Surface coats of pore tubules and olfactory sensory dendrites of a silkmoth revealed by cationic markers

Negatively charged surface coats have been demonstrated on the pore tubules and dendritic membranes of olfactory hairs of male Antheraea polyphemus silkmoths by application of the cationic markers lanthanum (La3+), ruthenium red (RR), and cationized ferritin (CF). Lanthanum and RR diffused readily into the apically opened hairs, whereas CF penetrated only for a relatively short distance. Deposits of the markers are distributed as follows: the inner surfaces of the hair walls are stained by RR and to a small degree by CF; the surfaces of the pore tubules and the dendritic membranes are stained by all three markers. The pore tubules have the strongest affinity for CF. The number of pore tubule-membrane contacts seems to be increased by the cationic dyes. The dendrites are often penetrated by RR, which forms deposits on the inner membrane leaflets, the cytoplasmic microtubules, and microfilaments, and by La3+, but never by CF. The observations provide support for the assumption that, first, the pore tubule-membrane contacts are formed via surface coats of both structures, possibly influenced by cations and, second, that the dendrites remain intact after pinching off the hair tips.     

Contacts of pore tubules and sensory dendrites in antennal chemosensilla of a silkmoth: Demonstration of a possible pathway for olfactory molecules

Antennal olfactory hairs of Antheraea polyphemus were investigated by means of transmission electron microscopy. Adequate preservation of dendrites and extracellular pore tubules is obtained by mechanical opening of the hair lumen and subsequent chemical fixation. The dendritic membrane has a cell coat. The dendrites contain microfilamentous structures in addition to their cytoplasmatic microtubules. The extracellular pore tubules traverse the hair cuticle and reach into the hair lumen for maximally 350 nm. Their diameter varies between 20 and 40 nm, depending on the preparation method. They consist of an electron-dense wall and an electron-lucent core. The wall has a helical substructure and is covered with a fuzzy coat. Contacts of pore tubules and dendritic membranes occur wherever dendrites are near the inner surface of the hair cuticle. Some of the pore tubules terminate approximately at right angles on the dendritic membrane, others lie against the membrane. The contact seems to be made via the surface coats of the tubules and the membrane. The structure of pore tubules which had been negatively stained with uranyl acetate is similar to the conventionally thin-sectioned material. The observations provide support for earlier assumptions that pore tubules are the pathways by which odor molecules reach the dendritic membrane.     

Lamellated outer dendritic segments of a chemoreceptor within wall-pore sensilla in the labial palp-pit organ of the butterfly,Pieris rapae L. (Insecta,Lepidoptera)

Summary The structure of the sensilla in the apical pit of the third segment of the labial palps in Pieris rapae was investigated in cryofixed and chemically fixed specimens. There is a field of about 80 club-shaped sensilla, 94% of which house a single sensory cell; 6% contain two sensory cells. All sensory cells are of the same type and are characterized by the structure of the dendritic outer segment. This consists of a proximal cylindrical and a distal lamellated section. The lamellae contain a lattice of longitudinally arranged microtubules. Filamentous strands connect the microtubules with the surface membrane of the lamellae. The surface area of the lamellated section amounts to about 40 m². Pores and pore tubules are present in the cuticular wall of the peg. Electrophysiological recordings show that the sensory cells are olfactory receptors, which react to a variety of complex plant odors and to the odor of conspecifics. It is shown that (a) the usual modality-specific characteristics of insect olfactory sensilla apply here also; (b) lamellation is not only a characteristic of thermoreceptors, but also of olfactory chemoreceptors; (c) there are pore tubules that are separated from the dendritic membranes by an extended dendritic sheath; and (d) in the labial palppit sensilla only the lamellated dendritic tip region may be involved in sensory transduction.Supported by the Deutsche Forschungsgemeinschaft (SFB 4/G1)     

Der Feinbau olfaktorischer Sensillen des Seidenspinners (Insecta,Lepidoptera)

