Activation of Mauthner neurons during prey capture

The Mauthner (M-) cells, a bilateral pair of medullary neurons in fish, initiate the characteristic “C-start” predatory escape response of teleosts. Similar movements have been described during hatching, social interactions, and feeding. M-cell firing, however, has not been correlated directly with these other behaviors. The objective of this study was to determine whether the M-cell, in addition to escape, plays a role in feeding.

1. Goldfish were chronically implanted with electrodes positioned near the axon cap of one of the two M-cells. Subsequently, M-cell activity was monitored for up to 8 days while fish were surface feeding on live crickets.
2. The M-cell fires and the fish performs a C-shaped flexion in association with the terminal phase of prey capture. Thus, the M-cell is active in the context of at least two behaviors, predator escape and prey capture, and may be considered a part of behaviorally shared neural circuitry.
3. For the goldfish, Mauthner-initiated flexions during feeding rapidly remove the prey from the water's surface and minimizes the fish's own susceptibility to surface predation. Other species may possess a diverse repertoire of Mauthner-mediated feeding behaviors that depend on their adaptive specializations for predation. Moreover, group competition between predators and their prey may have facilitated a “neural arms race” for M-cell morphology and physiology.

     

Functional development in the mauthner cell system of embryos and larvae of the zebra fish

In the embryonic zebra fish as early as 40 hr after fertilization, the Mauthner cells (M-cells) initiate an escape response, elicited by tactile-vibrational stimulation. The initial part of this behavior is similar to the acoustic startle reflex seen during the larval stage which begins at 96 hr. The embryonic response is directional and is followed by a series of strong tail flexures which are more pronounced than those during swimming. In the embryo the M-cell fired at the beginning of the response and rarely fired again during subsequent contractions; in our experiments the M-cell did not mediate iterative movements of the tail. The M-cell system is probably involved in evoked hatching behavior, as the tactile response is sufficient to rupture the egg membrane and allow the animal to escape. The M-cell sometimes fired spontaneously, which suggests that it might function also in spontaneous hatching behavior which occurs in the absence of phasic stimulation. At 48 hr the M-cell has morphologically mature synapses on its soma and dendrites, but its cytoplasm is relatively undifferentiated; it has few oriented neurofilaments and no distinct axon hillock. During these stages the extracellular M-spike is longer in duration and smaller in amplitude than at later times when the cell is more mature morphologically. Our data suggest that long-term inhibitory control of the M-cell system begins to function at about the time of hatching. At this time the cell is morphologically mature and is richly supplied with synaptic endings over its soma and dendrites.     

Ultrastructural studies on early events in fertilization of the bivalveLaternula limicola

Summary Ultrastructural studies on sperm-egg interaction at the time of fertilization inLaternula limicola were performed. The temporary-acrosome did not change morphologically while the sperm passed through the egg investments. At the onset of sperm entrance into the egg, however, the temporary-acrosome and mitochondria were eliminated from the sperm. Afterwards the sperm was engulfed by the egg surface without membrane fusion of the gametes. After entry the sperm nucleus was surrounded by four membranes: the plasma membranes of the egg and of the sperm, and the membranes of the sperm nuclear envelope. As the sperm nucleus differentiated into the male pronucleus, the plasma membranes of both the sperm and egg were initially vesiculated, then dispersed into the egg cytoplasm. Finally, the sperm nuclear envelope changed into the male pronuclear membrane accompanying sperm chromatin dispersion.     

