Thesis (MScEng (Mechanical and Mechatronic Engineering))--Stellenbosch University, 2008. / A fluoroscopy-guided needle-positioning system is designed and tested as a first prototype for aiding urologists in gaining fast, accurate and repeatable kidney calyx access during a PCNL procedure while also reducing radiation exposure of the people involved. Image guidance is realized by modelling the fluoroscopic system as an adapted pinhole camera model and utilizing stereo vision principles on a stereo image pair. Calibration, distortion correction and image processing algorithms are implemented on images of a designed calibration object. Thereafter the resulting variables are used in the targeting of the calyx with the aid of a graphical user interface. The required relative translation and rotation of the needle from its current position to the target is calculated and the system is adjusted accordingly. Using digital cameras, needle placement accuracies of 2.5 mm is achieved within the calibrated volume in a simulated environment. Similar results are achieved in the surgery room environment using the fluoroscopic system. Successful needle access in two porcine kidney calyxes concluded the testing

L'excitotoxicité est un mécanisme pathologique qui résulte généralement du relargage massif de glutamate par les neurones en souffrance et qui conduit à la destruction des réseaux neuronaux ainsi qu'à des pertes cellulaires qui peuvent sévèrement impacter les fonctions cognitives et motrices. Au niveau du système nerveux central la séquence d'évènements qui supportent ce processus a bien été décrite et sert aujourd'hui de base au développement d'approches thérapeutiques ciblés pour limiter les conséquences du phénomène. Au niveau de l'oreille interne, les processus excitotoxiques pourraient également être impliqués dans différents types d'atteintes des fonctions auditives et vestibulaires tels que les traumas acoustiques, les surdités brusques, la neurite vestibulaire ou encore les vertiges d'origines ischémiques. Bien que les mécanismes moléculaires qui supportent ce type d'atteinte aient été bien explorés au niveau cochléaire, ils restent peu documentés au niveau du vestibule. De récentes études réalisées par notre équipe ont mis en évidence la participation des récepteurs du glutamate à la neurotransmission vestibulaire en conditions normales et ont également démontré les conséquences histologiques et fonctionnelles d'une atteinte excitotoxique du vestibule. Elles ont par ailleurs mis en évidence le fort potentiel de réparation spontané des synapses vestibulaires après déafférentation. Dans le présent travail de thèse, nous avons voulu identifier les effecteurs cellulaires qui contrôlent les phénomènes de déafférentation et de réafférentation. Dans ce but, nous avons mis au point un modèle d'étude original, qui permet un suivi histologique et fonctionnel de la séquence des évènements biologiques mis en jeu dans ces processus. En combinant des approches en immunohistochimie, microscopie électronique, électrophysiologie moléculaire et pharmacologie nous démontrons que les récepteurs glutamatergiques de types AMPA et NMDA sont tous deux impliqués dans les processus de déafférentation. Les récepteurs NMDA sont également essentiels au processus de réparation synaptique. Les résultats de ce travail apportent une nouvelle lumière sur le rôle des récepteurs du glutamate dans le processus de lésion/réparation des synapses vestibulaires. Cette nouvelle donne pourrait impacter directement sur les stratégies pharmacologiques de protection en cours de développement dans le domaine de la pathologie vestibulaire, mais également par extension à celui de la pathologie auditive. En outre le modèle de culture organotypique de tranche d'organe vestibulaire pourrait trouver application pour le screening de nouveaux composés à propriétés protectrices ou régénératrices. / Excitotoxicity is a pathological mechanism that usually results from the massive release of glutamate by suffering neurons and that leads to destruction of neural networks as well as cell losses that may severely impact cognitive and motor functions. In the central nervous system the sequence of events that supports this process has been extensively studied and is now the basis for the development of targeted therapeutic approaches to limit the consequences of the phenomenon. In the inner ear, excitotoxic damages may also support in different types of auditory and vestibular disorders such as acoustic traumas, sudden hearing loss, vestibular neuritis or dizziness of ischemic origin. Though the molecular mechanisms that support this type of injury have been well explored at cochlear level, they remain poorly documented in the vestibule. Recent studies by our team have confirmed the involvement of glutamate receptors in the vestibular calyx neurotransmission in normal conditions and also showed histological and functional consequences of excitotoxic damage in the vestibule. They also revealed the potential for spontaneous repair of the vestibular synapses after deafferentation. In present thesis, we aimed at identifying the cellular effectors that control the phenomena of deafferentation and reafferentation. For this purpose, we developed an original study model, which allows tracking histological and functional assessment of biological events involved in these processes. By combining approaches in immunohistochemistry, electron microscopy, molecular electrophysiology and pharmacology we demonstrate that both AMPA and NMDA type glutamate receptors are mainly involved in the process of deafferentation. NMDA receptors are also essential for synaptic repair process. The results of this work provide a new light on the role of glutamate receptors in the process of injury / repair of vestibular synapses. The novel observations could directly impact on the ongoing pharmacological protection strategies in the field of the vestibular pathology, and by extension to that of the hearing pathology. In addition, the organotypic culture model of vestibular organ slices may find application for screening new compounds with protective or regenerative properties.

