Inducing dendritic growth in cultured sympathetic neurons.

Atefeh Ghogha, Donald A. Bruun, Pamela J Lein

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The shape of the dendritic arbor determines the total synaptic input a neuron can receive (1-3), and influences the types and distribution of these inputs (4-6). Altered patterns of dendritic growth and plasticity are associated with impaired neurobehavioral function in experimental models (7), and are thought to contribute to clinical symptoms observed in both neurodevelopmental disorders (8-10) and neurodegenerative diseases (11-13). Such observations underscore the functional importance of precisely regulating dendritic morphology, and suggest that identifying mechanisms that control dendritic growth will not only advance understanding of how neuronal connectivity is regulated during normal development, but may also provide insight on novel therapeutic strategies for diverse neurological diseases. Mechanistic studies of dendritic growth would be greatly facilitated by the availability of a model system that allows neurons to be experimentally switched from a state in which they do not extend dendrites to one in which they elaborate a dendritic arbor comparable to that of their in vivo counterparts. Primary cultures of sympathetic neurons dissociated from the superior cervical ganglia (SCG) of perinatal rodents provide such a model. When cultured in defined medium in the absence of serum and ganglionic glial cells, sympathetic neurons extend a single process which is axonal, and this unipolar state persists for weeks to months in culture (14,15). However, the addition of either bone morphogenetic protein-7 (BMP-7) (16,17) or Matrigel (18) to the culture medium triggers these neurons to extend multiple processes that meet the morphologic, biochemical and functional criteria for dendrites. Sympathetic neurons dissociated from the SCG of perinatal rodents and grown under defined conditions are a homogenous population of neurons (19) that respond uniformly to the dendrite-promoting activity of Matrigel, BMP-7 and other BMPs of the decapentaplegic (dpp) and 60A subfamilies (17,18,20,21). Importantly, Matrigel- and BMP-induced dendrite formation occurs in the absence of changes in cell survival or axonal growth (17,18). Here, we describe how to set up dissociated cultures of sympathetic neurons derived from the SCG of perinatal rats so that they are responsive to the selective dendrite-promoting activity of Matrigel or BMPs.

Original languageEnglish (US)
JournalJournal of visualized experiments : JoVE
Issue number61
StatePublished - 2012

Fingerprint

Neurons
Dendrites
Growth
Superior Cervical Ganglion
Bone Morphogenetic Protein 7
Rodentia
Bone
Neurodegenerative diseases
Proteins
Neuroglia
Neurodegenerative Diseases
Plasticity
Culture Media
Rats
Cell Survival
Theoretical Models
Cells
Availability
matrigel
Serum

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Inducing dendritic growth in cultured sympathetic neurons. / Ghogha, Atefeh; Bruun, Donald A.; Lein, Pamela J.

In: Journal of visualized experiments : JoVE, No. 61, 2012.

Research output: Contribution to journalArticle

@article{7fd084bfee6946b883d0100fbf2a4ebe,
title = "Inducing dendritic growth in cultured sympathetic neurons.",
abstract = "The shape of the dendritic arbor determines the total synaptic input a neuron can receive (1-3), and influences the types and distribution of these inputs (4-6). Altered patterns of dendritic growth and plasticity are associated with impaired neurobehavioral function in experimental models (7), and are thought to contribute to clinical symptoms observed in both neurodevelopmental disorders (8-10) and neurodegenerative diseases (11-13). Such observations underscore the functional importance of precisely regulating dendritic morphology, and suggest that identifying mechanisms that control dendritic growth will not only advance understanding of how neuronal connectivity is regulated during normal development, but may also provide insight on novel therapeutic strategies for diverse neurological diseases. Mechanistic studies of dendritic growth would be greatly facilitated by the availability of a model system that allows neurons to be experimentally switched from a state in which they do not extend dendrites to one in which they elaborate a dendritic arbor comparable to that of their in vivo counterparts. Primary cultures of sympathetic neurons dissociated from the superior cervical ganglia (SCG) of perinatal rodents provide such a model. When cultured in defined medium in the absence of serum and ganglionic glial cells, sympathetic neurons extend a single process which is axonal, and this unipolar state persists for weeks to months in culture (14,15). However, the addition of either bone morphogenetic protein-7 (BMP-7) (16,17) or Matrigel (18) to the culture medium triggers these neurons to extend multiple processes that meet the morphologic, biochemical and functional criteria for dendrites. Sympathetic neurons dissociated from the SCG of perinatal rodents and grown under defined conditions are a homogenous population of neurons (19) that respond uniformly to the dendrite-promoting activity of Matrigel, BMP-7 and other BMPs of the decapentaplegic (dpp) and 60A subfamilies (17,18,20,21). Importantly, Matrigel- and BMP-induced dendrite formation occurs in the absence of changes in cell survival or axonal growth (17,18). Here, we describe how to set up dissociated cultures of sympathetic neurons derived from the SCG of perinatal rats so that they are responsive to the selective dendrite-promoting activity of Matrigel or BMPs.",
author = "Atefeh Ghogha and Bruun, {Donald A.} and Lein, {Pamela J}",
year = "2012",
language = "English (US)",
journal = "Journal of Visualized Experiments",
issn = "1940-087X",
publisher = "MYJoVE Corporation",
number = "61",

}

TY - JOUR

T1 - Inducing dendritic growth in cultured sympathetic neurons.

