The role of under-investigated connexins in musculoskeletal tissue: a brief review with emphasis on bone tissue / El papel de las escasamente investigadas conexinas en el tejido músculo-esquelético: una breve revisión con énfasis en el tejido óseo

Connexins (Cxs) are a family of transmembrane proteins that form gap junctions and hemi-channels, which mediate cell-cell communication between neighboring cells and the respective extracellular milieu in different tissues. Most tissues and cell types throughout the body express one or more Cx proteins, highlighting its importance in regulating cell growth, differentiation, adhesion, migration, cell death and others. Moreover, Cx can propagate intracellular signals through its C-terminus domain, and thus function beyond a mere channel. Cx43 is the most highly expressed and most well studied Cx in bone and musculoskeletal tissues, although Cx40, Cx45, Cx46 and more recently, the Cx37 have been described in bone tissue, along with Cx26, Cx32 and Cx39 in other musculoskeletal tissues. Here, we discuss the basic structure of gap junctions and the role of the Cxs in musculoskeletal tissue, with special focus on Cx37. (AU)

Las conexinas (Cxs) son una familia de proteínas transmembrana que forman uniones en hendidura y hemicanales encargados de mediar la comunicación entre células vecinas y el respectivo medio extracelular en diferentes tejidos. La mayoría de los tejidos y células expresan una o más proteínas conexina, jugando un papel importante en la regulación de la proliferación celular, diferenciación, adhesión, migración y muerte celular, entre otras funciones. Además de actuar como un canal, las conexinas pueden propagar señales intracelulares a través del dominio C-terminal. La Cx43 es la conexina mas expresada y mejor estudiada en el tejido óseo y el músculo, aunque las Cx40, Cx45, Cx46, y mas recientemente Cx37, son también detectadas en el hueso. A su vez la expresión de la Cx26, Cx32 y Cx39 ha sido observada en otros tejidos músculoesqueléticos. En este manuscrito describimos la estructura básica de las uniones tipo gap y el papel que las Cxs, y en especial la Cx37, tienen en tejidos músculo-esqueléticos. (AU)

Importância das comunicações intercelulares para o desenvolvimento de folículos ovarianos / Intercellular communications in ovarian follicles

Durante a foliculogênese em mamíferos, ocorre um longo e complexo processo no qual o oócito adquire a competência necessária para a fecundação. Nesse processo ocorre uma comunicação metabólica bidirecional entre os oócitos e as células somáticas dentro do folículo que garante substratos para o oócito em desenvolvimento. Essa comunicação é mediada pelas junções celulares (junções comunicantes e junções aderentes) presentes nas projeções transzonais. As junções celulares e moléculas de adesão são responsáveis principalmente por promover a adesão entre as células foliculares; mas podem atuar em vias de sinalização celular e na regulação da transcrição gênica nas células somáticas e oócitos. Além disso, as junções comunicantes (junções gap) são canais intermembranares que intermediam a comunicação entre essas células através da passagem de pequenas moléculas. Essas junções comunicantes são compostas por proteínas denominadas conexinas; as conexinas 37 e 43 são as predominantes nos folículos ovarianos. Dessa forma, o conhecimento acerca das junções celulares é de extrema importância para o estudo da foliculogênese. A presente revisão teve como objetivo abordar os principais tipos de junções celulares existentes entre as células foliculares, com destaque para as junções gap e as principais proteínas de membranas (conexinas) presentes nos diferentes estágios do desenvolvimento folicular.

During the mammalian folliculogenesis, a long and complex process occurs, which the oocyte acquires the necessary competence for fecundation. In this process there is a metabolic bidirectional communication among the oocyte and somatic cells inside the follicle, which provides substrates for the oocyte developmental competence. This communication is mediated by cellular junctions (occlusions, adherens and gap junctions) localized in the transzonal projections. Cellular junctions and adhesion mollecules are responsable mainly for promoving the adhesion among follicular cells, however they can act in cellular signaling pathways and in regulation of genic transcription in the follicular cells and oocyte. Moreover, the communication junctions (gap junctions) are intermembrane channels that intermediate the communication among these cells through the passage of small molecules. These gap junctions are composed by connexins, of which the connexins 37 and 43 are the most frequently found in the ovarian follicle. Thus, knowledge of these cellular junctions are of great importance for studying the folliculogenesis process. The aim of this review was to report the main types of cellular junctions localized among the follicular cells, especially the gap junctions and the main membrane proteins (connexins) found in different stages of the follicular development.