Espécies da tribo Dipterygeae apresentam morfologia floral interessante, em especial o cálice, bem diferente dos demais membros de Papilionoideae. Representantes desta tribo exibem cálice assimétrico, constituído por duas sépalas adaxiais muito grandes e livres, e três sépalas unidas na base, com lobos abaxiais em forma de pequenos dentes. Assim, este trabalho visa a elucidar a origem deste tipo de cálice, por meio do estudo do desenvolvimento floral dos três gêneros da tribo, representados pelas espécies Dipteryx alata Vogel, Pterodon pubescens (Benth.) Benth. e Taralea oppositifolia Aubl.. Pretende-se também fornecer dados de desenvolvimento floral com valores taxonômico e evolutivo potenciais para o grupo. Botões florais e flores foram dissecados e preparados para análises de superfície em microscopia eletrônica de varredura e anatômica em microscopia de luz. As flores das espécies estão organizadas em inflorescências racemosas indeterminadas, são perfeitas e zigomorfas. A sequencia de surgimento dos verticilos florais é semelhante nas três espécies: bractéolas, sépalas, pétalas, carpelo + primeiros primórdios de estames antessépalos, e estames antepétalos. Dois primórdios de bractéolas são iniciados assincronicamente em D. alata e simultaneamente em P. pubescens e T. oppositifolia; estas bractéolas recobrem o meristema floral; a iniciação dos cinco primórdios de sépalas é unidirecional modificada em D. alata, helicoidal em P. pubescens e sequencial/sequencial modificada em T. oppositifolia; a iniciação dos cinco primórdios de pétalas é simultânea; o primórdio carpelar surge concomitante à iniciação dos primeiros primórdios de estames antessépalos; a iniciação dos estames antessépalos é em ordem unidirecional tendendo para verticilada; os cinco estames antepétalos surgem por último. A fenda carpelar surge do lado adaxial, sendo orientada para a direita ou para a esquerda, no lado oposto ao de iniciação do estame antepétalo adaxial. O androceu é assimétrico. Os filetes fundem-se devido ao meristema intercalar, mas não completamente, formando uma bainha aberta ao redor do carpelo. O carpelo é central com estigma truncado em D. alata, capitado em P. pubescens e puntiforme em T. oppositifolia. O tipo de cálice é elucidado nos estádios intermediários do desenvolvimento floral: a sépala abaxial e as laterais não se alongam, fundem-se, resultando em três lobos diminutos; já as adaxiais se alargam muito e alongam, permanecendo livres. O crescimento avantajado das sépalas adaxiais decorre do aumento do número de camadas e volume das células epidérmicas e subepidérmicas marginais. As flores possuem um apêndice em suas bractéolas, sépalas abaxiais e laterais, pétalas e anteras, onde se localiza um canal secretor. Em T. oppositifolia estes apêndices não ocorrem nas bractéolas e anteras. Além de canais, cavidades secretoras estão presentes no mesofilo das bractéolas, nas margens das sépalas e no apêndice das anteras. As alas exibem esculturas do tipo lamelado em D. alata e T. oppositifolia e lunado em P. pubescens. A presença de um canal secretor no apêndice dos órgãos florais constitui uma provável sinapomorfia para Dipterygeae. Caracteres com valor diagnóstico para as espécies são: (1) iniciação dos primórdios de bractéolas, (2) iniciação dos primórdios de sépalas, (3) proeminência do apêndice da antera, (4) tipo de escultura presente nas alas, (5) formato do estigma, (6) comprimento do estipe, (7) pilosidade do