A key role of TRPC channels in the regulation of electromechanical activity of the developing heart.

Aims It is well established that dysfunction of voltage-dependent ion channels results in arrhythmias and conduction disturbances in the foetal and adult heart. However, the involvement of voltage-insensitive cationic TRPC (transient receptor potential canonical) channels remains unclear. We assessed the hypothesis that TRPC channels play a crucial role in the spontaneous activity of the developing heart.Methods and results TRPC isoforms were investigated in isolated hearts obtained from 4-day-old chick embryos. Using RT-PCR, western blotting and co-immunoprecipitation, we report for the first time that TRPC1, 3, 4, 5, 6, and 7 isoforms are expressed at the mRNA and protein levels and that they can form a macromolecular complex with the alpha 1C subunit of the L-type voltage-gated calcium channel (Cav1.2) in atria and ventricle. Using ex vivo electrocardiograms, electrograms of isolated atria and ventricle and ventricular mechanograms, we found that inhibition of TRPC channels by SKF-96365 leads to negative chrono-, dromo-, and inotropic effects, prolongs the QT interval, and provokes first-and second-degree atrioventricular blocks. Pyr3, a specific antagonist of TRPC3, affected essentially atrioventricular conduction. On the other hand, specific blockade of the L-type calcium channel with nifedipine rapidly stopped ventricular contractile activity without affecting rhythmic electrical activity.Conclusions These results give new insights into the key role that TRPC channels, via interaction with the Cav1.2 channel, play in regulation of cardiac pacemaking, conduction, ventricular activity, and contractility during cardiogenesis.

Differential expression and hypoxic regulation of adenosine receptors and TRPC channels in the developing heart.

Objectives: To characterize the modifications of gene expression of adenosine receptors (AR), TRPC channels, HIF-1α and iNOS during the early cardiogenesis in response to chronic hypoxia exposure. Methods: 4-day-old chick embryos were subjected in ovo to 6H, 12H and 24H of hypoxia (10% O2). The mRNA expression was quantified by RT-qPCR. Results: The targeted genes were found to be expressed at mRNA level with a differential expression pattern within the heart. Hypoxia has no significant effect on mRNA expression of ARs, TRPCs channels and iNOS within the heart. By contrast, HIF-1α mRNA expression shows a tendency to be down-regulated by hypoxia. Conclusion: These results suggest that an intrauterine oxygen lack does not significantly affect expression of genes involved in adenosine signaling and in calcium handling by store operated channels (TRPC).

Identification des canaux TRPC impliqués dans la potentialisation à long terme des interneurones de la région CA1 de l'hippocampe chez le rat

Le réseau neuronal de l’hippocampe joue un rôle central dans la mémoire en modifiant de façon durable l’efficacité de ses synapses. Dans les interneurones de la couche oriens/alveus (O/A), l’induction de la potentialisation à long terme (PLT) requiert les courants postsynaptiques excitateurs évoqués par les récepteurs métabotropes du glutamate de sous-type 1a (CPSEmGluR1a) et l’entrée subséquente de Ca2+ via des canaux de la famille des transient receptor potential (TRP). Le but de ce projet était d’identifier les canaux TRP responsables des CPSEmGluR1a et d’explorer les mécanismes moléculaires régulant leur ouverture. Nous avons déterminé par des enregistrements électrophysiologiques que les CPSEmGluR1a étaient spécifiques aux interneurones O/A et qu’ils étaient indépendants de la phospholipase C. Nous avons ensuite examiné l’expression des TRPC et leur interaction avec mGluR1a par les techniques de RT-PCR, d’immunofluorescence et de co-immunoprécipitation. Nos résultats montrent que TRPC1 et mGluR1a s’associent dans l’hippocampe et que ces deux protéines sont présentes dans les dendrites des interneurones O/A. En revanche, TRPC4 ne semble s’associer à mGluR1a qu’en système recombinant et leur colocalisation paraît limitée au corps cellulaire. Finalement, nous avons procédé à des enregistrements d’interneurones dans lesquels l’expression des TRPC a été sélectivement supprimée par la transfection d’ARN interférant et avons ainsi démontré que TRPC1, mais non TRPC4, est une sous-unité obligatoire du canal responsable des CPSEmGluR1a. Ces travaux ont permis de mieux comprendre les mécanismes moléculaires à la base de la transmission synaptique des interneurones O/A et de mettre en évidence un rôle potentiel de TRPC1 dans la PLT.

