The SH3-binding domain of Cx43 participates in loop/tail interactions critical for Cx43-hemichannel activity

Connexin 43 (Cx43) hemichannels establish local signaling networks via the release of ATP and other molecules, but their excessive opening may result in cell death. Hence, the activity of Cx43-hemichannels ought to be critically controlled. This involves interactions between the C-terminal tail (CT) and the cytoplasmic loop (CL), more particularly the L2 domain within CL. Previous work revealed an important role for the last nine amino acids of the Cx43 CT by targeting the L2 domain, as these nine amino acids were sufficient to restore the activity of CT-truncated Cx43-hemichannels. However, we discovered that deletion of the last 19 amino acids of the CT only partially lowered the binding to the L2 domain, indicating that a second L2-binding region is present in the CT. We here provide evidence that the SH3-binding domain is another CT region that targets the L2 domain. At the functional level, the SH3-binding domain was able to restore the activity of CT-truncated Cx43-hemichannels and alleviate the inhibition of full-length Cx43-hemichannels by high intracellular Ca²⁺ concentration ([Ca²⁺]_i) as demonstrated by various approaches including patch clamp studies of unitary Cx43-hemichannel activity. Finally, we show that in full-length Cx43-hemichannels, deletion of either the SH3-binding domain or the CT9 region suppresses the hemichannel activity, while deletion of both domains completely annihilates the hemichannel activity. These results demonstrate that the Cx43 SH3-binding domain, in addition to the CT9 region, critically controls hemichannel activity at high [Ca²⁺]_i, which may be involved in pathological hemichannel opening.     

Lack ofα-amylase in horse serum

Riassunto Mediante gel-cromatografia si è confermata la presenza di -glucosidasi e l'assenza di -amilasi nel siero di cavallo. La -amilasi è anche assente nelle ghiandole salivari ma presente nel pancreas dello stesso animale.     

Prevention of cisplatin-induced ototoxicity by the inhibition of gap junctional intercellular communication in auditory cells

Cis-diamminedichloroplatinum (cisplatin) is an effective chemotherapeutic drug for cancer therapy. However, most patients treated with cisplatin are at a high risk of ototoxicity, which causes severe hearing loss. Inspired by the “Good Samaritan effect” or “bystander effect” from gap junction coupling, we investigated the role of gap junctions in cisplatin-induced ototoxicity as a potential therapeutic method. We showed that connexin 43 (Cx43) was highly expressed in House Ear Institute-Organ of Corti 1 (HEI-OC1) cells, mediating cell–cell communication. The viability of HEI-OC1 cells was greatly decreased by cisplatin treatment, and cisplatin-treated HEI-OC1 cells showed lower Cx43 expression compared to that of untreated HEI-OC1 cells. In particular, high accumulation of Cx43 was observed around the nucleus of cisplatin-treated cells, whereas scattered punctuate expression of Cx43 was observed in the cytoplasm and membrane in normal cells, suggesting that cisplatin may interrupt the normal gap junction communication by inhibiting the trafficking of Cx43 to cell membranes in HEI-OC1 cells. Interestingly, we found that the inhibition of gap junction activity reduced cisplatin-induced apoptosis of auditory hair cells. Cx43 siRNA- or 18α-GA-treated HEI-OC1 cells showed higher cell viability compared to control HEI-OC1 cells during cisplatin treatment; this was also supported by fluorescence recovery after photobleaching studies. Inhibition of gap junction activity reduced recovery of calcein acetoxymethyl ester fluorescence compared to control cells. Additionally, analysis of the mechanisms involved demonstrated that highly activate extracellular signal-regulated kinase and protein kinase B, combined with inhibition of gap junctions may promote cell viability during cisplatin treatment.     