Zusammenfassung Die Sensilla (S.) trichodea und S. basiconica auf den Antennen des Seidenspinners,Bombyx mori, wurden nach chemischer Fixierung, Gefriersubstitution und Gefrierätzung im Transmissionselektronenmikroskop untersucht. Es lassen sich fünf Typen von Sensillen unterscheiden, deren olfaktorische Funktion aus elektrophysiologischen Versuchen bekannt ist, mit Ausnahme des letzten Typs.Lange S. trichodea undhalblange S. trichodea I sind jeweils von zwei Sinneszellen innerviert, deren Rezeptorfortsätze (Dendriten) im wesentlichen unverzweigt bleiben. Diegroßen S. basiconica haben meist drei, diekleinen S. basiconica nur eine Sinneszelle; die Dendriten dieser Rezeptorzellen verzweigen sich büschelförmig beim Eintritt in das Haarlumen. Erstmals wird ein Zwischentyp beschrieben: diehalblangen S. trichodea II ähneln hinsichtlich der Innervation den S. basiconica, sind aber wegen der Form und Größe des Sinneshaars als S. trichodea zu klassifizieren. Für jeden Typ werden die Abmessungen der Rezeptorfortsätze sowie die Zahl und Verteilung der reizleitenden Poren und Porentubuli in der Haarwand angegeben.Auf der männlichen Antenne sind die langen S. trichodea am zahlreichsten; sie enthalten hochempfindliche Sexuallockstoffrezeptoren (mittlere Anzahl pro Antenne: 17 000 Sensillen mit 34 000 Sinneszellen). Beim Weibchen sind diese Sensillen in der Zahl auf etwa 35% reduziert und mit Sinneszellen anderer Spezifität und geringerer Empfindlichkeit ausgerüstet. Die beiden Dendriten der langen S. trichodea unterscheiden sich bei beiden Geschlechtern stark im mittleren Durchmesser und der Anzahl der cytoplasmatischen Mikrotubuli; beim Männchen reichen beide bis zur Haarspitze, beim Weibchen endet der dünnere Fortsatz bereits im proximalen Haardrittel. Die Cuticula der Sinneshaare ist von Poren durchbrochen (: 2–7 Poren/²; : 2–5 Poren/²), die stets in der Nähe von charakteristischen Stufen in der Haaroberfläche münden. Jeder Porenkanal führt in ca. fünf Porentubuli, die bis ins Haarlumen reichen und dort enden, zum Teil in Kontakt mit der Rezeptormembran der Dendriten. Die Häufigkeit solcher Tubulus-Membrankontakte ist in distalen Haarabschnitten größer als in proximalen. Der dickere Dendrit weist etwa viermal so viel Kontakte auf wie der dünnere. Die beiden Rezeptorzellen dieser Sensillen stellen funktionell verschiedene Reaktionstypen dar, was mit den beobachteten morphologischen Unterschieden zusam menhängen dürfte.Die S. basiconica haben 20 Poren pro ² ihrer Oberfläche und 12–23 Porentubuli pro Pore; dadurch erreichen oder übertreffen sie die viel größeren S. trichodea in der Gesamtzahl der Porentubuli pro Sinneshaar. Auf den S. trichodea steigt die Zahl der Poren pro Oberflächeneinheit zur Spitze hin stetig an, während die Zahl der Poren pro Haarlängeneinheit einen konstanten Wert annimmt. Eine Hypothese über die Morphogenese dieser Verteilung wird aufgestellt.Die funktionelle Bedeutung der äußeren Epicuticulaschichten und der Porentubulussysteme für die Reizleitung wird diskutiert. Ausgehend von beliebigen Orten ihres Auftreffens auf dem Sinneshaar können die Duftmoleküle zunächst durch zweidimensionale Diffusion entlang der Haaroberfläche zu den Poren gelangen und anschließend durch eindimensionale Diffusion über Porenkanäle und Porentubuli die Rezeptormembran erreichen. Die berechneten Diffusionszeiten sind kürzer als die bekannten Rezeptorlatenzen; die Reizleitung kann also durch Diffusion hinreichend erklärt werden und erfordert keine kompliziertere Hypothese.

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The fine structure of olfactory sensilla in the silk moth (insecta, lepidoptera)

