Search

In Drosophila sensory organ development, the balance of activities between proneural genes and repressor genes defines a proneural cluster as a population of competent cells for neural development. In this study, we report the isolation and analysis of the tamou (tam) gene that encodes a cell-cell junction-associated protein, which is homologous to mammalian ZO-1, a member of the membrane-associated guanylate kinase homolog family. The tam mutation reduces the transcription of a repressor gene, extramacrochaetae, and causes enlargement of a proneural cluster where supernumerary precursor cells emerge, resulting in extra mechanosensory organs in the fly. These results suggest that the membrane-associated Tam protein is involved in the signaling pathway that activates emc expression.

The ontogeny of cutaneous sensory nerve organs is described in higher vertebrates, and includes the lamellated corpuscles of Meissner, Pacini and Herbst, and the Merkel cell-neurite complex with bird Merkel and Grandry corpuscles, and mammalian Merkel cells. The main common feature is that for most corpuscles there is an inside-out order of assembly around the nerve ending which is present from the beginning of end-organ ontogeny. The exception is the mammalian Merkel cell which is present in the epidermis before the entrance of nerve fibers, and could play a promotional role in the development of skin innervation. The developmental origin of Herbst and Merkel corpuscles in birds is reported as demonstrated using embryological experiments with cell markers. Conclusions are that inner bulb cells of Herbst corpuscles and bird Merkel cells are of neural crest origin, whereas other cells (inner space and capsular cells for Herbst corpuscle and capsular cells for Merkel corpuscles) are provided by the local mesenchyme. The question of the ontogeny of mammalian Merkel cells is discussed in relation to the two debated hypothesis of epidermal and neural crest origins. Morphogenetic interactions during the development of cutaneous sensory end organs are also discussed.

We have isolated a novel class of gain-of-function mutations at the Bearded (Brd) locus which specifically affect the development of adult sensory organs in Drosophila. These Brd alleles cause bristle multiplication and bristle loss phenotypes resembling those described for the neurogenic genes Notch (N) and Delta (Dl). We have found that supernumerary sensory organ precursor (SOP) cells develop in the proneural clusters of Brd mutant imaginal discs; like normal SOPs, these are dependent on the function of the proneural genes achaete and scute, and express elevated levels of ac protein. At cuticular positions exhibiting the Brd bristle loss phenotype, we have found that the progeny of the multiplied SOPs develop aberrantly, in that neurons and thecogen (sheath) cells appear but not trichogen (shaft) and tormogen (socket) cells. This appears to represent a transformation of the pIIa secondary precursor cell within the SOP lineage to a pIIb secondary precursor cell fate. These results suggest that Brd gain-of-function alleles interfere with Notch pathway-dependent cell-cell interactions at two distinct stages of adult sensory organ development. We have also identified enhancers and suppressors of the Brd dominant phenotypes; these include both previously characterized mutations and alleles of apparently novel loci. Finally, we have found that Brd null mutants are viable and exhibit no mutant phenotypes, suggesting that Brd may be a component of an overlapping function.

A prominent element of the early larval pigment pattern in the salamander Ambystoma tigrinum tigrinum (family Ambystomatidae) is a horizontal stripe over the lateral surface of the myotomes where otherwise abundant, neural crest-derived melanophores are not found. This study examines the formation of this "melanophore-free region". When the trunk lateral lines were ablated (by removing cranial lateral line placodes), the melanophore-free region did not form; instead, melanophores populated the middle of the flank and the distribution of yellow, neural chest-derived zanthophores was perturbed. Time-lapse videomicrography demonstrated that during normal development, the melanophore-free region is established because melanophores retreat from the midbody lateral line primordium as it migrates caudally along the inner side of the epidermis. Melanophores do not repopulate the middle of the flank after primordium migration and heterochronic grafting experiments suggest that extracellular factors contribute to maintaining the melanophore-free region during these later stages. Finally, photographic series, microsurgical manipulations, electron microscopy, and staining for molecules of the extracellular matrix (peanut agglutinin-binding components, tenascin, chondroitin sulfate proteoglycans, fibronectin, laminin) suggest that several factors contribute to establishing and maintaining the melanophore-free region, including steric effects of the lateral lines, interactions between melanophores and xanthophores, lateral line-dependent alterations of the subepidermal basement membrane, and a general elaboration of the extracellular matrix. Lateral line effects on melanophores are inferred to be a shared, ancestral feature of pigment pattern development for the families Ambystomatidae and Salamandridae (D.M. Parichy, Dev. Biol. 174, 265-282. 1996). The results of this study thus provide insights into a phylogenetically primitive mechanism for stripe formation, and a context for interpreting evolutionary innovations in pattern-forming mechanisms.

In many species of salamanders, pigment cells derived from the neural crest give rise to a horizontal stripe pattern in hatchling larvae. A defining element of these horizontal stripe patterns is a region over the middle of the myotomes that is relatively free of melanophores. This study shows that formation of a "melanophore-free region" and horizontal stripe pattern in Ambystoma tigrinum tigrinum (family Ambystomatidae) correlates with the development of the trunk lateral line sensory system. Moreover, prevention of lateral line development results in greater densities of melanophores in the middle of the flank, essentially eliminating the melanophore-free region in this taxon. A phylogenetic survey also revealed that ablation of the lateral lines has qualitatively similar effects on melanophores in seven of eight additional taxa (Ambystomatidae: A. barbouri, A. maculatum, A. talpoideum; Salamandridae: Notophthalmus viridescens, Pleurodeles waltl, Taricha granulosa, T. rivularis). In Taricha torosa, however, a superficially similar melanophore-free region forms prior to lateral line development, and ablation of the lateral lines does not perturb the horizontal stripe pattern. Finally, heterospecific grafting experiments demonstrated that T. torosa lateral lines are competent to generate a melanophore-free region, and T. torosa melanophores are competent to respond to cues associated with the lateral lines. These results indicate that lateral line-dependent pattern-forming mechanisms are common and probably ancestral within the families Ambystomatidae and Salamandridae and suggest that these ancestral mechanisms have been retained in T. torosa as redundant, lateral line-dependent mechanisms for stripe formation have evolved.

