Biosynthesis and secretion of pituitary hormones: dynamics and regulation.

Production and secretion of hormones by the pituitary involve highly orchestrated intracellular transport and sorting steps. Hormone precursors are routed through a series of compartments before being packaged in secretory granules. These highly dynamic carriers play crucial roles in both prohormone processing and peptide exocytosis. We have employed the ACTH-secreting AtT-20 cell line to study the membrane sorting events that confer functionality (prohormone activation and regulated exocytosis) to these secretory carriers. The unique ability of granules to promote prohormone processing is attributed to their acidic interior. Using a novel avidin-targeted fluorescence ratio imaging technique, we have found that the trans-Golgi of live AtT-20 cells maintains a mildly acidic (approximately pH 6.2) interior. Budding of secretory granules causes the lumen to acidify to

Biogenesis of regulated exocytotic carriers in neuroendocrine cells.

Ca(2+)-triggered exocytosis is a hallmark of neurosecretory granules, but the cellular pathway leading to the assembly of these regulated exocytotic carriers is poorly understood. Here we used the pituitary AtT-20 cell line to study the biogenesis of regulated exocytotic carriers involved in peptide hormone secretion. We show that immature secretory granules (ISGs) freshly budded from the trans-Golgi network (TGN) exhibit characteristics of unregulated exocytotic carriers. During a subsequent maturation period they undergo an important switch to become regulated exocytotic carriers. We have identified a novel sorting pathway responsible for this transition. The SNARE proteins, VAMP4 and synaptotagmin IV (Syt IV), enter ISGs initially but are sorted away during maturation. Sorting is achieved by vesicle budding from the ISGs, because it can be inhibited by brefeldin A (BFA). Inhibition of this sorting pathway with BFA arrested the maturing granules in a state that responded poorly to stimuli, suggesting that the transition to regulated exocytotic carriers requires the removal of a putative inhibitor. In support of this, we found that overexpression of Syt IV reduced the stimulus-responsiveness of maturing granules. We conclude that secretory granules undergo a switch from unregulated to regulated secretory carriers during biogenesis. The existence of such a switch may provide a mechanism for cells to modulate their secretory activities under different physiological conditions.

Localization of the apex of the vagina: implications for radiation therapy planning.

PURPOSE: To evaluate (a) the displacement of the vaginal apex by a rod during radiation therapy simulation for gynecologic malignancy and (b) apical localization with implanted radiopaque markers. MATERIALS AND METHODS: Metallic markers were implanted in the cervix or vaginal cuff in nine patients with cervical or endometrial carcinoma who underwent irradiation. In all but one patient, radiographs were obtained with and then without the vaginal rod. Displacement of the markers relative to bone landmarks was measured. The total displacement was the square root of the sum of the squares of displacement in each axial direction. RESULTS: All patients showed displacement of the cervical markers by the vaginal rod (mean total displacement, 1.9 cm; range, 0.6-3.6 cm). The greatest displacement was cephalic (mean, 1.5 cm; range, 0.5-2.4 cm). Anteroposterior displacement occurred in all patients but was not as predictable as cephalic displacement. Displacement was anterior in five of the eight patients, posterior in three patients, and lateral in four patients. CONCLUSION: Displacement of the vaginal apex and/or cervix with placement of the vaginal rod during simulation was marked in all patients. Use of implanted cervical markers to localize the vaginal apex or the cervix during simulation is more accurate than use of a vaginal rod.

Solitary osteochondroma of the cervical spine.

OBJECTIVE: To discuss a case of cervical spine osteochondroma found incidentally during evaluation of a rib injury. To demonstrate diagnostic imaging, including plain film radiography, radionuclide bone scan, computed tomography and magnetic resonance imaging. CLINICAL FEATURES: This article presents the second reported case of osteochondroma associated with the C6 vertebral body. The classic imaging features of exostosis and a cartilaginous cap were identified. The osteochondroma was confluent with marrow of the C6 articular pillar. A biopsy was performed for confirmation of the lesion's histological character. INTERVENTION: Surgical resection was performed to remove the osteochondroma. CONCLUSION: Clinical examination, radiological evaluation and advanced diagnostic imaging helped detect and characterize an incidental cervical spine mass.

skn-1, a maternally expressed gene required to specify the fate of ventral blastomeres in the early C. elegans embryo.

By the 4-cell stage of C. elegans embryogenesis, a ventral blastomere, called EMS, is already committed to producing pharyngeal and intestinal cell types. Recessive, maternal-effect mutations in the gene skn-1 prevent EMS from producing both pharyngeal and intestinal cells. In skn-1 mutant embryos, EMS instead produces hypodermal cells and body wall muscle cells, much like its sister blastomere. Genetic analysis suggests that the skn-1 gene product is also required post-embryonically for development of the intestine. We have cloned and sequenced the skn-1 gene and describe sequence similarities to the basic regions of bZIP transcription factors. We propose that the maternally expressed skn-1 gene product acts to specify the fate of the EMS blastomere.

BK virus and JC virus shed during pregnancy have predominantly archetypal regulatory regions.

Twenty-three BK virus and JC virus DNA samples obtained from urine of pregnant women had almost exclusively archetypal transcriptional control regions. Rearrangements characteristic of laboratory strains are apparently not required for reactivation in humans. Unexpectedly, alignment shows that many elements identified previously in the BK virus enhancer are conserved in the JC virus archetype.