Summary The sensilla (s). trichodea and s. basiconica on the antennae of the silk moth,Bombyx mori, were studied under the transmission electron microscope. Chemical fixation, freeze substitution and freeze etching methods were used. The following results have been obtained: Five sensillum types were distinguished, the olfactory function of which is known from electrophysiological recordings, except for the last one. Thelong s. trichodea and themedium-sized s. trichodea I are innervated by two sensory cells which have essentially unbranched receptor processes (dendrites). Commonly thelarge s. basiconica contain three sensory cells, thesmall s. basiconica only one; the dendrite of these receptor cells branch multiply when entering the hair lumen. For the first time an intermediate type has been described: themedium-sized s. trichodea II, which resemble the s. basiconica in their branching innervation, but must be classified as s. trichodea because of the form and size of the sense hair. For each type, the dimensions of the receptor processes, as well as the number and distribution of the stimulus conducting pores and pore tubules in the hair wall are noted.On the male antenna the long s. trichodea are most abundant; they contain the highly sensitive sex pheromone receptors (mean number per antenna: 17 000 sensilla with 34 000 sense cells). In the female these sensilla are reduced in number to about 35% and supplied with receptor cells of different specificity and lower sensitivity. In both sexes, the two dendrites of the long s. trichodea differ markedly in their mean diameter, and the number of cytoplasmatic microtubules. In the male moth both receptor processes reach the hair tip, whereas in the female the thinner one invades only the proximal third of the sense hair. The cuticle of the hair wall is perforated by pores (: 2–7 pores per ²; : 2–5 pores per ²), which mostly open to the outside near to characteristic steps in the hair surface. Each pore canal leads into about five pore tubules, which proceed towards the hair lumen, where they end, partly in contact with the receptor membrane of the dendrites. Distal parts of the sense hairs show such tubule-membrane contacts more frequently than proximal regions. The number of contacts counted on the thicker dendrite is about four times greater than on the thinner one. In these sensilla, the two receptor cells constitute functionally different reaction types, which may relate to the observed morphological differences.The s. basiconica have about 20 pores per ² of the hair surface, and 12–23 pore tubules per pore: thus, these sensilla have the same or even a greater number of pore tubules per sensillum than the much larger s. trichodea. In the s. trichodea the number of pores per unit surface increases steadily towars the hair tip, while the number of pores per unit length of the hairs soon reaches a constant value. A hypothesis about the morphogenesis of this distribution is given.The functional significance of the epicuticular surface layers and of the pore tubule systems is discussed under the aspect of stimulus conduction. Starting from the site of impact anywhere on the sense hair, odour molecules may diffuse two-dimensionally along the hair surface to the pores, and then proceed by one-dimensional diffusion through pore canals and pore tubules until they eventually reach the receptor membrane at the end of a tubule. The calculated conduction times are shorter than the known receptor latencies; thus, the transport mechanism can be explained by diffusion and does not need a more complex hypothesis.

Ich danke Frl. B. Müller für ihre stete, sorgfältige Hilfe, Prof. L. Bachmann und Dr. W. Schmitt von der TU München für die Benützungsmöglichkeit der Gefrierätzanlage und ihren erfahrenen Rat, Dr. G. Adam von der Universität Konstanz sowie meinen Seewiesener Kollegen, Dr. W. A. Kafka, Dr. K.-E. Kaissling und Dr. E. Priesner, für viele anregende Diskussionen und konstruktive Kritik.     

Hair regeneration during moulting in the spiderCiniflo similis (Araneae,Dictynidae)

Summary The mechanoreceptive and chemoreceptive hairs on the legs of the cribellate spiderCiniflo similis were examined during the moulting cycle. In mechanoreceptive hairs the new hair shaft is formed around the extended dentrites, which emerge from near the tip of the newly forming hair and continue to the old sensillum within the extended dendritic sheath. Thus there is no ecdysial canal in the base of the hair shaft as found in insect mechanoreceptive hairs. The dendritic connection with the old hair is maintained until shortly before ecdysis by which time new tubular bodies have developed in the same dendrites at the base of the new hair. In chemoreceptive sensilla the new hair shaft is also formed around the elongated outer segment of the dendrites (19 chemosensitive and 2 mechanosensitive). The two mechanosensitive dendrites develop new tubular bodies at the base of the hair. As ecdysis occurs the old dendritic sheath and dendrites are snapped off at the tip of the new hair but the pore remains open. The ultrastructural evidence indicates that the roles of the three main enveloping cells are as follows: The dendritic sheath cell secretes the dendritic sheath, the middle enveloping cell forms the hair shaft while the outer enveloping cell forms the socket. This pattern corresponds closely to that observed in insecta sensilla. The extreme length of the chemoreceptive dendrites during moulting is mentioned in connection with receptor function. The unique multi-layered nature of the middle enveloping cell is seen as a device for the formation of regularly occurring rows of small spines on the shaft of the hair.     

