X-Ray Fluorescence Microscopy Reveals Accumulation and Secretion of Discrete Intracellular Zinc Pools in the Lactating Mouse Mammary Gland
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{"title"=>"X-ray fluorescence microscopy reveals accumulation and secretion of discrete intracellular zinc pools in the lactating mouse mammary gland", "type"=>"journal", "authors"=>[{"first_name"=>"Nicholas", "last_name"=>"McCormick", "scopus_author_id"=>"36446357200"}, {"first_name"=>"Vanessa", "last_name"=>"Velasquez", "scopus_author_id"=>"36446916600"}, {"first_name"=>"Lydia", "last_name"=>"Finney", "scopus_author_id"=>"7003957879"}, {"first_name"=>"Stefan", "last_name"=>"Vogt", "scopus_author_id"=>"7101896529"}, {"first_name"=>"Shannon L.", "last_name"=>"Kelleher", "scopus_author_id"=>"27169261400"}], "year"=>2010, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-77956214419", "pui"=>"359293466", "doi"=>"10.1371/journal.pone.0011078", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "sgr"=>"77956214419", "pmid"=>"20552032"}, "id"=>"e12f6d0f-f559-33c0-ae00-d4ea5700093c", "abstract"=>"BACKGROUND: The mammary gland is responsible for the transfer of a tremendous amount of zinc ( approximately 1-3 mg zinc/day) from maternal circulation into milk during lactation to support the growth and development of the offspring. When this process is compromised, severe zinc deficiency compromises neuronal development and immune function and increases infant morbidity and/or mortality. It remains unclear as to how the lactating mammary gland dynamically integrates zinc import from maternal circulation with the enormous amount of zinc that is secreted into milk.\n\nMETHODOLOGY/PRINCIPAL FINDINGS: Herein we utilized X-ray fluorescence microscopy (XFM) which allowed for the visualization and quantification of the process of zinc transfer through the mammary gland of the lactating mouse. Our data illustrate that a large amount of zinc first accumulates in the mammary gland during lactation. Interestingly, this zinc is not cytosolic, but accumulated in large, discrete sub-cellular compartments. These zinc pools were then redistributed to small intracellular vesicles destined for secretion in a prolactin-responsive manner. Confocal microscopy identified mitochondria and the Golgi apparatus as the sub-cellular compartments which accumulate zinc; however, zinc pools in the Golgi apparatus, but not mitochondria are redistributed to vesicles destined for secretion during lactation.\n\nCONCLUSIONS/SIGNIFICANCE: Our data directly implicate the Golgi apparatus in providing a large, mobilizable zinc storage pool to assist in providing for the tremendous amount of zinc that is secreted into milk. Interestingly, our study also provides compelling evidence that mitochondrial zinc pools expand in the mammary gland during lactation which we speculate may play a role in regulating mammary gland function.", "link"=>"http://www.mendeley.com/research/xray-fluorescence-microscopy-reveals-accumulation-secretion-discrete-intracellular-zinc-pools-lactat", "reader_count"=>31, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>3, "Researcher"=>7, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>1, "Student > Master"=>6, "Other"=>1, "Student > Bachelor"=>1, "Professor"=>4}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>3, "Researcher"=>7, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>1, "Student > Master"=>6, "Other"=>1, "Student > Bachelor"=>1, "Professor"=>4}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>6, "Materials Science"=>1, "Agricultural and Biological Sciences"=>19, "Physics and Astronomy"=>1, "Chemistry"=>3}, "reader_count_by_subdiscipline"=>{"Materials Science"=>{"Materials Science"=>1}, "Chemistry"=>{"Chemistry"=>3}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>19}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>6}, "Unspecified"=>{"Unspecified"=>1}}, "reader_count_by_country"=>{"Czech Republic"=>1, "United States"=>1, "Brazil"=>1, "Mexico"=>2, "Spain"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/845847"], "description"=>"<p>Representative confocal micrographs illustrate that FluoZin-3 (green) and Bodipy TR (red) are largely co-localized (merge, yellow) in non-secreting mammary epithelial cells (panel A). Mammary epithelial cells were treated with prolactin and cortisol for 24 h differentiate to a secreting phenotype (panel B). Confocal micrographs of FluoZin-3 (green) and Bodipy TR (red) in secreting mammary epithelial cells illustrate partial co-localization (merge, yellow) of “labile” Zn and the Golgi apparatus in hormonally treated cells. Moreover, distinct non-Golgi apparatus-associated Zn pools were detected in secreting cells.</p>", "links"=>[], "tags"=>["zn", "pools", "golgi"], "article_id"=>516300, "categories"=>["Cell Biology", "Medicine"], "users"=>["Nicholas McCormick", "Vanessa Velasquez", "Lydia Finney", "Stefan Vogt", "Shannon L. Kelleher"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011078.g006", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Labile_Zn_pools_are_partially_associated_with_the_Golgi_apparatus_/516300", "title"=>"Labile Zn pools are partially associated with the Golgi apparatus.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 01:03:52"}
