How to Wire the Diaphragm: Wholemount Staining Methods to Analyze Mammalian Respiratory Innervation
Direct or indirect impairment of breathing in humans by diseases or environmental factors can either cause long-term disability and pain, or can ultimately result in death. Automatic respiratory centers in the brainstem control the highly structured process of breathing and signal to a specialized g...
Saved in:
Published in | Methods in molecular biology (Clifton, N.J.) Vol. 1668; p. 177 |
---|---|
Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
United States
2017
|
Subjects | |
Online Access | Get more information |
Cover
Loading…
Abstract | Direct or indirect impairment of breathing in humans by diseases or environmental factors can either cause long-term disability and pain, or can ultimately result in death. Automatic respiratory centers in the brainstem control the highly structured process of breathing and signal to a specialized group of motor neurons in the cervical spinal cord that constitute the phrenic nerves. In mammals, the thoracic diaphragm separates the thorax from the abdomen and adopts the function of the primary respiratory musculature. Faithful innervation by the phrenic nerves is a prerequisite for correct functionality of this highly specialized musculature and thus, ultimately, the viability of the entire organism.To analyze the effects of diseases and genetic defects responsible for deleterious or lethal respiratory phenotypes, accurate imaging of respiratory innervation during embryonic development, e.g., in genetically modified mouse models enables the characterization of specific marker genes and pathways that underlie appropriate wiring of the diaphragm. Among the different available immunostaining techniques, wholemount staining methods provide the advantage of clear and faithful three-dimensional information about the location of the antigens of interest. In comparison to routine histological techniques, however, the researcher has to deal with technical challenges, such as antibody penetration, the stability and availability of the antigen, and clearing of the relevant tissue, and the need to be equipped with state-of-the-art microscope equipment.In this methodological chapter, we explain and share our expertise concerning wholemount processing of mouse embryos and thoracic diaphragms for the analysis of mammalian respiratory innervation. |
---|---|
AbstractList | Direct or indirect impairment of breathing in humans by diseases or environmental factors can either cause long-term disability and pain, or can ultimately result in death. Automatic respiratory centers in the brainstem control the highly structured process of breathing and signal to a specialized group of motor neurons in the cervical spinal cord that constitute the phrenic nerves. In mammals, the thoracic diaphragm separates the thorax from the abdomen and adopts the function of the primary respiratory musculature. Faithful innervation by the phrenic nerves is a prerequisite for correct functionality of this highly specialized musculature and thus, ultimately, the viability of the entire organism.To analyze the effects of diseases and genetic defects responsible for deleterious or lethal respiratory phenotypes, accurate imaging of respiratory innervation during embryonic