The hair-peg organs of the shore crab,Carcinus maenas (Crustacea,Decapoda): Ultrastructure and functional properties of sensilla sensitive to changes in seawater concentration

Summary The hair-peg organs of the shore crab, Carcinus maenas, are modified hair-sensilla. A small hair shaft (peg) is surrounded by a tuft of solid cuticular bristles (hairs). Each hair-peg organ is innervated by 6 sensory neurons, 2 of which have scolopidial (type-I) dendrites. The outer segments of all dendrites pass through a cuticular canal extending to the articulated hair base in which the 2 type-I dendrites terminate. The other 4 (type-II) dendrites reach the clavate tip of the hair shaft and have access to a terminal pore and a large sickle-shaped aperture. Three inner and 8–12 outer enveloping cells belong to a hair-peg organ. The innermost enveloping cell contains a scolopale, which has desmosomal connections to the ciliary rootlets of the type-I dendrites. An inner and an outer sensillum lymph space are present. The ultrastructural features of the dendrites and the cuticular apparatus indicate that the hair-peg organs are bimodal sensilla, comprising 2 mechano- and 4 chemosensitive sensory neurons. Extracellular recordings from the leg nerve indicate that the chemosensitive neurons of the hair-peg organs respond to changes in seawater concentration in the physiological range of Carcinus maenas.Supported by the Deutsche Forschungsgemeinschaft (SFB 45/A1; W. Gnatzy)     

The development of taste and tactile hairs in the pharate fly Protophormia terraenovae (Diptera,Calliphoridae) and in the embryonal cricket Acheta domestica (Orthopteroidea,Ensifera)

Summary 1. The development of taste hairs and tactile hairs of the fly Protophormia terraenovae is described using light microscope, scanning, and transmission electron microscope methods.2. The development of taste hairs proceeds in the same way on tarsi, labella, and wings. First the dendritic outer segments of ciliary origin become visible above the hypodermal cell surface [2 days after pupariation (AP) at 19° C]; then the dendritic sheath starts growing out and finally the trichogen process follows. In a typical intermediate stage (stage C) the distal sections of the dendrites float freely in the fluid surrounding the pharate adult. The more proximal sections are enclosed by the dendritic sheath around which the trichogen process is wrapped (4 days AP). The protruding dendrites disappear when the cuticle starts being deposited on the fully grown trichogen process, and the sheath vanishes later (9–10 days AP or 1 day before eclosion). The development is discussed with respect to the known structural organization of the adult hair.3. In the tactile hairs the single dendrite which grows outwards is completely covered by the dendritic sheath and lies beside the trichogen process [stage C(m)].4. The taste and tactile hair development proceeds in the same way on legs isolated from the pupa after disc eversion in an artificial medium containing ecdysterone.5. To check that both these patterns of development are widespread the development of taste and tactile hairs of the first instar cercus of the cricket Acheta domestica was studied with the light microscope: Both hair types pass through identical early stages.     

Antennal sensilla ofNeomysis integer (leach)

Summary The most frequent type of the hair sensilla on the antennae ofNeomysis integer is investigated by electron microscopic methods. The cellular properties of the sensilla are compared with those of other arthropods in order to detect possible homologies.The hairs are innervated by 2, 3, 6, 8, 9, or 10 sensory cells. The dendrites show an inner and outer dendritic segment. Five or six enveloping cells belong to a sensillum. In intermoult stage, processes of all the enveloping cells except the innermost one extend into the hair shaft. The sensory hairs possess only a single liquor cavity, which morphologically is homologous to the inner lymph cavity of insect sensilla. Around the liquor cavity, a supporting structure is located which seems to be identical to the scolopale of chordotonal organs. The six-to tenfold-innervated hairs possess two groups of differently structured dendrites which are regularly arranged on opposite sides of the liquor cavity. The outer dendritic segments are enclosed in a dendritic sheath. It is secreted by the innermost enveloping cell (= dendritic sheath cell of insect sensilla). All the outer dendritic segments terminate in the distal region of the hair shaft which shows a pore at its tip. The possible function of the sensilla is discussed. The double and triple-innervated hairs are considered to be mechano-receptors, whereas the sensilla associated with six to ten sensory cells might be mechano-chemoreceptors.     

Atomic-force microscopy on the olfactory dendrites of the silkmoths Antheraea polyphemus and A. pernyi

Olfactory transduction is thought to occur in the outer dendritic membrane of insect olfactory receptor neurons. Electrophysiological studies have indicated that the outer dendritic membrane has non-specific cation channels and inositol-triphosphate-dependent Ca²⁺ channels. The presence of such channels is further supported by the observation that pheromone-stimulated dendrites take up cobalt. However, to date, there is no structural evidence for these channels. Therefore, in order to search for putative ion channels, we have imaged the membrane of the olfactory dendrites in the scanning electron microscope (SEM) and the atomic-force microscope (AFM), after extruding the dendrites out of the olfactory hairs and fixing them on plastic coverslips. With the aid of the SEM, we could see the beaded structure of the dendrite but no fine structural details, as the membrane was sputtered with gold. With the use of the contact mode of the AFM, we could see “pores” that were deeper than 3 nm and with a diameter of about 15 nm. The density of the “pores” was approximately 20/µm² or 10?000 pores per thick dendrite. We believe these to be putative ion channels based on indirect evidence.     

