Multiple-input, multiple-output (MIMO) techniques have been proposed to meet the needs for higher data rates and lower delays in future wireless communication systems. The downlink capacity of multiuser MIMO systems is achieved when the system transmits to several users simultaneously. Frequently, many more users request service than the transmitter can simultaneously support. Thus, the transmitter requires a scheduling algorithm for the users, which must balance the goals of increasing throughput, reducing multiuser interference, lowering delays, ensuring fairness and quality of service (QoS), etc. In this thesis, we investigate the application of genetic algorithms (GAs) to perform scheduling in multiuser MIMO systems. GAs are a fast, suboptimal, low-complexity method of solving optimization problems, such as the maximization of a scheduling metric, and can handle arbitrary functions and QoS constraints. We first examine a system that transmits using capacity-achieving dirty paper coding (DPC). Our proposed GA structure both selects users and determines their encoding order for DPC, which affects the rates they receive. Our GA can also schedule users independently on different carriers of a multi-carrier system. We demonstrate that the GA performance is close to that of an optimal exhaustive search, but at a greatly reduced complexity. We further show that the GA convergence time can be significantly reduced by tuning the values of its parameters. While DPC is capacity-achieving, it is also very complex. Thus, we also investigate GA scheduling with two linear precoding schemes, block diagonalization and successive zero-forcing. We compare the complexity and performance of the GA with "greedy" scheduling algorithms, and find the GA is more complex, but performs better at higher signal-to-noise ratios (SNRs) and smaller user pool sizes. Both algorithms are near-optimal, yet much less complex than an exhaustive search. We also propose hybrid greedy-genetic algorithms to gain benefits from both types of algorithms. Lastly, we propose an improved method of optimizing the transmit covariance matrices for successive zero-forcing. Our algorithm significantly improves upon the performance of the existing method at medium to high SNRs, and, unlike the existing method, can maximize a weighted sum rate, which is important for fairness and QoS considerations. / Communications

Spacecraft missions such as spacecraft docking and formation flying require high precision relative position and attitude data. Although Global Positioining Systems can provide this capability near the earth, deep space missions require the use of alternative technologies. One such technology is the vision-based navigation (VISNAV) sensor system developed at Texas A&M University. VISNAV comprises an electro-optical sensor combined with light sources or beacons. This patented sensor has an analog detector in the focal plane with a rise time of a few microseconds. Accuracies better than one part in 2000 of the field of view have been obtained. This research presents a new approach involving simultaneous activation of beacons with frequency division multiplexing as part of the VISNAV sensor system. In addition, it discusses the synchronous demodulation process using digital heterodyning and decimating filter banks on a low-power fixed point DSP, which improves the accuracy of the sensor measurements and the reliability of the system. This research also presents an optimal and computationally efficient six-degree-of-freedom estimation algorithm using a new measurement model based on the attitude representation of Modified Rodrigues Parameters.

Multiuser and cooperative orthogonal frequency division multiplexing(OFDM) systems are being actively researched and adopted in wireless standards, owing to their advantages of robustness to multipath fading, modularity, and ability to achieve high data rates. In OFDM based systems, perfect frequency and timing synchronization is essential to maintain orthogonality among the subcarriers at the receiver. In multiuser OFDM on the uplink, timing offsets (TOs) and/or carrier frequency offsets (CFOs) of different users, caused due to path delay differences between different users, Doppler and/or poor oscillator alignment, can destroy orthogonality among subcarriers at the receiver. This results in multiuser interference (MUI)and consequent performance degradation. In this thesis, we are concerned with the analysis and mitigation of the effect of large CFOs and TOs in multiuser OFDM systems, including uplink orthogonal frequency division multiple access (OFDMA),uplink single-carrier frequency division multiple access(SC-FDMA), and cooperative OFDM. Uplink OFDMA: In the first part of this thesis, we analytically quantify the effect of large CFOs and TOs on the signal-to-interference plus noise ratio(SINR) and uncoded bit error rate(BER) performance of uplink OFDMA on Rayleigh and Rician fading channels, and show analytical results to closely match with simulation results. Such an SINR/BER analysis for uplink OFDMA in the presence of both large CFOs as well as TOs has not been reported before. We also propose interference cancelling(IC) receivers to mitigate the performance