The antennal system of the moth Manduca sexta is a useful model for studies of the development of olfactory glomeruli, the complex synaptic structures that typically underlie the initial processing of olfactory input in vertebrates and invertebrates. In this review, we summarize cellular events in the construction of glomeruli in Manduca and highlight experiments that reveal factors that influence glomerulus development. By methodically manipulating each of various cell types, both neuronal and glial, that contribute to glomerular architecture, we have found that: olfactory receptor axons lay a template for developing glomeruli, stabilization of the template by glial cells is necessary to permit subsequent steps in development of the glomeruli, and the hormone that regulates adult development causes production of adequate numbers of glial cells. Neither electrical activity nor the presence of a serotonin-containing neuron that persists throughout development is required for a glomerular pattern to develop; these factors might, however, influence the synaptic organization of individual glomeruli.

The development of the olfactory sense organs on the antenna of the fruit fly Drosophila utilises mechanisms distinct from those used in the rest of the adult peripheral nervous system. Lozenge (lz) is the only locus hither-to identified as required for the development of antennal sense organs. In addition to effects on the antenna, mutations in lz also affect the development of the eye and maxillary palp. We have used the readily-scored eye-phenotype in a temperature sensitive lz allele to screen for dominant modifiers of phenotypes at this locus. We analyse the phenotypes of both intragenic and extragenic modifiers. Our results reinforce the view from developmental studies that lz functions in eye and antennal development in distinct ways.

The neurogenic Notch locus of Drosophila encodes a receptor necessary for cell fate decisions within equivalence groups, such as proneural clusters. Specification of alternate fates within clusters results from inhibitory communication among cells having comparable neural fate potential. Genetically, Hairless (H) acts as an antagonist of most neurogenic genes and may insulate neural precursor cells from inhibition. H function is required for commitment to the bristle sensory organ precursor (SOP) cell fate and for daughter cell fates. Using Notch gain-of-function alleles and conditional expression of an activated Notch transgene, we show that enhanced signaling produces H-like loss-of-function phenotypes by suppressing bristle SOP cell specification or by causing an H-like transformation of sensillum daughter cell fates. Furthermore, adults carrying Notch gain of function and H alleles exhibit synergistic enhancement of mutant phenotypes. Over-expression of an H+ transgene product suppressed virtually all phenotypes generated by Notch gain-of-function genotypes. Phenotypes resulting from over-expression of the H+ transgene were blocked by the Notch gain-of-function products, indicating a balance between Notch and H activity. The results suggest that H insulates SOP cells from inhibition and indicate that H activity is suppressed by Notch signaling.

Each olfactory (antennal) lobe of the moth Manduca sexta contains a single serotonin (5-HT) immunoreactive neuron whose processes form tufted arbors in the olfactory glomeruli. To extend our present understanding of the intercellular interactions involved in glomerulus development to the level of an individual, identified antennal lobe neuron, we first studied the morphological development of the 5-HT neuron in the presence and absence of receptor axons. Development of the neuron's glomerular tufts depends, as it does in the case of other multiglomerular neurons, on the presence of receptor axons. Processes of the 5-HT neuron are excluded from the region in which the initial steps of glomerulus construction occur and thus cannot provide a physical scaffolding on which the array of glomeruli is organized. Because the neuron's processes are present in the antennal lobe neuropil throughout postembryonic development, 5-HT could provide signals that influence the pattern of development in the lobe. By surgically producing 5-HT-depleted antennal lobes, we also tested the importance of 5-HT in the construction of olfactory glomeruli. Even in the apparent absence of 5-HT, the glomerular array initiated by the receptor axons was histologically normal, glial cells migrated to form glomerular borders, and receptor axons formed terminal branches in their normal region within each glomerulus. In some cases, 5-HT-immunoreactive processes from abnormal sources entered the lobe and formed the tufted intraglomerular branches typical of most antennal lobe neurons, suggesting that local cues strongly influence the branching patterns of developing antennal lobe neurons.

The neurogenic genes of Drosophila act during many different times and places during development. It is thought their role is to repress cell fate within a group of equivalent cells and thus allow the singling out of discrete numbers of precursors. Amongst the genes at the neurogenic locus, Enhancer of split is a family of seven related genes that encode proteins containing the basic helix-loop-helix motif characteristic of transcriptional regulators. Previous functional analyses of these genes have relied on deletions which eliminate many other genes. We have ectopically expressed two of the Enhancer of split basic helix-loop-helix genes, m5 and m8, to test their effect on the determination of the precursor cells of adult sensory organs. Ectopic expression of m5 or m8 before bristle precursor division results in loss of sensory bristles from all parts of the adult fly. Ectopic expression after bristle precursor division produces bristles with aberrant cuticular structures. We have also tested the effect of reducing Enhancer of split gene function using mitotic recombination and show that this de-represses the neural fate and produces supernumerary sensory bristle neurons. We conclude that the Enhancer of split basic helix-loop-helix genes inhibit neural fate during the selection of neural precursors, and that they also play a role in restricting the neuronal fate to one of the four progeny cells of the bristle precursor.