Synthesis and pharmacological properties of [1-N4-dimethyl-asparagine]-angiotensin II

Summary [1-N⁴-Dimethyl-asparagine]-angiotensin II was synthesized by Merrifield's solid-phase procedure. The analogue gave rat blood pressure about 70% relative potency to Hypertensin (Ciba). Rabbit aorta strips gave intrinsic activity _E=1, a PD₂ of 6.92±0.09 and an affinity relative to [Asn¹]-angiotensin II of 6.5%.The editors do not hold themselves responsible for the opinions expressed in the authors' brief reports. — Les auteurs sont seuls responsables des opinions exprimées dans ces brèves communications. — Für die Kurzmitteilungen ist ausschliesslich der Autor verantwortlich. — Per le brevi comunicazioni è responsabile solo l'autore. — . — Solo los autores son responsables de las opiniones expresadas en estas comunicationes breves.Acknowledgment. The authors wish to express their appreciation to Dr Emanuel Escher, School of Medicine, Centre Hospitalier Universitaire, Sherbrooke, Quebec, for his help in providing the biological data and critical comments.     

Lead potentiation of endotoxin lethality in rats: Lack of effect of kininase inhibition

Summary Although lead and SQ₂₀₈₈₁ are potent in vitro inhibitors of kininase II activity, SQ₂₀₈₈₁ does not alter the sensitivity of rats to endotoxin. These results indicate that marked changes in plasma kininase activity do not contribute to endotoxin morbility and that kininase inhibition is not the mechanism whereby lead ions sensitize rats to endotoxin.This investigation was supported by the Naval Medical Research and Development Command, NNMC, Department of the Navy, Research Task No. MR041.20.01.0435. The opinions and assertions contained herein are the private ones of the author, and are not to be construed as official or reflecting the views of the Navy Department or the naval service at large.The experiments reported herein were conducted according to the principles set forth in the Guide for the Care and Use of Laboratory Animals, Institute of Laboratory Animal Resources, National Research Council, DHEW Pub. No. (NIH) 74-23.     

Further morphological and biochemical studies on normal and hybrid embryos of sea urchins

Zusammenfassung (1) Es wurde bei den SeeigelartenParacentrotus ,Arbacia undSphaerechinus sowie den drei BastardenPar ×Arb ,Arb ×Par undPar ×Sphaer für eine Reihe von Entwicklungsstadien die Anzahl der Kerne pro Keim bestimmt (Tabelle). Der morphogenetischen Hemmung der Bastarde entsprechen herabgesetzte Kernzahlen und parallel eine Herabsetzung der DNS-Synthese. (2) Es wurde bei den genannten reinen Arten und den Bastarden derUmsatz der RNS bestimmt. Er geht in allen Fällen, bei den reinen Arten wie bei den Bastarden, der DNS- Synthese parallel.

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Our work was supported by a grant from the Schweizerischer Nationalfonds zur Förderung der wissenschaftlichen Forschung. We express our sincere thanks to the Nationalfonds and also to the authorities at the Zoological Station at Naples for generous help and supply of materials and facilities. — This paper is thankfully dedicated byF. Baltzer toF. E. Lehmann for his kind hospitality in the Bernese Institute.     

The therapeutic potential of GPR43: a novel role in modulating metabolic health

GPR43 is a receptor for short-chain fatty acids. Preliminary data suggest a putative role for GPR43 in regulating systemic health via processes including inflammation, carcinogenesis, gastrointestinal function, and adipogenesis. GPR43 is involved in secretion of gastrointestinal peptides, which regulate appetite and gastrointestinal motility. This suggests GPR43 may have a role in weight control. Moreover, GPR43 regulates plasma lipid profile and inflammatory processes, which further indicates that GPR43 could have the ability to modulate the etiology and pathogenesis of metabolic diseases such as obesity, type 2 diabetes mellitus, and cardiovascular disease. This review summarizes the current evidence regarding the ability of GPR43 to mediate both systemic and tissue specific functions and how GPR43 may be modulated in the treatment of metabolic disease.     

