Characteristic distribution of gap junctions in rat lacrimal gland in vivo and reconstruction of a gap junction in an in vitro model

We investigated localization of the gap junction in rat lacrimal gland in vivo and in vitro using electron microscopy and immunostaining with anti‐connexin32 (Cx32) monoclonal antibody (HAM8). In immunofluorescence study of lacrimal gland tissues, Cx32 protein appeared to exist not only at the inter...

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Bibliographic Details
Published in:Journal of electron microscopy 2002-01, Vol.51 (1), p.35-44
Main Authors: Takayama, Tetsuya, Tatsukawa, Shuji, Kitamura, Hirokazu, Ina, Keisuke, Nakatsuka, Kazuo, Fujikura, Yoshihisa
Format: Article
Language:English
Subjects:
Animals
Antibodies, Monoclonal - metabolism
Cells, Cultured
connexin32
Connexins - immunology
Fluorescent Antibody Technique, Indirect - methods
Frozen Sections
gap junction
Gap Junction beta-1 Protein
Gap Junctions - chemistry
Gap Junctions - metabolism
Gap Junctions - ultrastructure
Lacrimal Apparatus - chemistry
Lacrimal Apparatus - cytology
Lacrimal Apparatus - ultrastructure
Male
Microscopy, Immunoelectron - methods
morphology
rat lacrimal gland
Rats
Rats, Sprague-Dawley
reflexive gap junction
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Summary:We investigated localization of the gap junction in rat lacrimal gland in vivo and in vitro using electron microscopy and immunostaining with anti‐connexin32 (Cx32) monoclonal antibody (HAM8). In immunofluorescence study of lacrimal gland tissues, Cx32 protein appeared to exist not only at the intercellular borders of acinar cells, but also in the basal regions, where there apparently was no contact with adjacent acinar cells. Thin sectioning and immunoelectron microscopy revealed that lacrimal acinar cells formed autocellular gap junctions (reflexive gap junctions) in the basal regions and intercellular gap junctions with adjacent acinar cell membranes. In immunofluorescence study of primary culture, Cx32 protein was found on the free surfaces of isolated acinar cells at the early stage of culture. With culturing time, cell aggregates were formed. We observed Cx32 immunoreactivity between acinar cells in these aggregates, but not on their free surface. Electron microscopic study confirmed that these aggregates possessed intercellular gap junctions and morphologically differentiated acinar‐like structures. However, reflexive gap junctions were not observed in these aggregates. In conclusion, lacrimal acinar cells form intercellular and reflexive gap junctions in vivo. On the other hand, the existence of an acinar‐like structure and intercellular gap junctions indicates that acinar cells differentiated in vitro morphologically. The cells may communicate with each other through these junctions, organizing themselves into an acinar cell network in an in vitro situation.
ISSN:0022-0744
1477-9986
2050-5701
DOI:10.1093/jmicro/51.1.35