Effect of magnesium ion on human osteoblast activity

Magnesium, a promising biodegradable metal, has been reported in several studies to increase bone formation. Although there is some information regarding the concentrations of magnesium ions that affect bone remodeling at a cellular level, little is known about the effect of magnesium ions on cell gap junctions. Therefore, this study aimed to systematically investigate the effects of different concentrations of magnesium on bone cells, and further evaluate its effect on gap junctions of osteoblasts. Cultures of normal human osteoblasts were treated with magnesium ions at concentrations of 1, 2 and 3 mM, for 24, 48 and 72 h. The effects of magnesium ions on viability and function of normal human osteoblasts and on gap junction intercellular communication (GJIC) in osteoblasts were investigated. Magnesium ions induced significant (P<0.05) increases in cell viability, alkaline phosphate activity and osteocalcin levels of human osteoblasts. These stimulatory actions were positively associated with the concentration of magnesium and the time of exposure. Furthermore, the GJIC of osteoblasts was significantly promoted by magnesium ions. In conclusion, this study demonstrated that magnesium ions induced the activity of osteoblasts by enhancing GJIC between cells, and influenced bone formation. These findings may contribute to a better understanding of the influence of magnesium on bone remodeling and to the advance of its application in clinical practice.

Neuropeptide Y, substance P, and human bone morphogenetic protein 2 stimulate human osteoblast osteogenic activity by enhancing gap junction intercellular communication

Bone homeostasis seems to be controlled by delicate and subtle “cross talk” between the nervous system and “osteo-neuromediators” that control bone remodeling. The purpose of this study was to evaluate the effect of interactions between neuropeptides and human bone morphogenetic protein 2 (hBMP2) on human osteoblasts. We also investigated the effects of neuropeptides and hBMP2 on gap junction intercellular communication (GJIC). Osteoblasts were treated with neuropeptide Y (NPY), substance P (SP), or hBMP2 at three concentrations. At various intervals after treatment, cell viability was measured by the MTT assay. In addition, cellular alkaline phosphatase (ALP) activity and osteocalcin were determined by colorimetric assay and radioimmunoassay, respectively. The effects of NPY, SP and hBMP on GJIC were determined by laser scanning confocal microscopy. The viability of cells treated with neuropeptides and hBMP2 increased significantly in a time-dependent manner, but was inversely associated with the concentration of the treatments. ALP activity and osteocalcin were both reduced in osteoblasts exposed to the combination of neuropeptides and hBMP2. The GJIC of osteoblasts was significantly increased by the neuropeptides and hBMP2. These results suggest that osteoblast activity is increased by neuropeptides and hBMP2 through increased GJIC. Identification of the GJIC-mediated signal transduction capable of modulating the cellular activities of bone cells represents a novel approach to studying the biology of skeletal innervation.

A low-protein diet during pregnancy prevents modifications in intercellular communication proteins in rat islets

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O tabagismo está associado com a remodelação de junções comunicantes no coração de ratos: explicação do paradoxo dos fumantes? / Smoking is associated with remodeling of gap junction in the rat heart: smoker's paradox explanation?

FUNDAMENTO: Em estudo anterior, utilizando o modelo de ratos, a exposição à fumaça do cigarro durante 5 semanas aumentou a sobrevida após IAM, apesar da idade similar e tamanho do infarto entre fumantes e não fumantes, e da ausência de reperfusão. OBJETIVO: Dessa forma, o presente estudo teve como objetivo analisar os efeitos da exposição à fumaça do cigarro sobre a intensidade, distribuição ou fosforilação da conexina 43 no coração de ratos. MÉTODOS: Ratos Wistar, pesando 100 g, foram distribuídos aleatoriamente em 2 grupos: 1) Controle (n = 25); 2) Expostos à fumaça do cigarro (ETS), n = 23. Depois de 5 semanas, foram conduzidas análise morfométrica do ventrículo esquerdo, imuno-histoquímica e Western blot para conexina 43 (Cx43). RESULTADOS: A fração do volume de colágeno, as áreas transversais e o peso ventricular não foram estatisticamente diferentes entre os grupos controle e ETS. O grupo ETS apresentou uma coloração de menor intensidade da Cx43 em discos intercalados (Controle: 2,32 ± 0,19; ETS: 1,73 ± 0,18; p = 0,04). A distribuição da Cx43 em discos intercalados não diferiu entre os grupos (Controle: 3,73 ± 0,12; ETS: 3,20 ± 0,17; p = 0,18). Os ratos do grupo ETS mostraram um nível maior de forma desfosforilada da Cx43 (Controle: 0,45 ± 0,11; ETS: 0,90 ± 0,11; p = 0,03). Por outro lado, o Cx43 total não diferiu entre os grupos de controle e ETS (Controle: 0,75 ± 0,19; ETS: 0,93 ± 0,27; p = 0,58). CONCLUSÃO: A exposição à fumaça do cigarro resultou na remodelação das junções comunicantes cardíacas, caracterizada por alterações na quantidade e fosforilação da Cx43 em corações de ratos. Essa constatação pode explicar o paradoxo dos fumantes observado em alguns estudos.