Population growth rate of the parasitic rotifer Proales gigantea,and susceptibility to parasitization in the snail Lymnaea acuminata at different stages of embryonic development

Developing eggs of the host snail Lymnaea acuminata were experimentally parasitized with the parasitic rotifer Proales gigantea to study the population growth rate of the parasite within the snail egg capsule and the susceptibility of the host eggs at different stages of embryonic development. The population growth rate of P. gigantea was 0.46 ± 0.07 individual^–1 day^–1 at the ambient temperature of 18–22 °C. Snail eggs were most susceptible to rotifer attack during the initial stages of development, becoming progressively more resistant after the hippo stage. Yet, regardless of the stage of development, the host embryo was doomed to die without hatching even if one individual rotifer gained entry inside the egg capsule. The presence of P. gigantea within the parasitized egg capsules or in the mucilage had no effect on the developmental rates and hatching success of non-parasitized eggs within the same egg mass.     

Analyses of<Emphasis Type="Italic"> cis</Emphasis> -acting elements that affect the transposition of<Emphasis Type="Italic"> Mos1 mariner</Emphasis> transposons in vivo

The left (5) inverted terminal repeat (ITR) of the Mos1 mariner transposable element was altered by site-directed mutagenesis so that it exactly matched the nucleotide sequence of the right (3) ITR. The effects on the transposition frequency resulting from the use of two 3 ITRs, as well as those caused by the deletion of internal portions of the Mos1 element, were evaluated using plasmid-based transposition assays in Escherichia coli and Aedes aegypti. Donor constructs that utilized two 3 ITRs transposed with greater frequency in E. coli than did donor constructs with the wild-type ITR configuration. The lack of all but 10 bp of the internal sequence of Mos1 did not significantly affect the transposition frequency of a wild-type ITR donor. However, the lack of these internal sequences in a donor construct that utilized two 3 ITRs resulted in a further increase in transposition frequency. Conversely, the use of a donor construct with two 3 ITRs did not result in a significant increase in transposition in Ae. aegypti. Furthermore, deletion of a large portion of the internal Mos1 sequence resulted in the loss of transposition activity in the mosquito. The results of this study indicate the possible presence of a negative regulator of transposition located within the internal sequence, and suggest that the putative negative regulatory element may act to inhibit binding of the transposase to the left ITR. The results also indicate that host factors which are absent in E. coli, influence Mos1 transposition in Ae. aegypti.Communicated by G. P. Georgiev     

Interspecific variation in temperature dependence of egg development of five congeneric stonefly species (Protonemura Kempny, 1898, Nemouridae,Plecoptera)

Embryonal development of the five congeners Protonemura auberti Illies, 1954, P. hrabei Rauser, 1956, P. meyeri (Pictet, 1841), P. nitida (Stephens, 1835), and P. praecox (Morton, 1894) was studied under various laboratory temperatures and different photoperiods.Mean number of eggs in field collected batches was between 470 (P. praecox) and 1211 (P. auberti). Spring species had smaller egg batches than autumn species (Table 1). Mean hatching success in the laboratory was 50–100% at 2–18 °C. In most species hatching success decreased slightly with increasing temperature (Figs. 1a-e). None of the eggs incubated at 24 °C developed. Hatching pattern followed an asymmetric frequency distribution. In general, the hatching periods were the shorter the higher the incubation temperature.Embryonic development of all five species was inversely temperature dependent (Figs. 2a-e), and well described by a power law relationship (Figs. 3a-e). Interspecific differences in incubation periods were notable at nearly all temperatures (Fig. 4). There was a distinct interspecific sequence in length of incubation period (with steps of about 4 days), which was the same as can be seen in the flight periods: The later the species flies the longer the incubation period. Temperature fluctuations and variations in photoperiod had no influence on incubation and hatching periods or hatching success.The thermal demand of the egg stage neither explains the recent geographical distribution of the Protonemura species, nor does it directly correspond to the field temperatures common during their egg development. However, it is optimal in respect to resource partitioning between the five species, with the consequence of temporal displacement of life cycles.Derived from Brittain's (in press) proposal to compare the two constants a and b of the regressions describing the temperature dependence of embryonal development, a new index (Integral Development Time, IDT) indicating the thermal demand was created for easier comparison of numerous species (Table 2). Evaluation of the IDT for various species of Plecoptera (Fig. 5) suggests that species belonging to the family group Systellognatha generally have higher thermal requirements in the egg stage than species belonging to the Euholognatha.     

