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Metabolic consequences of interleukin-6 challenge in developing neurons and astroglia.

Brown JA, Sherrod SD, Goodwin CR, Brewer B, Yang L, Garbett KA, Li D, McLean JA, Wikswo JP, Mirnics K - J Neuroinflammation (2014)

Bottom Line: While several proteins have been identified as having some link to these developmental disorders, their prevalence is still small and their causative role, if any, is not well understood.Our results revealed that 1) the use of this technology, due to its superb media volume:cell volume ratio, is ideally suited for analysis of cell-type-specific exometabolome signatures; 2) developing neurons have low secretory activity at baseline, while astroglia show strong metabolic activity; 3) both neurons and astroglia respond to IL-6 exposure in a cell type-specific fashion; 4) the astroglial response to IL-6 stimulation is predominantly characterized by increased levels of metabolites, while neurons mostly depress their metabolic activity; and 5) disturbances in glycerophospholipid metabolism and tryptophan/kynurenine metabolite secretion are two putative mechanisms by which IL-6 affects the developing nervous system.Our findings are potentially critical for understanding the mechanism by which IL-6 disrupts brain function, and they provide information about the molecular cascade that links maternal immune activation to developmental brain disorders.

View Article: PubMed Central - PubMed

Affiliation: Department of Psychiatry, 465 21st Avenue South, Vanderbilt University, Nashville, TN, 37232, USA. jacquelyn.a.brown@vanderbilt.edu.

ABSTRACT

Background: Maternal immune activation and subsequent interleukin-6 (IL-6) induction disrupt normal brain development and predispose the offspring to developing autism and schizophrenia. While several proteins have been identified as having some link to these developmental disorders, their prevalence is still small and their causative role, if any, is not well understood. However, understanding the metabolic consequences of environmental predisposing factors could shed light on disorders such as autism and schizophrenia.

Methods: To gain a better understanding of the metabolic consequences of IL-6 exposure on developing central nervous system (CNS) cells, we separately exposed developing neuron and astroglia cultures to IL-6 for 2 hours while collecting effluent from our gravity-fed microfluidic chambers. By coupling microfluidic technologies to ultra-performance liquid chromatography-ion mobility-mass spectrometry (UPLC-IM-MS), we were able to characterize the metabolic response of these CNS cells to a narrow window of IL-6 exposure.

Results: Our results revealed that 1) the use of this technology, due to its superb media volume:cell volume ratio, is ideally suited for analysis of cell-type-specific exometabolome signatures; 2) developing neurons have low secretory activity at baseline, while astroglia show strong metabolic activity; 3) both neurons and astroglia respond to IL-6 exposure in a cell type-specific fashion; 4) the astroglial response to IL-6 stimulation is predominantly characterized by increased levels of metabolites, while neurons mostly depress their metabolic activity; and 5) disturbances in glycerophospholipid metabolism and tryptophan/kynurenine metabolite secretion are two putative mechanisms by which IL-6 affects the developing nervous system.

Conclusions: Our findings are potentially critical for understanding the mechanism by which IL-6 disrupts brain function, and they provide information about the molecular cascade that links maternal immune activation to developmental brain disorders.

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Related in: MedlinePlus

Experimental design and analysis strategy. (A) Microfluidic devices or bioreactors were seeded with neurons or astroglia for 6 days prior to IL-6 exposure. (B) Illustration of the sample preparation method for bioreactor perfusate samples (control ± IL-6, astroglia ± IL-6, and neurons ± IL-6). Samples were prepared for exometabolomic analyses and analysis performed using an ultra-performance liquid chromatography-ion mobility-mass spectrometer.
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Fig1: Experimental design and analysis strategy. (A) Microfluidic devices or bioreactors were seeded with neurons or astroglia for 6 days prior to IL-6 exposure. (B) Illustration of the sample preparation method for bioreactor perfusate samples (control ± IL-6, astroglia ± IL-6, and neurons ± IL-6). Samples were prepared for exometabolomic analyses and analysis performed using an ultra-performance liquid chromatography-ion mobility-mass spectrometer.

Mentions: Microfluidic devices consisted of four separate microchannels, each having an inlet and outlet channel and one cell culture chamber region (Figure 1A). The devices were designed to reduce flow velocity by expanding the cell culture chamber. The larger cell culture chamber, with dimensions of 5,400 μm (length) × 800 μm (width) × 200 μm (height), facilitates cell attachment and has a total volume of approximately 0.86 μL. A PDMS wall separates the four chambers, ensuring that no cross talk occurs among cell culture regions.Figure 1


Metabolic consequences of interleukin-6 challenge in developing neurons and astroglia.