ovário, (8) curvatura do estilete, (9) posição da nervura central e (10) tipo de fruto. / Representatives of the tribe Dipterygeae exhibit interesting floral features, in special considering the calyx, which is quite different from that exhibited by other papilionoids. The calyx is asymmetric, constituted by two adaxial very large free sepals, and three abaxial unite sepals with teeth-like lobes. Thus, this work aims to to elucidate the developmental vias of this type of calyx, by studying the floral development and morphology of three genera of the tribe, represented by Dipteryx alata Vogel, Pterodon pubescens (Benth.) Benth. and Taralea oppositifolia Aubl. Also, we intend to provide other data on floral development with taxonomic and evolutionary values. Buds and flowers were dissected and prepared for surface and anatomical analyses, under scanning electron and light microscopy, respectively. The flowers of all species are arranged in indeterminate racemes, are zygomorphic and perfect. The initiation of floral whorls is similar in the three species: bracteoles, sepals, petals, carpel + antesepalous stamens, and antepetalous stamens. The floral apex initiates two bracteoles asynchronously in D. alata and simultaneously in P. pubescens and T. oppositifolia; these bracteoles cover the floral apex; the order of initiation of five sepal is modified unidirectional in D. alata, helical in P. pubescens and sequential or modified sequential in T. oppositifolia; the five petals initiate in a simultaneous order; the initiation of the carpel is concomitant to that of the antesepalous abaxial stamens; antesepalous stamens initiates in tendencies towards whorled order, and the five antepetalous stamens arise later. A central carpel with truncate stigma occurs in D. alata, capitate stigma in P. pubescens and punctiform stigma in T. oppositifolia. The carpel cleft of all species arises in the adaxial side and is positioned to the right or the left opposite to the antepetalous adaxial stamen primordium. The androecium is asymmetric. The filaments fuse due to intercalary meristematic activity, but not completely, forming an open sheath around the carpel. The different type of calyx is elucidate by observing the intermediary stages of floral development: the abaxial and lateral sepals do not elongate, fuse, resulting in a united structure with three lower lobes, while the adaxial sepals enlarge and elongate, remaining free. The great growth of adaxial sepals results of the increasing in the number of layers and volume of marginal epidermal and subepidermal cells. Bracteoles, abaxial and lateral sepals, petals and anthers are appendiculate. In T. oppositifolia these appendices do not occur in bracteoles and anthers. One secretory canal is observed within each appendix. Besides secretory canals, secretory cavities are found in the mesophyll of bracteoles, in the sepal margins and in the anther appendices. The wings show sculptures of lamellate type in D. alata and T. oppositifolia and of lunate type in P. pubescens. The presence of secretory canal in the floral appendices can be considered a synapomorphy for the tribe Dipterygeae. Diagnostic characters for the species are: (1) order of bracteole initiation, (2) order of sepal initiation, (3) anther appendix salience, (4) pattern of wing sculpture, (5) stigma shape, (6) stipe length, (7) ovary hairiness, (8) style bend, (6) position of the central rib, and (7) type of fruit.

