodal mechanosensory/gustatory), campaniformia (cuticular stress-receptors), and a variety of multiporous olfactory sensilla. Previously undescribed types are gustatory and mechanosensory receptors in the “apical cone complex” of Rhyacophiloidea, and poreless sensilla, each containing one single receptor cell with a lamellated outer dendritic segment. The ultrastructural similarities between these and the sensilla basiconica found in the labial palp pit organs (LPO) of Lepidoptera is discussed.

Fine structure of olfactory sensilla in myriapods and arachnids.

Structural features of various types of olfactory sensilla are reviewed. 1) Sensilla basiconica which differ in form and size are found on the antennae of centipedes and millipedes. Their walls show longitudinal slits or grooves that either open into the sensillum lumen or do not penetrate the cuticle. In other such sensilla the outer surface is pierced by pores and the inner surface grooved and pocketed. These sensilla are innervated by one to six sensory cells. Their unbranched outer dendritic segments extend to the tip of the sensillum. The sensory cells are surrounded by two or three sheath cells which terminate at the sensillum base or form a continuous tube around the entire length of the outer dendritic segments. 2) Temporal organs of centipedes are located between the insertion of the antenna and the ocelli. These sensilla consist of a shallow cuticular ring with a central sensory plate made up by a layer of unperforated cuticle or a capsule with a mushroom-shaped structure inside formed by fibrous-looking cuticle. A dozen sensory cells with unbranched outer dendritic segments innervate each sensillum. They extend toward the sensory cuticle and pass just below it. Numerous sheath cell processes run parallel to the outer dendritic segments up to the sensory cuticle. 3) Thread-like flagella of Pauropoda are found on the antennae. They possess a flexible unperforated cuticular wall. These sensilla contain nine sensory cells surrounded by several sheath cells which form a continuous cytoplasmic tube around the outer dendritic segments. 4) Single-walled sensilla with numerous plugged pores penetrating the cuticular wall occur on the tarsus of the first leg in ticks. Each sensillum is innervated by 4-15 sensory cells. Three sheath cells terminate in the base of the sensillum. 5) Double-walled sensilla with spoke canals are found on the first tarsus of ticks. Their shaft is longitudinally grooved. Pore canals lead inward from the bottom of the grooves and open into vase-shaped chambers. From its base these canals extend into the lumen of the sensillum which contains unbranched outer dendritic segments of 1-2 sensory cells. 6) Single-walled sensilla with pore openings occur on the distal tarsal segments of the first leg of whip spiders. These sensilla are innervated by 40-45 sensory cells. Their unbranched outer dendritic segments fill the shaft lumen and extend partly into the wall pores. Microvillus-shaped sheath cell processes line the inner surface of the cuticular wall.(ABSTRACT TRUNCATED AT 400 WORDS)

Ecto-ATPase/phosphatase activity in the olfactory sensilla of the spiny lobster, Panulirus argus: localization and characterization.

Electrophysiological studies have shown that the olfactory organ (antennule) of the spiny lobster, Panulirus argus, has chemoreceptors that are selectively excited by adenine nucleotides in seawater. Biochemical studies have revealed that these same nucleotides can be rapidly dephosphorylated by ectoenzymes associated with the olfactory sensilla (aesthetascs). In this study the distribution of ecto-ATPase/phosphatase activity within aesthetascs was determined cytochemically and the nature of the adenine-nucleotide dephosphorylating activity was dissected biochemically. Cytochemically, the distribution of ATP-dephosphorylating activity was similar to that shown previously for AMP and beta-glycerol phosphate; i.e., cerium phosphate reaction product was specifically localized to the transitional zone where the sensory dendrites develop cilia and branch to form the outer dendritic segments. Unlike the dephosphorylation of AMP and beta-glycerol phosphate, Mg2+ or Ca2+ was required for ecto-ATPase/phosphatase activity. Biochemical measures of both AMP- and ATP-dephosphorylating activity within aesthetascs corroborated the cytochemical evidence that these activities are localized to the transitional zone. A major portion of the AMP dephosphorylation (about 67%) derives from nonspecific alkaline phosphatase activity that is insensitive to levamisole and L-bromotetramisole. In contrast, nonspecific phosphatase activity accounted for a much smaller part of the ATP dephosphorylation (about 15%). Ectoenzymatic activity in the transitional zone may be an important means of removing excitatory/inhibitory nucleotides from this region.

Male-specific structures in the olfactory system of mysids (Mysidacea; Crustacea)

A sexual dimorphism in the olfactory system of mysids is displayed in male-specific sensilla and neuropid areas. The sensilla are found on the first antenna, either on a small appendage, called the lobus masculinus, or on a structure within the peduncle. In both cases there is a muscular arrangement that is involved in the movement of sensilla-bearing parts. Each sensillum is innervated by one sensory cell whose dendrite develops two unbranched outer dendritic segments; due to their morphology a chemoreceptive function is suggested. The afferents from the bipolar sensory cells converge onto a small neuropil area, found in males only. The neuropil is located anteriorly to each of the paired olfactory lobes.

Differentiation of the thermo-/hygrosensitive (no-pore) sensilla on the antenna of Antheraea pernyi (Lepidoptera, Saturniidae): a study of cryofixed material

The morphogenesis of the thermo- and hygro-sensitive sensilla styloconica of Antheraea pernyi was studied, exclusively by cryomethods, during the second half of pupal development. The three major processes taking place during this period are (1) the differentiation of the dendritic outer segments of the sensory cells, especially of the lamellated type-2 receptor, (2) the formation of the receptor-lymph cavities, (3) the formation of tubular structures of unknown function in the inner receptor-lymph cavity, and (4) the elongation of the dendrite sheath. The formation of lamellae in the type-2 dendritic outer segment is achieved by the enfolding of its originally cylindrical cytoplasmic membrane. Autocellular junctions, previously described in the sensilla of adult animals, are found to join the forming lamellae. Close similarities between the junctions and smooth septate junctions are demonstrated. Both the extensive inner and outer receptor-lymph cavities are formed by invagination and folding of the apical cytoplasmic membranes of the three enveloping cells. Formation starts at the most apical projection of the cells and proceeds in a proximal direction. Up to 4-μm-long tubular structures appear, exclusively in developmental stages, in the inner receptor-lymph cavity. They are composed of plasma membranes whose inner surface is studded with regularly spaced electron-dense particles. Contacts with the cytoplasmic membrane of the innermost enveloping cell demonstrate that the structures are composed of lipid membranes. During elongation of the dendrite sheath, which in these sensilla is apically attached to the hair wall, an ∼2-μm-long growth-zone is observed at its proximal end. By addition of sheath-forming material to the growth-zone, the latter continuously moves proximally until the sheath is completed.