  • {"files"=>["https://ndownloader.figshare.com/files/845193"], "description"=>"<p>Mouse mammary gland (20 µm) was visualized by phase contrast microscopy (20× magnification) and analyzed using XFM. The alveoli lumen (L) is surrounded by a single layer of mammary epithelial cells (MEC). The corresponding element (phosphorus, P; zinc, Zn) and its minimum and maximum threshold values in micrograms per square centimeter are given above each image. The rainbow-colored scale bar reflects the signal intensity measured as micrograms per square centimeter in each pixel, with darker pixels representing areas of low concentration and brighter pixels representing areas of increasing concentration. A scale bar (20 µm) is shown below the elemental maps.</p>", "links"=>[], "tags"=>["mammary", "gland", "non-lactating", "lactating"], "article_id"=>515624, "categories"=>["Cell Biology", "Medicine"], "users"=>["Nicholas McCormick", "Vanessa Velasquez", "Lydia Finney", "Stefan Vogt", "Shannon L. Kelleher"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011078.g001", "stats"=>{"downloads"=>0, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_XFM_analysis_of_the_mammary_gland_from_non_lactating_and_lactating_mice_/515624", "title"=>"XFM analysis of the mammary gland from non-lactating and lactating mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 00:59:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/845353"], "description"=>"<p>Mouse mammary gland (20 µm) was analyzed using XFM. The corresponding element (phosphorus, P; zinc, Zn) and its minimum and maximum threshold values in micrograms per square centimeter are given above each image. The rainbow-colored scale bar reflects the signal intensity measured as micrograms per square centimeter in each pixel, with darker pixels representing areas of low concentration and brighter pixels representing areas of increasing concentration. A scale bar (10 µm) is shown below the elemental maps. Zinc and phosphorus maps of ∼1–4 mammary cells in the mammary gland from a mouse that had suckled her offspring for 30 min (suckled; used as a baseline), a suckled mouse, 60 min post-suckling (Post-suckled; reflecting Zn accumulation), a mouse injected with bromocriptine (Bromocriptine; used to eliminate prolactin secretion) and a mouse injected with haloperidol (Haloperidol; used to increase prolactin secretion). The red circles on the Zn maps denote corresponding nuclei.</p>", "links"=>[], "tags"=>["intracellular", "zn", "pools", "mammary", "gland"], "article_id"=>515796, "categories"=>["Cell Biology", "Medicine"], "users"=>["Nicholas McCormick", "Vanessa Velasquez", "Lydia Finney", "Stefan Vogt", "Shannon L. Kelleher"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011078.g002", "stats"=>{"downloads"=>1, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_XFM_analysis_of_intracellular_Zn_pools_in_mouse_mammary_gland_during_lactation_/515796", "title"=>"XFM analysis of intracellular Zn pools in mouse mammary gland during lactation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 01:01:03"}
  • {"files"=>["https://ndownloader.figshare.com/files/845482"], "description"=>"<p>(A) Representative confocal images to illustrate vesicular and mitochondrial Zn pools detected using FluoZin-3 (green) and RhodZin-3 (red), respectively, in mammary cells (HC11). Scale bar (5 µm). (B) Mammary cells were treated with prolactin (1 µM) for 24 h and the accumulation of Zn in vesicular and mitochondrial Zn pools were detected by fluorometry using FluoZin-3 (green) and RhodZin-3 (red), respectively. Data represent mean fluorescence ± standard deviation, n = 6 samples/treatment. Asterisk indicates a significant effect of treatment (p<0.001).</p>", "links"=>[], "tags"=>["lactogenic", "prolactin", "increases", "vesicular", "mitochondrial", "zn", "pools", "mammary"], "article_id"=>515918, "categories"=>["Cell Biology", "Medicine"], "users"=>["Nicholas McCormick", "Vanessa Velasquez", "Lydia Finney", "Stefan Vogt", "Shannon L. Kelleher"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011078.g003", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_lactogenic_hormone_prolactin_increases_vesicular_and_mitochondrial_Zn_pools_in_mammary_cells_/515918", "title"=>"The lactogenic hormone prolactin increases vesicular and mitochondrial Zn pools in mammary cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 01:01:44"}