development, e.g., in genetically modified mouse models enables the characterization of specific marker genes and pathways that underlie appropriate wiring of the diaphragm. Among the different available immunostaining techniques, wholemount staining methods provide the advantage of clear and faithful three-dimensional information about the location of the antigens of interest. In comparison to routine histological techniques, however, the researcher has to deal with technical challenges, such as antibody penetration, the stability and availability of the antigen, and clearing of the relevant tissue, and the need to be equipped with state-of-the-art microscope equipment.In this methodological chapter, we explain and share our expertise concerning wholemount processing of mouse embryos and thoracic diaphragms for the analysis of mammalian respiratory innervation. |
Author | Alberton, Paolo Huettl, Rosa-Eva Huber, Andrea B Saller, Maximilian Michael |
Author_xml | – sequence: 1 givenname: Maximilian Michael surname: Saller fullname: Saller, Maximilian Michael email: maximilian.saller@med.uni-muenchen.de organization: Experimental Surgery and Regenerative Medicine (ExperiMed), Department of General, Trauma and Reconstructive Surgery, Ludwig-Maximilians-University (LMU), Nußbaumstraße 20, 80336, Munich, Germany. maximilian.saller@med.uni-muenchen.de – sequence: 2 givenname: Paolo surname: Alberton fullname: Alberton, Paolo organization: Experimental Surgery and Regenerative Medicine (ExperiMed), Department of General, Trauma and Reconstructive Surgery, Ludwig-Maximilians-University (LMU), Nußbaumstraße 20, 80336, Munich, Germany – sequence: 3 givenname: Andrea B surname: Huber fullname: Huber, Andrea B organization: Department of Biosystems Science and Engineering (D-BSSE), Swiss Federal Institute of Technology, Zurich (ETHZ), Mattenstrasse 26, 4058, Basel, Switzerland – sequence: 4 givenname: Rosa-Eva surname: Huettl fullname: Huettl, Rosa-Eva organization: Department of Stress Neurobiology and Neurogenetics, Max-Planck-Institute of Psychiatry, Kraepelinstr. 2, 80804, Munich, Germany |
BackLink | https://www.ncbi.nlm.nih.gov/pubmed/28842910$$D View this record in MEDLINE/PubMed |
BookMark | eNo1j8lOwzAURS0EogP8AUL-AYMnGptdVYZWaoXEoC6rl-S1MUrsyHFB4espAjb3rs7VuSNy7INHQi4EvxKcZ9c2M0wwbZVlmTSKmY1QR2QorOZswqUdkFHXvXOuMyX1KRlIY7S0gg9JMQ-fNAW6dhFpqpDeOWirCLvmlq6rUGMT9j7RlwTOO7-jK0xVKLsfZOqh7r-QrqBpoHbg6TN2rYuQQuzpwnuMH5Bc8GfkZAt1h-d_PSZvD_evszlbPj0uZtMl2x1cElNbO9GYm0LlUmhttJBC2UKVyA9pCsgzUCYTN6ByM7GYIZYABnOJW6HLQo7J5e9uu88bLDdtdA3EfvP_Vn4DCZ9ZUA |
ContentType | Journal Article |
DBID | CGR CUY CVF ECM EIF NPM |
DOI | 10.1007/978-1-4939-7283-8_13 |
DatabaseName | Medline MEDLINE MEDLINE (Ovid) MEDLINE MEDLINE PubMed |
DatabaseTitle | MEDLINE Medline Complete MEDLINE with Full Text PubMed MEDLINE (Ovid) |
DatabaseTitleList | MEDLINE |
Database_xml | – sequence: 1 dbid: NPM name: PubMed url: https://proxy.k.utb.cz/login?url=http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed sourceTypes: Index Database – sequence: 2 dbid: EIF name: MEDLINE url: https://proxy.k.utb.cz/login?url=https://www.webofscience.com/wos/medline/basic-search sourceTypes: Index Database |
DeliveryMethod | no_fulltext_linktorsrc |
Discipline | Biology |
EISSN | 1940-6029 |
ExternalDocumentID | 28842910 |