Activation of Transient Receptor Potential Canonical 3 (TRPC3)-mediated Ca2+ Entry by A1 Adenosine Receptor in Cardiomyocytes Disturbs Atrioventricular Conduction.

Although the activation of the A(1)-subtype of the adenosine receptors (A(1)AR) is arrhythmogenic in the developing heart, little is known about the underlying downstream mechanisms. The aim of this study was to determine to what extent the transient receptor potential canonical (TRPC) channel 3, functioning as receptor-operated channel (ROC), contributes to the A(1)AR-induced conduction disturbances. Using embryonic atrial and ventricular myocytes obtained from 4-day-old chick embryos, we found that the specific activation of A(1)AR by CCPA induced sarcolemmal Ca(2+) entry. However, A(1)AR stimulation did not induce Ca(2+) release from the sarcoplasmic reticulum. Specific blockade of TRPC3 activity by Pyr3, by a dominant negative of TRPC3 construct, or inhibition of phospholipase Cs and PKCs strongly inhibited the A(1)AR-enhanced Ca(2+) entry. Ca(2+) entry through TRPC3 was activated by the 1,2-diacylglycerol (DAG) analog OAG via PKC-independent and -dependent mechanisms in atrial and ventricular myocytes, respectively. In parallel, inhibition of the atypical PKCζ by myristoylated PKCζ pseudosubstrate inhibitor significantly decreased the A(1)AR-enhanced Ca(2+) entry in both types of myocytes. Additionally, electrocardiography showed that inhibition of TRPC3 channel suppressed transient A(1)AR-induced conduction disturbances in the embryonic heart. Our data showing that A(1)AR activation subtly mediates a proarrhythmic Ca(2+) entry through TRPC3-encoded ROC by stimulating the phospholipase C/DAG/PKC cascade provide evidence for a novel pathway whereby Ca(2+) entry and cardiac function are altered. Thus, the A(1)AR-TRPC3 axis may represent a potential therapeutic target.

Store-operated Ca2+ Entry Mediated by Orai1 and TRPC1 Participates to Insulin Secretion in Rat β-Cells.

Store-operated Ca(2+) channels (SOCs) are voltage-independent Ca(2+) channels activated upon depletion of the endoplasmic reticulum Ca(2+) stores. Early studies suggest the contribution of such channels to Ca(2+) homeostasis in insulin-secreting pancreatic β-cells. However, their composition and contribution to glucose-stimulated insulin secretion (GSIS) remains unclear. In this study, endoplasmic reticulum Ca(2+) depletion triggered by acetylcholine (ACh) or thapsigargin stimulated the formation of a ternary complex composed of Orai1, TRPC1, and STIM1, the key proteins involved in the formation of SOCs. Ca(2+) imaging further revealed that Orai1 and TRPC1 are required to form functional SOCs and that these channels are activated by STIM1 in response to thapsigargin or ACh. Pharmacological SOCs inhibition or dominant negative blockade of Orai1 or TRPC1 using the specific pore mutants Orai1-E106D and TRPC1-F562A impaired GSIS in rat β-cells and fully blocked the potentiating effect of ACh on secretion. In contrast, pharmacological or dominant negative blockade of TRPC3 had no effect on extracellular Ca(2+) entry and GSIS. Finally, we observed that prolonged exposure to supraphysiological glucose concentration impaired SOCs function without altering the expression levels of STIM1, Orai1, and TRPC1. We conclude that Orai1 and TRPC1, which form SOCs regulated by STIM1, play a key role in the effect of ACh on GSIS, a process that may be impaired in type 2 diabetes.

