Immunohistochemical visualization of mouse interneuron subtypes [v1; ref status: indexed, http://f1000r.es/4em]
The activity of excitatory neurons is controlled by a small, but highly diverse population of inhibitory interneurons. These cells show a high level of physiological, morphological and neurochemical heterogeneity, and play highly specific roles in neuronal circuits. In the mammalian hippocampus, the...
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| Format: | Article |
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F1000 Research Ltd
2014-10-01
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| Online Access: | http://f1000research.com/articles/3-242/v1 |
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| author | Simon Molgaard Maj Ulrichsen Simon Boggild Marie-Louise Holm Christian Vaegter Jens Nyengaard Simon Glerup |
| author_facet | Simon Molgaard Maj Ulrichsen Simon Boggild Marie-Louise Holm Christian Vaegter Jens Nyengaard Simon Glerup |
| author_sort | Simon Molgaard |
| collection | DOAJ |
| description | The activity of excitatory neurons is controlled by a small, but highly diverse population of inhibitory interneurons. These cells show a high level of physiological, morphological and neurochemical heterogeneity, and play highly specific roles in neuronal circuits. In the mammalian hippocampus, these are divided into 21 different subtypes of GABAergic interneurons based on their expression of different markers, morphology and their electrophysiological properties. Ideally, all can be marked using an antibody directed against the inhibitory neurotransmitter GABA, but parvalbumin, calbindin, somatostatin, and calretinin are also commonly used as markers to narrow down the specific interneuron subtype. Here, we describe a journey to find the necessary immunological reagents for studying GABAergic interneurons of the mouse hippocampus. Based on web searches there are several hundreds of different antibodies on the market directed against these four markers. Searches in the literature databases allowed us to narrow it down to a subset of antibodies most commonly used in publications. However, in our hands the most cited ones did not work for immunofluorescence stainings of formaldehyde fixed tissue sections and cultured hippocampal neurons, and we had to immunostain our way through thirteen different commercial antibodies before finally finding a suitable antibody for each of the four markers. The antibodies were evaluated based on signal-to-noise ratios as well as if positive cells were found in layers of the hippocampus where they have previously been described. Additionally, the antibodies were also tested on sections from mouse spinal cord with similar criteria for specificity of the antibodies. Using the antibodies with a high rating on pAbmAbs, stainings with high signal-to-noise ratios and location of the immunostained cells in accordance with the literature could be obtained, making these antibodies suitable choices for studying the GABAergic system. |
| first_indexed | 2024-12-11T08:52:18Z |
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| id | doaj.art-17e8ddf75e6643dda90318fc170e49af |
| institution | Directory Open Access Journal |
| issn | 2046-1402 |
| language | English |
| last_indexed | 2024-12-11T08:52:18Z |
| publishDate | 2014-10-01 |
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| spelling | doaj.art-17e8ddf75e6643dda90318fc170e49af2022-12-22T01:13:58ZengF1000 Research LtdF1000Research2046-14022014-10-01310.12688/f1000research.5349.15710Immunohistochemical visualization of mouse interneuron subtypes [v1; ref status: indexed, http://f1000r.es/4em]Simon Molgaard0Maj Ulrichsen1Simon Boggild2Marie-Louise Holm3Christian Vaegter4Jens Nyengaard5Simon Glerup6Danish Research Institute of Translational Neuroscience DANDRITE, Aarhus, 8000, DenmarkDanish Research Institute of Translational Neuroscience DANDRITE, Aarhus, 8000, DenmarkStereology and Electron Microscopy Laboratory, Department of Clinical institute, Aarhus University, Aarhus, 8000 C, DenmarkThe Lundbeck Foundation Research Center MIND, Department of Biomedicine, Aarhus University, Aarhus, 8000 C, DenmarkDanish Research Institute of Translational Neuroscience DANDRITE, Aarhus, 8000, DenmarkStereology and Electron Microscopy Laboratory, Department of Clinical institute, Aarhus University, Aarhus, 8000 C, DenmarkDanish Research Institute of Translational Neuroscience DANDRITE, Aarhus, 8000, DenmarkThe activity of excitatory neurons is controlled by a small, but highly diverse population of inhibitory interneurons. These cells show a high level of physiological, morphological and neurochemical heterogeneity, and play highly specific roles in neuronal circuits. In the mammalian hippocampus, these are divided into 21 different subtypes of GABAergic interneurons based on their expression of different markers, morphology and their electrophysiological properties. Ideally, all can be marked using an antibody directed against the inhibitory neurotransmitter GABA, but parvalbumin, calbindin, somatostatin, and calretinin are also commonly used as markers to narrow down the specific interneuron subtype. Here, we describe a journey to find the necessary immunological reagents for studying GABAergic interneurons of the mouse hippocampus. Based on web searches there are several hundreds of different antibodies on the market directed against these four markers. Searches in the literature databases allowed us to narrow it down to a subset of antibodies most commonly used in publications. However, in our hands the most cited ones did not work for immunofluorescence stainings of formaldehyde fixed tissue sections and cultured hippocampal neurons, and we had to immunostain our way through thirteen different commercial antibodies before finally finding a suitable antibody for each of the four markers. The antibodies were evaluated based on signal-to-noise ratios as well as if positive cells were found in layers of the hippocampus where they have previously been described. Additionally, the antibodies were also tested on sections from mouse spinal cord with similar criteria for specificity of the antibodies. Using the antibodies with a high rating on pAbmAbs, stainings with high signal-to-noise ratios and location of the immunostained cells in accordance with the literature could be obtained, making these antibodies suitable choices for studying the GABAergic system.http://f1000research.com/articles/3-242/v1Immunological Biomarkers |
| spellingShingle | Simon Molgaard Maj Ulrichsen Simon Boggild Marie-Louise Holm Christian Vaegter Jens Nyengaard Simon Glerup Immunohistochemical visualization of mouse interneuron subtypes [v1; ref status: indexed, http://f1000r.es/4em] F1000Research Immunological Biomarkers |
| title | Immunohistochemical visualization of mouse interneuron subtypes [v1; ref status: indexed, http://f1000r.es/4em] |
| title_full | Immunohistochemical visualization of mouse interneuron subtypes [v1; ref status: indexed, http://f1000r.es/4em] |
| title_fullStr | Immunohistochemical visualization of mouse interneuron subtypes [v1; ref status: indexed, http://f1000r.es/4em] |
| title_full_unstemmed | Immunohistochemical visualization of mouse interneuron subtypes [v1; ref status: indexed, http://f1000r.es/4em] |
| title_short | Immunohistochemical visualization of mouse interneuron subtypes [v1; ref status: indexed, http://f1000r.es/4em] |
| title_sort | immunohistochemical visualization of mouse interneuron subtypes v1 ref status indexed http f1000r es 4em |
| topic | Immunological Biomarkers |
| url | http://f1000research.com/articles/3-242/v1 |
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