Brown JA, Sherrod SD, Goodwin CR, Brewer B, Yang L, Garbett KA, Li D, McLean JA, Wikswo JP, Mirnics K - J Neuroinflammation (2014)

Experimental design and analysis strategy. (A) Microfluidic devices or bioreactors were seeded with neurons or astroglia for 6 days prior to IL-6 exposure. (B) Illustration of the sample preparation method for bioreactor perfusate samples (control ± IL-6, astroglia ± IL-6, and neurons ± IL-6). Samples were prepared for exometabolomic analyses and analysis performed using an ultra-performance liquid chromatography-ion mobility-mass spectrometer.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4233071&req=5

Fig1: Experimental design and analysis strategy. (A) Microfluidic devices or bioreactors were seeded with neurons or astroglia for 6 days prior to IL-6 exposure. (B) Illustration of the sample preparation method for bioreactor perfusate samples (control ± IL-6, astroglia ± IL-6, and neurons ± IL-6). Samples were prepared for exometabolomic analyses and analysis performed using an ultra-performance liquid chromatography-ion mobility-mass spectrometer.
Mentions: Microfluidic devices consisted of four separate microchannels, each having an inlet and outlet channel and one cell culture chamber region (Figure 1A). The devices were designed to reduce flow velocity by expanding the cell culture chamber. The larger cell culture chamber, with dimensions of 5,400 μm (length) × 800 μm (width) × 200 μm (height), facilitates cell attachment and has a total volume of approximately 0.86 μL. A PDMS wall separates the four chambers, ensuring that no cross talk occurs among cell culture regions.Figure 1

Bottom Line: While several proteins have been identified as having some link to these developmental disorders, their prevalence is still small and their causative role, if any, is not well understood.Our results revealed that 1) the use of this technology, due to its superb media volume:cell volume ratio, is ideally suited for analysis of cell-type-specific exometabolome signatures; 2) developing neurons have low secretory activity at baseline, while astroglia show strong metabolic activity; 3) both neurons and astroglia respond to IL-6 exposure in a cell type-specific fashion; 4) the astroglial response to IL-6 stimulation is predominantly characterized by increased levels of metabolites, while neurons mostly depress their metabolic activity; and 5) disturbances in glycerophospholipid metabolism and tryptophan/kynurenine metabolite secretion are two putative mechanisms by which IL-6 affects the developing nervous system.Our findings are potentially critical for understanding the mechanism by which IL-6 disrupts brain function, and they provide information about the molecular cascade that links maternal immune activation to developmental brain disorders.

View Article: PubMed Central - PubMed

Affiliation: Department of Psychiatry, 465 21st Avenue South, Vanderbilt University, Nashville, TN, 37232, USA. jacquelyn.a.brown@vanderbilt.edu.

ABSTRACT

Background: Maternal immune activation and subsequent interleukin-6 (IL-6) induction disrupt normal brain development and predispose the offspring to developing autism and schizophrenia. While several proteins have been identified as having some link to these developmental disorders, their prevalence is still small and their causative role, if any, is not well understood. However, understanding the metabolic consequences of environmental predisposing factors could shed light on disorders such as autism and schizophrenia.

Methods: To gain a better understanding of the metabolic consequences of IL-6 exposure on developing central nervous system (CNS) cells, we separately exposed developing neuron and astroglia cultures to IL-6 for 2 hours while collecting effluent from our gravity-fed microfluidic chambers. By coupling microfluidic technologies to ultra-performance liquid chromatography-ion mobility-mass spectrometry (UPLC-IM-MS), we were able to characterize the metabolic response of these CNS cells to a narrow window of IL-6 exposure.

Results: Our results revealed that 1) the use of this technology, due to its superb media volume:cell volume ratio, is ideally suited for analysis of cell-type-specific exometabolome signatures; 2) developing neurons have low secretory activity at baseline, while astroglia show strong metabolic activity; 3) both neurons and astroglia respond to IL-6 exposure in a cell type-specific fashion; 4) the astroglial response to IL-6 stimulation is predominantly characterized by increased levels of metabolites, while neurons mostly depress their metabolic activity; and 5) disturbances in glycerophospholipid metabolism and tryptophan/kynurenine metabolite secretion are two putative mechanisms by which IL-6 affects the developing nervous system.

Conclusions: Our findings are potentially critical for understanding the mechanism by which IL-6 disrupts brain function, and they provide information about the molecular cascade that links maternal immune activation to developmental brain disorders.

Show MeSH
Related in: MedlinePlus