In situ microwave fixation provides an instantaneous snapshot of the brain metabolome

Jelena A. Juras; Madison B. Webb; Lyndsay E.A. Young; Kia H. Markussen; Tara R. Hawkinson; Michael D. Buoncristiani; Kayli E. Bolton; Peyton T. Coburn; Meredith I. Williams; Lisa P.Y. Sun; William C. Sanders; Ronald C. Bruntz; Lindsey R. Conroy; Chi Wang; Matthew S. Gentry; Bret N. Smith; Ramon C. Sun

doi:10.1016/j.crmeth.2023.100455

In situ microwave fixation provides an instantaneous snapshot of the brain metabolome

Jelena A. Juras, Madison B. Webb, Lyndsay E.A. Young, Kia H. Markussen, Tara R. Hawkinson, Michael D. Buoncristiani, Kayli E. Bolton, Peyton T. Coburn, Meredith I. Williams, Lisa P.Y. Sun, William C. Sanders, Ronald C. Bruntz, Lindsey R. Conroy, Chi Wang, Matthew S. Gentry, Bret N. Smith, Ramon C. Sun

Research output: Contribution to journal › Article › peer-review

5 Scopus citations

Abstract

Brain glucose metabolism is highly heterogeneous among brain regions and continues postmortem. In particular, we demonstrate exhaustion of glycogen and glucose and an increase in lactate production during conventional rapid brain resection and preservation by liquid nitrogen. In contrast, we show that these postmortem changes are not observed with simultaneous animal sacrifice and in situ fixation with focused, high-power microwave. We further employ microwave fixation to define brain glucose metabolism in the mouse model of streptozotocin-induced type 1 diabetes. Using both total pool and isotope tracing analyses, we identified global glucose hypometabolism in multiple brain regions, evidenced by reduced ¹³C enrichment into glycogen, glycolysis, and the tricarboxylic acid (TCA) cycle. Reduced glucose metabolism correlated with a marked decrease in GLUT2 expression and several metabolic enzymes in unique brain regions. In conclusion, our study supports the incorporation of microwave fixation for more accurate studies of brain metabolism in rodent models.

Original language	English
Article number	100455
Journal	Cell Reports Methods
Volume	3
Issue number	4
DOIs	https://doi.org/10.1016/j.crmeth.2023.100455
State	Published - Apr 24 2023

Bibliographical note

Publisher Copyright:
© 2023 The Author(s)

Funding

This study was supported by National Institutes of Health (NIH) grants R35 NS116824 (M.S.G.), P01 NS097197 (M.S.G.), R01 AG066653 (R.C.S.), and R01 CA266004 (R.C.S.); NIH NIDDK R01 DK122811 (B.N.S.); NIH NINDS R01 NS092552 (B.N.S.); NIH/NCI F99CA264165 (L.E.A.Y.); NIH/NCI training grant T32CA165990 (L.R.C.); and a V-Scholar grant (R.C.S.).

Funders	Funder number
National Institutes of Health (NIH)	R01 CA266004, R01 AG066653, R35 NS116824, P01 NS097197
National Childhood Cancer Registry – National Cancer Institute	F99CA264165, T32CA165990
National Institute of Diabetes and Digestive and Kidney Diseases	R01 DK122811
National Institute of Neurological Disorders and Stroke	R01 NS092552

Keywords

CP: Metabolism
brain metabolism
isotope tracing
microwave fixation
regional metabolism
type 1 diabetes

ASJC Scopus subject areas

Biotechnology
Biochemistry
Biochemistry, Genetics and Molecular Biology (miscellaneous)
Genetics
Radiology Nuclear Medicine and imaging
Computer Science Applications

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.crmeth.2023.100455

Cite this

Juras, J. A., Webb, M. B., Young, L. E. A., Markussen, K. H., Hawkinson, T. R., Buoncristiani, M. D., Bolton, K. E., Coburn, P. T., Williams, M. I., Sun, L. P. Y., Sanders, W. C., Bruntz, R. C., Conroy, L. R., Wang, C., Gentry, M. S., Smith, B. N., & Sun, R. C. (2023). In situ microwave fixation provides an instantaneous snapshot of the brain metabolome. Cell Reports Methods, 3(4), Article 100455. https://doi.org/10.1016/j.crmeth.2023.100455