Structure and differentiation of the sensilla of the ventral sensory field on the maxillary palps ofPeriplaneta americana (Insecta,Blattodea), paying special attention to the ciliogenesis of the sensory cells

Summary The embryonic development of palpal contact chemosensitive sensilla was studied from 42% of development up to the hatching of the larvae. Ciliogenesis of the sensory cells can be observed at the earliest stages investigated. A complex consisting of two basal bodies and a cap-like ciliary vesicle is localized in the dendritic inner segment. It migrates apically and fuses with the cytoplasmic membrane. At the same time, microtubule doublets of the distal basal body elongate, thus generating the dendritic outer segment. Furthermore, the typical accessory structures of a motile cilium are formed. Although the central pair of microtubules is lacking, the dendritic outer segment can be considered as a modified motile cilium. At about 84% of development the hair structure starts to be formed. Whereas the socket is generated by the tormogen cell, the trichogen cell produces the hair shaft and terminal porus. The dendrite sheath, which rises above the newly formed hair, is attached apically to the embryonic cuticle forming an irregular pore. In larvae and imagines, the inner surface of the dendrite sheath is highly differentiated. A range of circular ledges and filamentous structures wrapping around the dendritic outer segments can be distinguished. These may have a stabilizing function. Furthermore, in cryofixed specimens, the dendritic outer segments possess regularly spaced swellings which are about 1 m in length and about 0.5 m in diameter. Their functional significance is still unclear.     

Are the funnel-canal organs the “campaniform sensilla” of the shore crab Carcinus maenas (Crustacea,Decapoda)?

Summary The topography of the funnel-canal organs of Carcinus maenas (Decapoda, Crustacea) and their stimulus-receiving cuticular and sensory apparatus were studied in the light and electron microscopes.About 4000 funnel-canal organs are situated within the exoskeleton of Carcinus. Almost all of them are on the distal segments of the walking legs, in particular on the epicuticular cap at the tip of the dactyl. They were not found to be arranged in groups or sensilla fields, and no sex-specific differences were observed.Characteristic features of the funnel-canal organs are as follows: (a) There is a terminal pore (0.5×0.8 m diameter) in the cuticle, at the tip of a small projection. It is closed by a plug of electron-dense material. (b) The terminal sections of the dendrites are enclosed in a dendritic sheath up to ca. 10 m below the pore. (c) The dendrites, 3–24 in number, end below the plug; none of the dendrites exhibits a tubular body; two of the dendrites are distinguished from the others by the greater number of microtubules in their outer segments.The structural characteristics, in particular the gustatory pore and the number of dendrites, are typical of bimodal receptors in arthropods. In such receptors, as in the contact chemoreceptors of insects and arachnids, mechano-and chemosensitive sensory cells are combined.This interpretation of the function of the funnel-canal organs is supported by electrophysiological data of other authors.The morphological parameters we find for the funnelcanal organs, in comparison with those of insect campaniform sensilla, provide clear evidence against the reclassification of the funnel-canal organs as crustacean campaniform organs proposed by Shelton and Laverack (1968).Supported by the Deutsche Forschungsgemeinschaft (W.G., SFB 45/A1)We thank Professor Dr. F.G. Barth for valuable discussion and Mr. K. Grommet for drawing the Figs. 1 and 6c     

The structure of bimodal chemo-, thermo-, and hygroreceptive sensilla on the antenna of Locusta migratoria