  • {"files"=>["https://ndownloader.figshare.com/files/845996"], "description"=>"<p>Representative confocal micrographs of p58 (Golgi apparatus marker) visualized with Alexa Fluor® 568-conjugated anti-mouse IgG in untreated (p58 −BFA) and Brefeldin A-treated (p58 +BFA) mammary epithelial cells. Images verify that Brefeldin A treatment disrupts the Golgi apparatus in mammary epithelial cells. Representative confocal micrographs of FluoZin-3 in untreated cells (FluoZin-3 −BFA) and cells treated with Brefeldin A (FluoZin-3 +BFA). Images illustrate that the distribution of “labile” Zn pools is diffuse by Brefeldin A treatment (inset) indicating that “labile” Zn is associated with the Golgi apparatus.</p>", "links"=>[], "tags"=>["zn", "pools", "disrupted", "brefledin"], "article_id"=>516441, "categories"=>["Cell Biology", "Medicine"], "users"=>["Nicholas McCormick", "Vanessa Velasquez", "Lydia Finney", "Stefan Vogt", "Shannon L. Kelleher"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011078.g007", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Labile_Zn_pools_are_disrupted_by_Brefledin_A_treatment_/516441", "title"=>"Labile Zn pools are disrupted by Brefledin A treatment.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 01:04:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/845721"], "description"=>"<p>Representative confocal micrographs of mammary epithelial cells treated with FluoZin-3 (green) and ER Tracker (red) which illustrate that “labile” Zn pools do not exist within the endoplasmic reticulum and are, in fact, associated with a distinct compartment proximal to the endoplasmic reticulum (merge).</p>", "links"=>[], "tags"=>["zn", "pools", "compartment", "proximal", "endoplasmic"], "article_id"=>516165, "categories"=>["Cell Biology", "Medicine"], "users"=>["Nicholas McCormick", "Vanessa Velasquez", "Lydia Finney", "Stefan Vogt", "Shannon L. Kelleher"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011078.g005", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Labile_Zn_pools_are_associated_with_a_distinct_compartment_directly_proximal_to_the_endoplasmic_reticulum_/516165", "title"=>"Labile Zn pools are associated with a distinct compartment directly proximal to the endoplasmic reticulum.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 01:03:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/845572"], "description"=>"<p>Representative confocal micrographs of lysosomes detected with Lysotracker Green in untreated cells (Lysotracker) and cells treated with chloroquine diphosphate (Lysotracker +CLQ).. Images verify that chloroquine diphosphate disrupts lysosomes in HC11 cells. Representative confocal micrographs of “labile” Zn pools detected using FluoZin-3 in untreated cells (FluoZin-3) and cells treated with chloroquine diphosphate (FluoZin-3 +CLQ). Images illustrate that the distribution of “labile” Zn pools in mammary cells is not affected by lysosomal disruption.</p>", "links"=>[], "tags"=>["zn", "pools"], "article_id"=>516019, "categories"=>["Cell Biology", "Medicine"], "users"=>["Nicholas McCormick", "Vanessa Velasquez", "Lydia Finney", "Stefan Vogt", "Shannon L. Kelleher"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011078.g004", "stats"=>{"downloads"=>1, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Labile_Zn_pools_are_not_associated_with_lysosomes_/516019", "title"=>"Labile Zn pools are not associated with lysosomes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 01:02:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/846135"], "description"=>"<p>Mammary cells were treated with prolactin for 24 h to differentiate to a secreting phenotype. Following differentiation, cells were stimulated with prolactin and changes in “labile” Zn pools were quantified by fluorometry using FluoZin-3. Data represent mean fluorescence ± standard deviation, n = 6–8 samples/treatment. Asterisk denotes a significant effect of prolactin on FluoZin-3 fluorescence, p<0.001.</p>", "links"=>[], "tags"=>["stimulates", "mobilization", "vesicular", "zn", "pools", "secreting", "mammary"], "article_id"=>516580, "categories"=>["Cell Biology", "Medicine"], "users"=>["Nicholas McCormick", "Vanessa Velasquez", "Lydia Finney", "Stefan Vogt", "Shannon L. Kelleher"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0011078.g008", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Prolactin_stimulates_the_mobilization_of_vesicular_Zn_pools_in_secreting_mammary_cells_/516580", "title"=>"Prolactin stimulates the mobilization of vesicular Zn pools in secreting mammary cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 01:05:28"}

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