degradation caused due to large CFOs and TOs of different users. SC-FDMA versus OFDMA: An issue with uplink OFDMA is its high peak-to-average power ratio(PAPR).Uplink SC-FDMA is proposed in the standards as a good low-PAPR alternative to uplink OFDMA; e.g., SC-FDMA has been adopted in the uplink of 3GPP LTE. A comparative investigation of uplink SC-FDMA and OFDMA from a sensitivity to large CFOs and TOs view point has not been reported in the literature. Consequently, in the second part of the thesis, we carry out a comparative study of the sensitivity of SC-FDMA and OFDMA schemes to large CFOs and TOs of different users on the uplink. Our results show that while SC-FDMA achieves better performance due to its inherent frequency diversity advantage compared to OFDMA in the case of perfect synchronization, its performance can get worse than that of OFDMA in the presence of large CFOs and TOs. We further show that use of low-complexity multistage IC techniques, with the knowledge of CFOs and TOs of different users at the receiver, can restore the performance advantage of SC-FDMA over OFDMA. Cooperative OFDM: Cooperative OFDM is becoming popular because of its ability to provide spatial diversity in systems where each node has only one antenna. In most studies on cooperative communications, perfect time synchronization among cooperating nodes is assumed. This implies that the transmissions from different cooperating nodes reach the destination receiver in orthogonal time slots. In practice, however, due to imperfect time synchronization, orthogonality among different nodes’ signals at the destination receiver can be lost, causing inter-symbol interference(ISI).In the third part of the thesis, we investigate cooperative OFDM communications using amplify-and-forward(AF) protocol at the relay, in the presence of imperfect timing synchronization. We derive analytical expressions for the ISI as function of timing offset for cooperative OFDM with AF protocol, and propose an IC receiver to mitigate the effects of timing offset induced ISI.

El món de les comunicacions per la xarxa elèctrica (CXE) pot ser dividit en tres grans tipus: CXE en baix voltatge (CXE-BV), en mig voltatge (CXE-MV) i en alt voltatge (CXE-AV). En aquests últims anys, les CXE-BV han atret una gran expectació, ja que les seves capacitats han fet d'aquesta tecnologia una bona opció com alternativa pel bucle local d'accés i pel desplegament de xarxes d'àrea local, focalitzades aquestes últimes en l'entorn domèstic. A més, les CXE-BV inclouen un conjunt d'aplicacions de baixa velocitat orientades a l'operador, com la lectura automàtica de comptadors (LAC), distribució de càrrega, facturació dinàmica, etc. Per altra banda, les CXE-MV i CXE-AV, històricament lligades a tasques de telecontrol i teleprotecció, s'estan començant a considerar com un canal de comunicacions eficient i fiable. El desenvolupament de sistemes digitals i els esforços d'estandardització estan fent d'aquests canals un medi atractiu per a que els operadors elèctrics ofereixin serveis de comunicacions, ja que no necessiten invertir en infraestructura perquè la xarxa elèctrica ja està desplegada.En aquesta Tesi s'introduiran i es comentaran les particularitats de les tres xarxes elèctriques, després, es mostraran al lector les solucions tecnològiques existents pels canals de BV basats en la norma Europea CENELEC així com pels canals d'AV, mostrant que els sistemes actuals de LAC ofereixen una diversitat freqüencial molt baixa i que els mòdems CXE-AV estan ancorats en estendards antiquats.Aquest treball es mou per les tres topologies de la xarxa, particularment, en aplicacions orientades a la banda CENELEC, en mesura i modelat de canal, i en mesura i disseny del nivell físic per sistemes CXE-BV, CXE-MV i CXE-AV respectivament. Els sistemes actuals que exploten la banda CENELEC ofereixen mecanismes d'explotació de la diversitat freqüencial del canal molt limitats o nuls, donant lloc a una baixa robustesa en front a interferències i soroll de fons acolorit. Aquest treball proposa un esquema de modulació multiportadora que, mantenint una complexitat baixa, ofereix unes altes prestacions permetent un bon nivell d'explotació de la selectivitat freqüencial. Per al que a CXE-MV respecta, aquesta Tesi desenvolupa un model de canal determinístic-estadístic pels anells urbans de distribució de potència i, finalment, en sistemes CXE-AV, aquest treball proposa, basat en mesures de canal i proves de camp, un nivell físic de banda ampla capaç de incrementar la velocitat de comunicació mentre manté una baixa densitat espectral de potència limitant així la interferència a altres sistemes.PARAULES CLAU: Power line communications (PLC), low voltage (LV), medium voltage (MV), high voltage (HV), automatic meter reading (AMR), orthogonal frequency division multiplexing (OFDM), multicarrier spread spectrum (MC-SS), communication system design, channel measurements, channel modeling, scattering parameters. / El mundo de las comunicaciones por la red eléctrica (CRE) puede ser dividido en tres grandes tipos: CRE en bajo voltaje (CRE-BV), en medio voltaje (CRE-MV) y en alto voltaje (CRE-AV). En estos últimos años, las CRE-BV han atraído una gran expectación, ya que sus