Summary Coeloconic sensilla on the antenna of Locusta migratoria were investigated electrophysiologically, labeled and then examined under the scanning or transmission electron microscope. They can be categorized into two main morphological types: 1) sensilla with wall pores and two concentric cuticular walls (wp-dw sensilla), and 2) sensilla lacking wall pores (np-sensilla).Hygroreceptors were observed only in np-sensilla. Olfactory receptors were observed in wp-dw sensilla, but not in np-sensilla. Cold-sensitive units were found in both types. Wp-dw sensilla with a cold-sensitive unit also manifest a fatty acid olfactory receptor. This cold-sensitive unit appears to be less sensitive to drops in temperature than the cold receptor combined with hygroreceptors in np-sensilla. Recordings from both types revealed up to three or sometimes four units that could be distinguished on the basis of impulse amplitude.The number of np-sensilla on each antennal segment ranges from 0 to 6. These sensilla are pegs 4–5 m long with a terminal swelling. The dendritic outer segments of two sensory cells fit tightly into these pegs. The dendrite of the third sensory cell ends at its base. All three are up to 1 m thick, unbranched, and densely filled with microtubules. The cuticular wall and socket of the peg are characterized by (1) an electron-dense inner layer surrounding the dendrites and continuous with electron dense clusters at the base of the peg, and (2) clefts in the cuticle at the insertion of the peg.The present observations favor the hypothesis that insect sensilla combining the triad of moist air, dry air and cold receptors generally lack pores. Specifical, though still alternative roles in stimulus transduction can be suggested for substructures demonstrated in the present and previous electron microscopic investigations.Supported by the Deutsche Forschungsgemeinschaft (SFB 4:G1/D)     

Fine structure of the tip of the labellar taste hair of the blow flies,Phormia regina (Mg.) and Calliphora vicina R.-D. (Diptera,Calliphoridae)

Summary The labellar taste hairs of the blow flies, Phormia regina and Calliphora vicina, have an opening mechanism at the tip which consists of two stump cuticular prongs and a funnel-like cuticular pouch. Opening and folding of these structures are regulated by the pressure within the dendrite-free lumen of the hair. The extrusion of viscous substance at the tip of the taste hair is possible through spongy cuticle and one pore in each prong; it seems likewise to depend on the pressure within the dendrite-free lumen and results in regional collapsing of this lumen. Described and discussed are: The cuticle and pores of the structures at the hair tip, pore filaments which extend from the dendrites, and the number and arrangement of the dendrites.This work was supported by a grant from the 7USDA, Entomology Research Division, Beltsville, Md., and the grant GB-13500 from the National Science Foundation.We thank Dr. J. F. Worley, USDA, Plant Science Research Division, Beltsville, Md., for his collaboration in fluorescence microscopy.     

Morphogenesis of the antenna of the male silkmoth. Antheraea polyphemus, III. Development of olfactory sensilla and the properties of hair-forming cells

During adult development of the male silkmoth Antheraea polyphemus, the anlagen of olfactory sensilla arise within the first 2 days post-apolysis in the antennal epidermis (stage 1-3). Approximately on the second day, the primary dendrites as well as the axons grow out from the sensory neurons (stage 4). The trichogen cells start to grow apical processes approximately on the third day, and these hair-forming 'sprouts' reach their definite length around the ninth day (stages 5-6). Then the secretion of cuticle begins, the cuticulin layer having formed on day 10 (stage 7a). The primary dendrites are shed, the inner dendritic segments as well as the thecogen cells retract from the prospective hair bases, and the inner tormogen cells degenerate around days 10/11 (stage 7b). The hair shafts of the basiconic sensilla are completed around days 12/13 (stage 7c), and those of the trichoid sensilla around days 14/15 (stage 7d). The trichogen sprouts retract from the hairs after having finished cuticle formation, and the outer dendritic segments grow out into the hairs: in the basiconic sensilla directly through, and in the trichoid sensilla alongside, the sprouts. The trichogen sprouts contain numerous parallel-running microtubules. Besides their cytoskeletal function, these are most probably involved in the transport of membrane vesicles. During the phase of cuticle deposition, large numbers of vesicles are transported anterogradely from the cell bodies into the sprouts, where they fuse with the apical cell membrane and release their electron-dense contents (most probably cuticle precursors) to the outside. As the cuticle grows in thickness, the surface area of the sprouts is reduced by endocytosis of coated vesicles. When finally the sprouts retract from the completed hairs, the number of endocytotic vesicles is further increased and numerous membrane cisterns seem to be transported retrogradely along the microtubules to the cell bodies. Here the membrane material will most probably be used again in the formation of the sensillum lymph cavities. Thus, the trichogen cells are characterized by an intensive membrane recycling. The sensillum lymph cavities develop between days 16-20 (stage 8), mainly via apical invaginations of the trichogen cells. The imago emerges on day 21.     

Die Feinstruktur der tarsalen Schmeckhaare der Fliege Phormia terraenovae Rob.-Desv.

