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Tubulin increases with lactation in rat and guinea pig mammary alveolar cells

Fluorescence and EM immunogold localization

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Summary

Changes in mammary gland tubulin were studied immunocytochemically during transition from late pregnancy to lactation. Indirect immunofluorescence was used to localize tubulin in mammary glands from late pregnant, early lactating and peak lactating guinea pigs. Whole rabbit antiserum against guinea pig brain tubulin and affinity-purified antibody indicated increases in alveolar cell tubulin content from late pregnancy through peak lactation coincident with the development of lactation. Only alveolar cells displayed high, specific fluorescence or underwent a developmental increase. Tubulin was concentrated apically, in association with secretory structures. In a second study comparing mammary tissues from 18 days pregnant and 10 days lactating rats, EM immunogold was used with three commercial antitubulins ranging from a rabbit polyclonal antiserum against chick brain MTs to a monoclonal mouse anti-alpha tubulin. Gold particle counts indicated 2- to 5- fold tubulin increases in alveolar cells with lactation and development of an apicobasal (high apical) tubulin gradient. Variations among the three anti-tubulins is discussed. The results confirmed previous observations of whole gland tubulin increases based on colchicine binding assays and localized the site of the increase primarily in alveolar cells.

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Abbreviations

EM:

electron microscope-(ic)

GAM:

goat anti-mouse

GAR:

goat anti-rabbit

Ig:

immunoglobulin

MC:

monoclonal

MT:

microtubule

PAGE:

polyacrylamide gel electrophoresis

PIPES:

1,4-piperazine diethane sulfonic acid

PBS:

phosphate buffered saline

PC:

polyclonal

Rb:

rabbit

SDS:

sodium dodecyl sulfate

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Authors and Affiliations

  1. Research Resources Center, m/c 937, University of Illinois at Chicago, Box 6998, 60680, Chicago, IL, USA

    R. F. Loizzi

  2. Department of Physiology and Biophysics, University of Illinois at Chicago, Chicago, Illinois

    R. F. Loizzi & DeRen Shao

Authors
  1. R. F. Loizzi
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  2. DeRen Shao
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Dedicated to Professor Stuart Patton on the occasion of his 70th birthday.

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Loizzi, R.F., Shao, D. Tubulin increases with lactation in rat and guinea pig mammary alveolar cells. Protoplasma 159, 129–143 (1990). https://doi.org/10.1007/BF01322596

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  • DOI: https://doi.org/10.1007/BF01322596

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