The formation of ommatidia in the compound eyes and sensilla on the antennae of the honeybee was followed and the development of their sensory neurons was traced using an antiserum against taurine as a marker. Taurine-like immunoreactivity (Tau-IR) is expressed in sensory neurons of several modalities, namely visual, olfactory, gustatory, and mechanosensory. Staining intensity is very high in the larva and in the first half of the pupal stage and gradually decreases towards the end of metamorphosis. In the photoreceptor cells of the compound eyes, Tau-IR can be detected from the fifth larval instar onwards, prior to differentiation of other components of the ommatidium. Already in the midstage larvae, when the antennal primordia of the adult still lie within the peripodial cavity, a few presumably mechanosensory neurons are labelled in the pedicellus of the developing antenna. The majority of the antennal sensory neurons which are located on the flagellum start to exhibit Tau-IR upon pupation, long before any cuticular specializations such as sensory hairs or plates are detectable. All known types of antennal sensilla were identified and it could be shown that all of them are innervated by Tau-IR sensory neurons. Thus, taurine immunocytochemistry can be applied as a useful label for developing sensory neurons. Functional implications of taurine during development are discussed.

The olfactory sensilla on the antenna of adult Drosophila melanogaster develop during the first 36 hr after pupariation, from their anlagen in the cephalic disc. We have used tissue-specific beta-galactosidase expression in the enhancer trap strain A101.IF3 and the monoclonal antibody 22C10 as sensory cell markers, as well as the lineage tracer 5-bromo-2'-deoxyuridine (BrdU), to describe this process. The development of an olfactory sensillum begins with the selection of a "founder cell" (FC). These cells are distinct in that they possess large apically located nuclei revealed by beta-galactosidase expression in A101.IF3. In the following 6 hr, a few cells neighboring the FC also start expressing beta-galactosidase and together comprise a group. Cells of this group, denoted a "presensillum-cluster" (PSC), undergo at least one round of replication and give rise to all of the cells of a sensillum. A subset of the cells within each PSC and, later, all the sensory neurons are recognized by MAb22C10. The antennae of the mutant lozenge3 (lz3) lack all basiconic and some trichoid sensilla. The mutation apparently affects early steps in sensillum development and many of the FCs fail to form. Those that are present, however, proceed to form mature olfactory sensilla. Therefore, we conclude that the selection of an FC is the first step in olfactory sense organ development. Our study reveals novel aspects of sensory development in Drosophila.

We studied the birth times of neurons of labellar taste sensilla in blowflies using incorporation of the thymidine analogue 5-bromodeoxyuridine (BrdU) as an indicator of birth time. We found that one of the two main sensillum types, the taste papillae, arise according to a clear spatial gradient of birth times, whereas the other sensillum type, taste hairs, arise without any apparent spatial ordering. Within each sensillum type, there was a strong tendency for either all or none of the neurons to have incorporated BrdU. Among those rare sensilla in which only some of the neurons incorporated BrdU, there were clear patterns of the distribution of labeled and unlabeled neurons per sensillum. These results suggest that subsets of the neurons of a sensillum are siblings, and thus argue against the possibility that the several neurons of a sensillum arise from a single stem cell precursor through repeated asymmetrical divisions.

The formation of sense organs in Drosophila involves a series of choices, each of which depends on the co-ordinated activity of a small battery of genes. Two essential steps of this process have been extensively studied over the past few years: the determination of neural precursor cells, and their diversification. In both cases, the choices are dichotomous, and each choice reflects the fact that a specific control gene is or is not expressed. This principle is illustrated in the case of the genes "cut" and "poxn", the expression of which controls the type of sense organ that a given precursor will form.

In Drosophila imaginal discs, the spatially restricted activities of the achaete (ac) and scute (sc) proteins, which are transcriptional activators of the basic-helix-loop-helix class, define proneural clusters (PNCs) of potential sensory organ precursor (SOP) cells. Here, we report the identification of several genes that are direct downstream targets of ac-sc activation, as judged by the following criteria. The genes are expressed in the PNCs of the wing imaginal disc in an ac-sc-dependent manner; the proximal promoter regions of all of these genes contain one or two high-affinity ac-sc binding sites, which define the novel consensus GCAGGTG(T/G)NNNYY; where tested, these binding sites are required in vivo for PNC expression of promoter-reporter fusion genes. Interestingly, these ac-sc target genes, including Bearded, Enhancer of split m7, Enhancer of split m8, and scabrous, are all known or believed to function in the selection of a single SOP from each PNC, a process mediated by inhibitory cell-cell interactions. Thus, one of the earliest steps in adult peripheral neurogenesis is the direct activation by proneural proteins of genes involved in restricting the expression of the SOP cell fate.

A family of neural RNA-binding proteins has recently been described in both vertebrates and invertebrates. We have identified a new member of this family, the Drosophila musashi (msi) locus, which is required for development of adult external sensory organs (sensilla). In contrast with wild-type sensilla, which contain two outer support cells, the msi mutation typically results in the appearance of extra outer support cells. The msi putative RNA-binding protein is localized to the nucleus and appears to be expressed in all cells in each sensillum and predominantly in neurons during embryogenesis. We propose that the msi protein regulates sensillum development by controlling the expression of target genes at the posttranscriptional level.

