Gene Transfer into Neurons from Hippocampal Slices: Comparison of Recombinant Semliki Forest Virus, Adenovirus, Adeno-Associated Virus, Lentivirus, and Measles Virus

Ehrengruber, Markus U.; Hennou, Sonia; Büeler, Hansruedi; Naim, Hussein Y.; Déglon, Nicole; Lundström, Kenneth

doi:10.1006/mcne.2001.0982

Cited by 123 publications

(115 citation statements)

References 52 publications

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“…In accordance with previous reports about the inert nature of GFP on neuronal activity (18,60), green AAV-GFP-infected neurons resembled control cells in their electrophysiological characteristic, indicating that the change of neuronal function in CMV-infected neurons is attributable to the virus and not due to the expression of GFP.…”

Section: Induction Of Intrinsic Antiviral Response By Neurons and Glisupporting

confidence: 91%

Cytomegalovirus InducesInterferon-Stimulated Gene Expression and Is Attenuated by Interferon in the DevelopingBrain

Pol¹,

Robek

Ghosh³

et al. 2007

J Virol

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Cytomegalovirus (CMV) is considered the most common infectious agent causing permanent neurological dysfunction in the developing brain. We have previously shown that CMV infects developing brain cells more easily than it infects mature brain cells and that this preference is independent of the host B-and T-cell responses. In the present study, we examined the innate antiviral defenses against mouse (m) and human ( Cytomegalovirus (CMV), a double-stranded DNA virus of the betaherpesvirus family, is considered the most common infectious agent that causes permanent neurological dysfunction of the developing brain (2, 62). Human CMV (hCMV) infections in the developing brain can lead to mental retardation, epilepsy, microcephaly, microgyria, hydrocephalus, and deafness (4, 21, 37). The developing brain is particularly sensitive to CMV infection (6,29,54,55,58) due to both the immature state of the systemic immune system and a preference of CMV for developing glia and neurons (61). CMV in the mature brain is less of a concern, except in immunocompromised individuals (22,23,32).Outside the brain, interferon (IFN) has been reported to limit some CMV infections (27,30,66). A number of reports have suggested that brain cells differ from cells of other organs relative to innate and systemic antiviral immune responses. The brain has some characteristics of an immunologically privileged region. For instance, although most cells outside the brain express major histocompatibility complex (MHC) class I molecules, neurons in the brain either do not express MHC class I or do so at a substantially reduced level (45). This may be beneficial due to the fact that neurons do not replicate, and so a vigorous attack by the immune system on virally infected postmitotic neurons may cause more problems than the virus itself. In the brain, functional receptors for IFN-␥ are expressed in all cells, but they are expressed at different levels, and actions of IFN-␣/␤ on microglia, astrocytes, and neurons have also been described (12,13,31,43). Most cells, including astrocytes and microglia, are able to produce IFN-␣/␤ as an initial nonspecific immune response against virus infection (52, 64); IFN-␥ is released by activated T lymphocytes as part of the specific immune response. The intrinsic IFN system provides the first line of defense against viral infections, including systemic CMV infections, and can delay viral replication allowing the activation of the systemic adaptive immune responses.One explanation for the increased pathogenesis in the developing brain compared to that in the mature brain is that the systemic immune system is too immature during development to fight CMV infection. T and B lymphocytes of the adaptive immune system, as well as other cells of the systemic immune system, including natural killer cells and macrophages/monocytes, are involved in combating CMV infections (2,5,8,9,10,20,46,49), and the efficacies of these systems increase with development. Another mechanism that might underlie the susceptibility of developing...