capacidades han hecho de esta tecnología una buena opción como alternativa para el bucle local de acceso y para el despliegue de redes de área local, focalizadas estas últimas en el entorno doméstico. Además, las CRE-BV incluyen un conjunto de aplicaciones de baja velocidad orientadas al operador como la lectura automática de contadores (LAC), distribución de carga, facturación dinámica, etc. Por otro lado, las CRE-MV y CRE-AV, históricamente ligadas a tareas de telecontrol y teleprotección, se están empezando a considerar como un canal de comunicaciones eficiente y fiable. El desarrollo de sistemas digitales y los esfuerzos de estandarización están haciendo de estos canales un medio atractivo para que los operadores eléctricos ofrezcan servicios de comunicaciones, ya que no necesitan invertir en infraestructura porque la red eléctrica ya está desplegada.En esta Tesis se introducirán y se comentarán las particularidades de las tres redes eléctricas, luego, se mostrarán al lector las soluciones tecnológicas existentes para los canales de BV basados en la norma Europea CENELEC así como para los canales de AV, mostrando que los sistemas actuales de LAC ofrecen una diversidad frecuencial muy baja y que los módems CRE-AV están anclados en estándares anticuados.Este trabajo se mueve por las tres topologías de red, particularmente, en aplicaciones orientadas a la banda CENELEC, en medida y modelado de canal, y en medida y diseño del nivel físico para sistemas CRE-BV, CRE-MV y CRE-AV respectivamente. Los sistemas actuales que explotan la banda CENELEC ofrecen mecanismos de explotación de la diversidad frecuencial del canal muy limitados o nulos, dando lugar a una escasa robustez frente a interferencias y ruido de fondo coloreado. Este trabajo propone un esquema de modulación multiportadora que, manteniendo una complejidad baja, ofrece unas altas prestaciones permitiendo un buen nivel de explotación de la selectividad frecuencial. Por lo que a CRE-MV respecta, esta Tesis desarrolla un modelo de canal determinístico-estadístico para los anillos urbanos de distribución de potencia y, finalmente, en sistemas de CRE-AV, este trabajo propone, basado en medidas de canal y pruebas de campo, un nivel físico de banda ancha capaz de incrementar la velocidad de comunicación mientras mantiene una baja densidad espectral de potencia limitando así la interferencia a otros sistemas.PALABRAS CLAVE: Power line communications (PLC), low voltage (LV), medium voltage (MV), high voltage (HV), automatic meter reading (AMR), orthogonal frequency division multiplexing (OFDM), multicarrier spread spectrum (MC-SS), communication system design, channel measurements, channel modeling, scattering parameters. / The world of Power line communications (PLC) can be divided into three main types: low voltage PLC (LV-PLC), medium voltage PLC (MV-PLC) and high voltage PLC (HV-PLC). These last years, LV-PLC has attracted a great expectation since its wideband capabilities has made this technology a suitable choice for last-mile access and in-home communications. Moreover, LV-PLC also includes a utility oriented low frequency and low speed applications, such as automatic meter reading (AMR), load distribution, dynamic billing and so on. On the other hand, MV-PLC and HV-PLC, historically oriented to teleprotection and telecontrol tasks, are being considered as a reliable communication channel. The development of digital equipment and the standardization efforts are making those channels an attractive medium for electrical utilities telecommunications services, since the network, as well as in LV-PLC, is already deployed. In this PhD dissertation, the three different PLC topologies are reviewed and the different communications techniques in such channels exposed. Then, a deep technological review of existing AMR solutions for the European CENELEC band, as well as HV-PLC systems is given, showing that existing AMR systems deliver low frequency diversity and HV-PLC systems are anchored in old fashioned standards.This work walks around the three topologies, specifically, CENELEC band utility oriented applications, channel measurement and modeling and channel measurement and physical layer design, regarding LV-PLC, MV-PLC and HV-PLC respectively. Existing CENELEC compliant systems deliver low or none frequency diversity mechanisms, yielding in a low robustness against colored noise and interference. This work propose a multicarrier based physical layer approach that, while keeping the complexity low, delivers high performance allowing a great level of frequency diversity. Focusing on MV-PLC, a hybrid deterministic-statistical channel model for urban underground rings is developed and, finally, in HV-PLC systems, this work proposes, based on measurements and field tests, a wideband physical layer in order to increase data rate while keeping low both the power spectral density and possible interference to other systems.KEYWORDS: Power line communications (PLC), low voltage (LV), medium voltage (MV), high voltage (HV), automatic meter reading (AMR), orthogonal frequency division multiplexing (OFDM), multicarrier spread spectrum (MC-SS), communication system design, channel measurements, channel modeling, scattering parameters.

