Differential Metabolic Profiles in Rat Retrosplenial Cortex, Cingulate Cortex and Medial Prefrontal Cortex: Relationship with Cytoarchitecture and Functional Implications
Hui Zhang1 and Hao Lei1

1National Center of Magnetic Resonance in Wuhan, State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, Wuhan, China, People's Republic of

Synopsis

In this study, we measured regional neurochemical variations in rat prelimbic cortex (PrL)/infralimbic cortex (IL), cingulate cortex (Cg) and retrosplenial cortex (RSC) with in vivo 1H-MRS at 7T. It was found that the regional metabolic variations follow cytoarchitectural/receptor-architectonical organization in these brain regions.

Introduction

Medial prefrontal cortex (mPFC) and cingulate cortex are important nodes of brain networks known to be involved in many psychiatric disorders [1]. A recent in vivo 1H-MRS study has demonstrated that regional variations in glutamate (Glu) and γ-aminobutyric acid (GABA) concentrations in human cingulate cortex may reflect receptor-architectonical and functional segregation of the cingulate cortex along the rostral-caudal axis [2]. To further elucidate the relationship between the metabolic profile and cytoarchitectural/receptor-architectonical organization in the prefrontal and cingulate cortex, we measured regional neurochemical variations in rat prelimbic cortex (PrL)/infralimbic cortex (IL), cingulate cortex (Cg) and retrosplenial cortex (RSC) with in vivo 1H-MRS at 7 T.

Materials and methods

Eleven adult male Sprague-Dawley rats (about 3.6 months old, weighting 525±32 g) were used. All spectra were acquired on a 7 T/20 cm Bruker Biospec scanner with a volume coil for transmission and a quadrature surface coil for detection. The animals were anesthetized with 1.8-2.5% isoflurane. Localized spectra were acquired from PrL+IL (2.5 mm×2.4 mm×2.0 mm), Cg (2.3 mm×2.0 mm×2.5 mm) and RSC (2.0 mm×2.0 mm×2.5 mm) of each animal with a PRESS sequence (Fig. 1), VAPOR water suppression, TR/TE 4000/15 ms, spectral bandwidth 4 kHz, 2048 data points and 512 averages. LCModel was used for quantification, and only the results with fitting CRLBs less than 20% were reported. Paired t-test was used for statistical analysis. Bonferroni correction was applied for multiple comparisons among different brain regions. A corrected p<0.05 was considered to be statistically significant.

Results

Figure 1 shows the voxels for in vivo 1H-MRS, representative spectra acquired and the corresponding LCModel fits. Figure 2 plots quantitative regional metabolic variations. RSC had the highest tNAA/tCr, but the lowest Glu/tCr and Glu/tNAA among the three regions. PrL+IL had the highest Ins/tCr and Gln/tCr. The three regions showed similar Glu/Gln.

Discussion

The observation that the RSC had higher tNAA/tCr is consistent with the results from a previous in vivo 1H-MRS study on mice brain [3]. Although the three brain regions measured had similar density of total neurons [4], the RSC is known to have higher density of neuropil (i.e., apical dendrites of layer V pyramidal neurons) than the other two regions [4, 5]. This might explain the higher level of tNAA observed in the region. The rostrocaudal gradients of Glu/tCr and Glu/tNAA corroborated with the known distribution of N-methyl-D-aspartate (NMDA) receptors in rat brain [6]. For example, the RSC had the lowest Glu/tCr and Glu/tNAA, and was also reported to have the lowest NMDA-sensitive L-glutamate binding sites among the three brain region [6]. RSC had a prominent granular layer (i.e., layer IV), while the other two brain regions did not have [4]. The local excitatory/inhibitory network within the RSC and the external excitatory disinhibition inputs of RSC could be different from those in the Cg and PrL/IL [7]. Our results are in line with these previous findings. Ins and Gln are metabolites mainly located in glia cells [8, 9]. Regional variations in these metabolites are consistent with the previous finding that the density of s-100β positive glial cells in the PrL/IL is higher than that in the Cg [10]. Glu/Gln did not show significant regional variations, perhaps indicating that the overall neuronal-glial interaction is similar among these brain regions.

Acknowledgements

Supported by National Basic Research Program of China (2011CB707802), and Natural Science Foundation of China (21221064 and 81000598).

References

[1]Lu H et al, Proc Natl Acad Sci U S A, 2012, 109: 3979-84. [2]Dow W et al, J Neurosci, 2013, 33:12698-704. [3]Kulak A et al, J Neurochem, 2010, 115: 1466-77. [4]Vogt BA et al, J Comp Neuro, 1981, 195: 603-25. [5]Monaghan DT et al, Brain Res, 1984, 324: 160-4. [6]Monaghan DT et al, J Neurosci, 1985, 5: 2909-19. [7]Li Q et al, J Neurosci, 2002, 22: 3070-80. [8]Brand A et al, Dev Neurosci, 1993, 15: 289-98. [9]Urenjak J et al, J Neurosci, 1993, 13: 981-9. [10]Gosselin RD et al, Neuroscience, 2009, 15: 915-25.

Figures

Figure 1. Localized 1H spectra from rat PrL+IL, Cg and RSC. Raw spectra are shown in black solid lines; LCModel fits to the raw spectra are shown in red solid lines. tNAA: N-acetyl aspartate + N-acetyl aspartylglutamate; tCr: creatine + phosphocreatine; Gln: glutamine; Ins: myo-inositol.

Figure 2. Regional neurochemical variations. *: p<0.05, compared to another brain region and #: p<0.05 compared to the other two brain regions, paired t-tests with Bonferroni correction for multiple comparisons.



Proc. Intl. Soc. Mag. Reson. Med. 24 (2016)
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