Zusammenfassung Die Schmeckhaare der Fliege Phormia terraenovae enthalten fünf Rezeptorzellen (vier Kontaktchemo- und eine Mechanorezeptorzelle), die von zwei Hüllzellen unter Aussparung von Lymphräumen (Vakuole) umgeben sind. Die Rezeptorzellen gliedern sich in Axon, Zellkörper und Dendrit, der aus Innenglied, Übergangsabschnitt und Außenglied besteht. Der Übergangsabschnitt hat die 9+0-Struktur der modifizierten Cilien mit zwei Basalkörpern. Im Plasma der Außenglieder sind nur Mikrotubuli zu erkennen. Die Außenglieder der Kontaktchemorezeptoren ziehen durch die röhrenförmige Dendritenscheide und den Kanal I des Haarschafts bis zu einem Porus in der Haarspitze, durch den sie mit den Schmeckstoffen in Kontakt kommen (Ort der Primärprozesse). Das Außenglied der Mechanorezeptorzelle enthält einen tubulären Körper und setzt an der Haarbasis an. Die Hüllzelle II (tormogene Zelle) bildet zum großen Sensillenlymphraum hin ein stark gefaltetes Membransystem aus. Der Feinbau wird in Beziehung zur Funktion der Schmeckhaare gesetzt.

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The fine structure of the tarsal taste hairs of the blowfly, Phormia terraenovae Rob.-Desv.

Summary The tarsal taste hairs of the blowfly contain five receptor cells: four contact chemoreceptors and one mechanoreceptor. These cells are surrounded concentrically by two sheath cells with fluid-filled spaces (vacuoles) between them. The receptor cell parts are the axon, the cell body, and the dendrite. The latter has an inner segment, a transitional region, and an outer segment. The transitional region shows the 9+0 configuration of a modified cilium with two basal bodies. In the outer segment only microtubules are found. The outer segments of contact chemoreceptors pass through the tube-like dendrite sheath, then through canal I of the hair shaft, to reach the pore at the tip of the hair. The outer segment of the mechanoreceptor contains a tubular body and terminates at the hair base. The sheath cell II (tormogene cell) forms a folded membrane system toward the main liquor space. The fine structure of the tarsal taste hairs is discussed in relation to their function.

Den Herren Dr. W. Pflumm, Prof. Dr. U. Thurm, Prof. Dr. E. Schnepf und Dr. K. E. Kaissling sei für anregende Diskussionen und der Deutschen Forschungsgemeinschaft für eine Sachbeihilfe gedankt. Frau R. Föhr danken wir für ausgezeichnete technische Mitarbeit.     

Pores from mitochondrial outer membranes of yeast and a porin-deficient yeast mutant: A comparison

Reconstitution experiments were performed on lipid bilayer membranes in the presence of purified mitochondrial porin from yeast and of detergent-solubilized mitochondrial outer membranes of a porin-free yeast mutant. The addition of the porin resulted in a strong increase of the membrane conductance, which was caused by the formation of ion-permeable channels in the membranes. Yeast porin has a single-channel conductance of 4.2 nS in 1 M KCl. In the open state it behaves as a general diffusion pore with an effective diameter of 1.7 nm and possesses properties similar to other mitochondrial porins. Surprisingly, the membrane conductance also increased in the presence of detergent extracts of the mitochondrial outer membrane of the mutant. Single-channel recordings of lipid bilayer membranes in the presence of small concentration of the mutant membranes suggested that this membrane also contained a pore. The reconstituted pores had a single-channel conductance of 2.0 nS in 1 M KCl and the characteristics of general diffusion pores with an estimated effective diameter of 1.2 nm. This means that the pores present in the mitochondrial outer membranes of the yeast mutant have a much smaller effective diameter than normal mitochondrial porins. Zero-current membrane potential measurements suggested that the second mitochondrial porin is slightly cation-selective, while yeast porin is slightly anion-selective in the open state but highly cation-selective in the closed state. The possible role of these pores in the metabolism of mitochondria is discussed.     