The auditory brainstem is comprised of a number of synapses specialized for the transmission of high-fidelity synaptic signals. Within the first three postnatal weeks, these pathways acquire the ability to process high-frequency signals without compromising timing information. However, little is known regarding developmental adaptations which confer this ability. Situated in the sound localization pathway, the calyx of Held-medial nucleus of the trapezoid body synapse provides an ideal model for investigating such adaptations as both the pre- and postsynaptic neurons are accessible to electrophysiological experimentation. Using this synapse, we have shown herein that the spatial coupling between voltage-gated calcium channels (VGCCs) and synaptic vesicles (SVs) tightens during development. Immature synapses use a loosely-coupled arrangement of many N- and P/Q-type VGCCs (“microdomain” modality) while mature synapses use a tightly-coupled arrangement of fewer P/Q-type VGCCs, to release SVs (“nanodomain” modality). As a consequence of this tightening, synaptic delay (SD) shortens. By fluorescence- and electron microscopy of SVs near active zones, we further identified the filamentous protein septin 5 as a molecular substrate, differentiating the two release modalities, which may act as a spatial barrier separating VGCCs and SVs in immature synapses. Finally, we have demonstrated that changes in release modality affect the nature of short-term plasticity observed at this synapse. Using trains of action potentials as presynaptic voltage-commands, we showed that, downstream of calcium influx, the microdomain modality promotes short-term facilitation in excitatory postsynaptic currents (IEPSC), and calcium-dependent decreases in SD, with these being absent in synapses employing the nanodomain modality. In contrast, we found that as a result of depletion of SVs, short-term depression of IEPSC dominates in synapses using the nanodomain modality, and correlates with calcium-dependent increases in SD. These findings imply that the type of release modality has a significant impact on the strength and timing of synaptic responses. The microdomain modality imparts greater dynamic range in timing and strength, but does so at the cost of efficiency and fidelity, while the nanodomain modality is a key accomplishment consolidating the high-fidelity abilities of this synapse.

The auditory brainstem is comprised of a number of synapses specialized for the transmission of high-fidelity synaptic signals. Within the first three postnatal weeks, these pathways acquire the ability to process high-frequency signals without compromising timing information. However, little is known regarding developmental adaptations which confer this ability. Situated in the sound localization pathway, the calyx of Held-medial nucleus of the trapezoid body synapse provides an ideal model for investigating such adaptations as both the pre- and postsynaptic neurons are accessible to electrophysiological experimentation. Using this synapse, we have shown herein that the spatial coupling between voltage-gated calcium channels (VGCCs) and synaptic vesicles (SVs) tightens during development. Immature synapses use a loosely-coupled arrangement of many N- and P/Q-type VGCCs (“microdomain” modality) while mature synapses use a tightly-coupled arrangement of fewer P/Q-type VGCCs, to release SVs (“nanodomain” modality). As a consequence of this tightening, synaptic delay (SD) shortens. By fluorescence- and electron microscopy of SVs near active zones, we further identified the filamentous protein septin 5 as a molecular substrate, differentiating the two release modalities, which may act as a spatial barrier separating VGCCs and SVs in immature synapses. Finally, we have demonstrated that changes in release modality affect the nature of short-term plasticity observed at this synapse. Using trains of action potentials as presynaptic voltage-commands, we showed that, downstream of calcium influx, the microdomain modality promotes short-term facilitation in excitatory postsynaptic currents (IEPSC), and calcium-dependent decreases in SD, with these being absent in synapses employing the nanodomain modality. In contrast, we found that as a result of depletion of SVs, short-term depression of IEPSC dominates in synapses using the nanodomain modality, and correlates with calcium-dependent increases in SD. These findings imply that the type of release modality has a significant impact on the strength and timing of synaptic responses. The microdomain modality imparts greater dynamic range in timing and strength, but does so at the cost of efficiency and fidelity, while the nanodomain modality is a key accomplishment consolidating the high-fidelity abilities of this synapse.

Short-term synaptic plasticity results from use-dependent activity, lasts on the timescale of milliseconds to minutes, and is thought to underlie working memory and neuronal information processing. Here, we focus on two forms of short-term plasticity: 1) post-tetanic potentiation (PTP), which is induced by high-frequency stimulation, and 2) presynaptic ionotropic receptor-activated synaptic enhancement, which can be produced by the activation of presynaptic glycine receptors. Potentiation of evoked and spontaneous responses is thought to arise from elevations in presynaptic residual Ca2+, which activates one or more molecular targets to increase neurotransmitter release. However, the Ca2+ sensor protein has not yet been identified. The overall goal of this work is to elucidate the Ca2+-dependent mechanisms of short-term plasticity.

