In vivo neural activity measured with FM 1-43 in rat mesenteric microcirculation

D. J. Meyer, J. T. Cunningham, M. J. Sullivan, K. S. Curtis, J. P. Vuchetich, M. Hay

Research output: Contribution to journalArticle

Abstract

To date, local neural control of microcirculation has been investigated in an indirect manner due to inability to visualize or directly record activity of small terminal nerve fibers in vivo. Thus, the present study was designed to measure neural activity associated with intact mesenteric microcirculation in vivo. First, in order to determine the location of sympathetic microcirculatory innervation, we employed dopamine-β-hydroxylase (DBH) immunofluorescence in paraformaldahyde fixed rat mesenteries. Most DBH-positive fibers were noted to course along with microvessels down to diameters as small as 8 μm. Nevertheless, not all microvessels were associated with DBH-positive fibers, and visa versa. To visualize neural activity associated with mesenteric micorcirculation in vivo, the fluorescent dye FM 1-43 was employed. Rats were anesthetized with Inactin and their mesentery was exteriorized for viewing of the microcirculation. The mesentery was supervised with 10 μM FM 1-43 for 2 minutes followed by a 4 minute wash with Ringers and 4 minutes monitoring of FM 1-43 fluorescence decay. This procedure was repeated following 2 minutes exposure to 1% lidocaine. The pattern of FM 1-43 fluorescence was similar to the pattern of DBH staining, indicating uptake of the dye by sympathetic neurons. FM 1-43 fluorescence decay rate is proportional to the rate of neural exocytotic activity. Prior to lidocaine exposure, FM 1-43 fluorescence at single neural boutons decayed rapidly. This decay was significantly attenuated by lidocaine. Thus, FM 1-43 fluorescence may be used to monitor neuronal exocytotic activity in an in vivo microcirculatory preparation.

Original languageEnglish
JournalFASEB Journal
Volume12
Issue number5
StatePublished - 20 Mar 1998

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Microcirculation
Rats
Fluorescence
Mesentery
Lidocaine
Microvessels
Fibers
Mixed Function Oxygenases
FM1 43
Fluorescent Dyes
Nerve Fibers
Neurons
Fluorescent Antibody Technique
Dopamine
Coloring Agents
Staining and Labeling
Monitoring

Cite this

Meyer, D. J., Cunningham, J. T., Sullivan, M. J., Curtis, K. S., Vuchetich, J. P., & Hay, M. (1998). In vivo neural activity measured with FM 1-43 in rat mesenteric microcirculation. FASEB Journal, 12(5).
Meyer, D. J. ; Cunningham, J. T. ; Sullivan, M. J. ; Curtis, K. S. ; Vuchetich, J. P. ; Hay, M. / In vivo neural activity measured with FM 1-43 in rat mesenteric microcirculation. In: FASEB Journal. 1998 ; Vol. 12, No. 5.
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Meyer, DJ, Cunningham, JT, Sullivan, MJ, Curtis, KS, Vuchetich, JP & Hay, M 1998, 'In vivo neural activity measured with FM 1-43 in rat mesenteric microcirculation', FASEB Journal, vol. 12, no. 5.

In vivo neural activity measured with FM 1-43 in rat mesenteric microcirculation. / Meyer, D. J.; Cunningham, J. T.; Sullivan, M. J.; Curtis, K. S.; Vuchetich, J. P.; Hay, M.

In: FASEB Journal, Vol. 12, No. 5, 20.03.1998.

Research output: Contribution to journalArticle

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AU - Meyer, D. J.

AU - Cunningham, J. T.

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AU - Hay, M.

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AB - To date, local neural control of microcirculation has been investigated in an indirect manner due to inability to visualize or directly record activity of small terminal nerve fibers in vivo. Thus, the present study was designed to measure neural activity associated with intact mesenteric microcirculation in vivo. First, in order to determine the location of sympathetic microcirculatory innervation, we employed dopamine-β-hydroxylase (DBH) immunofluorescence in paraformaldahyde fixed rat mesenteries. Most DBH-positive fibers were noted to course along with microvessels down to diameters as small as 8 μm. Nevertheless, not all microvessels were associated with DBH-positive fibers, and visa versa. To visualize neural activity associated with mesenteric micorcirculation in vivo, the fluorescent dye FM 1-43 was employed. Rats were anesthetized with Inactin and their mesentery was exteriorized for viewing of the microcirculation. The mesentery was supervised with 10 μM FM 1-43 for 2 minutes followed by a 4 minute wash with Ringers and 4 minutes monitoring of FM 1-43 fluorescence decay. This procedure was repeated following 2 minutes exposure to 1% lidocaine. The pattern of FM 1-43 fluorescence was similar to the pattern of DBH staining, indicating uptake of the dye by sympathetic neurons. FM 1-43 fluorescence decay rate is proportional to the rate of neural exocytotic activity. Prior to lidocaine exposure, FM 1-43 fluorescence at single neural boutons decayed rapidly. This decay was significantly attenuated by lidocaine. Thus, FM 1-43 fluorescence may be used to monitor neuronal exocytotic activity in an in vivo microcirculatory preparation.

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Meyer DJ, Cunningham JT, Sullivan MJ, Curtis KS, Vuchetich JP, Hay M. In vivo neural activity measured with FM 1-43 in rat mesenteric microcirculation. FASEB Journal. 1998 Mar 20;12(5).