Fine structure of the trichomycete,Harpella melusinae,from black-fly guts

Summary The fungus, Harpella melusinae, attached to the peritrophic membrane of black-fly larvae, was fixed in glutaraldehyde-acrolein-osmium and in KMnO₄. The holdfast does not penetrate the peritrophic membrane, and is comprised of many finger-like outgrowths (digits) surrounded by secretions of cementing substance that arise from a specialized basal part of the thallus. Undifferentiated cells contain many organelles, membranes, and lipid droplets commonly found in other phycomycetes, but in addition contain large concentrations of electron-opaque material with vesicles and tubules. The cytoplasm also has crystals in membrane-bound vesicles, myelin-like aggregations of membranes, and large nuclei with associated semi-circular plaques. Septa with small pores like bordered pits, plugged with an electron-opaque material, resemble those of other Harpellales and Linderina (Mucorales). As a spore grows outward, 4 appendages are formed within the generative cell, attached to the base of the spore body, and form a spiral between the wall and plasmalemma. Forming vescles transport material from the cytoplasm to the growing appendages, resulting in a periodic structure of alternating dark and light bands. Mature spores have a thick, wavy (in cross section) inner wall and a loose outer wall, and are separated from the generative cell by a septum with a pore.     

Fine structure of the stretch receptor in the bursa copulatrix of the butterfly,Pieris rapae crucivora

Summary A pair of multipolar stretch-receptive neurons were found in the bursa copulatrix of the female cabbage white butterfly, Pieris rapae crucivora. The cell body of each neuron, about 10 m in diameter, lies on the edge of the muscular region in the antero-lateral wall of the corpus bursae. No special accessory structure, such as a receptor muscle, is associated with the neuron. The several dendrites extend radially into the muscle layer. The dendrites are ensheathed except for their terminal tips, and, on their course, they anchor repeatedly on the epithelial cells or the muscle fibers in such a manner that their basement membranes fuse together. While the ensheathed dendrite is usually 0.1–0.2 m in diameter, it often forms 1–2 m varicosities especially at anchor sites, so that it looks like a varicose, or beaded, chain. The varicosities contain a number of mitochondria, but only microtubules are found in the fine interconnecting parts of the dendrite. The naked dendritic tips terminate in the basement membrane of the epithelial cell. The varicosities, as well as naked tips, seem to be important for stimulus transduction in the sensory cell of this type.     

The innervation and chemical sensitivity of single aesthetasc hairs

Summary Each aesthetasc hair of the lateral antennule of the California spiny lobsterPanulirus interruptus (Randall) is shown by light and scanning electron microscopy to be innervated by a basally situated cluster of sensory neurons encased in a glial sheath which isolates each cluster from those of other hairs (Figs. 1, 3, 4). The dendrites of these neurons penetrate the aesthetasc hairs and their axons extend to the central nervous system. Extracellular recordings with suction electrodes from the axons of single neuronal clusters were used to determine the responsiveness of individual hairs to a spectrum of amino acids, amines, amides, carbohydrates, carboxylic acids, nucleotides, and a tripeptide (Tables 1, 2, Figs. 6, 8). Randomly selected hairs from the antennules of juvenile, and male and female adult lobsters were shown to be broadly sensitive to a variety of stimuli and are homogeneous in their breadth of responsiveness (Figs. 5, 7). Cluster analysis does not reveal distinct chemoreceptive hair types based on their response spectra, suggesting that the receptor populations of single hairs are uniformly competent to respond to diverse chemical stimuli (Figs. 6, 8). Further, the sensitivity profile of aesthetascs to these stimuli correlates well with behavioral responses ofPanulirus interruptus to these same stimuli (Tables 1, 2).Abbreviation ² Chi-squared     

Ultrastructural Aspects of the Somatic Cortex and Contractile Vacuole of the Ciliate,Ichthyophthirius multifiliis Fouquet1

The trophont stage in the life cycle of Ichthyophthirius multifiliis was studied in the electron microscope. Surface ridges contain up to 24 ridge microtubules, disposed as a ribbon. Kinetosomes show the classic morphology of 9 triplets of microtubules. Associated with each kinetosome is a kinetodesmal fibril, originating in proximity to triplets 5, 6, and 7, and having a 30 nm periodicity; 3 to 5 postciliary microtubules, originating between triplets 8 and 9; and up to 3 transverse microtubules, originating at triplet 4, as well as a parasomal sac. Each cell is partially enclosed by a system of 3 “unit” membranes: the outer limiting membrane, and the outer and inner alveolar membranes. The last two membranes define the alveolar sac. Mucocysts, each with a dense core, are present in large numbers. The contractile vacuole system includes the contractile vacuole, associated tubules and vesicles, injection canals, a discharge canal, and a pore. Microtubules abound in the walls of the contractile vacuole, injection and discharge canals, and in the region of the pores, where both ring and radial microtubular arrangements are noted. The ultrastructure suggests that I. multifiliis is more closely related to Tetrahymena pyriformis than to Paramecium aurelia.