Lateral line sensory receptors and their cranial nerves in axolotls arise from a dorsolateral series of placodes, including the octaval placode, that gives rise to the inner ear and the octaval nerve. Anterodorsal and anteroventral placodes occur rostral to the octaval placode and give rise to anterodorsal and anteroventral lateral line nerves and electroreceptors and mechanoreceptors of the snout, cheek, and lower jaw. Middle, supratemporal, and posterior placodes occur caudal to the octaval placode and give rise to similarly named lateral line nerves, electroreceptors and mechanoreceptors of the occipital region of the head, and trunk neuromasts. All placodes, except the posterior placode, elongate, forming sensory ridges, following the genesis of sensory ganglia. Primary mechanoreceptor primordia begin to form within the central zone of the sensory ridges at stage 36; primary electroreceptor primordia originate within the lateral zones of these ridges at stage 38. The first primary mechanoreceptors erupt during stage 37; all primary mechanoreceptors have erupted at hatching (stage 41). Primary electroreceptors begin to erupt at stage 43. Secondary mechanoreceptor primordia begin to form in 1-week-old larvae and erupt 1-2 weeks later. Secondary electroreceptor primordia also begin to form in 1-week-old larvae and continue until clusters of two to five electroreceptors are formed. The developmental stages thought to characterize lateral line placodes in the earliest gnathostomes suggest that this ancestral ontogeny has been truncated in modern amphibians, and ontogenetic mechanisms underlying placodal differentiation are suggested.

Graviceptor (rhopalium) development in Aurelia aurita ephyrae which developed on Earth and in space during the nine-day NASA SLS-1 mission was compared. The space-developed ephyrae made graviceptors which were morphologically similar to those of their ground-based controls. Rhopalia of both groups developed statocysts with statoliths, ocelli, ciliated mechanoreceptor cells, and immature touch-plates with one type of hair cell. The number of rhopalia formed per arm of ephyrae of both groups revealed no significant differences. The number of statoliths formed per rhopalium was statistically higher in ephyrae which were induced to form in space with iodine than in L(Launch)+8h controls. Statolith numbers were not significantly different between Earth-formed control ephyrae and those formed from polyps induced on Earth and then sent into space 24h and 48h later. Statolith loss from rhopalia was significantly enhanced in the space-maintained ephyrae in ASW as compared to their controls. Ephyrae formed through thyroxine treatment and those maintained in thyroxine in space had statolith numbers comparable to thyroxine-treated controls. Pulsing abnormalities seen in some space-developed ephyrae suggest that some space-formed ephyrae may have developed abnormal rhopalia because normal rhopalia development and function is necessary for normal pulsing.

The central organization of antennular motoneurons in the brain of the spiny lobster, Panulirus argus, was analyzed by combining biocytin backfills with serial reconstructions of the antennular nerves and the brain. Eighty-nine to 99 antennular motoneurons occur in each hemibrain. The somata of the motoneurons are distributed in a consistent pattern in two complex soma clusters, the ventral paired mediolateral cluster of the deutocerebrum and the dorsal unpaired median cluster of the tritocerebrum. The motoneurons arborize ipsilaterally in the lateral and median antennular neuropils and the tegumentary neuropil. The backfills indicate a minimum of five morphological types of motoneurons with different arborization patterns. The innervation pattern of the motoneurons, together with previously reported innervation patterns of antennular sensory afferents, suggest that the lateral antennular neuropil is a lower motor center driving local antennular reflexes in response to chemical and mechanical stimulation of the antennule, whereas the median antennular neuropil is a lower motor center for equilibrium responses.

During adult metamorphosis, the moth olfactory neurons and their glia-like support cells pass through a coordinated and synchronous development. By 60% of development, the olfactory system is anatomically complete, but functional maturation does not occur until about 90% of development. Maturation is characterized by the onset of odorant sensitivity in the sensory neurons and the expression of certain antennal-specific proteins including odorant binding proteins (OBPs) and odorant degrading enzymes (ODEs). The OBPs have been cloned and sequenced, and are thus useful models for investigating the molecular mechanisms coordinating final maturation of the developing olfactory system. The ecdysteroid hormones have been observed to regulate many cellular level neuronal changes during adult metamorphosis. In particular, the late pupal decline in ecdysteroids is known to influence programmed death of nerves and muscles at the end of metamorphosis. Experiments are presented here which indicate that this decline in ecdysteroids also induces the expression of the OBPs. Normal OBP expression occurs 35-40 h before adult emergence. In culture, OBP expression could be induced at least 90 h before adult emergence by the premature removal of ecdysteroid. This premature expression was blocked by culturing tissue in the presence of the biologically active ecdysteroid 20-hydroxyecdysone. These findings suggest that maturation of the olfactory system is regulated by the decline in ecdysteroids, and support the view that olfactory development, in general, may be coordinated by changing levels of pupal ecdysteroids.

Different types of sense organs are present on the larva of Drosophila. Several genes that specify the type of sense organ that will form at a particular position have been recently identified. Here we review the functional and molecular analyses of these genes, and summarize the evidence which supports a role in the choice of which type of organ will be formed. Most or all of these genes are required for the appropriate specification of adult as well as larval sense organs, suggesting that the larval and adult systems share many gene requirements. Interestingly, the specifying genes identified so far in the peripheral nervous system are also expressed in subsets of cells in the central nervous system, where they might have similar roles.