Sensory information is processed and encoded by neural networks. In order to understand how the nervous system is able to rapidly integrate and store sensory information, knowledge of the connections and properties of the neurons in these circuits is required. The fruit fly Drosophila melanogaster provides a particularly powerful species to investigate the neural circuits of the olfactory system because in addition to possessing a simple olfactory system amenable to circuit analysis, a host of genetic reagents are available, including the GAL4-UAS system for targeted gene expression. The mushroom bodies, paired structures historically implicated in olfactory learning and memory, receive olfactory information at the mushroom body calyx from second-order olfactory projection neurons (PNs). Within the calyx, individual PN axonal boutons are surrounded by dendritic arborizations from intrinsic Kenyon cells (KCs) and each tiny cluster constitutes a single microglomerulus. Cells that connect the calyx with other areas of the brain, extrinsic neurons (ENs), also contribute to microglomeruli. Most of these contain the neurotransmitter, GABA, and are presumed to be inhibitory. In this study, the synaptic characteristics, neural circuits, and plasticity of calycal cells have been investigated using a combination of serial section electron and confocal microscopy. The findings reveal several new features of the circuits in the calyx: 1) The calyx contains three ultrastructurally distinct types of PN boutons that are heterogeneous in shape and exhibit subtle differences in synaptic densities. 2) All PN boutons form both ribbon and non-ribbon synapses, and from their smaller size and fewer postsynaptic partners, non-ribbon synapses may possibly become converted to ribbon synapses after activity; the olfactory signal may then be transmitted more strongly and efficiently at ribbon synapses. 3) PN boutons with an electron-dense cytoplasm have the most ribbon synapses per unit area of membrane as well as the highest ratio of ribbon to non-ribbon synapses, and thus may be more active and efficient than other boutons. 4) KC neurites are not exclusively postsynaptic in the calyx and can form occasional ribbon synapses, the functional interpretation of which awaits identification of their postsynaptic partners and vesicle contents. 5) Each PN bouton may contribute input to a single dendritic KC claw at about three presynaptic sites. For the postsynaptic side, a single claw receives input from individual presynaptic sites that must be highly redundant. 6) There may be important processing of the olfactory signal by local circuits formed by ENs in the calyx; ENs form synaptic connections with PNs, KCs, and other ENs. 7) Extensive serial synapses link EN terminals into a network, presumed to be GABAergic and inhibitory, that extends between microglomeruli and may be autaptic. 8) The structure and synaptic connectivity of microglomeruli may undergo changes after adult emergence. 9) vGAT and GAD1-GAL4 lines drive ectopic expression of marker genes in KCs and are not reliable reporters of GABA-positive cells. 10) Previously identified calycal ENs (MB-C1, MB-C2/C3, MB-CP1) are not immunopositive for GAD1, a marker of GABA-containing cells. 11) A network of ENs expressing a GABA phenotype differently innervates anatomically and functionally discrete areas of the honeybee calyx, and in addition the density of innervation may change with alterations in age and/or experience.

L'homéostasie glutamatergique dans les fentes synaptiques régule la neurotransmission et préserve de l'excitotoxicité. Cela est particulièrement important dans l'oreille interne où il y a une libération soutenue de neurotransmetteur. Pour la plupart des cellules ciliées cochléaires et vestibulaires, la clairance du glutamate est assurée par les transporteurs du glutamate EAAT1 (GLAST) exprimés par les cellules de soutien. Un tel mécanisme n'est pas possible pour les cellules ciliées vestibulaires de type I car leur terminaison synaptique en calice empêche tout accès à la fente synaptique. Nous avons donc postulé qu'un ou plusieurs transporteurs du glutamate devaient être présents au niveau des cellules ciliées de type I ou du calice ou des deux.Grâce à des enregistrements électrophysiologiques, nous avons démontré qu'un courant anionique induit par le glutamate et bloqué par le DL-TBOA est présent dans les cellules ciliées de type I. Les techniques d'hybridation in situ et d'immunohistochimie ont révélé la présence d'EAAT4 et EAAT5. Ces deux