BACKGROUND: In a previous study utilizing the rat model, exposure to tobacco smoke for 5 weeks increased survival after AMI, despite similar age and infarct size between the smokers and nonsmokers, and absence of reperfusion. OBJECTIVE: Thus, this study aimed to analyze the effects of exposure to tobacco smoke on intensity, distribution or phosphorylation of connexin 43 in the rat heart. METHODS: Wistar rats weighing 100 g were randomly allocated into 2 groups: 1) Control (n = 25); 2) Exposed to tobacco smoke (ETS), n = 23. After 5 weeks, left ventricular morphometric analysis, immunohisthochemistry and western blotting for connexin 43 (Cx43) were performed. RESULTS: Collagen volume fraction, cross-sectional areas, and ventricular weight were not statistically different between control and ETS. ETS showed lower stain intensity of Cx43 at intercalated disks (Control: 2.32 ± 0.19; ETS: 1.73 ± 0.18; p = 0.04). The distribution of CX43 at intercalated disks did not differ between the groups (Control: 3.73 ± 0.12; ETS: 3.20 ± 0.17; p = 0.18). ETS rats showed higher levels of dephosphorylated form of Cx43 (Control: 0.45 ± 0.11; ETS: 0.90 ± 0.11; p = 0.03). On the other hand, total Cx43 did not differ between control and ETS groups (Control: 0.75 ± 0.19; ETS: 0.93 ± 0.27; p = 0.58). CONCLUSION: Exposure to tobacco smoke resulted in cardiac gap junction remodeling, characterized by alterations in the quantity and phosphorylation of the Cx43, in rats hearts. This finding could explain the smoker's paradox observed in some studies.

Papel das sinapses elétricas em crises epilépticas / The role of eletrical synapses in epileptic seizures

INTRODUÇÃO: No sistema nervoso central a comunicação entre neurônios se realiza através de estruturas denominadas sinapses: elétricas ou químicas. As sinapses elétricas são formadas pela aproximação das membranas plasmáticas de dois neurônios formando estruturas chamadas junções comunicantes (gap junctions, do inglês). As junções comunicantes são compostas por seis subunidades da proteína conexina de cada membrana, formando poros que comunicam o citoplasma de células adjacentes e permitem a passagem de íons e pequenas moléculas. OBJETIVOS: A presente revisão pretende descrever e discutir os principais resultados que apontam para uma importante relação entre junções comunicantes e sincronia neuronal durante crises epilépticas. RESULTADOS E CONCLUSÃO: Quando um neurônio é despolarizado, este tipo de comunicação permite a rápida transferência iônica entre as células, promovendo alta sincronia neuronal. Recentemente, o papel das junções comunicantes na geração e propagação de descargas epilépticas tem sido estudado através do uso de diferentes modelos experimentais in vivo, in vitro e in silico (modelos computacionais).

INTRODUCTION: In the central nervous system, neuronal communication is accomplished by structures called synapses: electrical or chemical. Electrical synapses are formed by the apposition of plasmatic membranes at gap junctions and the interaction of connexin subunits from two neurons. At this site, connexin complexes create intercellular pores that communicate the cytoplasm of adjacent neurons and allow free flow of ions and small molecules. OBJECTIVE: In this review, we will present and discuss recent results showing the possible involvement of electrical synapses in the neuronal hypersynchronization during epileptic seizures. RESULTS AND CONCLUSION: When a neuron is depolarized, ions flow very rapidly from one cell to the other promoting high neuronal synchrony. More recently, the role of gap junctions in the generation and propagation of epileptic discharges has been investigated using combined approaches of in vivo, in vitro and in silico (computational) models.