The cut locus is both necessary and sufficient to specify the identity of a class of sensory organs in Drosophila embryos. It is also expressed in and required for the development of a number of other embryonic tissues, such as the central nervous system, the Malpighian tubules and the tracheal system. We here describe the expression of cut in the precursors of adult sensory organs. We also show that cut is expressed in cells of the prospective wing margin and correlate the wing margin phenotype caused by two cut mutations with altered cut expression patterns. Finally, we observe cut-expressing cells in other adult tissues, including Malpighian tubules, muscles, the central nervous system and ovarian follicle cells.

Drosophila uses different olfactory organs at different developmental stages. The larval and adult olfactory organs are morphologically dissimilar and have different developmental origins: the antenno-maxillary complex (AMC), which houses the larval olfactory organ, is histolyzed during metamorphosis; the third antennal segment--the principal adult olfactory organ--derives from an imaginal disc. A screen for genes expressed in both larval and adult olfactory organs, but in relatively few other tissues, has been carried out. Seven enhancer trap lines showing reporter gene expression in both the larval AMC and in certain subsets of the adult antenna are described. The antennal staining pattern of one line shows a striking change over the first few days of adult life, with a time course comparable to that of the development of sexual maturity. A pronounced sexual dimorphism in antennal staining pattern is seen in another line. Some staining patterns resemble the patterns of certain classes of antennal sensilla; others show expression restricted to only a small number of cells. Some lines also show expression associated with other chemosensory organs at either the larval or adult stage, including the maxillary palps, labellum, and anterior wing margin. One line, which also shows staining in the male reproductive tract, is male sterile. The significance of these results is considered in terms of (1) the molecular organization of the olfactory system; (2) the recruitment of olfactory genes for use in two developmental contexts; (3) the sharing of genes among different sensory modalities; (4) the role of olfaction in sexual behavior; and (5) posteclosional changes in the olfactory system.

The mechanosensory bristles of adult Drosophila are composed of four cells that, in most cases, are progeny of a single sensory organ precursor (SOP) cell. Two sister cells in this lineage, the trichogen and tormogen, produce the external shaft and socket of the bristle, respectively. Loss-of-function mutations of Hairless (H) confer two distinct mutant phenotypes on adult bristles. The bristle loss phenotype results from the failure to specify and/or execute the SOP cell fate; the double socket phenotype results from the transformation of the trichogen (shaft) cell into a second tormogen (socket) cell. We have found that the H gene encodes a novel basic protein with a predicted molecular mass of 109 kD. Basal levels of expression of a transgene (P[Hs-H]) in which the H protein-coding region is under the control of the Hsp70 promoter are sufficient to provide full rescue of H mutant phenotypes. Heat shock treatment of P[Hs-H] transgenic animals as late larvae and early pupae produces a tormogen-to-trichogen (double shaft) cell fate transformation, as well as bristle multiplication and loss phenotypes very similar to those caused by loss-of-function mutations in the neurogenic gene Notch. Our results indicate that the SOP cell fate requires H to antagonize the activity of the neurogenic group of genes and that the expression of distinct cell fates by the trichogen/tormogen sister cell pair depends on an asymmetry in their levels of H+ activity or in their thresholds for response to H.

The scalloped (sd) gene of Drosophila melanogaster was initially characterized by mutants affecting structures on the wing of the adult fly. The sequence of a cDNA clone of the gene reveals a predicted protein sequence homologous to that of a human transcriptional enhancer factor, TEF-1 (68% identity). The homology includes a sequence motif, the TEA domain, that was shown previously to be a DNA-binding domain of TEF-1. An sd enhancer trap strain expresses the reporter gene in a subset of neuroblasts in the central nervous system and in the peripheral sense organs of the embryo. The reporter gene is later expressed in specific regions of the imaginal discs, including regions of the wing disc destined to become structures defective in viable sd mutants. Later still, expression in the adult brain is restricted to subsets of cells, some in regions involved in the processing of gustatory information. These observations indicate that the sd gene encodes a transcription factor that functions in the regulation of cell-specific gene expression during Drosophila development, particularly in the differentiation of the nervous system.

The imaginal leg disc of Phormia regina contains eight neurons that arise during embryogenesis. Five of the neurons are associated with Keilin's organ, and of these five, two persist to the adult fly. Two new neurons arise at about the time of pupariation and flank each of these persisting neurons, forming two triplets of cells. Both triplets can be followed throughout metamorphosis; in the late pupa they are situated anteriorly and posteriorly at the tip of the fifth tarsomere. Two triplets of cuticular specializations are found at corresponding positions in the adult fly, each consisting of two campaniform sensilla and a trichoid hair. The central member of each set of sensilla, a campaniform sensillum, is associated with the persisting cell.

Neurons within the precursor of the adult leg, the imaginal disc, innervate a larval sense organ, Keilin's organ. Electron microscopical investigations of first instar larvae show that five dendrites end at the organ: three insert at the bases of the three hairs of the organ and two end against the cuticle, without any apparent cuticular specialization. In third instar larvae, the imaginal leg discs invaginate into the body cavity, and only four of the dendrites (the outer segments of which become greatly elongated) remain in contact with Keilin's organ. The axons of the neurons that supply Keilin's organ project into a ventral neuropile region of the central nervous system, with a pattern that resembles the projections of other larval sensilla. Electrical activity can be recorded from neurons of the imaginal disc in response to mechanical stimulation.