transporteurs du glutamate, qui pourraient êtres à l'origine des courants enregistrés, sont exprimés par les cellules ciliées de type I et de type II. De plus, des expériences de RT-PCR et de microscopie électronique ont confirmé ces résultats et suggéré que ces transporteurs pourraient aussi être exprimés postsynaptiquement par le calice. Ces travaux de thèse montrent qu'EAAT4 et EAAT5, considérés respectivement comme spécifiques des tissus cérébelleux et rétiniens, ont une distribution plus large. Ces résultats posent la question des rôles potentiels de ces transporteurs dans l'homéostasie glutamatergique vestibulaire. / Glutamate homeostasis in synaptic clefts shape neurotransmission and prevent excitotoxicity. This may be particularly important in the inner ear where there is a continually high rate of neurotransmitter release. In the case of most cochlear and vestibular hair cells, clearance involves the diffusion of glutamate to supporting cells, where it is taken up by EAAT1 (GLAST), a glial glutamate transporter. A similar mechanism is unlikely to work in vestibular type I hair cells because the presence of calyx endings separates supporting cells from the synaptic zone. Based on this arrangement, we postulated that a glutamate transporter must be present in the type I hair cell, the calyx ending, or both. Using whole-cell patch-clamp recordings, we demonstrated that a glutamate-activated anion current blocked by DL-TBOA is expressed in type I hair cells. In situ hybridization and immunohistochemistry revealed that EAAT4 and EAAT5, two glutamate transporters that could support the anion current, are expressed in both type I and type II hair cells. Furthermore, RT-PCR and immunogold investigations confirmed those results and added that although preferentially expressed presynaptically, the transporters may also be present in the postsynaptic calyx membrane. Previously thought to be exclusively expressed in the cerebellum and retina respectively, this thesis work shows that EAAT4 and EAAT5 have a wider distribution. The potential role of these transporters in the glutamatergic homeostasis of the calyx synapse is then discussed.

Espécies da tribo Dipterygeae apresentam morfologia floral interessante, em especial o cálice, bem diferente dos demais membros de Papilionoideae. Representantes desta tribo exibem cálice assimétrico, constituído por duas sépalas adaxiais muito grandes e livres, e três sépalas unidas na base, com lobos abaxiais em forma de pequenos dentes. Assim, este trabalho visa a elucidar a origem deste tipo de cálice, por meio do estudo do desenvolvimento floral dos três gêneros da tribo, representados pelas espécies Dipteryx alata Vogel, Pterodon pubescens (Benth.) Benth. e Taralea oppositifolia Aubl.. Pretende-se também fornecer dados de desenvolvimento floral com valores taxonômico e evolutivo potenciais para o grupo. Botões florais e flores foram dissecados e preparados para análises de superfície em microscopia eletrônica de varredura e anatômica em microscopia de luz. As flores das espécies estão organizadas em inflorescências racemosas indeterminadas, são perfeitas e zigomorfas. A sequencia de surgimento dos verticilos florais é semelhante nas três espécies: bractéolas, sépalas, pétalas, carpelo + primeiros primórdios de estames antessépalos, e estames antepétalos. Dois primórdios de bractéolas são iniciados assincronicamente em D. alata e simultaneamente em P. pubescens e T. oppositifolia; estas bractéolas recobrem o meristema floral; a iniciação dos cinco primórdios de sépalas é unidirecional modificada em D. alata, helicoidal em P. pubescens e sequencial/sequencial modificada em T. oppositifolia; a iniciação dos cinco primórdios de pétalas é simultânea; o primórdio carpelar surge concomitante à iniciação dos primeiros primórdios de estames antessépalos; a iniciação dos estames antessépalos é em ordem unidirecional tendendo para verticilada; os cinco estames antepétalos surgem por último. A fenda carpelar surge do lado adaxial, sendo orientada para a direita ou para a esquerda, no lado oposto ao de iniciação do estame antepétalo adaxial. O androceu é assimétrico. Os filetes fundem-se devido ao meristema intercalar, mas não completamente, formando uma bainha aberta ao redor do carpelo. O carpelo é central com estigma truncado em D. alata, capitado em P. pubescens e puntiforme em T. oppositifolia. O tipo de cálice é elucidado nos estádios intermediários do desenvolvimento floral: a sépala abaxial e as laterais não se alongam, fundem-se, resultando em três