Unresponsiveness to cannabinoids and reduced addictive effects of opiates in CB1 receptor knockout mice

The function of the central cannabinoid receptor (CB1) was investigated by invalidating its gene. Mutant mice did not respond to cannabinoid drugs, demonstrating the exclusive role of the CB1 receptor in mediating analgesia, reinforcement, hypothermia, hypolocomotion, and hypotension. The acute effects of opiates were unaffected, but the reinforcing properties of morphine and the severity of the withdrawal syndrome were strongly reduced. These observations suggest that the CB1 receptor is involved in the motivational properties of opiates and in the development of physical dependence and extend the concept of an interconnected role of CB1 and opiate receptors in the brain areas mediating addictive behavior.     

Cell surface regulators of complement, 5I2 antigen, and CD59, in the rat eye and adnexal tissues

PURPOSE: Cell surface complement regulatory proteins have been identified in high levels in ocular tissues, but no experimental model is available for examining their physiological roles. To develop such a model, the distribution of 5I2 antigen, a protein possessing the functions of the human decay-accelerating factor (DAF [CD55]) and membrane cofactor protein (MCP [CD46]), and rat inhibitory protein (CD59), the homologue of the human membrane inhibitor of reactive lysis (MIRL[CD59]) were characterized in the rat eye and ocular adnexal structures. METHODS: After euthanasia of female Wistar rats, followed by orbital exenteration, eyelids and orbital tissue including the lacrimal gland were separated from the globes and immediately snap-frozen in liquid nitrogen at -70 degrees C. Tissues then were sectioned at -20 degrees C and examined immunohistochemically for 5I2 antigen and rat CD59. RESULTS: Both molecules were found to be present in high levels in multiple sites. Corneal and conjunctival epithelia showed moderate to intense labeling for both regulators. Fibroblasts in the corneal stroma, conjunctiva, and sclera labeled similarly. Corneal endothelial cells showed intense labeling for rat CD59 but not for 5I2 antigen. The iris and ciliary body showed intense labeling for both proteins. The retina showed labeling at multiple levels, with that of rat CD59 being more intense than that of 5I2 antigen. The lacrimal gland labeled for both regulators. Vessels, muscle, and nerves in the orbit labeled intensely for both antigens. In the eyelid, conjunctiva, sebaceous glands, and muscle and nerve tissues labeled moderately to intensely for both molecules, whereas skin epithelium labeled less intensely. CONCLUSIONS: 5I2 antigen and rat CD59 are expressed in high levels and distributed similarly in the rat eye and lacrimal gland to DAF, MCP, and MIRL in the human eye and lacrimal gland. These findings establish the rat ocular surface as a model for studying the role of cell surface complement regulators in this site. This first identification of copious expression of these proteins in eyelid structures, which also participate in protection of the ocular surface, further suggests an important role for surface complement regulatory proteins in this location.     

The concept of synergism in radiobiology]

We have demonstrated previously that pemphigus vulgaris (PV)-IgG induces activation of phospholipase C (PLC), production of inositol 1,4,5-trisphosphate, and a rapid transient increase in [Ca2+]i in cultured human keratinocytes, leading to secretion of plasminogen activator and cell-cell detachment in cell culture. In the current study, to examine the involvement of protein kinase C (PKC) in the mechanism of blister formation in PV, we studied the PV-IgG-induced translocation of PKC isozymes from the cytosol to the particulate/cytoskeleton (p/c) fractions and the activation of PKC in human keratinocytes. Cells cultured in Eagle's minimum essential medium were incubated with PV-IgGs for 30 s, 1 min, 5 min, or 30 min. PV-IgG binding to the cell surface antigen (desmoglein III) induced translocation of PKC-alpha from the cytosol to the p/c fractions within 30 s, with a peak at 1 min that lasted at least 30 min. PKC-delta also was translocated within 1 min and reached a peak at 5 min but was reduced to basal levels at 30 min. Alternatively, PKC-eta translocation to the p/c fraction was induced slowly, taking more than 5 min, and was reduced to approximately half-maximum at 30 min, whereas PKC-zeta translocation reached a maximum at 30 s, rapidly returning to baseline by 5 min after PV-IgG stimulation. The total PKC activity in the p/c fraction also was increased after PV-IgG exposure, peaked at 1 min, and was sustained for at least 30 min. These findings suggest that a unique activation profile of PKC isomers may be involved in mediating the intracellular signaling events induced by PV-IgG binding to desmoglein III in cultured human keratinocytes.     

Interfollicular fibroblasts in the human thyroid gland: recognition of a CD34 positive stromal cell network communicated by gap junctions and terminated by autonomic nerve endings

While epithelial structure and functions have been substantially investigated in many organs, the mesenchymal elements have received less attention. Compared with follicular epithelial cells, there are a few morphological studies on the stroma of human thyroid gland. In order to characterize more fully and assess its possible functions, 15 samples of surgical and autopsy human thyroid tissue were studied by classical histology, immunohistochemistry, transmission electron microscopy, electron microscopic immunohistochemistry, and scanning electron microscopy. In human thyroid gland, the interfollicular connective tissue surrounding the follicles contained collagenous matrix, fibroblasts, unmyelinated nerve fibers with Schwann cells, small blood vessels, lymphatics, lymphocytes, plasma cells, macrophages, and mast cells. At the ultrastructural level, gap junctions between the cytoplasmic processes of interfollicular fibroblasts constituted a novel observation. Immunohistochemistry using a monoclonal antibody against Cx43 confirmed the distribution of gap junctions between stromal fibroblastic cells, which was compatible with the ultrastructural findings. The frequent and intimate association of fibroblastic processes with nerve terminals was also shown. Interfollicular stromal fibroblasts also stained with CD34. The main constituent of the human thyroid stromal tissue was a CD34 positive reticular network involving fibroblasts, mononuclear cells and nerve terminals. It represents a highly ordered stroma, with potential structural and functional similarities to the stroma of bone marrow (Yamazaki and Allen, 1990).