TR(i)P, TR(i)P, hooray : new functions for TRPC2 and TRPC3

Kalciumjonen påverkar många aspekter av cellbiologi, från fertiliseringen av äggcellen till tillväxt och differentiering av alla typer av celler. Störningar i hur kalciumjonerna transporteras in och friges i cellerna kan ha stor betydelse för hur sjukdomar framskrider. På grund av detta är det viktigt att studera hur kalciumtransportörer såsom kalciumkanaler fungerar och hur de påverkar cellernas funktioner. I avhandlingen har klassiska transient receptorpotential (TRPC) katjonkanaler studerats. Dessa kanaler är permeabla för kalcium och aktiveras nedströms av G-proteinkopplade receptorer. Resultaten som presenteras i avhandlingen visar på nya funktioner för TRPC2- och TRPC3-kanalerna. Utöver TRPC3-kanalens roll i plasmamembranen, där den medverkar i receptorreglerat kalciuminflöde, tyder våra resultat på att TRPC3 har en potentiell intracellulär roll. Vid överuttryck av kanalen lokaliserade kanalen till endoplasmatiska nätverket där kanalen friger intracellulärt kalcium. TRPC2 har en viktig funktion i möss där kanalen reglerar hur djuret reagerar på feromoner. Våra resultat tyder på att TRPC2 är involverad i autokrin sfingosin-1-fosfat (S1P)-signalering. S1P som produceras i cellerna transporterades ut från cellerna varpå det aktiverade S1P2-receptorer på plasmamembranen. Diacylglycerol som bildas som en följd av aktivering av S1P2-receptorn aktiverade i sin tur inflöde av extracellulärt kalcium. Våra resultat indikerar att TRPC2 är kanalen som förmedlar detta kalciuminflöde. I sköldkörtelceller studerades nedströms effekter för TRPC2. Kanalen reglerade negativt cAMP-produktionen i cellerna vilket hämmar extracellulärt signalreglerat kinas 1/2 (ERK1/2). När uttrycket för TRPC2 minskades i cellerna, ökades produktionen av cAMP och fosforyleringen av ERK1/2, vilket resulterade i ökat uttryck för receptorn för sköldkörtelstimulerande hormon. Slutligen, presenterar vi resultat som tyder på att TRPC2 är viktigt för sekretion av prekursorn till sköldkörtelhormon.

Role of the canonical transient receptor potential channel 2 (TRPC2) in thyroid cell function

Kalciumjonen reglerar flera processer i celler såsom transkribering av gener, celldelning, cellernas rörlighet och celldöd. Därför har cellerna utvecklat många mekanismer för att reglera den intracellulära kalciumkoncentrationen. Kalciumkanaler spelar en viktig roll i denna regleringsprocess. TRPC-kanalerna (eng. canonical transient receptor potential) är en familj av jonkanaler med sju medlemmar (TRPC1-7) vars regleringsmekanismer och fysiologiska roller är varierande. TRPC2-kanalens fysiologiska signifikans, samt hur kanalen regleras, är dåligt karakteriserad. För första gången, rapporterar vi närvaron av TRPC2 kanalen i råttans sköldkörtelceller samt primära sköldkörtelceller från råtta. Hos gnagare har TRPC2 antagits vara exklusivt uttryckt i det vomeronasala organet. För att undersöka den fysiologiska betydelsen av kanalen, har vi utvecklat stabila celler med nedreglerat TRPC2 (shTRPC2) m.h.a. shRNA-teknik. Nedreglering av TRPC2 resulterade i stora skillnader i flera viktiga cellulära funktioner och i regleringen av sköldkörtelcellernas cellsignalering. Nedreglering av TRPC2 orsakade minskad agonist-beroende frigivning av kalcium från det endoplasmatiska nätverket, samt minskat agonist-beroende inflöde av extracellulärt kalcium, men ökade det basala kalciuminflödet. Uttrycket av PKCβ1 och PKCδ, SERCA-aktiviteten och kalciumhalten i det endoplasmatiska nätverket minskade i shTRPC2 celler. Kommunikation mellan kalcium- och cAMP-signalering påvisades vara TRPC2-beroende, vilket visades reglera uttrycket av TSH-receptorn. Vi undersökte också betydelsen av TRPC2 kanalen i reglering av sköldkörtelcellers proliferation, migration, vidhäftning och invasion; processer som alla var dämpade i shTRPC2 celler. Samamnfattningsvis påvisade dessa resultat en ny och viktig fysiologisk betydelse för TRPC2 kanalerna.

Sildenafil decreases rat tracheal hyperresponsiveness to carbachol and changes canonical transient receptor potential gene expression after antigen challenge