Genre | Journal Article |
GroupedDBID | --- 29M 53G ACGFS ALMA_UNASSIGNED_HOLDINGS CGR CUY CVF ECM EIF F5P NPM P2P RSU SPO UDS WH7 ZGI |
ID | FETCH-LOGICAL-g291t-3f964eb8c3b21448412139c3de09c38cab7a38715a3b869e7eedaa8eb2ef14dc2 |
IngestDate | Fri May 24 00:01:50 EDT 2024 |
IsPeerReviewed | false |
IsScholarly | true |
Keywords | Muscle development Diaphragm Phrenic nerve Axon guidance Fasciculation |
Language | English |
LinkModel | OpenURL |
MergedId | FETCHMERGED-LOGICAL-g291t-3f964eb8c3b21448412139c3de09c38cab7a38715a3b869e7eedaa8eb2ef14dc2 |
PMID | 28842910 |
ParticipantIDs | pubmed_primary_28842910 |
PublicationCentury | 2000 |
PublicationDate | 2017-00-00 |
PublicationDateYYYYMMDD | 2017-01-01 |
PublicationDate_xml | – year: 2017 text: 2017-00-00 |
PublicationDecade | 2010 |
PublicationPlace | United States |
PublicationPlace_xml | – name: United States |
PublicationTitle | Methods in molecular biology (Clifton, N.J.) |
PublicationTitleAlternate | Methods Mol Biol |
PublicationYear | 2017 |
SSID | ssj0047324 |
Score | 2.2170577 |
Snippet | Direct or indirect impairment of breathing in humans by diseases or environmental factors can either cause long-term disability and pain, or can ultimately... |
SourceID | pubmed |
SourceType | Index Database |
StartPage | 177 |
SubjectTerms | Animals Axon Fasciculation Axon Guidance Cell Adhesion Molecules - metabolism Diaphragm - chemistry Diaphragm - innervation Embryo, Mammalian Fluorescent Dyes - chemistry Mice Motor Neurons - metabolism Muscle Development Optical Imaging Phrenic Nerve - growth & development Staining and Labeling - methods Thorax - chemistry Thorax - innervation |
Title | How to Wire the Diaphragm: Wholemount Staining Methods to Analyze Mammalian Respiratory Innervation |
URI | https://www.ncbi.nlm.nih.gov/pubmed/28842910 |
Volume | 1668 |
hasFullText | |
inHoldings | 1 |
isFullTextHit | |
isPrint | |
link | http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwnV3da9swEBdpy2Avpfvu2g097C2oOLFjy3srbUdWSClby_oWTopUAnNSqFu2_lH7G3enj1h03dj2IoyFZOH7Wbo7392PsXejGR7bVlphbJmJAkwpwKLVCpmVlHk5KB3X4eSkHJ8Xxxeji17vRxK1dNOqPX33YF7J_0gV76FcKUv2HyS7mhRv4DXKF1uUMLZ_JWPHB7d0EaxOgTycA8oGLp3z_Qsx3zZEBUEapeOB6E8cX7Qr6uCqkdwR81DTeF_Hp-Sv-0di5LrtpBY5n8L4-aLfRF7dfqzj5P8H2zamcB3vJW6GzxBzDifwbd7M3RPTmH0CHdXbCsNPASftMKcC6TaFX0LHFD2-MW371UeIX4NAsyD1YgyqxIth_M5bUx5CFtYVt-bSc-6E3XXgGV9-2fW7QA-0h-u8FhUqTUJOfZZrAoSrxiFhKCUewz6c9s-992pxx641tlZJIgo5Id-QP_eLCnXTJDnzoeVQ6ekwxT0zxqkzZ1tsM9ghfN-D6gnrmcVT9sgzk35_xjRCi7dLTtDiCC2-gtZ73gGLR2DxAAwaEoDFV8DiCbB4Aqzn7PzD0dnBWAQ-DnGJK25FbuuyMErqXFGhPVlQOcBa5zOTYSs1qApyNMBHkCtZ1qZC_QtAGjU0dlDM9PAFW18sF-YV4zY3mZYZ1BnYgmoO1tZUWpfSDE2hpN1mL_3bmV75oivT-N5e_7Znhz3ukLXLNix-5eYNqoyteusk9RP7pGtR |
link.rule.ids | 780 |
linkProvider | National Library of Medicine |
openUrl | ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=How+to+Wire+the+Diaphragm%3A+Wholemount+Staining+Methods+to+Analyze+Mammalian+Respiratory+Innervation&rft.jtitle=Methods+in+molecular+biology+%28Clifton%2C+N.J.%29&rft.au=Saller%2C+Maximilian+Michael&rft.au=Alberton%2C+Paolo&rft.au=Huber%2C+Andrea+B&rft.au=Huettl%2C+Rosa-Eva&rft.date=2017-01-01&rft.eissn=1940-6029&rft.volume=1668&rft.spage=177&rft_id=info:doi/10.1007%2F978-1-4939-7283-8_13&rft_id=info%3Apmid%2F28842910&rft_id=info%3Apmid%2F28842910&rft.externalDocID=28842910 |