@article{pop00207,
abstract = {The subcellular localization of mRNA sequences encoding neuropeptides in neuropeptidergic cells of the pond snail Lymnaea stagnalis was investigated at the electron microscopic (EM) level by non-radioactive in situ hybridization. Various classes of probes specific for 28S rRNA and for the ovulation hormone (caudodorsal cell hormone; CDCH) mRNA were labeled with biotin or digoxigenin and were detected after hybridization with gold-labeled antibodies. Hybridizations were performed on ultra-thin sections of both Lowicryl-embedded and frozen cerebral ganglia, and a comparison demonstrated that most intense hybridization signals with an acceptable preservation of morphology were obtained with ultra-thin cryosections. Addition of 0.1{\%} glutaraldehyde to the formaldehyde fixative improved the morphology, but on Lowicryl sections this added fixative resulted in a decrease of label intensity. A variety of probes, including plasmids, PCR products, and oligonucleotides, were used and all provided good results, although the use of oligonucleotides on Lowicryl sections resulted in decreased gold labeling. The gold particles were found mainly associated with rough endoplasmic reticulum (RER) but were also observed in lysosomal structures. Finally, the in situ hybridization method presented in this study proved to be compatible with the immunocytochemical detection of the caudodorsal cell hormone, as demonstrated by double labeling experiments.},
annote = {Query date: 2020-06-29 13:05:30},
author = {Dirks, Roeland W. and {Van Dorp}, A. G.M. and {Van Minnen}, J. and Fransen, J. A.M. and {Van der Ploeg}, M. and Raap, A. K.},
doi = {10.1177/40.11.1431053},
issn = {0022-1554},
journal = {Journal of Histochemistry {\&} Cytochemistry},
keywords = {Biotin,Cryosections,Digoxigenin,Electron microscopy,In situ hybridization,Lowicryl,Lymnaea stagnalis,mRNA},
month = {nov},
number = {11},
pages = {1647--1657},
publisher = {journals.sagepub.com},
title = {{Electron microscopic detection of RNA sequences by non-radioactive in situ hybridization in the mollusk Lymnaea stagnalis.}},
url = {https://journals.sagepub.com/doi/abs/10.1177/40.11.1431053 http://journals.sagepub.com/doi/10.1177/40.11.1431053},
volume = {40},
year = {1992}
}

{"_id":"gqSxJn9jzjx5XZkH9","bibbaseid":"dirks-vandorp-vanminnen-fransen-vanderploeg-raap-electronmicroscopicdetectionofrnasequencesbynonradioactiveinsituhybridizationinthemollusklymnaeastagnalis-1992","author_short":["Dirks, R. W.","Van Dorp, A. G.","Van Minnen, J.","Fransen, J. A.","Van der Ploeg, M.","Raap, A. K."],"bibdata":{"bibtype":"article","type":"article","abstract":"The subcellular localization of mRNA sequences encoding neuropeptides in neuropeptidergic cells of the pond snail Lymnaea stagnalis was investigated at the electron microscopic (EM) level by non-radioactive in situ hybridization. Various classes of probes specific for 28S rRNA and for the ovulation hormone (caudodorsal cell hormone; CDCH) mRNA were labeled with biotin or digoxigenin and were detected after hybridization with gold-labeled antibodies. Hybridizations were performed on ultra-thin sections of both Lowicryl-embedded and frozen cerebral ganglia, and a comparison demonstrated that most intense hybridization signals with an acceptable preservation of morphology were obtained with ultra-thin cryosections. Addition of 0.1% glutaraldehyde to the formaldehyde fixative improved the morphology, but on Lowicryl sections this added fixative resulted in a decrease of label intensity. A variety of probes, including plasmids, PCR products, and oligonucleotides, were used and all provided good results, although the use of oligonucleotides on Lowicryl sections resulted in decreased gold labeling. The gold particles were found mainly associated with rough endoplasmic reticulum (RER) but were also observed in lysosomal structures. Finally, the in situ hybridization method presented in this study proved to be compatible with the immunocytochemical detection of the caudodorsal cell hormone, as