Inhibition of type-5 phosphodiesterase by sildenafil decreases capacitative Ca2+ entry mediated by transient receptor potential proteins (TRPs) in the pulmonary artery. These families of channels, especially the canonical TRP (TRPC) subfamily, may be involved in the development of bronchial hyperresponsiveness, a hallmark of asthma. In the present study, we evaluated i) the effects of sildenafil on tracheal rings of rats subjected to antigen challenge, ii) whether the extent of TRPC gene expression may be modified by antigen challenge, and iii) whether inhibition of type-5 phosphodiesterase (PDE5) may alter TRPC gene expression after antigen challenge. Sildenafil (0.1 µM to 0.6 mM) fully relaxed carbachol-induced contractions in isolated tracheal rings prepared from naive male Wistar rats (250-300 g) by activating the NO-cGMP-K+ channel pathway. Rats sensitized to antigen by intraperitoneal injections of ovalbumin were subjected to antigen challenge by ovalbumin inhalation, and their tracheal rings were used to study the effects of sildenafil, which more effectively inhibited contractions induced by either carbachol (10 µM) or extracellular Ca2+ restoration after thapsigargin (1 µM) treatment. Antigen challenge increased the expression of the TRPC1 and TRPC4 genes but not the expression of the TRPC5 and TRPC6 genes. Applied before the antigen challenge, sildenafil increased the gene expression, which was evaluated by RT-PCR, of TRPC1 and TRPC6, decreased TRPC5 expression, and was inert against TRPC4. Thus, we conclude that PDE5 inhibition is involved in the development of an airway hyperresponsive phenotype in rats after antigen challenge by altering TRPC gene expression.

Stromal interaction molecule 1 and its role in the regulation, proliferation and protein expression in follicular ML-1 thyroid cancer cells

Calcium (Ca2+) is involved in the regulation of variety of cellular functions including hallmarks of cancer development such as cellular migration and cellular proliferation. Store-operated calcium entry (SOCE) is a central mechanism in cellular calcium signaling and in maintaining the cellular calcium balance. Stromal interaction molecule 1(STIM1) has been identified as an important constituent of SOCE. In this thesis , the STIM1 proteins are studied for their importance in cellular processes and their effects on the expression of S1P1, S1P2, S1P3, VEGFR-2, and TRPC-1 in follicular ML-1 thyroid cancer cells. The results show the importance of STIM1 proteins in SOCE in these cells. The SOCE is significantly reduced in the STIM1 knockdown cells. The results also show the importance of STIM1 proteins in the expression of S1P2 and VEGFR-2 in these cells, as knockdown of STIM1 was shown to upregulate the expression of S1P2 and VEGFR-2. The migration and proliferation is also considerably reduced in the cells in which STIM1 has been knocked down showing the significance of STIM1 in the migration and proliferation in these cells.

Increased Activation of Stromal Interaction Molecule-1/Orai-1 in Aorta From Hypertensive Rats A Novel Insight Into Vascular Dysfunction

Disturbances in the regulation of cytosolic calcium (Ca(2+)) concentration play a key role in the vascular dysfunction associated with arterial hypertension. Stromal interaction molecules (STIMs) and Orai proteins represent a novel mechanism to control store-operated Ca(2+) entry. Although STIMs act as Ca(2+) sensors for the intracellular Ca(2+) stores, Orai is the putative pore-forming component of Ca(2+) release-activated Ca(2+) channels at the plasma membrane. We hypothesized that augmented activation of Ca(2+) release-activated Ca(2+)/Orai-1, through enhanced activity of STIM-1, plays a role in increased basal tonus and vascular reactivity in hypertensive animals. Endothelium-denuded aortic rings from Wistar-Kyoto and stroke-prone spontaneously hypertensive rats were used to evaluate contractions because of Ca(2+) influx. Depletion of intracellular Ca(2+) stores, which induces Ca(2+) release-activated Ca(2+) activation, was performed by placing arteries in Ca(2+) free-EGTA buffer. The addition of the Ca(2+) regular buffer produced greater contractions in aortas from stroke-prone spontaneously hypertensive rats versus Wistar-Kyoto rats. Thapsigargin (10 mu mol/L), an inhibitor of the sarcoplasmic reticulum Ca(2+) ATPase, further increased these contractions, especially in stroke-prone spontaneously hypertensive rat aorta. Addition of the Ca(2+) release-activated Ca(2+) channel inhibitors 2-aminoethoxydiphenyl borate (100 mu mol/L) or gadolinium (100 mu mol/L), as well as neutralizing antibodies to STIM-1 or Orai-1, abolished thapsigargin-increased contraction and the differences in spontaneous tone between the groups. Expression of Orai-1 and STIM-1 proteins was increased in aorta from stroke-prone spontaneously hypertensive rats when compared with Wistar-Kyoto rats. These results support the hypothesis that both Orai-1 and STIM-1 contribute to abnormal vascular function in hypertension. Augmented activation of STIM-1/Orai-1 may represent the mechanism that leads to impaired control of intracellular Ca(2+) levels in hypertension. (Hypertension. 2009; 53[part 2]: 409-416.)