Particle transport by benthic invertebrates: its role in egg bank dynamics

The ecological and evolutionary dynamics of zooplankton is in part a function of the numbers and ages of dispausing eggs hatching from aquatic sediments. Successful recruitment from this egg bank must depend upon the eggs being present at or near the sediment surface. Often, however, zooplankton diapausing eggs are found as deep as 15 to 30 cm in the mud. Bioturbation may provide a mechanism for the regular return of buried eggs to the sediment surface. A substantial portion of the population of the copepod, Diaptomus sanguineus, living in Bullhead Pond, a small lake in Rhode Island, USA, is present as diapausing eggs. To study the role of bioturbation in egg-bank dynamics, we introduced polystyrene beads, the same size and specific gravity as copepod eggs, at two depths in large-diameter sediment chambers in the laboratory. Treatments included chambers with natural and reduced densities of benthos. Consistent with other studies, our results show that the joint activities of tubificid oligochaetes and chironomid larvae are responsible for bidirectional (up and down) transport of beads in the top 2 cm of the sediment. We observed no bead movement below this depth. Thus, eggs in the top two centimeters of sediment in this lake are exposed with some regularity to conditions that stimulate hatching at the sediment-water interface. In Bullhead Pond, these eggs have a mean age of 12.2 years (based on ²¹⁰Pb-dating). Eggs buried more deeply will only be returned to the sediment surface by relatively rare, localized disturbances. This return of old eggs to the surface affects ecological and evolutionary dynamics in a complex way.     

Synthesis of hexaploid (AABBCC) somatic hybrids: a bridging material for transfer of ‘tour’ cytoplasmic male sterility to different Brassica species

Most of the alloplasmic cytoplasmic male sterility (CMS) systems are known to be associated with a number of floral abnormalities that result from nuclear-cytoplasmic incompatibilities. One such system, tour, which is derived from Brassica tournefortii, induces additional floral abnormalities and causes chlorosis in Brassica spp. While the restorer for this CMS has been reported to be present in B. napus, in B. juncea, where the abnormalities are more pronounced, no restorer has yet been identified. Rectification of these floral abnormalities through mitochondrial recombinations and chloroplast replacement might result in the improvement of this CMS system. As organelle recombinations can possibly be achieved only by somatic cell hybridization, fusion experiments were carried out between hygromycin-resistant B. juncea AABB carrying tour cytoplasm and phosphinotricin-resistant, normal B. oleracea CC to generate AABBCC hexaploid somatic hybrids. The presence of selectable marker genes facilitated the selection of hybrids in large numbers. The resulting hybrids showed wide variation in floral morphology and organelle composition. Regenerants with normal, male-sterile flowers having recombinant tour-or oleracea-type mitochondria and oleracea-type chloroplasts were obtained. Hybrids with male-fertile flowers were also obtained that had recombined tour mitochondria. The AABBCC hexaploid hybrids synthesized in the present study were successfully utilized as a bridging material for transferring variability in the organelle genome simultaneously to all the digenomic Brassica species, and all of these hybrids are now being stabilized through repeated backcrosses to the allopolyploid crop brassicas.     

Degradation of benzene compounds by yeasts in acidic soils

Attemps were made to demonstrate the role of yeasts in the degradation of benzene compounds under natural soil conditions. Yeasts were isolated from acidic sandy soil supplied with benzene compounds. For this purpose the slant culture method was used. Growth on the benzene compounds took place on solid growth media at 10°C. Several yeast species were isolated: Leucosporidium scottii, Rhodotorula aurantiaca, Rhodotorula mucilaginosa, Trichosporon dulcitum, Trichosporon moniliiforme and Schizoblastosporion starkeyi-henricii. Cryptococcus humicolus and Cryptococcus laurentii were isolated from liquid enrichment cultures. All these strains assimilated several benzene compounds in pure culture.Cresol removal from contaminated soil was speeded up by inoculation with Rhodotorula aurantiaca G36. It was demonstrated that this yeast utilized this compound in competition with the soil microflora.     