The somatotopically and functionally organized electrosensory system of gymnotiform teleosts provides a model for the study of the formation of ordered nerve connections. This paper describes the development of the major electrosensory nuclei within the hind- and midbrain. All three main electrosensory nuclei--the electrosensory lateral line lobe (ELL), dorsal torus semicircularis (torus), and tectum--grow by adding cells at their caudolateral borders. Toral and tectal germinal zones arise from lateral ventricular outpocketings that either completely or partially close by maturity. In the ELL before day 5 postspawning, germinal cells form from an initial periventricular germinal zone, then migrate to the caudolateral border of the hindbrain and begin dividing. The ELL grows from two main germinal zones, one for the medial segment, and one for the three lateral tuberous segments. Within each ELL germinal zone, newly formed cells arise from two areas: granular cells arise from a ventral subzone, pyramidal cells are generated more dorsally. Granular cells remain in situ, whereas pyramidal cells may migrate rostromedially. Cells begin differentiating as soon as they are formed. Spherical and pyramidal cells send ascending axons into the internal plexiform layer by day 14-18 and the ELL gradually begins to assume its mature laminar appearance. The ELL grows caudally, preceding the caudal lobe of the cerebellum, which will eventually lie over and fuse with it. Primary electrosensory afferents enter the ELL by day 6; incoming afferents form four fascicles within the ELL, suggesting the formation of separate ELL segments. Unlabelled projections between labelled fields from a single nerve branch filled with HRP on day 7 suggest that somatotopic order is already present at this early age. In the periphery, receptor addition is unordered, occurring along nerve branch pathways. Meanwhile the ELL adds cells in an orderly fashion at its caudolateral border. This suggests that primary afferents shift position caudally with growth to maintain their somatotopic relationships. Because all three central nuclei are in topographic register and grow by adding cells caudally, during growth ELL efferents to the torus and toral efferents to the tectum may utilize passive mechanisms, such as fiber-fiber interactions, to guide axons.

Sex-pheromone components released by Manduca sexta females are detected solely by male-specific olfactory receptor neurons that innervate long sensilla trichodea on the male antennae. To facilitate studies of the development and physiology of these receptor cells, we have produced primary in vitro cultures of cells dissociated from pupal male antennae. These cultures comprise several morphological types of cells, 2 of which have been characterized immunocytochemically with a pair of monoclonal antibodies that were shown previously to recognize certain antigens in olfactory receptor neurons at defined stages of development. The good correlation between in vivo and in vitro expression of these antigens suggests that the immunocytochemically recognized cells are olfactory receptor neurons that follow at least partially their normal course of differentiation in vitro.

During development of Xenopus laevis, the supraorbital lateral line system (i.e. the parietal and supraorbital lines of organs and the anterior auditory group of organs) is all derived from a single primordium located in the ear region of the epidermis. The primordium elongates first by active movement along the dorsal margin of the eye. Individual primary organs are then formed by progressive fragmentation of the streak-like primordium. After fragmentation, passive displacement of the organs due to skin growth seems to play the main role in altering the arrangement of the line system. Transplantation experiments confirmed that non-placodal epidermal cells are not incorporated into the developing system. The active elongation of the primordium is due to cell multiplication, and not due to cell rearrangement or change in cell shape or size. Cell multiplication is not confined to a growth zone, but dividing cells are randomly distributed throughout the primordium. All cells of a primordium have to change position during its elongation.

The sensory development of 22 kittens was observed for 12 weeks. Vision-related responses began as a palpebral reflex at 1.8 days and continued as a light blink response at 5.7 days (mean values). The eyes unsealed between days 6 and 17 and the pupillary reflex was well developed within a few days. Depth perception began at 13.2 days mean. Auditory development progressed rapidly after the opening of the external canals between days 7 and 14. Pain and rooting reflexes were present at birth, while auriculonasocephalic and Galant's reflexes were highly variable.

Cell multiplication was studied during development of the supraorbital lateral line system in Xenopus laevis. The increase in cell number is biphasic. The first phase extends from the beginning of primordial elongation to the end of primary organ formation. Cell number increases linearly during this interval. Throughout this phase, a constant number of cells is in S phase of the cell cycle at a given time, despite a more than 10-fold increase in total cell number. After their formation, the number of the primary organs remains essentially constant. The individual primary organs are not clones of cells. Different organs grow at different rates, and become more and more heterogeneous in size. The second phase which is correlated with accessory organ formation is characterized by an elevated growth rate. This phase was not studied in detail. If developing larvae are starved, growth is normal up to completion of the first growth phase but is arrested at this point. The frequency distribution of the sizes of such growth-arrested organs approximates a binominal distribution. From its characteristics, a detailed model of cell proliferation and organ formation can be deduced: cell multiplication occurs through asymmetrically dividing stem cells, which become allocated to the forming organs at random and go through a fixed number of cell divisions.

The organization of the peripheral electrosensory system of the cheek was studied in an age-graded series of Sternopygus dariensis in Nissl-stained sections and silver-stained whole mounts of skin. As in other gymnotoids, both ampullary and tuberous electroreceptors are present. Small fish have only one ampullary organ or tuberous organ per axon, and the number of receptor organs per axon increases with age in both ampullary and tuberous systems. Large fish may have up to ten tuberous organs per axon, although the distribution of tuberous organs per axon is bimodal with one peak occurring at a single receptor organ per axon and the other peak shifting upward in relation to the age of the fish. The ampullary system adds receptor organs at a faster rate and a large fish may have 20 ampullary organs per axon. With increasing size, the number of sensory receptor cells in each organ remains constant for both types of electroreceptors. Evidence is presented for addition of new electroreceptor units by de novo production in small fish and increases in the number of organs in existing electroreceptor units by division of previously formed organs in medium-sized and large fish. As the surface area of the skin increases with growth, the density of electroreceptor units decreases and, although new receptor organs are still being added to existing receptor units, no generation of new receptor units occurs in medium-sized to large fish.