Employing an error control code is one of the techniques to reduce the Peak-to-Average Power Ratio (PAPR) in an Orthogonal Frequency Division Multiplexing system; a well known class of such codes being the cosets of Reed-Muller codes. In this thesis, classes of such coset-codes of arbitrary linear codes are considered. It has been proved that the size of such a code can be doubled with marginal/no increase in the PAPR. Conditions for employing this method iteratively have been enunciated. In fact this method has enabled to get the optimal coset-codes. The PAPR of the coset-codes of the extended codes is obtained from the PAPR of the corresponding coset-codes of the parent code. Utility of a special type of lengthening is established in PAPR studies

This work deals with the design of MT transmit, MR receive antenna MIMO (Multiple Input Multiple Output) communication system where the transmitter performs a linear operation on data. This linear precoding model includes systems which involve signal shaping for achieving higher data rates, uncoded MIMO Multicarrier and Single-Carrier systems and, the more recent, MIMO-OFDM (Orthogonal Frequency Division Multiplexing) systems employing full diversity Space-Frequency codes. The objective of this work is to design diversity centric and rate centric linear precoded MIMO systems whose performance is better than the existing designs. In particular, we consider MIMO-OFDM systems, Zero Padded MIMO systems and MIMO systems with limited rate feedback. Design of full diversity MIMO-OFDM systems of rate symbol per channel use (1 s/ pcu) : In literature, MIMO-OFDM systems exploiting full diversity at a rate of 1 s/ pcu are based on a few specific Space-Frequency (SF)/ Space-Time-Frequency (STF) codes. In this work, we devise a general parameterized framework for the design of MIMO-OFDM systems employing full diversity STF codes of rate 1 s/ pcu. This framework unifies all existing designs and provides tools for the design of new systems with interesting properties and superior performance. Apart from rate and diversity, the parameters of the framework are designed for a low complexity receiver. The parameters of the framework usually depend on the channel characteristics (number of multipath, Delay Profile (DP)). When channel characteristics are available at the transmitter, a procedure to optimize the performance of STF codes is provided. The resulting codes are termed as DP optimized codes. Designs obtained using the optimization are illustrated and their performance is shown to be better than the existing ones. To cater to the scenarios where channel characteristics are not available at the transmitter, a complete characterization of a class of full diversity DP Independent (DPI) STF codes is provided. These codes exploit full diversity on channels with a given number of multipath irrespective of their characteristics. Design of DP optimized STF codes and DPI codes from the same framework highlights the flexibility of the framework. Design of Zero Padded (ZP) MIMO systems : While the MIMO-OFDM transmitter needs to precode data for exploiting channel induced multipath diversity, ZP MIMO systems with ML receivers are shown to exploit multipath diversity without any precoding. However, the receiver complexity of such systems is enormous and hence a study ZP MIMO system with linear receivers is undertaken. Central to this study involves devising low complexity receivers and deriving the diversity gain of linear receivers. Reduced complexity receiver implementations are presented for two classes of precoding schemes. An upper bound on the diversity gain of linear receivers is evaluated for certain precoding schemes. For uncoded systems operating on a channel of length L, this bound is shown to be MRL_MT +1 for uncoded transmissions, i.e, such systems tend to exploit receiver and multipath diversities. On the other hand, MIMO-OFDM systems designed earlier have to trade diversity with receiver complexity. These observations motivate us to use ZP MIMO systems with linear receivers for channels with large delay spread when receiver complexity is at a premium. Design examples highlighting the attractiveness of ZP systems when employed on channels with large delay spread are also presented. Efficient design of MIMO systems with limited feedback : Literature presents a number of works that consider the design of MIMO systems with partial feedback. The works that consider feedback of complete CSI, however, do not provide for an efficient system design. In this work, we consider two schemes, Correlation matrix feedback and Channel information feedback that convey complete CSI to the transmitter. This CSI is perturbed due to various impairments. A perturbation analysis is carried out to study the variations in mutual information for each of the proposed schemes. For ergodic channels, this analysis is used to design a MIMO system with a limited rate feedback. Using a codebook based approach, vector quantizers are designed to minimize the loss in ergodic capacity for each of the proposed schemes. The efficiency of the design stems from the ability to obtain closed-form expression for centroids during the iterative vector quantizer design. The performance of designed vector quantizers compare favorably with the existing designs. The vector quantizer design for channel information feedback is robust in the sense that the same codebook can be used across all operating SNR. Use of vector quantizers for improving the outage performance is also presented.