Gap junction reduction in cardiomyocytes following transforming growth factor-â treatment and Trypanosoma cruzi infection

Gap junction connexin-43 (Cx43) molecules are responsible for electrical impulse conduction in the heart and are affected by transforming growth factor-â (TGF-â). This cytokine increases during Trypanosoma cruzi infection, modulating fibrosis and the parasite cell cycle. We studied Cx43 expression in cardiomyocytes exposed or not to TGF-â T. cruzi, or SB-431542, an inhibitor of TGF-â receptor type I (ALK-5). Cx43 expression was also examined in hearts with dilated cardiopathy from chronic Chagas disease patients, in which TGF-â signalling had been shown previously to be highly activated. We demonstrated that TGF-â treatment induced disorganised gap junctions in non-infected cardiomyocytes, leading to a punctate, diffuse and non-uniform Cx43 staining. A similar pattern was detected in T. cruzi-infected cardiomyocytes concomitant with high TGF-â secretion. Both results were reversed if the cells were incubated with SB-431542. Similar tests were performed using human chronic chagasic patients and we confirmed a down-regulation of Cx43 expression, an altered distribution of plaques in the heart and a significant reduction in the number and length of Cx43 plaques, which correlated negatively with cardiomegaly. We conclude that elevated TGF-â levels during T. cruzi infection promote heart fibrosis and disorganise gap junctions, possibly contributing to abnormal impulse conduction and arrhythmia that characterise severe cardiopathy in Chagas disease.

Como a célula óssea reconhece o estímulo mecânico? / How do bone cells sense mechanical loading?

Sob a influência da gravidade, o tecido ósseo sofre maior ou menor deformação de acordo com a intensidade das atividades da vida diária. Sabe-se que as atividades que resultam em impacto são as que mais estimulam a osteogênese e assim reduzem a perda de massa óssea. Conhecer como as células ósseas reconhecem a deformação mecânica imposta ao osso e iniciam uma série de reações bioquímicas em cadeia é de fundamental importância para o desenvolvimento de práticas terapêuticas e preventivas na atividade ortopédica. Ainda há um longo caminho para o entendimento de todo esse processo, mas o conhecimento atual progrediu bastante e há pesquisas com finalidade terapêutica. O sinal mecânico para ser transformado em biológico (mecanotransdução) deve ser amplificado no nível celular pela estrutura histológica do tecido ósseo, gerando tensões em proteínas da membrana celular (integrinas) e alterando a estrutura espacial dessas proteínas. Essa alteração ativa ligações entre elas e o citoesqueleto, originando as adesões focais, locais onde proteínas citoplasmáticas são recrutadas para facilitar as reações bioquímicas. A quinase de adesão focal (FAK) é a principal delas, sendo autoativada após sofrer alteração estrutural pelas integrinas. A FAK ativada incita reações em cascata, resultando na ativação da ERK-1/2 e da Akt, proteínas que, junto com a FAK, regulam a produção da massa óssea. Acredita-se que o osteócito seja a célula óssea responsável por reconhecer o estímulo mecânico e transmiti-lo aos osteoblastos e osteoclastos. Canais iônicos e gap junctions são cogitados como meios de comunicação intercelular para a transmissão bioquímica do estímulo mecânico. Esses eventos ocorrem continuamente no tecido ósseo e regulam a remodelação óssea.

Influenced by gravidity, bone tissue experiences stronger or lighter deformation according to the strength of the activities of daily life. Activities resulting in impact are particularly known to stimulate osteogenesis, thus reducing bone mass loss. Knowing how bone cells recognize the mechanical deformation imposed to the bone and trigger a series of biochemical chain reactions is of crucial importance for the development of therapeutic and preventive practices in orthopaedic activity. There is still a long way to run until we can understand the whole process, but current knowledge has shown a strong progression, with researches being conducted focused on therapies. For a mechanical sign to be transformed into a biological one (mechanotransduction), it must be amplified at cell level by the histological structure of bone tissue, producing tensions in cell membrane proteins (integrins) and changing their spatial structure. Such change activates bindings between these and the cytoskeleton, producing focal adhesions, where cytoplasmatic proteins are recruited to enable easier biochemical reactions. Focal adhesion kinase (FAK) is the most important one being self-activated when its structure is changed by integrins. Activated FAK triggers a cascade of reactions, resulting in the activation of ERK-1/2 and Akt, which are proteins that, together with FAK, regulate the production of bone mass. Osteocytes are believed to be the mechanosensor cells of the bone and to transmit the mechanical deformation to osteoblasts and osteoclasts. Ionic channels and gap junctions are considered as intercellular communication means for biochemical transmission of a mechanical stimulus. These events occur continuously on bone tissue and regulate bone remodeling.

