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      Long extensions with varicosity-like structures in gonadotrope Lh cells facilitate clustering in medaka pituitary culture

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          Abstract

          Accumulating evidence indicates that some pituitary cell types are organized in complex networks in both mammals and fish. In this study, we have further investigated the previously described cellular extensions formed by the medaka ( Oryzias latipes) luteinizing hormone gonadotropes (Lh cells). Extensions, several cell diameters long, with varicosity-like swellings, were common both in vitro and in vivo. Some extensions approached other Lh cells, while others were in close contact with blood vessels in vivo. Gnrh further stimulated extension development in vitro. Two types of extensions with different characteristics could be distinguished, and were classified as major or minor according to size, origin and cytoskeleton protein dependance. The varicosity-like swellings appeared on the major extensions and were dependent on both microtubules and actin filaments. Immunofluorescence revealed that Lhβ protein was mainly located in these swellings and at the extremity of the extensions. We then investigated whether these extensions contribute to network formation and clustering, by following their development in primary cultures. During the first two days in culture, the Lh cells grew long extensions that with time physically attached to other cells. Successively, tight cell clusters formed as cell somas that were connected via extensions migrated towards each other, while shortening their extensions. Laser photolysis of caged Ca 2+ showed that Ca 2+ signals originating in the soma propagated from the soma along the major extensions, being particularly visible in each swelling. Moreover, the Ca 2+ signal could be transferred between densely clustered cells (sharing soma-soma border), but was not transferred via extensions to the connected cell. In summary, Lh gonadotropes in medaka display a complex cellular structure of hormone-containing extensions that are sensitive to Gnrh, and may be used for clustering and possibly hormone release, but do not seem to contribute to communication between cells themselves.

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          Fiji is a distribution of the popular open-source software ImageJ focused on biological-image analysis. Fiji uses modern software engineering practices to combine powerful software libraries with a broad range of scripting languages to enable rapid prototyping of image-processing algorithms. Fiji facilitates the transformation of new algorithms into ImageJ plugins that can be shared with end users through an integrated update system. We propose Fiji as a platform for productive collaboration between computer science and biology research communities.
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            BoxPlotR: a web tool for generation of box plots.

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              The lamellipodium: where motility begins

              Lamellipodia, filopodia and membrane ruffles are essential for cell motility, the organization of membrane domains, phagocytosis and the development of substrate adhesions. Their formation relies on the regulated recruitment of molecular scaffolds to their tips (to harness and localize actin polymerization), coupled to the coordinated organization of actin filaments into lamella networks and bundled arrays. Their turnover requires further molecular complexes for the disassembly and recycling of lamellipodium components. Here, we give a spatial inventory of the many molecular players in this dynamic domain of the actin cytoskeleton in order to highlight the open questions and the challenges ahead.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: InvestigationRole: Project administrationRole: Writing – original draftRole: Writing – review & editing
                Role: InvestigationRole: Writing – original draftRole: Writing – review & editing
                Role: InvestigationRole: Writing – original draftRole: Writing – review & editing
                Role: InvestigationRole: Writing – original draft
                Role: Investigation
                Role: Funding acquisitionRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: Funding acquisitionRole: InvestigationRole: Writing – original draftRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS One
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                28 January 2021
                2021
                : 16
                : 1
                Affiliations
                [1 ] Faculty of Health and Welfare, Østfold University College, Halden, Norway
                [2 ] Physiology Unit, Faculty of Veterinary Medicine, Norwegian University of Life Sciences, Oslo, Norway
                [3 ] Department of Biosciences, Faculty of Natural Sciences, University of Oslo, Oslo, Norway
                [4 ] Department of Oral Biology, Faculty of Dentistry, University of Oslo, Oslo, Norway
                Universite de Rouen, FRANCE
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Article
                PONE-D-20-05785
                10.1371/journal.pone.0245462
                7842944
                33507913
                © 2021 Grønlien et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                Page count
                Figures: 9, Tables: 0, Pages: 23
                Product
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100005416, Norges Forskningsråd;
                Award ID: 184851
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100005416, Norges Forskningsråd;
                Award ID: 191825
                Award Recipient :
                Supported by grants from the Norwegian Research Council ( https://www.forskningsradet.no/en/): 184851 (FAW) and 191825 (TMH) The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Anatomy
                Endocrine System
                Pituitary Gland
                Medicine and Health Sciences
                Anatomy
                Endocrine System
                Pituitary Gland
                Biology and Life Sciences
                Anatomy
                Nervous System
                Neuroanatomy
                Pituitary Gland
                Medicine and Health Sciences
                Anatomy
                Nervous System
                Neuroanatomy
                Pituitary Gland
                Biology and Life Sciences
                Neuroscience
                Neuroanatomy
                Pituitary Gland
                Biology and Life Sciences
                Cell Biology
                Cellular Structures and Organelles
                Cytoskeleton
                Research and Analysis Methods
                Biological Cultures
                Cell Cultures
                Research and Analysis Methods
                Imaging Techniques
                Fluorescence Imaging
                Biology and Life Sciences
                Organisms
                Eukaryota
                Animals
                Vertebrates
                Amniotes
                Mammals
                Biology and Life Sciences
                Zoology
                Animals
                Vertebrates
                Amniotes
                Mammals
                Biology and Life Sciences
                Anatomy
                Cardiovascular Anatomy
                Blood Vessels
                Medicine and Health Sciences
                Anatomy
                Cardiovascular Anatomy
                Blood Vessels
                Biology and Life Sciences
                Biochemistry
                Hormones
                Research and Analysis Methods
                Biological Cultures
                Cell Culturing Techniques
                Primary Cell Culture
                Custom metadata
                The data underlying this study is accessible at http://dx.doi.org/10.17632/2vmwpzr3wb.1.

                Uncategorized

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