lobos diminutos; já as adaxiais se alargam muito e alongam, permanecendo livres. O crescimento avantajado das sépalas adaxiais decorre do aumento do número de camadas e volume das células epidérmicas e subepidérmicas marginais. As flores possuem um apêndice em suas bractéolas, sépalas abaxiais e laterais, pétalas e anteras, onde se localiza um canal secretor. Em T. oppositifolia estes apêndices não ocorrem nas bractéolas e anteras. Além de canais, cavidades secretoras estão presentes no mesofilo das bractéolas, nas margens das sépalas e no apêndice das anteras. As alas exibem esculturas do tipo lamelado em D. alata e T. oppositifolia e lunado em P. pubescens. A presença de um canal secretor no apêndice dos órgãos florais constitui uma provável sinapomorfia para Dipterygeae. Caracteres com valor diagnóstico para as espécies são: (1) iniciação dos primórdios de bractéolas, (2) iniciação dos primórdios de sépalas, (3) proeminência do apêndice da antera, (4) tipo de escultura presente nas alas, (5) formato do estigma, (6) comprimento do estipe, (7) pilosidade do ovário, (8) curvatura do estilete, (9) posição da nervura central e (10) tipo de fruto. / Representatives of the tribe Dipterygeae exhibit interesting floral features, in special considering the calyx, which is quite different from that exhibited by other papilionoids. The calyx is asymmetric, constituted by two adaxial very large free sepals, and three abaxial unite sepals with teeth-like lobes. Thus, this work aims to to elucidate the developmental vias of this type of calyx, by studying the floral development and morphology of three genera of the tribe, represented by Dipteryx alata Vogel, Pterodon pubescens (Benth.) Benth. and Taralea oppositifolia Aubl. Also, we intend to provide other data on floral development with taxonomic and evolutionary values. Buds and flowers were dissected and prepared for surface and anatomical analyses, under scanning electron and light microscopy, respectively. The flowers of all species are arranged in indeterminate racemes, are zygomorphic and perfect. The initiation of floral whorls is similar in the three species: bracteoles, sepals, petals, carpel + antesepalous stamens, and antepetalous stamens. The floral apex initiates two bracteoles asynchronously in D. alata and simultaneously in P. pubescens and T. oppositifolia; these bracteoles cover the floral apex; the order of initiation of five sepal is modified unidirectional in D. alata, helical in P. pubescens and sequential or modified sequential in T. oppositifolia; the five petals initiate in a simultaneous order; the initiation of the carpel is concomitant to that of the antesepalous abaxial stamens; antesepalous stamens initiates in tendencies towards whorled order, and the five antepetalous stamens arise later. A central carpel with truncate stigma occurs in D. alata, capitate stigma in P. pubescens and punctiform stigma in T. oppositifolia. The carpel cleft of all species arises in the adaxial side and is positioned to the right or the left opposite to the antepetalous adaxial stamen primordium. The androecium is asymmetric. The filaments fuse due to intercalary meristematic activity, but not completely, forming an open sheath around the carpel. The different type of calyx is elucidate by observing the intermediary stages of floral development: the abaxial and lateral sepals do not elongate, fuse, resulting in a united structure with three lower lobes, while the adaxial sepals enlarge and elongate, remaining free. The great growth of adaxial sepals results of the increasing in the number of layers and volume of marginal epidermal and subepidermal cells. Bracteoles, abaxial and lateral sepals, petals and anthers are appendiculate. In T. oppositifolia these appendices do not occur in bracteoles and anthers. One secretory canal is observed within each appendix. Besides secretory canals, secretory cavities are found in the mesophyll of bracteoles, in the sepal margins and in the anther appendices. The wings show sculptures of lamellate type in D. alata and T. oppositifolia and of lunate type in P. pubescens. The presence of secretory canal in the floral appendices can be considered a synapomorphy for the tribe Dipterygeae. Diagnostic characters for the species are: (1) order of bracteole initiation, (2) order of sepal initiation, (3) anther appendix salience, (4) pattern of wing sculpture, (5) stigma shape, (6) stipe length, (7) ovary hairiness, (8) style bend, (6) position of the central rib, and (7) type of fruit.

No description available.