Agrobacterium tumefasciens-mediated transformation of the aquatic fungus Blastocladiella emersonii

Agrobacterium tumefaciens is widely used for plant DNA transformation and more recently, has also been used to transform yeast, filamentous fungi and even human cells. Using this technique, we developed the first transformation protocol for the saprobic aquatic fungus Blastocladiella emersonii, a Blastocladiomycete localized at the base of fungal phylogenetic tree, which has been shown as a promising and interesting model of study of cellular function and differentiation. We constructed binary T-DNA vectors containing hygromycin phosphotransferase (hph) or enhanced green fluorescent protein (egfp) genes, under the control of Aspergillus nidulans trpC promoter and terminator sequences. 24 h of co-cultivation in induction medium (IM) agar plates, followed by transfer to PYG-agar plates containing cefotaxim to kill Agrobacterium tumefsciens and hygromycin to select transformants, resulted in growth and sporulation of resistant transformants. Genomic DNA from the pool o resistant zoospores were shown to contain T-DNA insertion as evidenced by PCR amplification of hph gene. Using a similar protocol we could also evidence the expression of enhanced green fluorescent protein (EGFP) in zoospores derived from transformed cells. This protocol can also open new perspectives for other non-transformable closely related fungi, like the Chytridiomycete class. (C) 2011 Elsevier Inc. All rights reserved.

Caracterização de genes de quitanases do entomopatógeno e acaricida Metarhizium anisopliae

O fungo entomopatogênico e acaricida Metarhizium anisopliae é patógeno de uma vasta gama de insetos, sendo extensivamente utilizado em experimentos, bem como, no controle efetivo de alguns insetos-praga. Seu potencial uso para o controle de carrapatos como Boophilus microplus é também considerável. O processo de infecção de M. anisopliae é o melhor caracterizado entre os fungos entomopatogênicos, e combina pressão mecânica, por diferenciação do apressório, síntese e secreção de enzimas hidrolíticas altamente reguladas como proteases e, provavelmente, quitinases e lipases. As quitinases em fungos também são importantes em processos que requerem digestão celular, como germinação, crescimento e ramificação das hifas e autólise, visto que a quitina é o maior constituinte da parede celular desses organismos, sendo um sistema altamente regulado. Objetivamos neste trabalho, obter mais informações sobre o sistema quitinolitico do fungo M. anisopliae var. anisopliae linhagem E6 durante o processo de infecção do hospedeiro ou na morfogênese e crescimento. Com o objetivo de analisarmos o gene chi2 de M. anisopliae E6, clonamos e caracterizamos sua seqüência genômica, incluindo a região flanqueadora 5’. O gene chi2 é interrompido por dois pequenos íntrons típicos, de 210 pb e 75 pb, respectivamente. A ORF do gene chi2 apresenta 1.545 pb e codifica uma proteína predita de 419 aminoácidos (denominada CHI2), com massa molecular estimada de 44 kDa. Um peptídeo sinal característico com sítio de clivagem no aminoácido V19 está presente. A forma madura dessa proteína tem uma massa molecular estimada de 42 kDa e um pI teórico de 4,8. Análise por Southern de DNA genômico indica cópia única de chi2 no genoma de M. anisopliae. A seqüência de consenso SXGG, correspondendo ao sítio de ligação à quitina, foi identificada e a seqüência NGFDFDIE, que compõem o domínio catalítico de quitinases, está presente em CHI2. A construção de uma árvore filogenética determinou que a quitinase CHI2 pertence a um grupo diferente daquele da CHIT42 a qual provavelmente não está envolvida na patogenicidade. Uma análise in sílico da seqüência 5’ franqueadora do gene chi2 para determinação de possíveis elementos regulatórios foi efetuada. A regulação da transcrição dos genes chit1 e chi2 em M. ansisoplaie frente a diferentes fontes de carbono e em diferentes tempos de cultivo foi analisada. Os genes chit1 e chi2 apresentaram uma expressão tardia no fungo, a partir de 30 horas. O gene chi2 foi expresso majoritariamente em cultivos com quitina e sua expressão foi reprimida por glicose. O gene chit1 foi induzido em presença de fontes de carbono facilmente assimiláveis, como glicose e NAcGlc. Ambos os genes, chit1 e chi2, apresentaram alta expressão quando a fonte de carbono já estava exaurida e o fungo estava em autólise, sugerindo o requerimento dessas enzimas nessa fase. O cDNA do chit1 foi inserido em um vetor de expressão, em ambas orientações senso e antisenso, sob regulação do promotor do gene tef1α de M. anisopliae e o terminador do gene trpC de A. nidulans. Os transformantes com o gene chit1 na orientação senso mostraram superexpressão de atividade de quitinase e o transformante com o gene na orientação antisenso apresentou uma redução na atividade de quitinase. Também construímos quatro deleções na região flanqueadora 5’ do gene chit1 fusionadas com a proteína repórter SGFP, para localizar seqüências reguladoras no promotor e, destas construções, três foram transformadas em M. anisopliae.