RFLP mapping of three new loci for resistance genes to powdery mildew (Erysiphe graminis f. sp. hordei) in barley

Three new major, race-specific, resistance genes to powdery mildew (Erysiphe graminis f. sp. hordei) were identified in three barley lines, RS42-6*O, RS137-28*E, and HSY-78*A, derived from crosses with wild barley (Hordeum vulgare ssp. spontaneum). The resistance gene origining from wild barley in line RS42-6*O, showed a recessive mode of inheritance, whereas the other wild barley genes were (semi)-dominant. RFLP mapping of these three genes was performed in segregating F₂ populations. The recessive gene in line RS42-6*O, was localized on barley chromosome 1S (7HS), while the (semi)-dominant genes in lines RS137-28*E, and HSY-78*A, were localized on chromosomes 1L (7HL) and 7L (5HL), respectively. Closely linked RFLP clones mapped at distances between 2.6cM and 5.3 cM. Hitherto, specific loci for powdery mildew resistance in barley had not been located on these chromosomes. Furthermore, tests for linkage to the unlocalized resistance gene Mlp revealed free segregation. Therefore, these genes represent new loci and new designations are suggested: mlt (RS42-6*O), Mlf (RS137-28*E), and Mlj (HSY-78*A). Comparisons with mapped QTLs for mildew resistance were made and are discussed in the context of homoeology among the genomes of barley (H-vulgare), wheat (Triticum aestivum), and rye (Secale cereale). Duplications of RFLP bands detected in the neighbourhood of Mlf and mlt might indicate an evolutionary interrelationship to the Mla locus for mildew resistance.     

Somatic embryogenesis,plant regeneration and somaclonal variation in barley

In vitro culture of immature embryo and young leaf tissues was carried out with five cultivars of barley, Hordeum vulgare. Two cultivars (Albacete and Porthos) responded poorly from both types of explants, while the three others (Dissa, Golden Promise and Ingrid) produced a high frequency of embryogenic callus from these explants (25–60%). For Dissa and Ingrid, young leaf explants were slightly better than immature embryo explants for embryogenic callus induction, while immature embryo cultures of Golden Promise responded better than young leaf explants. Thus, there appears to be a significant genotype × explant interaction in the initiation of embryogenic callus in barley.Some phenotypic variants were detected among the regenerated plants of Golden Promise and Ingrid, most originating by epigenetic changes. Only in one case was the variant phenotype heritable, probably due to a mutation in the chloroplast DNA. Mitotic alteractions were not detected. Consequently, somaclonal variation did not appear to be a very frequent event in plants regenerated from 1- to 6- month-old cultures of barley.     

Neurons in a variety of molluscs react to antibodies raised against the VD1/RPD2 α-neuropeptide of the pond snail Lymnaea stagnalis

