We investigated the changes caused by microtubule disruption in cell contact-induced translocation of alkaline phosphatase (ALP) from the Golgi area to the plasma membrane in McA-RH 7777 cells. When the cells were treated with colchicine, the tubular structure of microtubules in the cytoplasm was lost. Colchicine treatment also resulted in the appearance of numerous dots containing mannosidase II (man II) throughout the cytoplasm. Moreover, ALP was distributed in small dots throughout the cytoplasm, as well as in all regions of the plasma membrane, although it was most concentrated at sites of intercellular contact. On the other hand, when the cells were incubated in basal medium after colchicine treatment, large spots containing ALP reappeared in the perinuclear cytoplasm more quickly than the accumulation of small dots containing man II. These findings suggest that colchicine causes disassembly of the Golgi complex into fragments, which scatter throughout the cytoplasm, but that it does not interfere with translocation of ALP to the plasma membrane. Furthermore, cytoplasmic ALP may be localized at sites other than the Golgi complex.
As an important member of the cyclooxygenase isoenzymes, cyclooxygenase-2 (COX-2) mainly catalyzes the first two steps in prostanoid synthesis. In mammalian animals, although COX-2 was thought to be rarely expressed in most normal tissues and was usually upregulated in a variety of epithelial tumors and inflammatory reactions, recently it was reported that COX-2 could localize in the epidermis as well as the pilosebaceous unit of the normal human and mouse skin. Until now, the function of COX-2 in normal skin has remained unknown. To investigate the possible roles of COX-2 in normal skin by RT-PCR and immunochemistry, we studied the expression pattern of COX-2 in hair cycle of the normal rat skin. The expression of COX-2 mRNA was detected in normal rat skin sample and was related to the hair follicle cycle. When the hair cycle entered catagen and telogen, COX-2 mRNA transcription in skin increased significantly. Furthermore, the location of COX-2 immunoreactivity showed that COX-2 protein is mainly concentrated in the epidermis and pilosebaceous unit. In the stratified epidermis, the strong COX-2 protein expression was detected in the suprabasal layers of epidermis in anagen and declined in catagen and telogen. In hair follicle, COX-2 protein was obviously expressed in the outer root sheath of the anagen hair follicle, and was barely detectable in catagen as well as telogen. In the sebaceous gland, the COX-2 protein expression became more intense in catagen and telogen, with an increase in sebaceous gland size. Our results suggested that COX-2 was not specific to some abnormal tissues and was indeed involved in the normal physiology of rat skin, such as the differentiation of epidermis, the morphogenesis of the hair follicle, the transformation of hair cycle stages, and the lipid production of the sebaceous gland.
The distribution of histo-blood group A type 1, 2 and 3 antigens was investigated using immunohistochemistry in normal human skin and extramammary Paget’s disease (EMPD). We used monoclonal antibodies (mAbs) Bioclone-A (BA) and AR-1, which react with histo-blood group A type 1/2, and type 3 antigens, respectively. We found that A type 1, 2 and 3 antigens were expressed in the upper layer of the epidermis. We also found that the duct cells of the eccrine glands expressed A type 1/2 antigens and A type 3 antigens regardless of secretor status. The dark cells of the eccrine glands expressed A type 1, 2 and 3 antigens from A blood group secretors, but not from non-secretors. Apocrine glands, hair follicles and sebaceous glands did not express these antigens. Since these antigens were localized in the eccrine glands, we examined the possibility of a skin tumor marker. Interestingly, 7 out of 16 extramammary Paget’s disease cases were immunopositive for these antigens. Six cases were accompanied by dermal invasion. Five cases without dermal invasion were immunonegative against these antigens. These results suggest that the expression of histo-blood group A antigens in EMPD are associated with a poor histopathological prognosis.
In response to injury, synapse alteration may occur earlier than the changes in the cell body of neurons. Although retinal ganglion cell death and thinning of the inner part of retina were found after acute high intraocular pressure (HIOP), the structural and functional changes of synapses in the retina remain unknown. In the present study, we investigated the protein and mRNA expression of synaptophysin (SYN), an important molecule closely related to synaptic activities, synaptogenesis and synaptic plasticity. In addition, we also studied the ultrastructural changes of the retinal synapses. We found that (1) synaptophysin was upregulated transiently at both protein and mRNA level following HIOP; (2) broadened distribution of synaptophysin protein was present within the outer nuclear layer at the early stage following HIOP; (3) in the outer nuclear layer bouton-like vesicle-containing structures were observed by electron microscopy. This data suggested that, besides degeneration, synapses in rat retina may undergo regenerative events following HIOP.