JOURNAL OF THE KYORIN MEDICAL SOCIETY

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The anterior pituitary gland comprises of the anterior and intermediate lobes. α-melanocyte-stimulating hormone (α-MSH) is released from melanotropes in the intermediate lobe. Amphibians can camouflage themselves under the control of α-MSH. However, in mammals, the physiological importance of α-MSH and the control of its secretion remain unclear. In examining the function of α-MSH, isolating the melanotropes for use in several in vitro assays is crucial. We previously developed a method for separating a particular cell population from the rat anterior lobe using an antibody against a cluster of differentiation (CD) proteins. In this study, we aimed to adapt this powerful separation system to isolate melanotropes from the intermediate lobe of adult rats. CD362 was detected in melanotropes of the rat intermediate lobe via immunohistochemistry and was identified as a novel marker. We then isolated melanotropes from the rat intermediate lobe using a pluriBead-cascade cell isolation system based on the anti-CD362 antibody-mediated binding of cells to beads of different sizes and their isolation through sieves of different mesh sizes. As a result, we successfully isolated melanotrope populations from the intermediate lobe, with 95% purity. These findings improve the understanding of melanotropes in the intermediate lobe of the pituitary gland.

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