In situ hybridization study of the distribution of choline acetyltransferase mRNA and its splice variants in the mouse brain and spinal cord

Trifonov, Stefan; Houtani, Takeshi; Hamada, Satoko; Kase, Masahiko; Maruyama, Masato; Sugimoto, Tetsuo

doi:10.1016/j.neuroscience.2008.12.054

Cited by 40 publications

(26 citation statements)

References 37 publications

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“…ISH data of the mouse spinal cord from the Allen Brain Atlas (http://www.brain-map.org/) demonstrate that the distribution of IMS images is reasonable compared with the localization of mRNA for ACh-related molecules such as ChAT, acetylcholinesterase (AChE), nicotinic receptor (nAChR), muscarinic receptor (mAChR), vesicular acetylcholine transporter (VAChT), and choline transporter (ChT). Many previous results matching our IMS data also match those of ISH and/or immunohistochemistry for ACh-related molecules including ChAT [42, 44, 46], AChE [47], nAChR [48], and VAChT [46, 49, 50]. Through its action on spinal cholinergic receptors, endogenous ACh participates not only in motor action but also in the setting of nociceptive thresholds and in the effect of clinically relevant analgesics; thus, the direct detection of ACh in the spinal cord will be useful for assessments such as evaluations of therapeutic strategies.…”

Section: Resultssupporting

confidence: 88%

Visualization of acetylcholine distribution in central nervous system tissue sections by tandem imaging mass spectrometry

et al. 2012

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Metabolite distribution imaging via imaging mass spectrometry (IMS) is an increasingly utilized tool in the field of neurochemistry. As most previous IMS studies analyzed the relative abundances of larger metabolite species, it is important to expand its application to smaller molecules, such as neurotransmitters. This study aimed to develop an IMS application to visualize neurotransmitter distribution in central nervous system tissue sections. Here, we raise two technical problems that must be resolved to achieve neurotransmitter imaging: (1) the lower concentrations of bioactive molecules, compared with those of membrane lipids, require higher sensitivity and/or signal-to-noise (S/N) ratios in signal detection, and (2) the molecular turnover of the neurotransmitters is rapid; thus, tissue preparation procedures should be performed carefully to minimize postmortem changes. We first evaluated intrinsic sensitivity and matrix interference using Matrix Assisted Laser Desorption/Ionization (MALDI) mass spectrometry (MS) to detect six neurotransmitters and chose acetylcholine (ACh) as a model for study. Next, we examined both single MS imaging and MS/MS imaging for ACh and found that via an ion transition from m/z 146 to m/z 87 in MS/MS imaging, ACh could be visualized with a high S/N ratio. Furthermore, we found that in situ freezing method of brain samples improved IMS data quality in terms of the number of effective pixels and the image contrast (i.e., the sensitivity and dynamic range). Therefore, by addressing the aforementioned problems, we demonstrated the tissue distribution of ACh, the most suitable molecular specimen for positive ion detection by IMS, to reveal its localization in central nervous system tissues.

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Section: Resultssupporting

confidence: 88%

Visualization of acetylcholine distribution in central nervous system tissue sections by tandem imaging mass spectrometry

et al. 2012

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“…In situ hybridization was performed according to the methods used by Wang and Yang (2010) and Trifonov et al (2009), with some modifications. Tissue sections were pre-hybridized at 58°C for 2h in 50% deionized formamide in 5× SSC (standard saline citrate, pH 7.0), which contained denatured salmon sperm DNA.…”

Section: In Situ Hybridizationmentioning

confidence: 99%

Expression characteristics of the SUMOylation genes SUMO-1 and Ubc9 in the developing testis and ovary of Chinese mitten crab, Eriocheir sinensis

Wang

Chen

et al. 2012

Gene

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“…Amino acid sequence of ChAT was elucidated after the isolation of the complete sequence of a cDNA encoding porcine and rat ChAT (11,14). This allowed the subsequent in situ hybridization studies that have revealed the expression of ChAT mRNAs in neurons of the central nervous system (83,97). However, the putative localization of ChAT mRNAs in the nerve terminals has not, so far, been analyzed by in situ hybridization.…”

Section: Localization Of Choline Acetyltrans-ferasementioning

confidence: 99%

Historical landmarks in the histochemistry of the cholinergic synapse: perspectives for future researches

Anglade¹,

Larabi-Godinot

2010

Biomed. Res.

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Nearly one hundred years ago, acetylcholine (ACh) was proposed as a chemical agent responsible for nerve transmission at the synapse, the junction area between one neuron and its target cell. Since it has been proved that ACh played, indeed, a major role in the functioning of the nerve system in the vertebrates, cholinergic nerve transmission became a basic field of study in neuroscience. The birth of histochemistry and its ulterior developments allowed in situ localization of the molecular agents related to the functioning of the cholinergic synapse. This report presents historical landmarks in the histochemistry of major cholinergic agents (acetylcholinesterase, nicotinic acetylcholine receptor, choline acetyltransferase, and ACh), a domain which has greatly contributed to the knowledge of the nerve system. It is emphasized that despite extraordinary progresses made in this field, basic problems, such as in situ localization of ACh, still remain to be solved.

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In situ hybridization study of the distribution of choline acetyltransferase mRNA and its splice variants in the mouse brain and spinal cord

Cited by 40 publications

References 37 publications

Visualization of acetylcholine distribution in central nervous system tissue sections by tandem imaging mass spectrometry

Visualization of acetylcholine distribution in central nervous system tissue sections by tandem imaging mass spectrometry

Expression characteristics of the SUMOylation genes SUMO-1 and Ubc9 in the developing testis and ovary of Chinese mitten crab, Eriocheir sinensis

Historical landmarks in the histochemistry of the cholinergic synapse: perspectives for future researches

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