Conexinas y sistema cardiovascular / Conexins and the cardiovascular system

Las comunicaciones intercelulares son eventos biológicos esenciales en los organismos multicelulares, asociadas con el control del crecimiento y la diferenciación celular, la apoptosis, las respuestas adaptativas de células diferenciadas y la sincronización de funciones celulares. Las uniones intercelulares, conocidas como uniones gap, estructuralmente constituidas por conexinas, tienen una participación activa en estos procesos. A nivel cardiovascular, la comunicación célula a célula es indispensable, en condiciones normales, para la embriogénesis cardíaca, la transmisión del impulso eléctrico, la sincronización de la actividad contráctil cardíaca, la transmisión de señales reflejas vasculares, entre otras funciones biológicas, mientras que en condiciones patológicas, a causa de mutaciones genéticas heredadas o adquiridas, participan en el desarrollo de cardiopatías congénitas, arritmogénesis y remodelación miocárdica. La investigación en genética y biología molecular seguramente encontrará recursos para la prevención y el tratamiento de las distintas cardiopatías en las que la comunicación intercelular tiene un papel fisiopatológico.

Gap junctions in hematopoietic stroma control proliferation and differentiation of blood cell precursors

Investigamos a comunicação intercelular mediada por junções comunicantes em um modelo in vitro de hematopoiese, usando uma linhagem celular murina de estroma de medula óssea, S-17, e culturas primárias de precursores hematopoiéticos murinos. As S-17 expressam diversas conexinas, sendo a principal a conexina43. A expressão de conexinas e a formação de canais funcionais são moduladas pela densidade das células de estroma. A transfecção de células S-17 com um vetor contendo seqüências senso ou anti-senso de conexina43 aumenta ou diminui, respectivamente, a síntese de conexina43 e o acoplamento intercelular. Nestas condições, a modulação da comunicação mediada pelas junções comunicantes modifica o padrão de crescimento das células do próprio estroma, bem como a capacidade do estroma para sustentar a hematopoiese. O aumento na expressão de conexina43 resulta em um retardo na diferenciação das células sanguíneas, e no aumento da produção de precursores hematopoiéticos, enquanto a diminuição na expressão da conexina43 resulta numa diferenciação acelerada dos precursores mielóides. Estes resultados sugerem que o acoplamento mediado por conexina nas células de estroma modula a razão entre proliferação e diferenciação dos precursores hematopoiéticos. Propomos portanto, que o aumento da comunicação mediada por junções comunicantes no estroma elicita uma produção aumentada de células imaturas de medula óssea, através de um retardo em sua diferenciação terminal, induzindo consequentemente um período de proliferação prolongado dos precursores hematopoiéticos.

Uniones en hendidura y su papel funcional en el tracto reproductor femenino

En el aparato reproductor femenino se expresan diferentes conexinas (Cxs), proteínas que forman canales de uniones en hendidura (CUH) entre células en contacto, permitiendo la coordinación de respuestas metabólicas y/o eléctricas de grupos celulares. Los CUH jugarían un papel relevante en el desarrollo de las células de la granulosa, ya que permiten la comunicación heteróloga entre el ovocito y las células del cúmulo manteniendo la detención meiótica. En la trompa de Falopio, los CUH coordinarían el batido ciliar del epitelio y la contracción muscular, facilitando el desplazamiento de los gametos y del embrión. En el útero, los CUH conectan a las células miometriales y también a las endometriales. El aumento de CUH durante el preparto permitiría la contracción uterina coordinada facilitando el trabajo de parto al término del embarazo. La expresión de las Cxs es regulada por hormonas, lo que explicaría el perfil de CUH presentes en los diversos tipos celulares del tracto genital en diferentes estadios fisiológicos del sistema reproductor.