It is well known that communication using multiple antennas provides high data rate and reliability. Coding across space and time is necessary to fully exploit the gains offered by multiple input multiple output (MIMO) systems. One such popular method of coding for MIMO systems is space-time block coding. In applications where the terminals do not have enough physical space to mount multiple antennas, relaying or cooperation between multiple single antenna terminals can help achieve spatial diversity in such scenarios as well. Relaying techniques can also help improve the range and reliability of communication. Recently it has been shown that certain space-time block codes (STBCs) can be employed in a distributed fashion in single antenna relay networks to extract the same benefits as in point to point MIMO systems. Such STBCs are called distributed STBCs. However an important practical issue with STBCs and DSTBCs is its associated high maximum likelihood (ML) decoding complexity. The central theme of this thesis is to systematically construct STBCs and DSTBCs applicable for various scenarios such that are amenable for low decoding complexity. The first part of this thesis provides constructions of high rate STBCs from crossed product algebras that are minimum mean squared error (MMSE) optimal, i.e., achieves the least symbol error rate under MMSE reception. Moreover several previous constructions of MMSE optimal STBCs are found to be special cases of the constructions in this thesis. It is well known that STBCs from orthogonal designs offer single symbol ML decoding along with full diversity but the rate of orthogonal designs fall exponentially with the number of transmit antennas. Thus it is evident that there exists a tradeoff between rate and ML decoding complexity of full diversity STBCs. In the second part of the thesis, a definition of rate of a STBC is proposed and the problem of optimal tradeoff between rate and ML decoding complexity is posed. An algebraic framework based on extended Clifford algebras is introduced to study the optimal tradeoff for a class of multi-symbol ML decodable STBCs called ‘Clifford unitary weight (CUW) STBCs’ which include orthogonal designs as a special case. Code constructions optimally meeting this tradeoff are also obtained using extended Clifford algebras. All CUW-STBCs achieve full diversity as well. The third part of this thesis focusses on constructing DSTBCs with low ML decoding complexity for two hop, amplify and forward based relay networks under various scenarios. The symbol synchronous, coherent case is first considered and conditions for a DSTBC to be multi-group ML decodable are first obtained. Then three new classes of four-group ML decodable full diversity DSTBCs are systematically constructed for arbitrary number of relays. Next the symbol synchronous non-coherent case is considered and full diversity, four group decodable distributed differential STBCs (DDSTBCs) are constructed for power of two number of relays. These DDSTBCs have the best error performance compared to all previous works along with low ML decoding complexity. For the symbol asynchronous, coherent case, a transmission scheme based on orthogonal frequency division multiplexing (OFDM) is proposed to mitigate the effects of timing errors at the relay nodes and sufficient conditions for a DSTBC to be applicable in this new transmission scheme are given. Many of the existing DSTBCs including the ones in this thesis are found to satisfy these sufficient conditions. As a further extension, differential encoding is combined with the proposed transmission scheme to arrive at a new transmission scheme that can achieve full diversity in symbol asynchronous, non-coherent relay networks with no knowledge of the timing errors at the relay nodes. The DDSTBCs in this thesis are proposed for application in the proposed transmission scheme for symbol asynchronous, non-coherent relay networks. As a parallel to the non-coherent schemes based on differential encoding, we also propose non-coherent schemes for symbol synchronous and symbol asynchronous relay networks that are based on training. This training based transmission scheme leverages existing coherent DSTBCs for non-coherent communication in relay networks. Simulations show that this training scheme when used along with the coherent DSTBCs in this thesis outperform the best known DDSTBCs in the literature. Finally, in the last part of the thesis, connections between multi-group ML decodable unitary weight (UW) STBCs and groups with real elements are established for the first time. Using this connection, we translate the necessary and sufficient conditions for multi-group ML decoding of UW-STBCs entirely in group theoretic terms. We discuss various examples of multi-group decodable UW-STBCs together with their associated groups and list the real elements involved. These examples include orthogonal designs, quasi-orthogonal designs among many others.

