Partial inhibition of the in vitro infection of adult mouse dorsal root ganglion neurons by rabies virus using nicotinic antagonists

Castellanos, Jaime E.; Castañeda, David R.; Velandia, Alvaro; Hurtado, Hernán

doi:10.1016/s0304-3940(97)00440-0

Cited by 17 publications

(10 citation statements)

References 25 publications

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“…One of the first receptors to be suggested was the nicotinic acetylcholine receptor (36), which is also found on cultured DRG neurons. Nicotinic antagonists partially (25%) block infection of DRG neurons (5), suggesting that this receptor could play a role in RV binding and infection. Most large cells express the ␣7 subunit; however, this is also found in a third of small DRG neurons (19), which is in contrast to the size profile we have established for infected DRG cells.…”

Section: Discussionmentioning

confidence: 99%

The Rabies Virus Glycoprotein Receptor p75 ^NTR Is Not Essential for Rabies Virus Infection

Tuffereau

Schmidt

Langevin

et al. 2007

J Virol

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Rabies virus glycoprotein (RVG) is known to be the only factor that mediates rabies infection. The neurotrophin receptor (p75 NTR ), through its cysteine-rich domain 1, is a specific receptor for RVG and neutralizes virus infectivity, but its role in virus infection has remained obscure. We used adult mouse dorsal root ganglion (DRG) neurons as a model to study the role of p75 NTR in RV infection of primary neurons. We show that RV infects around 20% of DRG neurons, of which more than 80% are p75 NTR positive, have large diameters, and are capsaicin insensitive. Surprisingly, RV binding and infection are absent in about half of the p75 NTRexpressing DRG neurons which have small diameters and are often capsaicin sensitive. This indicates that p75 NTR is not sufficient to mediate RV interaction in sensory neurons. The rate and specificity of neural infection are unchanged in RV-infected p75 NTRExonIV؊/؊ mice that lack all extracellular receptor domains and in wild-type mice infected with two independent RV mutants that lack p75 NTR binding. Accordingly, the mortality rate is unchanged in the absence of RV-p75 NTR interaction. We conclude that although p75 NTR is a receptor for soluble RVG in transfected cells of heterologous expression systems, an RVG-p75 NTR interaction is not necessary for RV infection of primary neurons. This means that other receptors are required to mediate RV infection in vivo and in vitro.Rabies causes severe progressive encephalitis, myelitis, and paralysis and affects more than 50,000 people worldwide each year (59). After onset of symptoms, the infection progresses relentlessly, and there is only one documented case of a nonvaccinated human patient who survived the disease (56). Rabies virus (RV) belongs to the genus Lyssavirus in the family Rhabdoviridae and is a simple, enveloped, nonsegmented, negative-strand RNA virus that encodes only five proteins (44). The glycoprotein (G) is known to be the only protein component of the viral envelope that mediates viral entry into host cells (44).RV is highly neurotropic, and after inoculation it is retrogradely transported by peripheral neurons before being passed on to second and higher-order neurons without being taken up by glia. It is this unique property of the virus that makes it a useful transsynaptic neuronal tracer over many synapses (28,52), which is dependent on RVG (17).Although several receptors have been suggested to be responsible for RV infection, none has been conclusively identified. In previous studies we adopted an expression cloning strategy using a soluble form of RVG as a ligand. That work identified the neurotrophin receptor p75 NTR as a unique interacting receptor (50). We have also shown that the glycoprotein from another lyssavirus genotype (EBL2) is also able to interact with p75 NTR (51). p75 NTR belongs to the tumor necrosis factor receptor family (11,14) and is the common receptor for all known mature neurotrophins as well immature proneurotrophins (35). In addition, several other ligands implicated in the patholo...

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Section: Discussionmentioning

confidence: 99%

The Rabies Virus Glycoprotein Receptor p75 ^NTR Is Not Essential for Rabies Virus Infection

Tuffereau

Schmidt

Langevin

et al. 2007

J Virol

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“…Nevertheless, they provide valuable information on the infectious cycle of RV in its host cell. Experiments were carried out using ganglionic neurons (Tsiang et al, 1983(Tsiang et al, , 1989Lycke and Tsiang, 1987;Castellanos et al, 1997Castellanos et al, , 2000 or neurons from CNS (Tsiang et al, , 1991. These cultures were used to study in vitro the infectabilit y of neurons under various infection conditions.…”

Section: Introductionmentioning

confidence: 99%

Rabies virus is not cytolytic for rat spinal motoneurons in vitro

Guigoni¹,

Coulon²

2002

J Neurovirol

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Cultures of puri ed rat embryonic spinal cord motoneurons were used to investigate the capacity of the neurons to survive rabies virus infection in vitro. In crude primary spinal cord cultures, neurons did not survive more than 2 days after rabies virus infection with the xed strain Challenge Virus Standard. In contrast, virus-infected puri ed motoneurons resisted cytolysis for at least 7 days, as also did infected motoneurons treated with conditioned medium sampled from rabies virus-infected crude spinal cord cultures. This survival rate was also observed when motoneurons were grown in the presence of astrocytes or broblasts and it was not dependent on the presence of growth factors in the culture medium. Moreover, terminal deoxynucleotidy l transferase-mediated dUTP nick end labeling experiments showed that only 30% of infected motoneurons were apoptotic after 7 days of infection. In vivo, despite the massive infection of the spinal cord in infected rat neonates, the moderate number of apoptotic cells in the ventral horn suggests that only a few motoneurons were affected by this mechanism of cell death. Morphometric analyses showed that motoneurons' axon elongated at a comparable rate in virus-infected and noninfected cultures, a sign of high metabolic activity maintained in rabies virus-infected motoneurons. In contrast, hippocampus neurons were susceptible to rabies virus infection, because 70% of infected neurons were destroyed within 3 days, a large proportion of them being apoptotic. These experiments suggest that spinal cord motoneurons consist in a neuronal population that survive rabies virus infection because the viral induction of apoptosis is delayed in these neurons. They suggest also that paralyses frequently observed in rabid animals could be the consequence of dysfunctions of the locomotor network or of the spinal cord motoneurons themselves, whose parameters could be studied in vitro. Journal of NeuroVirology (2002) 8, 306-317.