Functional interaction between TRPC1 channel and connexin-43 protein: a novel pathway underlying S1P action on skeletal myogenesis

We recently demonstrated that skeletal muscle differentiation induced by sphingosine 1-phosphate (S1P) requires gap junctions and transient receptor potential canonical 1 (TRPC1) channels. Here, we searched for the signaling pathway linking the channel activity with Cx43 expression/function, investigating the involvement of the Ca²⁺-sensitive protease, m-calpain, and its targets in S1P-induced C2C12 myoblast differentiation. Gene silencing and pharmacological inhibition of TRPC1 significantly reduced Cx43 up-regulation and Cx43/cytoskeletal interaction elicited by S1P. TRPC1-dependent functions were also required for the transient increase of m-calpain activity/expression and the subsequent decrease of PKCα levels. Remarkably, Cx43 expression in S1P-treated myoblasts was reduced by m-calpain-siRNA and enhanced by pharmacological inhibition of classical PKCs, stressing the relevance for calpain/PKCα axis in Cx43 protein remodeling. The contribution of this pathway in myogenesis was also investigated. In conclusion, these findings provide novel mechanisms by which S1P regulates myoblast differentiation and offer interesting therapeutic options to improve skeletal muscle regeneration.     

Monoamines in sympathetic ganglia studied with fluorescence microscopy

Zusammenfassung Sympathische Ganglien wurden mit einer spezifischen und besonders empfindlichen histochemischen Methode zum Nachweis gewisser Monoamine studiert. Die meisten Zellkörper der Ganglienzellen enthalten im Cytoplasma mehr oder weniger einer primären Monoamine, wahrscheinlich Noradrenalin. In prävertebralen Ganglien wurden auch adrenerge Nevenendigungen nachgewiesen, die synaptische Verbindungen mit den Ganglienzellen eingehen.

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This work has been supported by grants from the United States Public Health Service (NB 02854-03), the Foundation Therese and Johan Anderssons Minne, and Karolinska Institutet.     

The control of melanoblast differentiation in the periodic albino mutant ofXenopus

Summary Studies on the incidence of melanophores in older ventral trunk tissues and in isolated regions of periodic albino embryos ofXenopus suggest that melanin granule formation in mutant melanoblasts depends on an environmental contribution which arises at stage 43 in the endodermal tissues.     

Steroidal sapogenins. XXX stereochemistry of the side chain

Zusammenfassung Die optischen Drehungen einiger 20, 25d; 20, 25l; 20, 25d; und 20, 25-l-Sapogenine wurden bestimmt. Die ersten drei Serien gaben übereinstimmend linksdrehende Werte, aber die letztere Gruppe erwies sich als rechtsdrehend.Die Struktur der vier Serien der Sapogenine folgte aus der Analyse dieser Befunde. Der mögliche Mechanismus bei der Entstehung dieser Verbindungen aus Pseudosapogeninen wurde besprochen. Die Autoren gelangen zum Schluss, dass sterische Faktoren an C₂₀ und C₂₅ die Richtung der Ringschliessung beeinflussen.

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Steroidal sapogenins. XXX stereochemistry of the side chain

     

γ-Glutamyl-transpeptidase in lymphatic tissues ofMastomys natalensis during an infection withAcanthocheilonema viteae

Summary DuringAcanthocheilonema viteae infection, the specific activity of -glutamyltranspeptidase (-GT) increased in peritoneal exudate cells and bone marrow and decreased in lymphnodes ofMastomys natalensis throughout the course of infection. However, though there was an increase in specific activity of -GT in thymus and spleen during the prepatent phase ofA. viteae infection, the level either returned to normal or decreased during the latent phase of infection. A close correlation was observed between the host's immune status duringA. viteae infection and the level of -GT in lymphoid tissues.Communication No. 4535 from Central Drug Research Institute, Lucknow.     

An inheritable abnormal β-chain in rabbit haemoglobin

Riassunto Mediante tecniche cromatografiche ed elettroforetiche, è state trovata una globina anormale nella emoglobina di un coniglio. La mutazione globinica riguarda verosimilmente la catena. Questa emoglobina anormale è trasmessa ereditariamente.     

Connexins and pannexins in the skeleton: gap junctions,hemichannels and more

Regulation of bone homeostasis depends on the concerted actions of bone-forming osteoblasts and bone-resorbing osteoclasts, controlled by osteocytes, cells derived from osteoblasts surrounded by bone matrix. The control of differentiation, viability and function of bone cells relies on the presence of connexins. Connexin43 regulates the expression of genes required for osteoblast and osteoclast differentiation directly or by changing the levels of osteocytic genes, and connexin45 may oppose connexin43 actions in osteoblastic cells. Connexin37 is required for osteoclast differentiation and its deletion results in increased bone mass. Less is known on the role of connexins in cartilage, ligaments and tendons. Connexin43, connexin45, connexin32, connexin46 and connexin29 are expressed in chondrocytes, while connexin43 and connexin32 are expressed in ligaments and tendons. Similarly, although the expression of pannexin1, pannexin2 and pannexin3 has been demonstrated in bone and cartilage cells, their function in these tissues is not fully understood.     