Spacecraft missions such as spacecraft docking and formation flying require high precision relative position and attitude data. Although Global Positioining Systems can provide this capability near the earth, deep space missions require the use of alternative technologies. One such technology is the vision-based navigation (VISNAV) sensor system developed at Texas A&M University. VISNAV comprises an electro-optical sensor combined with light sources or beacons. This patented sensor has an analog detector in the focal plane with a rise time of a few microseconds. Accuracies better than one part in 2000 of the field of view have been obtained. This research presents a new approach involving simultaneous activation of beacons with frequency division multiplexing as part of the VISNAV sensor system. In addition, it discusses the synchronous demodulation process using digital heterodyning and decimating filter banks on a low-power fixed point DSP, which improves the accuracy of the sensor measurements and the reliability of the system. This research also presents an optimal and computationally efficient six-degree-of-freedom estimation algorithm using a new measurement model based on the attitude representation of Modified Rodrigues Parameters.

La communication cellulaire est un phénomène important pour le maintien de l’homéostasie des cellules. Au court des dernières années, cette sphère de recherche sur la signalisation cellulaire a connue des avancées importantes au niveau de l’identification des acteurs principaux impliqués dans la reconnaissance extracellulaire des signaux, ainsi que la compréhension des voies de signalisation engagées par les cellules pour répondre aux facteurs extracellulaires. Malgré ces nouvelles informations, les diverses interrelations moléculaires entre les acteurs ainsi que les voies de signalisation cellulaire, demeurent mal comprises. Le transfert d’énergie de résonance de bioluminescence (BRET) permet la mesure d’interactions protéiques et peut être utilisé dans deux configurations, le BRET480-YFP (connu aussi comme le BRET1) et le BRET400-GFP (connu aussi en tant que BRET2). Suite à l’oxydation de son substrat, la luciférase de renilla peut transférer son énergie à une protéine fluorescente, uniquement si elles sont à proximité l’une de l’autre (≤100Å). La combinaison dans un seul essai des BRET480-YFP et BRET400-GFP, a permis de suivre trois paires d’interactions, sur une même population cellulaire. Par contre, l’utilisation de deux substrats pour la réaction de bioluminescence rend impossible la mesure simultanée des différents signaux de BRET, pour ce trois nouvelles configurations de BRET ont été mises au point en utilisant des nouvelles protéines fluorescentes. Ainsi deux des nouvelles couleurs de BRET ayant des émissions résolues, le BRET400-BFP et le BRET400mAmetrine ont pu être combinées pour mesurer l’engagement par un RCPG d’une protéine G, ainsi que l’accumulation du second messager. La combinaison de ces BRET a également permis de révéler la formation d’un complexe entre le récepteur α2A adrénergique (α2AAR), Gαi1, le dimère Gβγ ainsi que la kinase des récepteurs couplés aux protéines G (GRK2), suite à l’activation du récepteur. De plus, seule l’entrée de GRK2 semble être en mesure de causer la désensibilisation du α2AAR, en s’intercalant entre Gαi1 et Gβγ. Par contre, la stabilisation de l’interaction entre α2AAR et la β-arrestine2 semble nécessiter l’activité kinase de GRK2. Une autre étude a révélé l’importance de différentes Gα pour la mobilisation du calcium, suite à l’activation du récepteur aux opioïdes de type delta (DOR). Suite à la surexpression de Gα de la famille Gαq, il a été possible de mesurer une influence de ces Gα sur la mobilisation du calcium. Toutefois, cette réponse calcique mesurée en présence des Gαq demeure sensible aux prétraitements à la toxine de Bordetella pertussis, qui inhibe sélectivement l’activité des Gαi. De plus, la co-expression de Gαi et Gαq permet de potentialiser la mobilisation de calcium, démontrant une interrelation entre ces deux familles de protéine Gα, pour la signalisation du DOR. Afin de démontrer l’interrelation directe, des expériences de BRET ont été réalisées entre différentes Gα. En plus de montrer la