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Journal of Histochemistry and Cytochemistry, Vol. 48, 569-578, April 2000, Copyright © 2000, The Histochemical Society, Inc.

TECHNICAL NOTE

A New Approach to Three-dimensional Reconstructed Imaging of Hormone-secreting Cells and Their Microvessel Environments in Rat Pituitary Glands by Confocal Laser Scanning Microscopy

J. Itoha, K. Kawaib, A. Serizawab, K. Yasumurad, K. Ogawae, and R.Y. Osamurac
a Laboratories for Structure and Function Research, Tokai University School of Medicine, Isehara, Japan
b Division of Diagnostic Pathology, Tokai University School of Medicine, Isehara, Japan
c Department of Pathology, Tokai University School of Medicine, Isehara, Japan
d Department of Surgery, NKK Hospital, Kawasaki, Japan
e Department of Diagnostic Radiology, NKK Hospital, Kawasaki, Japan

Correspondence to: J. Itoh, Laboratories for Structure and Function Research, Tokai U. School of Medicine, Bohseidai Isehara Kanagawa 259-1193, Japan.

There has been considerable interest in the relationship between hormone- secreting endocrine cells and their microvessels in human pituitary gland. However, microcirculatory networks have rarely been studied in three dimensions (3D). This study was designed to visualize and to reveal the relationship between hormone-secreting endocrine cells and their microvessel environment in 3D, using rat pituitary glands under various (hyper/hypo) experimental conditions by confocal laser scanning microscopy (CLSM). Female adult Wistar rats were used after bilateral adrenalectomy or ACTH administration for 2 weeks. Clear 3D reconstructed images of ACTH cells, the microvessel network and counterstained nuclei were obtained at a maximal focus depth of 1 mm by CLSM without any background noise. In the hyperfunctional state, slender cytoplasmic processes of hypertrophic stellate ACTH cells frequently extended to the microvessels. In the hypofunctional state, ACTH cells appeared atrophic and round with scanty cytoplasm, and cytoplasmic adhesions to microvessel network patterns were inconspicuous. Therefore, 3D reconstructed imaging by CLSM is a useful technique with which to investigate the microvessel environment of hormone-secreting cells and has the potential to reveal dynamic hormone-secreting pathways. (J Histochem Cytochem 48:569–577, 2000)

Key Words: 3D analysis, microvessel environments, hormone-secreting endocrine, cells, ACTH, pituitary gland, confocal laser scanning, microscopy, FITC-conjugated gelatin, injection, histochemistry, immunohistochemistry


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