The VD₁ and RPD₂ neurons of Lymnaea stagnalis innervate other central neurons, certain skin areas, the pneumostome area, and the auricle of the heart. Recently, a set of four (, , , ) neuropeptides produced by these giant neurons and by certain other central neurons has been characterized. Although alternative splicing of the preprohormone of these neurons yields at least 10 different neuropeptides, an affinity-purified antiserum directed against a domain common to all neuropeptides has previously been shown to be highly selective in staining VD₁, RPD₂ and other neurons that produce the preprohormone. Since the gene encoding the neuropeptides is structurally similar to that expressed in R15 of the marine opisthobranch Aplysia californica, we have used the affinity purified antiserum as a marker for VD₁/RPD₂-related systems in other molluscs. Immunopositive neurons and fibers are observed in the central nervous systems of all species studied (Achatina fulica, Anodonta sp., Aplysia brasiliana, A. californica, Bulinus truncatus, Cepea sp., Eobania vermiculata, Helix aspersa, H. pomatia, Limax maximus, Mytilus edulis, Nassarius reticulatus, Viviparus viviparus). Several medium-sized and small neurons and 1–4 giant neurons are found in the pulmonates and opisthobranchs. The giant neurons in pulmonates have locations in the subesophageal ganglion, axonal branching patterns, and terminal arborizations in the auricle of the heart; all these characteristics are similar to those of VD₁ and RPD₂. Double-labelling (Lucifer yellow injection, immunocytochemistry) confirms that the two giant neurons in Helix pomatia are Br and Br. The immunoreactive cells in A. fulica appear to include the VIN and PON neurons. The antiserum also stains cells that appear to be the R15 neurons in two Aplysia species. The small and medium-sized neurons are distributed widely over the central ganglia of opisthobranchs and pulmonates. In prosobranchs and bivalves, small neurons are found in the cerebral and abdominal ganglia. No evidence has been found for innervation of the heart in these latter groups but in M. edulis, immunoreactive terminals can be observed in the gill. The results suggest the evolutionary conservation of immunoreactive peptides and the neurons that produce them, and thus support and extend previous hypotheses regarding the homology of certain giant neurons across molluscan species.     

Glycerol-, inositol-, and reducing end hexose-containing oligosaccharides in human urine

Ten previously unreported oligosaccharides have been purified from the urines of human subjects using a combination of gel filtration, ion exchange, and thin-layer chromatographies. Their structures were determined by direct probe mass spectrometry, methylation analysis, and proton NMR spectroscopy of the permethylated oligosaccharide alditols.On the basis of composition, the oligosaccharides could be divided into three groups. Five oligosaccharides containing glycerol were characterized as glucosyl1-1glycerol; glucosyl1-1glycerol; galactosyl1-1glycerol; glucosyl-1-1(fucosyl-1-2)glycerol and/or fucosyl-1-1(glucosyl-1-2)glycerol; and glucosyl-1-1(galactosyl-1-2)glycerol or galactosyl-1-1(glucosyl-1-2)glycerol. Four inositol-containing oligosaccharides were characterized as galactosyl1 (fucosyl1)inositol,N-acetylgalactosaminyl1 (fucosyl1)inositol, fucosyl1-2galactosyl1 (N-acetylgalactosaminyl1)inositol and fucosyl1-2galactosyl1-4-N-acetylglucosaminyl1(N-acetylgalactosaminyl1)inositol. Finally, galactosyl1-3(fucosyl1-2)galactosyl1-6galactosyl1-4(fucosyl1-3)glucose, an oligosaccharide with glucose at its reducing end, was tentatively identified. The significance and possible origins of the carbohydrate structures are discussed.     

Local motion processing in the optic tectum of the Japanese toad,Bufo japonicus

Summary The results of previous behavioral studies can be so interpreted that the prey-catching behavior in the toad is elicited if there is a local motion restricted with-in a small part of the visual field, while it is suppressed if there is a global motion over a large part of the visual field. This has led us to design experiments to answer a specific question (yet a very essential one for understanding neural processes underlying this behavior): Are there local motion detectors in the toad's visual system that are not activated by global motion over a large part of the visual field but are activated by local motion confined within a smaller part of it? The present study showed that (1) the majority of the toad's tectal neurons exhibit properties of the local motion detectors as defined above, and (2) these properties can be explained from the receptive field structure revealed in the present experiments. Based on these results, we suggest that the tectal local motion detectors are essential for the detection and localization of small moving prey-objects in the natural environment while ignoring the large moving objects or the self-induced motion of the visual field.Abbreviations ERF excitatory receptive field - G1-5 group 1–5 neurons     

Chick growth and survival in gentoo penguins (Pygoscelis papua): effect of hatching asynchrony and variation in food supply