formation de complexes sélectifs entre les Gα, les expériences de BRET réalisées en parallèle d’analyses de séquences de Gα, ont également mis à jour un site de sélectivité d’interaction entre les Gα, l’hélice α4. Suite à la transposition de cette hélice α4 de Gα12 sur Gαi1, qui normalement n’interagissent pas, il a été possible de forcer l’interaction entre Gα12 et Gαi1, confirmant ainsi que cette hélice α contient l’information permettant une sélectivité d’interaction. Au cours de cette thèse, il a été possible de générer de nouvelles méthodes de mesure d’interactions protéiques qui permettent de multiplexer différents signaux, ce qui a permis de mettre à jour de nouvelles interactions entre divers effecteurs de la signalisation de RCGP / Cellular communication is an important phenomenon for the maintenance of cellular homeostasis. Recently, important progress has been made in the cell signalling research field concerning the identification of the major actors and the cellular pathways engaged in response to these extracellular factors. However, in spite of this new information, the interrelationships at the molecular level between the various cellular actors and the different signalling pathways remain badly understood. Bioluminescence resonance energy transfer (BRET) monitors interactions between proteins and can be used in two configurations, the BRET480-YFP (also known as BRET1) and the BRET400-GFP (also known as BRET2). Following oxidation of its substrate, renilla luciferase transfers its energy to a fluorescent protein, only if they are in close proximity (≤100Å). By combining the BRET480-YFP and BRET400-GFP in one assay, it is possible to follow three pair-wise interactions in the same cellular population. However, using two bioluminescence reaction substrates limits the possibility of measuring the different BRET signals simultaneously. In order to measure multiple BRET signals simultaneously, three new BRET configurations, based on the BRET400-GFP, were developed using fluorescent proteins with different emission wavelengths. Two of the new BRET colors which have resolved emission wavelengths, the BRET400-BFP and BRET400mAmetrine, were combined for measuring the heterotrimeric G protein engagement by the vasopressin V2 receptor, as well as the accumulation of the second messenger. Combining these new BRET techniques reveals for the first time the formation of a complex between the α2A adrenergic receptor (α2AAR), Gαi1, the Gβγ dimer and G protein-receptor kinase (GRK2) following receptor activation. Moreover, only the entry of GRK2 into the receptor complex is required for the α2AAR desensitization, by inserting between Gαi1 and Gβγ. On the other hand, the stabilization of the interaction between α2AAR and β-arrestin2 requires the kinase activity of GRK2. Another study revealed the importance of multiple Gα subunits for calcium mobilization induced upon activation of the delta opioid receptor (DOR). Gαq subfamily member overexpression altered the DOR-induced calcium mobilization, but this Gαq calcium mobilization remained sensitive to pre-treatement pertussis toxin, through selective inhibition of the activity of Gαi members. Moreover, Gαi and Gαq co-expression potentiated calcium mobilization, suggesting an interrelationship between these two Gα families in DOR signaling. This Gαi and Gαq interrelationship could result from the formation of a complex close to the receptor. In order to test this hypothesis, BRET experiments were performed, with the aim of measuring the presence of complexes between different Gα. In addition to demonstrating complex formation between Gα subunits, the BRET experiments in parallel with sequence analysis, also revealed a selective interaction site between the Gα, the α4 helix. By swapping the a4 helix of Gαi with the α4 helix of Gα12, which doesn’t normally interact with Gα12, it was possible to force the interaction between Gα12 and Gαi to confirm that this α helix contains information concerning the selectivity of interactions between Gα subunits. During this thesis, new methods were to detect protein interactions and multiplexing these methods allowed the detection of novel interactions between signalling effectors of GPCRs.

No description available.