Functional Magnetic Resonance Spectroscopy at 7 T in the Rat Barrel Cortex During Whisker Activation

Summary

After checking by blood-oxygen-level-dependent functional magnetic resonance imaging (BOLD fMRI) that the corresponding somatosensory barrel field cortex area (called S1BF) is correctly activated, the main goal of this study is to quantify lactate content fluctuations in the activated rat brains by localized proton magnetic resonance spectroscopy (¹H-MRS) at 7 T.

Abstract

Nuclear magnetic resonance (NMR) spectroscopy offers the opportunity to measure cerebral metabolite contents in vivo and noninvasively. Thanks to technological developments over the last decade and the increase in magnetic field strength, it is now possible to obtain good resolution spectra in vivo in the rat brain. Neuroenergetics (i.e., the study of brain metabolism) and, especially, metabolic interactions between the different cell types have attracted more and more interest in recent years. Among these metabolic interactions, the existence of a lactate shuttle between neurons and astrocytes is still debated. It is, thus, of great interest to perform functional proton magnetic resonance spectroscopy (¹H-MRS) in a rat model of brain activation and monitor lactate. However, the methyl lactate peak overlaps lipid resonance peaks and is difficult to quantify. The protocol described below allows metabolic and lactate fluctuations to be monitored in an activated brain area. Cerebral activation is obtained by whisker stimulation and ¹H-MRS is performed in the corresponding activated barrel cortex, whose area is detected using blood-oxygen-level-dependent functional magnetic resonance imaging (BOLD fMRI). All steps are fully described: the choice of anesthetics, coils, and sequences, achieving efficient whisker stimulation directly in the magnet, and data processing.

Introduction

The brain possesses intrinsic mechanisms that allow the regulation of its major substrate (i.e., glucose), both for its contribution and its utilization, depending on variations in local cerebral activity. Although glucose is the main energy substrate for the brain, experiments performed in recent years have shown that lactate, which is produced by the astrocytes, could be an efficient energy substrate for the neurons. This raises the hypothesis of a lactate shuttle between astrocytes and neurons¹. Known as ANLS, for astrocyte-neuron lactate shuttle², the theory is still highly debated but has led to the proposa....

Protocol

All animal procedures were conducted in accordance with the Animal Experimentation Guidelines of the European Communities Council Directive of November 24, 1986 (86/609/EEC). The protocol met the ethical guidelines of the French Ministry of Agriculture and Forests and was approved by the local ethics committees (Comité d'éthique pour L'expérimentation Animale Bordeaux n°50112090-A).

NOTE: During the MR measurements, an adequate level of anesthesia and physiological monitoring (body temperature, respiratory rate) are indispensable requirements.

1. Animals

1. Use male Wistar rat....

Results

This protocol allows the quantification of metabolite fluctuations during cerebral activation, which is obtained by right whisker stimulation directly in the magnet.

In this study, the overall goal of BOLD fMRI was to check that the whisker stimulation was efficient, to visualize the activated S1BF area, and to correctly locate the voxel for ¹H-fMRS. The device built for whisker activation is efficient. Indeed, when r.......

Discussion

The barrel cortex, also called S1BF for the somatosensory cortex or barrel field, is a region within the cortical layer IV that can be observed using cytochrome c oxidase staining⁹, and its organization is well known since it has been largely described^10,11. One vibrissa is connected to one barrel, in which around 19,000 neurons are organized in a column¹². The whisker-to-barrel cortex pathway has several advantages.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
0.5 mL syringe with needle	Becton, Dickinson and Company, USA	2020-10	0.33 mm (29 G) x 12.7 mm
1H spectroscopy surface coil	Bruker, Ettlingen, Germany	T116344
7T Bruker Biospec system	Bruker, Ettlingen, Germany	70/20 USR
Arduino Uno based pulsing device	custom made
Atipamezole	Vétoquinol, S.A., France	V8335602	Antisedan, 4.28 mg
Breathing mask	custom made
Eye ointment	TVM laboratoire, France	40365	Ocry gel 10 g
Induction chamber	custom made		30x17x15 cm
Inlet flexible pipe	Gardena, Germany	1348-20	4.6-mm diameter, 3m long
Isoflurane pump, Model 100 series vaporizer, classic T3	Surgivet, Harvard Apparatus	WWV90TT	from OH 43017, U.S.A
Isoflurane, liquid for inhalation	Vertflurane, Virbac, France	QN01AB06	1000 mg/mL
KD Scientific syringe pump	KD sientific, Holliston, USA	Legato 110
LCModel software	LCModel Inc., Ontario, Canada	6.2
Medetomidine hydrochloride	Vétoquinol, S.A., France	QN05CM91	Domitor, 1 mg/mL
Micropore roll of adhesive plaster	3M micropore, Minnesota, United States	MI912
Micropore roll of adhesive plaster	3M micropore, Minnesota, United States	MI925
Monitoring system of physiologic parameter	SA Instruments, Inc, Stony Brook, NY, USA	Model 1025
NaCl	Fresenius Kabi, Germany	B05XA03	0.9 % 250 mL
Outlet flexible pipe	Gardena, Germany	1348-20	4.6-mm diameter, 4m long
Paravision software	Bruker, Ettlingen, Germany	6.0.1
Peripheral intravenous catheter	Terumo, Shibuya, Tokyo, Japon	SP500930S	22 G x 1", 0.85x25 mm, 35 mL/min
Rat head coil	Bruker, Ettlingen, Germany
Sodic heparin, injectable solution	Choai, Sanofi, Paris, France	B01AB01	5000 IU/mL
Solenoid control valves, plunger valve 2/2 way direct-acting	Burkert, Germany	3099939	Model type 6013
Terumo 2 ml syringe	Terumo, Shibuya, Tokyo, Japon	SY243	with 21 g x 5/8" needle
Terumo 5 mL syringe	Terumo, Shibuya, Tokyo, Japon	05SE1
Wistar RJ-Han rats	Janvier Laboratories, France