Synaptic rearrangement during development is a characteristic of the vertebrate nervous system and was thought to distinguish vertebrates from the invertebrates. However, examination of the wind-sensitive cercal sensory system of the cricket demonstrates that some identified synaptic connections systematically decrease in strength as an animal matures, while others increase in strength over the same period. Moreover, a single sensory neuron could increase the strength of its synaptic connection with one interneuron while decreasing the strength of its connection with another interneuron. Thus, rather than being a hallmark of the vertebrate nervous system, synaptic rearrangement is probably characteristic of the development of many if not all nervous systems.

The girdle epidermis of adult Mopalia muscosa secretes several types of structures, including calcareous spicules and innervated hairs. Newly metamorphosed chitons superficially resemble adult animals, but they lack the adult girdle ornaments, shell sculpture, and coloration. The morphogenesis of the adult girdle structures has not been described previously for any species. Juvenile Mopalia muscosa secrete hairs at metamorphosis, but it was not known if these hairs were sensory or if they were retained as the animals grew. I discovered that the hairs of juveniles become the tips of adult hairs. When juvenile hairs are detectable by light microscopy the sensory components already exist, suggesting that they are functional receptor organs. The other girdle ornaments of young juveniles, the primary calcareous spicules, are lost as the animal grows. I also demonstrated that the hairs are not uniquely innervated; the same sensory structures are produced in conjunction with other girdle ornaments on the marginal and ventral faces.

The postembryonic development of the antennal lobes of Periplaneta americana L. was examined with light- and electron-microscopical methods. There is no difference in the number of glomeruli and neurons in the antennal lobes of larval and adult animals. At hatching, the first larva already possesses the adult number of approximately 125 glomeruli and 500 to 560 deutocerebral neurons in the dorsolateral cell group of each antennal lobe. During postembryonic development the volume of the deutocerebral neurons increases three- to fourfold. The glomeruli of the first larva have about 7% of the volume of the corresponding adult glomeruli. Since number, pattern, and size ratio of glomeruli (with the exception of the macroglomerulus) are constant in all larval stages and adult animals, it is possible to identify individual glomeruli. During the whole postembryonic development the ordinary glomeruli show a continuous volume increase, which parallels the increase in antennal sensory input. The macroglomerulus develops by way of special growth of two to four neuropil units, but not before the last three to four larval stages and only in males. Its growth precedes the formation of antennal pheromone receptors during the final molt; these receptors are known to project into the macroglomerulus. The development of the macroglomerulus in the last larval stages of the male may be caused by a genetically fixed growth program of specific deutocerebral neurons.

The possibility that the exposure of the embryo to certain chemical substances can lead to behavioral disturbances is known from human epidemiological studies, e.g., in chronic poisoning with mercury and ethanol. Therefore, efforts are made to develop toxicological techniques with which new behavioral teratogens can be recognized. The review describes the most important experimental methods which are presently explored, and which are based on a rich body of knowledge accumulated by experimental psychologists. Most of the tests were developed with small animals, mostly with rats. They range from a rather straightforward determination of various reflexes to complex behavioral situations involving mechanical devices, operant conditioning techniques and procedures evaluating social behavior. In applying these methods in routine toxicology, it is important to remember, that many behavioral effects determined in newborn and adult animals are subtle. Moreover, they are influenced by a large variety of environmental factors affecting the health and the behavior of the mothers and of the offspring in the early and later phases of development. Therefore, the experiments must be conducted under highly standardized conditions and must be controlled rigorously. It is concluded that the best experimental strategy for the evaluation of potential behavioral teratogens is not yet established. Therefore, it would be premature to decide on a fixed protocol to be included in routine animal safety experiments for drugs and other chemical substances.

The morphology of the antennal hair-sensilla of Periplaneta americana, their distribution and frequency on the antennal flagellum have been examined by transmission- and scanning-electron microscopy. The types of sensilla were distinguished with respect to physiologically relevant criteria such as wall structure and number of sensory cells. Among the sensilla of the antenna of the adult male, long, single-walled sensilla with four sensory cells (types sw B), Probably responsible for reception of sexual pheromones, are most frequent, representing about 54% of the antennal sensilla. About half of these sensilla are newly-formed at the imaginal ecdysis; the other half are derived from the shorter type sw B sensilla of the nymphal antenna. Short type sw B sensilla are present in all larval stages of both sexes and in adult females as well. During the imaginal ecdysis of males, however, the length of these sensilla increases to double that found in nymphs. Dendritic branches also increase in number. During postembryonic development, the number of sensory fibers in the antennal flagellum increases nearly 20-fold, from 14,000 in the first larval instar to about 270,000 in the adult male. The greatest increase, approximately 90%, occurs during the last developmental stage.

