Tomoo Ohashi

Estrogen target cells during the early stage of medullary bone osteogenesis: Immunohistochemical detection of estrogen receptors in osteogenic cells of estrogen-treated male Japanese quail

SummaryThe localization of estrogen receptors (ER) in osteogenic cells during the early stage of medullary bone osteogenesis was studied immunohistochemically in the femurs of estrogen-treated male Japanese quail. Alkaline phosphatase (ALP) activity was used as a marker for osteogenic cells. ER immunostaining was observed in the nuclei of weak ALP-positive bone lining cells on the endosteal bone surface of nontreated birds. After 24 hours of estrogen treatment, nuclear immunostaining was detected in ALP-positive preosteoblasts on the endosteal bone surface. After 48 hours, the medullary bone appeared to some degree along the endosteal surface. ER immunostaining was observed in the nuclei of ALP-positive osteoblasts on the medullary bone surface. This study demonstrates that ER are present in osteogenic cells, and suggests that estrogen directly acts on medullary bone osteogenesis.

IMMUNOELECTRON MICROSCOPIC DEMONSTRATION OF ESTROGEN RECEPTORS IN OSTEOGENIC CELLS OF JAPANESE QUAIL

SummaryThe localization of estrogen receptors (ERs) in osteogenic cells was immunoelectron microscopically examined in the femurs of female and estrogen-treated male Japanese quail. An electron dense reaction product showing ER localization was observed in the nuclei of osteoblasts and immature osteocytes in the medullary bone of the female quail. However, reaction product was not seen in the osteoclasts. On the endosteal bone surface of male quail, nuclear reaction product was detected in bone lining cells. After 24 h of estrogen treatment, reaction product was observed in the nuclei of preosteoblasts on the endosteal bone surface. After 48 h, the medullary bone partly appeared along the endosteal surface. Nuclear reaction product was seen in osteoblasts on the medullary bone surface.

Effects of oestrogen and anti‐oestrogen on the cells of the endosteal surface of male Japanese quail

1. The endosteal cellular response occurring in the early stages of medullary bone formation was histologically examined in the femurs of male Japanese quail treated with oestrogen and antioestrogen. 2. The numbers of endosteal cells decreased, and preosteoblasts and osteoblasts increased, on the endosteal surface following oestrogen treatment. 3. After oestrogen and anti-oestrogen treatment the numbers of preosteoblasts and osteoblasts on the endosteal surface did not increase. However, endosteal cells slowly decreased in number and osteoclasts increased. 4. This study demonstrates that differentiation of osteoblasts is induced by oestrogen stimulation and suggests that osteoclasts occur by oestrogen inhibition on the endosteal surface.

Immunoelectron microscopic detection of estrogen target cells in the bone marrow of estrogen-treated male Japanese quail

The localization of estrogen receptors (ERs) in bone marrow cells was studied immunoelectron microscopically in the femurs of estrogen-treated male Japanese quail. The electron-dense reaction product showing ER localization was not detected in the bone marrow cells of the male quail. After 48 h of estrogen treatment, nuclear reaction product was observed in many mononuclear cells in the bone marrow. Most were located in the marrow stroma, and they were divided into undifferentiated and differentiated type cells, characterized by poorly and well-developed cytoplasm, respectively. Some ER-positive cells were situated in the hematopoietic tissue and had moderately developed cytoplasm. Thus, estrogen target cells may be present in the stroma and hematopoietic tissue of bone marrow.

Histochemical identification of oestrogen target cells in the medullary bone of laying hens

1. Oestrogen target cells in the medullary bone of laying hens were examined using a histochemical method with fluorescein isothiocyanate labelled oestradiol. 2. Specific fluorescence was found to occur strongly in osteoblasts and weakly in osteoclasts on the medullary bone surface. 3. This study suggests that osteoblasts on the medullary bone surface are oestrogen target cells.

Effects of estrogen on the proliferation and differentiation of osteogenic cells during the early stage of medullary bone formation in cultured quail bones

Quail bone fragments cultured in the medium containing 17β estradiol (E2) were examined morphologically. On the endosteal bone surface of bones cultured in serum-containing medium, preosteoblasts were observed and labeled by3H-thymidine after 24 h of culturing. After 48 h of culturing, medullary bones occasionally appeared along the endosteal surface, and their bone surfaces were covered with labeled osteoblasts. These osteoblasts were frequently found when added E2. On the endosteal bone surface of cultured bones in the serum-free medium alone, bone lining cells were observed and not labeled throughout the culture period. On the endosteal bone surface of bones cultured in the serum-free medium containing E2, labeled preosteoblasts were seen after 24 and 48 h of culturing. However, osteoblasts did not appear. These findings suggest that estrogen acts directly on the proliferation and differentiation of osteogenic cells.