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abstract = "Brain glucose metabolism is highly heterogeneous among brain regions and continues postmortem. In particular, we demonstrate exhaustion of glycogen and glucose and an increase in lactate production during conventional rapid brain resection and preservation by liquid nitrogen. In contrast, we show that these postmortem changes are not observed with simultaneous animal sacrifice and in situ fixation with focused, high-power microwave. We further employ microwave fixation to define brain glucose metabolism in the mouse model of streptozotocin-induced type 1 diabetes. Using both total pool and isotope tracing analyses, we identified global glucose hypometabolism in multiple brain regions, evidenced by reduced 13C enrichment into glycogen, glycolysis, and the tricarboxylic acid (TCA) cycle. Reduced glucose metabolism correlated with a marked decrease in GLUT2 expression and several metabolic enzymes in unique brain regions. In conclusion, our study supports the incorporation of microwave fixation for more accurate studies of brain metabolism in rodent models.",

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doi = "10.1016/j.crmeth.2023.100455",

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Juras, JA, Webb, MB, Young, LEA, Markussen, KH, Hawkinson, TR, Buoncristiani, MD, Bolton, KE, Coburn, PT, Williams, MI, Sun, LPY, Sanders, WC, Bruntz, RC, Conroy, LR, Wang, C, Gentry, MS, Smith, BN & Sun, RC 2023, 'In situ microwave fixation provides an instantaneous snapshot of the brain metabolome', Cell Reports Methods, vol. 3, no. 4, 100455. https://doi.org/10.1016/j.crmeth.2023.100455

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T1 - In situ microwave fixation provides an instantaneous snapshot of the brain metabolome

AU - Juras, Jelena A.

AU - Webb, Madison B.

AU - Young, Lyndsay E.A.

AU - Markussen, Kia H.

AU - Hawkinson, Tara R.

AU - Buoncristiani, Michael D.

AU - Bolton, Kayli E.

AU - Coburn, Peyton T.

AU - Williams, Meredith I.

AU - Sun, Lisa P.Y.

AU - Sanders, William C.

AU - Bruntz, Ronald C.

AU - Conroy, Lindsey R.

AU - Wang, Chi

AU - Gentry, Matthew S.

AU - Smith, Bret N.

AU - Sun, Ramon C.

PY - 2023/4/24

Y1 - 2023/4/24

N2 - Brain glucose metabolism is highly heterogeneous among brain regions and continues postmortem. In particular, we demonstrate exhaustion of glycogen and glucose and an increase in lactate production during conventional rapid brain resection and preservation by liquid nitrogen. In contrast, we show that these postmortem changes are not observed with simultaneous animal sacrifice and in situ fixation with focused, high-power microwave. We further employ microwave fixation to define brain glucose metabolism in the mouse model of streptozotocin-induced type 1 diabetes. Using both total pool and isotope tracing analyses, we identified global glucose hypometabolism in multiple brain regions, evidenced by reduced 13C enrichment into glycogen, glycolysis, and the tricarboxylic acid (TCA) cycle. Reduced glucose metabolism correlated with a marked decrease in GLUT2 expression and several metabolic enzymes in unique brain regions. In conclusion, our study supports the incorporation of microwave fixation for more accurate studies of brain metabolism in rodent models.

AB - Brain glucose metabolism is highly heterogeneous among brain regions and continues postmortem. In particular, we demonstrate exhaustion of glycogen and glucose and an increase in lactate production during conventional rapid brain resection and preservation by liquid nitrogen. In contrast, we show that these postmortem changes are not observed with simultaneous animal sacrifice and in situ fixation with focused, high-power microwave. We further employ microwave fixation to define brain glucose metabolism in the mouse model of streptozotocin-induced type 1 diabetes. Using both total pool and isotope tracing analyses, we identified global glucose hypometabolism in multiple brain regions, evidenced by reduced 13C enrichment into glycogen, glycolysis, and the tricarboxylic acid (TCA) cycle. Reduced glucose metabolism correlated with a marked decrease in GLUT2 expression and several metabolic enzymes in unique brain regions. In conclusion, our study supports the incorporation of microwave fixation for more accurate studies of brain metabolism in rodent models.

KW - CP: Metabolism

KW - brain metabolism

KW - isotope tracing

KW - microwave fixation

KW - regional metabolism

KW - type 1 diabetes

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In situ microwave fixation provides an instantaneous snapshot of the brain metabolome

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

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