Gap junctions in human hippocampal sclerosis

Um grande número de portadores de epilepsia do lobo temporal não obtém controle de suas crises com os medicamentos antiepilépticos atualmente disponíveis. Nesses casos de epilepsia fármaco-resistente uma possibilidade terapêutica é a ressecção cirúrgica do foco epiléptico. Estudos em preparações de fatias hipocampais de tecido epileptogênico proveniente da cirurgia de epilepsia tem mostrado a ocorrência de junções comunicantes (gap junctions) o que poderia estar relacionada com o hipersincronismo observado no foco epiléptico. Neste trabalho, nós mostramos estudos eletrofisiológicos in vitro feitos na formação hipocampal obtida de um paciente após uma cirurgia de epilepsia. População de espículas (Population spikes) de pequena amplitude foram registradas sugerindo uma lesão neuronal significante. As propriedades intrínsicas de membranas obtidas desses neurônios mostraram-se normais. A presença das junções comunicantes nas células granulares do giro dentado também foram observadas. Nossos achados sugerem que a presença de junções comunicantes no tecido epiléptico pode ter um papel importante na geração de descargas epileptiformes

Cell migration in the postnatal subventricular zone

New neurons are constantly added to the olfactory bulb of rodents from birth to adulthood. This accretion is not only dependent on sustained neurogenesis, but also on the migration of neuroblasts and immature neurons from the cortical and striatal subventricular zone (SVZ) to the olfactory bulb. Migration along this long tangential pathway, known as the rostral migratory stream (RMS), is in many ways opposite to the classical radial migration of immature neurons: it is faster, spans a longer distance, does not require radial glial guidance, and is not limited to postmitotic neurons. In recent years many molecules have been found to be expressed specifically in this pathway and to directly affect this migration. Soluble factors with inhibitory, attractive and inductive roles in migration have been described, as well as molecules mediating cell-to-cell and cell-substrate interactions. However, it is still unclear how the various molecules and cells interact to account for the special migratory behavior in the RMS. Here we will propose some candidate mechanisms for roles in initiating and stopping SVZ/RMS migration

Importância da junçäo comunicante na fisiologia sensorioneural da cóclea / Importance of gap junction in cochlear sensorioneural physiology

A membrana celular estabelece contato com outras células por meio de estruturas de junção especializada. Na fisiologia sensorioneural da cóclea, a junção comunicante ou gap junction é a de maior relevância. Pela sua estrutura protéica, em forma hexagonal, quando unida à proteína correspondente da membrana celular adjacente, forma um canal com alta difusão para íons com pequena massa molecular. Pela formação desse canal intercelular, as proteínas foram denominadas de conexinas, sendo sua produção regulada pelo gene conexina 26. Na cóclea, essas junções têm como função promover a remoção rápida de íons potássio das células ciliadas sensoriais a fim de se manter a capacidade auditiva. Mutações nesse gene levaria a alterações na estrutura protéica dessas junções prejudicando a remoção dos íons potássio das células ciliadas e impedindo, portanto, sua excitação a novos estímulos sonoros, resultando em perda auditiva. A alta proporção de deficiência auditiva sensorioneural não-sindrômica, quando ocasionada pela mutação no gene conexina 26, indica a relevância do estudo molecular desse gene na fisiologia da audição.

Gap junctions in the cardiovascular and immune systems

Gap junctions are clusters of intercellular channels directly connecting the cytoplasm of adjacent cells. These channels are formed by proteins named connexins and are present in all metazoan organisms where they serve diverse functions ranging from control of cell growth and differentiation to electric conduction in excitable tissues. In this overview we describe the presence of connexins in the cardiovascular and lympho-hematopoietic systems giving the reader a summary of the topics to be covered throughout this edition and a historical perspective of the discovery of gap junctions in the immune system

Gap junctions in cells of the immune system: structure, regulation and possible functional roles

Gap junction channels are sites of cytoplasmic communication between contacting cells. In vertebrates, they consist of protein subunits denoted connexins (Cxs) which are encoded by a gene family. According to their Cx composition, gap junction channels show different gating and permeability properties that define which ions and small molecules permeate them. Differences in Cx primary sequences suggest that channels composed of different Cxs are regulated differentially by intracellular pathways under specific physiological conditions. Functional roles of gap junction channels could be defined by the relative importance of permeant substances, resulting in coordination of electrical and/or metabolic cellular responses. Cells of the native and specific immune systems establish transient homo- and heterocellular contacts at various steps of the immune response. Morphological and functional studies reported during the last three decades have revealed that many intercellular contacts between cells in the immune response present gap junctions or "gap junction-like" structures. Partial characterization of the molecular composition of some of these plasma membrane structures and regulatory mechanisms that control them have been published recently. Studies designed to elucidate their physiological roles suggest that they might permit coordination of cellular events which favor the effective and timely response of the immune system