demonstrated by double labeling experiments.","annote":"Query date: 2020-06-29 13:05:30","author":[{"propositions":[],"lastnames":["Dirks"],"firstnames":["Roeland","W."],"suffixes":[]},{"propositions":[],"lastnames":["Van Dorp"],"firstnames":["A.","G.M."],"suffixes":[]},{"propositions":[],"lastnames":["Van Minnen"],"firstnames":["J."],"suffixes":[]},{"propositions":[],"lastnames":["Fransen"],"firstnames":["J.","A.M."],"suffixes":[]},{"propositions":[],"lastnames":["Van der Ploeg"],"firstnames":["M."],"suffixes":[]},{"propositions":[],"lastnames":["Raap"],"firstnames":["A.","K."],"suffixes":[]}],"doi":"10.1177/40.11.1431053","issn":"0022-1554","journal":"Journal of Histochemistry & Cytochemistry","keywords":"Biotin,Cryosections,Digoxigenin,Electron microscopy,In situ hybridization,Lowicryl,Lymnaea stagnalis,mRNA","month":"nov","number":"11","pages":"1647–1657","publisher":"journals.sagepub.com","title":"Electron microscopic detection of RNA sequences by non-radioactive in situ hybridization in the mollusk Lymnaea stagnalis.","url":"https://journals.sagepub.com/doi/abs/10.1177/40.11.1431053 http://journals.sagepub.com/doi/10.1177/40.11.1431053","volume":"40","year":"1992","bibtex":"@article{pop00207,\nabstract = {The subcellular localization of mRNA sequences encoding neuropeptides in neuropeptidergic cells of the pond snail Lymnaea stagnalis was investigated at the electron microscopic (EM) level by non-radioactive in situ hybridization. Various classes of probes specific for 28S rRNA and for the ovulation hormone (caudodorsal cell hormone; CDCH) mRNA were labeled with biotin or digoxigenin and were detected after hybridization with gold-labeled antibodies. Hybridizations were performed on ultra-thin sections of both Lowicryl-embedded and frozen cerebral ganglia, and a comparison demonstrated that most intense hybridization signals with an acceptable preservation of morphology were obtained with ultra-thin cryosections. Addition of 0.1{\\%} glutaraldehyde to the formaldehyde fixative improved the morphology, but on Lowicryl sections this added fixative resulted in a decrease of label intensity. A variety of probes, including plasmids, PCR products, and oligonucleotides, were used and all provided good results, although the use of oligonucleotides on Lowicryl sections resulted in decreased gold labeling. The gold particles were found mainly associated with rough endoplasmic reticulum (RER) but were also observed in lysosomal structures. Finally, the in situ hybridization method presented in this study proved to be compatible with the immunocytochemical detection of the caudodorsal cell hormone, as demonstrated by double labeling experiments.},\nannote = {Query date: 2020-06-29 13:05:30},\nauthor = {Dirks, Roeland W. and {Van Dorp}, A. G.M. and {Van Minnen}, J. and Fransen, J. A.M. and {Van der Ploeg}, M. and Raap, A. 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K."],"key":"pop00207","id":"pop00207","bibbaseid":"dirks-vandorp-vanminnen-fransen-vanderploeg-raap-electronmicroscopicdetectionofrnasequencesbynonradioactiveinsituhybridizationinthemollusklymnaeastagnalis-1992","role":"author","urls":{"Paper":"https://journals.sagepub.com/doi/abs/10.1177/40.11.1431053 http://journals.sagepub.com/doi/10.1177/40.11.1431053"},"keyword":["Biotin","Cryosections","Digoxigenin","Electron microscopy","In situ hybridization","Lowicryl","Lymnaea stagnalis","mRNA"],"metadata":{"authorlinks":{}},"html":""},"bibtype":"article","biburl":"http://lymnaea.org/data/lymnaeastagnalis-neuroscience.bib","dataSources":["aGHieW6wJ8vBRjMFD"],"keywords":["biotin","cryosections","digoxigenin","electron microscopy","in situ hybridization","lowicryl","lymnaea stagnalis","mrna"],"search_terms":["electron","microscopic","detection","rna","sequences","non","radioactive","situ","hybridization","mollusk","lymnaea","stagnalis","dirks","van dorp","van minnen","fransen","van der ploeg","raap"],"title":"Electron microscopic detection of RNA sequences by non-radioactive in situ hybridization in the mollusk Lymnaea stagnalis.","year":1992}