<abstract xmlns="http://www.w3.org/1999/xhtml"><p>Two sets of reactants for modelling neurogenesis (SRMN) were developed based on the designed and tested genetic structures of lentiviral vectors. SRMN-1 contains the genetic construct LVV-GFAP-GCaMP3 and is intended for cellspecific transduction in astroglia cells. SRMN-2 contains the genetic construct LVV-PRSx8-TN-XXL and is intended for the phenotype-specific transduction in neurons. The present study examined SRMN-1 and SRMN-2 samples and assessed their efficiency in vitro and in vivo in Norvegicus rats. Specificity to particular cell types for all SRMN samples exceeded 97%. The number of induced signalling cascades was determined via activation of intracellular ingsignalling cascades in neurons and astrocytes (purinergic receptors and β-adrenoceptors). The results demonstrated dynamic recording of fluorescent signals and a two-fold increase in intensity after addition of the activator in all samples. The experimental SRMN samples revealed successful and stable transfection of catecholaminergic neurons and astrocytes, data on transfection efficiency, specificity of the developed genetic structures of SRMN, and calcium dynamics in transfected neurons and astrocytes.</p><p>These results confirm the crucial role of astrocytes in ensuring neurogenesis. The results in pure cell culture (in vitro) were identical to the in vivo results in animals.</p></abstract>

Two sets of reactants for modelling neurogenesis (SRMN) were developed based on the designed and tested genetic structures of lentiviral vectors. SRMN-1 contains the genetic construct LVV-GFAP-GCaMP3 and is intended for cellspecific transduction in astroglia cells. SRMN-2 contains the genetic construct LVV-PRSx8-TN-XXL and is intended for the phenotype-specific transduction in neurons. The present study examined SRMN-1 and SRMN-2 samples and assessed their efficiency in vitro and in vivo in Norvegicus rats. Specificity to particular cell types for all SRMN samples exceeded 97%. The number of induced signalling cascades was determined via activation of intracellular ingsignalling cascades in neurons and astrocytes (purinergic receptors and β-adrenoceptors). The results demonstrated dynamic recording of fluorescent signals and a two-fold increase in intensity after addition of the activator in all samples. The experimental SRMN samples revealed successful and stable transfection of catecholaminergic neurons and astrocytes, data on transfection efficiency, specificity of the developed genetic structures of SRMN, and calcium dynamics in transfected neurons and astrocytes.These results confirm the crucial role of astrocytes in ensuring neurogenesis. The results in pure cell culture (in vitro) were identical to the in vivo results in animals.

Genetic Methods for Detecting Astrocytes, Neurons and Neurogenesis

Immanuel Kant Baltic Federal University (IKBFU)

I.M. Sechenov First Moscow State Medical University (Sechenov University)

N.I. Pirogov Russian National Research Medical University (RNRMU)

Experimental and Applied Biomedical Research (EABR)

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

{"article-title":"Genetic Methods for Detecting Astrocytes, Neurons and Neurogenesis"}

Two sets of reactants for modelling neurogenesis (SRMN) were developed based on the designed and tested genetic structures of lentiviral vectors. SRMN-1...

Genetic Methods for Detecting Astrocytes, Neurons and Neurogenesis

Published Online: Jul 17, 2018

Page range: 175 - 184

Received: Apr 23, 2018

Accepted: Apr 30, 2018

DOI: https://doi.org/10.2478/sjecr-2018-0013

Keywords
neurogenesis astrocytes, neurons, signalling cascades, genetic structure, viral vector

© 2018 Natalia Nikolaevna Shusharina et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Genetic Methods for Detecting Astrocytes, Neurons and Neurogenesis

Published Online: Jul 17, 2018

Page range: 175 - 184

Received: Apr 23, 2018

Accepted: Apr 30, 2018

DOI: https://doi.org/10.2478/sjecr-2018-0013

Keywordsneurogenesis astrocytes, neurons, signalling cascades, genetic structure, viral vector

© 2018 Natalia Nikolaevna Shusharina et al., published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Keywords
neurogenesis astrocytes, neurons, signalling cascades, genetic structure, viral vector