AU - Ghogha, Atefeh

AU - Bruun, Donald A.

AU - Lein, Pamela J

PY - 2012

Y1 - 2012

N2 - The shape of the dendritic arbor determines the total synaptic input a neuron can receive (1-3), and influences the types and distribution of these inputs (4-6). Altered patterns of dendritic growth and plasticity are associated with impaired neurobehavioral function in experimental models (7), and are thought to contribute to clinical symptoms observed in both neurodevelopmental disorders (8-10) and neurodegenerative diseases (11-13). Such observations underscore the functional importance of precisely regulating dendritic morphology, and suggest that identifying mechanisms that control dendritic growth will not only advance understanding of how neuronal connectivity is regulated during normal development, but may also provide insight on novel therapeutic strategies for diverse neurological diseases. Mechanistic studies of dendritic growth would be greatly facilitated by the availability of a model system that allows neurons to be experimentally switched from a state in which they do not extend dendrites to one in which they elaborate a dendritic arbor comparable to that of their in vivo counterparts. Primary cultures of sympathetic neurons dissociated from the superior cervical ganglia (SCG) of perinatal rodents provide such a model. When cultured in defined medium in the absence of serum and ganglionic glial cells, sympathetic neurons extend a single process which is axonal, and this unipolar state persists for weeks to months in culture (14,15). However, the addition of either bone morphogenetic protein-7 (BMP-7) (16,17) or Matrigel (18) to the culture medium triggers these neurons to extend multiple processes that meet the morphologic, biochemical and functional criteria for dendrites. Sympathetic neurons dissociated from the SCG of perinatal rodents and grown under defined conditions are a homogenous population of neurons (19) that respond uniformly to the dendrite-promoting activity of Matrigel, BMP-7 and other BMPs of the decapentaplegic (dpp) and 60A subfamilies (17,18,20,21). Importantly, Matrigel- and BMP-induced dendrite formation occurs in the absence of changes in cell survival or axonal growth (17,18). Here, we describe how to set up dissociated cultures of sympathetic neurons derived from the SCG of perinatal rats so that they are responsive to the selective dendrite-promoting activity of Matrigel or BMPs.

AB - The shape of the dendritic arbor determines the total synaptic input a neuron can receive (1-3), and influences the types and distribution of these inputs (4-6). Altered patterns of dendritic growth and plasticity are associated with impaired neurobehavioral function in experimental models (7), and are thought to contribute to clinical symptoms observed in both neurodevelopmental disorders (8-10) and neurodegenerative diseases (11-13). Such observations underscore the functional importance of precisely regulating dendritic morphology, and suggest that identifying mechanisms that control dendritic growth will not only advance understanding of how neuronal connectivity is regulated during normal development, but may also provide insight on novel therapeutic strategies for diverse neurological diseases. Mechanistic studies of dendritic growth would be greatly facilitated by the availability of a model system that allows neurons to be experimentally switched from a state in which they do not extend dendrites to one in which they elaborate a dendritic arbor comparable to that of their in vivo counterparts. Primary cultures of sympathetic neurons dissociated from the superior cervical ganglia (SCG) of perinatal rodents provide such a model. When cultured in defined medium in the absence of serum and ganglionic glial cells, sympathetic neurons extend a single process which is axonal, and this unipolar state persists for weeks to months in culture (14,15). However, the addition of either bone morphogenetic protein-7 (BMP-7) (16,17) or Matrigel (18) to the culture medium triggers these neurons to extend multiple processes that meet the morphologic, biochemical and functional criteria for dendrites. Sympathetic neurons dissociated from the SCG of perinatal rodents and grown under defined conditions are a homogenous population of neurons (19) that respond uniformly to the dendrite-promoting activity of Matrigel, BMP-7 and other BMPs of the decapentaplegic (dpp) and 60A subfamilies (17,18,20,21). Importantly, Matrigel- and BMP-induced dendrite formation occurs in the absence of changes in cell survival or axonal growth (17,18). Here, we describe how to set up dissociated cultures of sympathetic neurons derived from the SCG of perinatal rats so that they are responsive to the selective dendrite-promoting activity of Matrigel or BMPs.

UR - http://www.scopus.com/inward/record.url?scp=84863771453&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84863771453&partnerID=8YFLogxK

M3 - Article

JO - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

SN - 1940-087X

IS - 61

ER -