Gap junction modulation by extracellular signaling molecules: the thymus model

Gap junctions are intercellular channels which connect adjacent cells and allow direct exchange of molecules of low molecular weight between them. Such a communication has been described as fundamental in many systems due to its importance in coordination, proliferation and differentiation. Recently, it has been shown that gap junctional intercellular communication (GJIC) can be modulated by several extracellular soluble factors such as classical hormones, neurotransmitters, interleukins, growth factors and some paracrine substances. Herein, we discuss some aspects of the general modulation of GJIC by extracellular messenger molecules and more particularly the regulation of such communication in the thymus gland. Additionally, we discuss recent data concerning the study of different neuropeptides and hormones in the modulation of GJIC in thymic epithelial cells. We also suggest that the thymus may be viewed as a model to study the modulation of gap junction communication by different extracellular messengers involved in non-classical circuits, since this organ is under bidirectional neuroimmunoendocrine control

Regulation of gap junctions by protein phosphorylation

Gap junctions are constituted by intercellular channels and provide a pathway for transfer of ions and small molecules between adjacent cells of most tissues. The degree of intercellular coupling mediated by gap junctions depends on the number of gap junction channels and their activity may be a function of the state of phosphorylation of connexins, the structural subunit of gap junction channels. Protein phosphorylation has been proposed to control intercellular gap junctional communication at several steps from gene expression to protein degradation, including translational and post-translational modification of connexins (i.e., phosphorylation of the assembled channel acting as a gating mechanism) and assembly into and removal from the plasma membrane. Several connexins contain sites for phosphorylation for more than one protein kinase. These consensus sites vary between connexins and have been preferentially identified in the C-terminus. Changes in intercellular communication mediated by protein phosphorylation are believed to control various phsysiological tissue and cell functions as well as to be altered under pathological conditions. (AU)Gap junctions are constituted by intercellular channels and provide a pathway for transfer of ions and small molecules between adjacent cells of most tissues. The degree of intercellular coupling mediated by gap junctions depends on the number of gap junction channels and their activity may be a function of the state of phosphorylation of connexins, the structural subunit of gap junction channels. Protein phosphorylation has been proposed to control intercellular gap junctional communication at several steps from gene expression to protein degradation, including translational and post-translational modification of connexins (i.e., phosphorylation of the assembled channel acting as a gating mechanism) and assembly into and removal from the plasma membrane. Several connexins contain sites for phosphorylation for more than one protein kinase. These consensus sites vary between connexins and have been preferentially identified in the C-terminus. Changes in intercellular communication mediated by protein phosphorylation are believed to control various phsysiological tissue and cell functions as well as to be altered under pathological conditions.

Biphasic effects of gap junctional uncoupling agents on the propagation of retinal spreading depression

The spreading depression wave of Leäo (SD) propagates in the nervous tissue at 3-10 mm/min and is a accompanied by an increase of intracellular Ca²+ concentration and a decrease in the extracellular milieu. Recently the spread of Ca²+ waves with propagating velocities of the same order of magnitude has been detected in cultures of nerve cells. It has been suggested that these waves, which can be blocked by gap junctional agents, are related to SD. The present experiments describe some effects of heptanol and octanol (10µM to 10mM), well-known uncoupling agents of gap junctions, on the slow voltage changes and the velocity of propagation of Leäo's phenomenon in isolated chick retina: 1) a 50 to 700 µM concentration of these alcohols in the superfusate solutions increased the velocity, whereas high concentration (1 to 10 mM) decreased and subsequently halted the spread; 2) the recovery period of the slow voltage changes of SD was shorter in the presence of alcohol and its amplitude was larger during faster SD waves. These effects were observed in the retina during successive individual waves as well as in the course of circling SD. All of these effects were reversible, without any sign of damage to the retina. In the concentration range of 50 µM, methanol, ethanol, butanol and cyclo-hexanol had no effect on retinal SD. These data suggest the involvement of gap junctions in Leäo's wave. This involvement is complex and its functional effects together with other factors that influence the velocity of propagation of SD sre being investigated