Three-dimensional analysis of the developing pituitary gland in the mouse

Prenatal development of the mouse pituitary gland was analyzed in three-dimensions by using confocal laser-scanning microscopy. In any wholemount preparation of the fetal pituitary gland from day 10 to day 18, immunofluorescence of laminin was observed in the deeper regions of the organ and demarcated the boundary of the epithelial tissue from the surrounding mesenchyme. Three-dimensional capillary networks in the fetal pituitary gland at day 13, day 15, and day 18 were visualized clearly by perfusing the blood vessels with an fluorescein isothiocyanate-labelled gelatin solution. During days 12-14, cellular plates protruded from the anterior wall of the pituitary anlage and extended toward the base of the infundibulum. At the same time, the mesenchyme situated among the cellular plates separated from the surrounding mesenchyme except at the anteroventral portion of the anlage. In a posterior wall of the anlage, intercellular depositions of laminin were found among the epithelial cells, some of which reached the laminin sheet in the basement membrane of the anlage. From days 15-18, the pituitary gland enlarged, and the mesenchyme spread radially from the median anteroventral portion to the lateral, posterior, and dorsal regions and finally expanded throughout the entire organ. At the periphery, the mesenchyme in the pituitary gland connected with that surrounding the capsule. Laminin was still found in the intercellular spaces between the epithelial cells, and most of the intercellular depositions of laminin were combined with the laminin sheet in the epithelial basement membrane. An abundance of vessels entered the organ from its anteroventral portion, spread, and connected with one another throughout the gland, similar to the distribution of mesenchyme. Intercellular laminin would induce the polarization of the contacting epithelial cells, resulting in rearrangement of the epithelial cells. The mesenchyme might spread in the pituitary gland, providing the intercellular laminin a foothold, in order to cleave the epithelial cell mass into lobules. The development of capillary nets in the adenohypophysis corresponded well with that of the mesenchyme. Because laminin localizes mainly in the basement membrane in the fetal organs, immunostaining of laminin clearly indicates the boundary between epithelial tissue and the mesenchyme. Three-dimensional observation of laminin in wholemount preparation is very useful for studying morphogenesis.