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Section: Induction Of Intrinsic Antiviral Response By Neurons and Glisupporting

confidence: 91%

Cytomegalovirus InducesInterferon-Stimulated Gene Expression and Is Attenuated by Interferon in the DevelopingBrain

Pol¹,

Robek

Ghosh³

et al. 2007

J Virol

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“…12 SFV-based expression indicated that more than 90% of the GFP-positive cells were of neuronal origin. 13 Transduction of primary neurons and hippocampal slice cultures has allowed studies on protein transport and localization as well as functional activity, typically by electrophysiological recordings. 14 Moreover, alphavirus vectors have been applied for in vivo administration in the rodent brain.…”

Section: Neurosciencementioning

confidence: 99%

“…The first step towards gene therapy application was the demonstration of high transduction rates of various Large-scale production Drug screening, structural biology 9,10 Primary neurons Localization, electrophysiology 11 Hippocampal slices Localization, electrophysiology 12,13 In vivo, rat brain Localization, duration 16 Gene therapy Cancer vaccines Tumor protection, tumor regression 20 Tumor cell lines Transduction, cell killing 17,18 Intratumoral injection Tumor regression 23,24 Liposome-encapsulation Tumor-targeted gene delivery, therapy 28 18 Clearly, the infection rate was modest, although a bystander effect was observed for the therapeutic TK (thymidine kinase) after ganciclovir treatment. Another application of alphaviruses for cancer therapy has been vaccination with tumor antigens.…”

Section: Gene Therapymentioning

confidence: 99%

Biology and application of alphaviruses in gene therapy

Lundström¹

2005

Gene Ther

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“…A multitude of gene transfer methods for postmitotic neurons are now available, including different viral vectors (Ehrengruber et al, 2001) to make adenovirus, adeno-associated virus, Sindbis virus (Ehrengruber et al, 1999), Semliki Forest virus, measles virus and lentivirus (Zufferey et al, 1997). Each viral vector has its own specificity and advantages (Ehrengruber et al, 2001).…”

Section: Lentivirus Preparation and Injectionmentioning

confidence: 99%

“…Each viral vector has its own specificity and advantages (Ehrengruber et al, 2001). Here we describe the use of lentiviral vectors to transduce genes into CA1 pyramidal cells in hippocampal brain slices, primarily for long-term expression studies.…”

Section: Lentivirus Preparation and Injectionmentioning

confidence: 99%

Characterization of Oligophrenin‐1, a RhoGAP Lost in Patients Affected with Mental Retardation: Lentiviral Injection in Organotypic Brain Slice Cultures

Kasri

Govek

Aelst

2008

Methods in Enzymology

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Mutations in regulators and effectors of the Rho GTPases underlie various forms of mental retardation (MR). Among them, oligophrenin-1 (OPHN1), which encodes a Rho-GTPase activating protein (Rho-GAP), was one of the first Rho-linked MR gene identified. Upon characterization of OPHN1 in hippocampal brain slices, we obtained evidence for the requirement of OPHN1 in dendritic spine morphogenesis and synaptic function of CA1 pyramidal neurons. Organotypic hippocampal brain slice cultures are commonly used as a model system to investigate the morphology and synaptic function of neurons, mainly because they allow for the long-term examination of neurons in a preparation where the gross cellular architecture of the hippocampus is retained. In addition, the maintenance of the tri-synaptic circuitry in hippocampal slices enables the study of synaptic connections. Today, a multitude of gene transfer methods for postmitotic neurons in brain slices are available to easily manipulate and scrutinize the involvement of signaling molecules, such as Rho GTPases, in specific cellular processes in this system. This chapter covers techniques detailing the preparation and culturing of organotypic hippocampal brain slices, as well as the production and injection of lentivirus into brain slices.

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Gene Transfer into Neurons from Hippocampal Slices: Comparison of Recombinant Semliki Forest Virus, Adenovirus, Adeno-Associated Virus, Lentivirus, and Measles Virus

Cited by 123 publications

References 52 publications

Cytomegalovirus InducesInterferon-Stimulated Gene Expression and Is Attenuated by Interferon in the DevelopingBrain

Cytomegalovirus InducesInterferon-Stimulated Gene Expression and Is Attenuated by Interferon in the DevelopingBrain

Biology and application of alphaviruses in gene therapy

Characterization of Oligophrenin‐1, a RhoGAP Lost in Patients Affected with Mental Retardation: Lentiviral Injection in Organotypic Brain Slice Cultures

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