Role of TRP Channels in Mediating the Calcium Signaling Response of Brain Endothelial Cells to Mechanical Stretch

Traumatic brain injury (TBI) often results in disruption of the blood brain barrier (BBB), which is an integral component to maintaining the central nervous system homeostasis. Recently cytosolic calcium levels ([Ca2+]i), observed to elevate following TBI, have been shown to influence endothelial barrier integrity. However, the mechanism by which TBI-induced calcium signaling alters the endothelial barrier remains unknown. In the present study, an in vitro BBB model was utilized to address this issue. Exposure of cells to biaxial mechanical stretch, in the range expected for TBI, resulted in a rapid cytosolic calcium increase. Modulation of intracellular and extracellular Ca2+ reservoirs indicated that Ca2+ influx is the major contributor for the [Ca2+]i elevation. Application of pharmacological inhibitors was used to identify the calcium-permeable channels involved in the stretch-induced Ca2+ influx. Antagonist of transient receptor potential (TRP) channel subfamilies, TRPC and TRPP, demonstrated a reduction of the stretch-induced Ca2+ influx. RNA silencing directed at individual TRP channel subtypes revealed that TRPC1 and TRPP2 largely mediate the stretch-induced Ca2+ response. In addition, we found that nitric oxide (NO) levels increased as a result of mechanical stretch, and that inhibition of TRPC1 and TRPP2 abolished the elevated NO synthesis. Further, as myosin light chain (MLC) phosphorylation and actin cytoskeleton rearrangement are correlated with endothelial barrier disruption, we investigated the effect mechanical stretch had on the myosin-actin cytoskeleton. We found that phosphorylated MLC was increased significantly by 10 minutes post-stretch, and that inhibition of TRP channel activity or NO synthesis both abolished this effect. In addition, actin stress fibers formation significantly increased 2 minutes post-stretch, and was abolished by treatment with TRP channel inhibitors. These results suggest that, in brain endothelial cells, TRPC1 and TRPP2 are activated by TBI-mechanical stress and initiate actin-myosin contraction, which may lead to disruption of the BBB.

Specific association of the gene product of PKD2 with the TRPC1 channel

The function(s) of the genes (PKD1 and PKD2) responsible for the majority of cases of autosomal dominant polycystic kidney disease is unknown. While PKD1 encodes a large integral membrane protein containing several structural motifs found in known proteins involved in cell–cell or cell–matrix interactions, PKD2 has homology to PKD1 and the major subunit of the voltage-activated Ca2+ channels. We now describe sequence homology between PKD2 and various members of the mammalian transient receptor potential channel (TRPC) proteins, thought to be activated by G protein-coupled receptor activation and/or depletion of internal Ca2+ stores. We show that PKD2 can directly associate with TRPC1 but not TRPC3 in transfected cells and in vitro. This association is mediated by two distinct domains in PKD2. One domain involves a minimal region of 73 amino acids in the C-terminal cytoplasmic tail of PKD2 shown previously to constitute an interacting domain with PKD1. However, distinct residues within this region mediate specific interactions with TRPC1 or PKD1. The C-terminal domain is sufficient but not necessary for the PKD2–TRPC1 association. A more N-terminal domain located within transmembrane segments S2 and S5, including a putative pore helical region between S5 and S6, is also responsible for the association. Given the ability of the TRPC to form functional homo- and heteromultimeric complexes, these data provide evidence that PKD2 may be functionally related to TRPC proteins and suggest a possible role of PKD2 in modulating Ca2+ entry in response to G protein-coupled receptor activation and/or store depletion.

Modulation des neurones GABAergiques du mésencéphale ventral

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.