Total replacement of blood by an emulsion of fluorocarbon in the rat-water extravasation as a cause of failure

Summary After exchange transfusion by an emulsion of fluorocarbons (Fluosol 43) in rats, an increase in fluorocarbons and¹³¹I-labelled albumin was observed. These changes suggest a transfer of water from vascular to interstitial space possibly owing to the inability of the emulsion to reproduce the oncotic pressure of normal blood.     

Intrinsic disorder in proteins involved in amyotrophic lateral sclerosis

Five structurally and functionally different proteins, an enzyme superoxide dismutase 1 (SOD1), a TAR-DNA binding protein-43 (TDP-43), an RNA-binding protein FUS, a cofilin-binding protein C9orf72, and polypeptides generated as a result of its intronic hexanucleotide expansions, and to lesser degree actin-binding profilin-1 (PFN1), are considered to be the major drivers of amyotrophic lateral sclerosis. One of the features common to these proteins is the presence of significant levels of intrinsic disorder. The goal of this study is to consider these neurodegeneration-related proteins from the intrinsic disorder perspective. To this end, we employed a broad set of computational tools for intrinsic disorder analysis and conducted intensive literature search to gain information on the structural peculiarities of SOD1, TDP-43, FUS, C9orf72, and PFN1 and their intrinsic disorder predispositions, and the roles of intrinsic disorder in their normal and pathological functions.     

Disruption of mitochondrial function as the basis of the trypanocidal effect of trifluoperazine onTrypanosoma cruzi

Summary The tricyclic anti-calmodulin drug trifluoperazine (TFP) inhibited growth and motility of epimastigotes ofTrypanosoma cruzi, at concentrations lower than 100 M, and motility and infectivity of the bloodstream trypomastigote form at 200 M. Electron microscopy of TFP-treated epimastigotes showed that the major effect was at the mitochondrial level, with gross swelling and disorganization. The oligomycin-sensitive, mitochondrial ATPase was completely inhibited by 20 M TFP, and the same drug concentration caused a 60% decrease in intracellular ATP content. The results suggest that the trypanocidal effect of TFP may be related more to mitochondrial damage than to the well-known anticalmodulin effect of the drug.     

The plasma volume of the Wistar rat in relation to the body weight

Summary The plasma volume of 43 male Wistar rats, weighing between 140 and 350 g, was determined. A close linear relationship between plasma volume and body weight was found: plasma volume (ml)=0.0291×body weight (g)+2.54.Acknowledgment. The excellent assistance of Mr J.S. Bouwer in animal experiments is gratefully acknowledged.     

Untersuchungen über Bildung und Weitergabe von Drüsensekreten beiFormica (Hymenopt. Formicidae) mit Hilfe der Radioisotopenmethode

Summary Marking experiments with the tracer isotope P³² had the result that food-secretion of ants is mainly formed in the pharyngeal gland, then reaches the crop through the exit channel of the gland, where it is stored and released again, on regurgitation-stimuli, to and and larvae as well as workers.     

Cytogenetics of South American akodont rodents (Cricetidae). V. Segregation of chromosome No. 1 polymorphism inAkodon molinae

Summary Akodon molinae is polymorphic with 2n=42, 43, 44, where the metacentric autosome No. 1 is homologous to 2 acrocentrics 1a and 1b. Matings between 2n=43 heterozygotes 1/1a, 1b gave a surplus of 1/1 offspring, a moderate reduction of heterozygous and a strong reduction of homozygous 1a, 1b/1a, 1b offspring. The latter type also has a highly reduced fertility.This work was supported by grants from the Consejo Nacional de Investigaciones Científicas y Técnicas, the Comisión de Investigaciones Científicas de la Provincia de Buenos Aires and the Organización de Estados Americanos.