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“…En ambas técnicas, se logró hacer la detección de las zonas infectadas con un alto grado de especificidad, pero usando diluciones de anticuerpo diferentes, pues para la fluorescencia se usó la dilución de rutina (1:30) y la dilución que mejor se comportó en la técnica de peroxidasa fue de 1 :480. Resultados similares se reportaron en un trabajo anterior (6,7) y las dificultades fueron resueltas diluyendo también el anticuerpo primario. Los controles usados demuestran una buena sensibilidad y especificidad, de tal manera que la técnica de cortar en vibrátomo cerebros fijados podría ser útil en los casos en que los especímenes hayan sido fijados antes de las pruebas de confirmación de fluorescencia de improntas o inoculación en cerebros de ratón.…”

Section: Discussionunclassified

Uso de una técnica de inmunoperoxidasa para la detección de virus de rabia en cortes gruesos de cerebro

Castellanos¹,

Guayacán²,

Castañeda³

et al. 1998

biomedica

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ResumenCon el propósito de evaluar una técnica de avidina-peroxidasa biotinilada, se utilizó el conjugado antirrábico producido en el INS para la detección del virus de rabia en cortes gruesos de cerebros de ratón tratados con varios tipos de fijadores y con varias diluciones del conjugado. Los animales infectados experimentalmente fueron perfundidos vía intracardiaca con varios fijadores; los cerebros se recuperaron, se les hicieron cortes gruesos de 60 pm y se sometieron a la inmunodetección. Se encontró que en las diluciones normalmente usadas en fluorescencia se presentó un fuerte marcaje inespecífico sin mejorar la detección. En este trabajo, se presenta un protocolo fácil y rápido que pudiera ser útil para la obse~ación de muestras sospechosas. The use of an immunoperoxydase technique for the detection of rabies virus in thick brain slicesWith the intention of evaluating an avidine-peroxidase biotinilated technique, the antirabies' sera, produced in the INS, was used for the detection of the rabies' virus in mouse brain tissue, treated with various types of fixers and various antisera dilutions. The experimentally infected animals were killed by intra-aortic perfusion with various fixers; their brains were recovered, cut into 60 pm slices and submitted to immunodetection. It was found that the normally used fluorescent dilutions presented strong non-specific marking without improving detection. In this work a protocol which is fast and easy is presented, which could be useful for the observation of suspect samples.El diagnóstico histopatológico de la rabia se ha bral y células de Purkinje en el cerebelo (3). En basado por años en la detección de inclusiones 10s e~pecímenes en los que no es posible identípicas en el citoplasma de las neuronas infecta-tificar 10s corpúsculos de Negri, se hacen pruedas (cuerpos de Negri), que también pueden ser bas adicionales como la inoculación en ratón o evidenciadas en las preparaciones para la prueba de inmunofluorescencia, que son inmunofluorescencia (2). En los casos en los confirmatorias de la presencia del virus (3). que se evidencian las inclusiones, se encuen-La reacción con sueros antirrábicos acoplados a tran preferencialmente en las neuronas del fluoresceína se usa normalmente para hacer el hipocampo (cuerno de Ammon), corteza cere-diagnóstico en especímenes sospechosos y se

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Partial inhibition of the in vitro infection of adult mouse dorsal root ganglion neurons by rabies virus using nicotinic antagonists

Cited by 17 publications

References 25 publications

The Rabies Virus Glycoprotein Receptor p75 ^NTR Is Not Essential for Rabies Virus Infection

The Rabies Virus Glycoprotein Receptor p75 ^NTR Is Not Essential for Rabies Virus Infection

Rabies virus is not cytolytic for rat spinal motoneurons in vitro

Uso de una técnica de inmunoperoxidasa para la detección de virus de rabia en cortes gruesos de cerebro

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Partial inhibition of the in vitro infection of adult mouse dorsal root ganglion neurons by rabies virus using nicotinic antagonists

Cited by 17 publications

References 25 publications

The Rabies Virus Glycoprotein Receptor p75 NTR Is Not Essential for Rabies Virus Infection

The Rabies Virus Glycoprotein Receptor p75 NTR Is Not Essential for Rabies Virus Infection

Rabies virus is not cytolytic for rat spinal motoneurons in vitro

Uso de una técnica de inmunoperoxidasa para la detección de virus de rabia en cortes gruesos de cerebro

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The Rabies Virus Glycoprotein Receptor p75 ^NTR Is Not Essential for Rabies Virus Infection

The Rabies Virus Glycoprotein Receptor p75 ^NTR Is Not Essential for Rabies Virus Infection