Summary In the gentoo penguin, Pygoscelis papua, we examined the effects of intra-clutch egg size differences and hatching asynchrony on differential chick growth and survival (including post-fledging survival), in five years for which indices of food supply were available. An initial size hierarchy within-broods at hatching was due to hatching asynchrony not intra-clutch egg size differences. In 1988 only (a poor food year), the weight advantage gained by the first-hatched (A) chick persisted to the end of brooding (30 days), with more second-hatched (B) chicks dying. There was no difference between A- and B-chick weights at fledging (60 days) or in overall chick survival between synchronous and asynchronous broods in any year. Postfledging survival (measured in one year) was not related to fledging weight or hatching order. These results provide only partial support for the hypothesis that gentoo penguins operate a brood reduction strategy to optimise chick survival in years of low food availability. We suggest that hatching asynchrony in gentoo penguins may result from selection to keep the first egg warm as soon as it is laid, due to extreme low ambient temperatures.     

Rhythmicity of swimming activity in an axenic population of Paramecium multimicronucleatum

A unicellular organism, Paramecium, exhibits circadian rhythm activities in many physiological phenomena, i.e., mating reactivity, photoaccumulation in Paramecium bursaria and mating type reversals in Paramecium multimicronucleatum. In this study, we used an image-processing system to analyze swimming activity in a population of Paramecium multimicronucleatum cultured axenically under 12 h-light/12 h-dark cycles (LD 1212). Swimming behavior was recorded both under LD 1212 and constant darkness and images tracing the tracks of Paramecium were produced every 4 min. Swimming activity was represented by the occupied area by the tracks relative to the total observed area. It is high during daytime and low at night and exhibits a freerunning rhythm in constant darkness. Furthermore, criteria for two major components of swimming behavior, straight and circle swimming, were established and analyzed. The results indicate that swimming behavior alters depending on the time of day: straight swimming increased during the day and circling was dominant around dusk both under LD 1212 and constant darkness.Abbreviations ZT Zeitgeber Time - LD 1212 12h- light/12h-dark cycles - TF transversing frequency     

Molecular mapping of a new gene Rrs4 CI 11549 for resistance to barley scald (Rhynchosporium secalis)

Doubled haploid (DH) progeny from a cross between the scald susceptible barley (Hordeum vulgare L.) cultivar Ingrid and the resistant accession CI 11549 (Nigrinudum) was evaluated for resistance in the pathogen Rhynchosporium secalis (Oudem) J.J. Davis. Two linked and incompletely dominant loci confer resistance CI 11549 against isolate 4004. One is an allele at the complex Rrs1 locus on chromosome 3H close to the centromere; the other is located 22 cM distally on the long arm. The latter locus is designated Rrs4. In BC₃-lines into Ingrid from CI 2222 (another Nigrinudum) resistance seems governed by one locus close to the telomeric region of chromosome 7H, probably allelic to Rrs2. In neither case did we find any trace of the recessive gene rh8 reported to be present in Nigrinudum. Various resistance donors of Ethiopian origin designated as Nigrinudum, Jet or Abyssinian were identical to a great extent with respect to markers, but differed in resistance to different isolates of scald or in barley yellow dwarf virus (BYDV) resistance. The implications for their use as differentials in scald tests and screening of germplasm collections are discussed.     