In Asellus aquaticus certain distal antennular segments bear single sensilla referred to as aesthetascs. These show a proximal stem and a distal bulbous region. Depending on its position, each aesthetasc is innervated by either 50-60 or 70-80 bipolar sensory cells, the perikarya of which are situated within the pedunculus. Within the antennular segment the dendrites develop unbranched cilia (9 X 2 + 0 structure). The sensory cells are unusual in that mono- as well as biciliary dendrites are present within a single aesthetasc, the ratio of both types being correlated with the number of sensory cells. Cilia and receptor lymph cavity are enveloped by a set of 3-4 inner and 13-14 outer sheath cells, which terminate at the base of the sensillum, so that the delicate and poreless cuticle of the bulbous region encloses only outer segments within the receptor lymph fluid. A new molting type in arthropods is described in which the outer sheath cells alone build the new cuticle, whereas the inner sheath cells most probably have a protective function. A definition of aesthetascs is proposed based on fine-structural criteria. Functionally the sensilla are considered to be chemoreceptors. This assumption is confirmed by experiments with diluted vital dye as well as lanthanum showing that dissolved substances penetrate the poreless cuticle instantaneously.

The identities of two types of sensory organs in the body wall of Drosophila, namely the external sensory organs and the chordotonal organs, are under genetic control. Embryonic lethal mutations in the cut gene complex transform the external sensory organs into chordotonal organs. The neurons, as well as the support cells forming the external sensory structures, change their morphological and antigenic characteristics to those of chordotonal organs, providing genetic evidence that these two types of sensory organs are homologous. Similar transformations of external sensory organs are observed in adult mosaic flies. Analysis of mosaic larvae and flies suggests that the cut gene function is required either in or near external sensory organs in order for them to acquire their correct identity.

Enhancer of split (E(spl)) is one of a group of so-called neurogenic genes of Drosophila. We describe two different types of E(spl) alleles, dominant and recessive, which exert opposite effects on both central and peripheral nervous system development. The only extant dominant allele determines a reduction in the number of central neurons and peripheral sensilla; this phenotype is not reduced by a normal complement of wild-type alleles. Since animals carrying a triploidy for the wild-type locus develop similar defects, the dominant allele is probably the result of a gain-of-function mutation. Several recessive alleles, obtained as revertants of the dominant allele, are loss-of-function mutations and determine considerable neural hyperplasia. The present evidence suggests that neural defects of E(spl) mutants are due to defective segregation of neural and epidermal lineages, leading to neural commitment of less or of more cells than in the wild type, depending upon whether the animals carry the dominant or any of the recessive alleles, respectively. Therefore, E(spl) formally behaves as a gene switching between neural and epidermal pathways.

We have investigated the temporal pattern of appearance, cell lineage, and cytodifferentiation of selected sensory organs (sensilla) of adult Drosophila. This analysis was facilitated by the discovery that the monoclonal antibody 22C10 labels not only the neuron of the developing sensillum organ, but the accessory cells as well. The precursors of the macrochaetes and the recurved (chemosensory) bristles of the wing margin divide around and shortly after puparium formation, while those of the microchaetes and the stout and slender (mechanosensory) bristles of the wing margin divide between 9 h and 18 h after puparium formation (apf). The onset of sensillum differentiation follows the terminal precursor division within a few hours. Four of the cells in an individual microchaete organ are clonally related: A single first-order precursor cell divides to produce two second-order precursors; one of these divides into the neuron and thecogen cell, the other into the trichogen cell and tormogen cell. Along the anterior wing margin, two rounds of division generate the cells of the mechanosensory sensilla; here, no strict clonal relationship seems to exist between the cells of an individual sensillum. At the time of sensillum precursor division, many other, non-sensillum-producing cells within the notum and wing proliferate as well. This mitotic activity follows a spatially non-random pattern.

A consideration of head development in two species of Esox, lucius and americanus (ssp. vermiculatus) representing the two subgenera Esox and Kenozoa respectively, focused on the significance of the variations of the latero-sensory canal system, its associated bones, and other skeletal elements. In living forms only aspects of "regression" or specialization can be studied. Canals tend to be reduced to pit lines first at their termini but can be broken in their course. Pit lines range from nearly canals to surface structures, or even fail to develop. The number of neuromasts varies. Canal bones develop from two centers: neuromast related and deeper membranous centers which may have no relationship to neuromasts. Tooth-bearing and non-canal-related dermal bones have only membranous (original) centers. The number of neuromasts associated with a bone usually does not affect its development or form. In the case of the circumorbital bones, the extrascapulars, and the nasal, a one to one relationship has developed by regression--towards the development of the latero-sensory component only. The idea that reductions in bone number are commonly traceable to fusion is rejected although examples of fusion are know. Most bones that disappear are simply lost (no blastema or other evidence of their presence seen in development). The relationship between dermal bone and chondral bone is examined and there is evidence of the former giving rise to the latter. The ontogenic order of appearances shows a feeding (functional) correlation.

Several kinds of sensory organs (SOs) appear in stereotyped positions on the adult Drosophila cuticle. The generation of these SOs requires the activity of the achaete (ac) and scute (sc) genes. To investigate whether ac and sc also provide spatial information for the positioning of SOs, we have analyzed the patterns of expression of these genes in the wing imaginal disc around the time that SO precursors are being specified. We find that expression coincides with and is restricted to areas of the disc where these precursors are known to be located. In the loss-of-function sc mutant, sc RNA is depleted in a single area located in the region where the precursor for the suppressed macrochaeta should be found. Moreover, some, and probably all, SOs require expression of these genes to reach the earliest detectable differentiated state. These and other results presented here, together with the finding that expansion of the areas of ac and/or sc expression causes the development of ectopic SOs, indicate that ac and sc promote the determination of SO precursors and delimit the regions of the imaginal discs where they can develop.