The relationship between heat-stress and photobleaching in green and blue-green algae

Two characteristic temperatures were identified from measurements of the temperature dependence of O₂ evolution by Chlorella vulgaris and Anacystis nidulans: T₁, the threshold temperature for inhibition of O₂ evolution under saturating light conditions, and T₂, the upper temperature limit for O₂ evolution. Measurement of delayed light emission from photosystem II (PSII) showed that it passed through a maximum at T₁ and was virtually eliminated on heating the samples to T₂. Related changes were observed in low-temperature (77K) fluoresence emission spectra. Heat-stress had little effect on the absorption properties of the cells at temperatures below T₁ but incubation at higher temperatures, particularly under high-light conditions, resulted in extensive absorption losses. An analysis of these measurements suggests that this increased susceptibility to photobleaching is triggered by an inhibition of the flow of reducing equivalents from PSII that normally serves to protect the light-harvesting apparatus of the cells from photo-oxidation. Adaptation to higher growth temperatures resulted in increases in the values of T₁ and T₂ for Anacystis nidulans but not for Chlorella vulgaris.Abbreviations PSI photosystem I - PSII photosystem II - Chl a chlorophyll a - Chl b chlorophyll b - DCMU 3-(3 4 dichlorophenyl)-11-dimethylurea - PC plastocyanin - APC allophycocyanin CIW-DPB Publication No. 887.     

Mothers influence offspring body size through post-oviposition maternal effects in the redbacked salamander, Plethodon cinereus

In the terrestrial salamander (Plethodon cinereus), previous work has shown that mothers body size is positively correlated to offspring size at the time of hatching even after controlling for the effects of egg size. This study was designed to determine whether maternal body size affects offspring size via pre-oviposition factors (e.g., yolk quality, jelly coat composition, or maternal genes) or post-oviposition factors (e.g., parental care behaviors, parental modification of environment). Gravid females were captured and induced to lay eggs in experimental chambers in which the environment was standardized. Fifteen clutches were exchanged, or cross-fostered, between female pairs differing in body size. Ten females whose eggs were taken away and then returned served as controls for the crossing treatment. Foster mothers did not significantly differ from control mothers in the time spent with eggs, body position, or number of egg movements during brooding. Average egg mass measured midway through development was not significantly correlated to the body size of either the genetic or foster mother, but was correlated to pre-oviposition oocyte size. At hatching, offspring body length was positively correlated to egg size and the foster mothers body size. This correlation suggests that in P. cinereus post-oviposition maternal effects have a greater impact on offspring size than other maternal factors incorporated into the egg prior to oviposition. While our study showed that larger mothers moved their eggs less often and tended to spend more time in contact with their eggs, further work needs to be done to identify the specific mechanisms through which larger mothers influence the body size of their offspring. This is the first experimental demonstration of post-oviposition maternal effects for any amphibian with parental care.     

Immuno-electron microscopy of sorting and release of neuropeptides in Lymnaea stagnalis

Summary The cerebral caudodorsal cells of the pulmonate snail Lymnaea stagnalis control egg laying and egglaying behavior by releasing various peptides derived from two precursors. The biosynthesis, storage, intracellular breakdown and release of three caudodorsal cell peptides were studied by means of immuno-electron microscopy using antisera raised to fragments of these peptides: (1) Caudodorsal Cell Hormone-I (CDCH-I; derived from precursor I), (2) Caudodorsal Cell Hormone-II (CDCH-II; from precursor II), and (3) -Caudodorsal Cell Peptide ( CDCP; from both precursors). After affinity purification of the antisera, the specificity of the sera was confirmed with dotting immunobinding assays. From the ultrastructural immunocytochemical data it has been concluded that the precursor molecules are cleaved at the level of the Golgi apparatus after which the C-terminal parts (containing CDCP) and N-terminal parts (containing CDCH-I or CDCH-II) are sorted and preferentially packaged into large electron-dense granules (MD 150 nm), respectively. Very probably, the content of the large electron-dense granules is degraded within the cell body. The immunoreactivity of the secretory granules increases during discharge from the Golgi apparatus, indicating further processing. At least a portion of the secretory granules contains all three peptides, as shown by double and triple immunopositive stainings whereas other granules appear to contain only one or two of these peptides. The caudodorsal cells release multiple peptides via exocytosis from neurohemal axon terminals into the hemolymph and from blindly ending axon collaterals into the intercellular space of the cerebral commissure (nonsynaptic release).