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SUMMARYMost gastrointestinal stromal tumors (GISTs) develop due to gain-of-function mutations in the tyrosine kinase,KIT. We recently showed that mutant KIT mislocalizes to the Golgi area and initiates uncontrolled signaling. However, the molecular mechanisms underlying its Golgi retention remain unknown. Here, we show that protein kinase D2 (PKD2) is activated by the mutant, which causes KIT’s Golgi retention. In PKD2-inhibited cells, KIT migrates from the Golgi region to lysosomes and subsequently undergoes degradation. Importantly, delocalized KIT is unable to trigger downstream activation. In the Golgi area, KIT activates the PKD2-phosphatidylinositol 4-kinaseIIIβ (PKD2-PI4KIIIβ) pathway through phospholipase γ2 (PLCγ2) to generate a PI4P-rich membrane domain, where the AP1-GGA1 complex is aberrantly recruited. Disruption of any factors in this cascade results in KIT release from the Golgi region, indicating that these PKD2-related pathways are responsible for the Golgi retention of KIT. Our findings unveil the molecular mechanisms underlying KIT mislocalization and provide evidence for a new strategy for inhibition of oncogenic signaling.

We have proposed and demonstrated a high-Tc SQUID system for detecting metallic contaminants in foodstuffs. There is a demand for the development of systems for detecting not only magnetic materials but also non-magnetic materials such as Cu and aluminium in foodstuffs to ensure food safety. The system consists of a SQUID magnetometer, an excitation coil and a permanent magnet. For a non-magnetic sample, an AC magnetic field is applied during detection to induce an eddy current in the sample. For a magnetizable sample, a strong magnetic field is applied to the sample prior to the detection attempt. We were able to detect a stainless steel ball with a diameter of 0.1 mm and a Cu ball less than 1 mm in diameter, for example.

Stem cells in developing tissues give rise to the multiple specialized cell types that make up the heart, lung, skin, and other tissues. For scientists, it is important to know the stages of stem cell development. We propose a new method to obtain knowledge of the cell development stages by using a high-Tc SQUID magnetometer. In the first step of the research, we used Wister rat myocardial cells incubated for 12–14 days in a culture dish as a sample, which were not derived from stem cells. With the cells we tried to measure the signal using a SQUID magnetometer. We were able to measure the magnetic signals, which may be generated by ion transport through the cell membrane. A beat signal with a period of 0.6 s was observed. The peak-to-peak value was about 400 pT. This showed good agreement with the result of microscopic observation.

We have developed a new type of superconducting interference device (SQUID) microscope. A direct-coupled SQUID magnetometer with a high-μ metal needle was used and the substrate was machined to create a dimple for the needle at the centre of the pick-up loop. One end of the needle penetrated through the superconducting pick-up loop in a vacuum; the needle was fixed in the vacuum window with the other end at room temperature in the outside atmosphere. Several kinds of simulation using a Maxwell simulator were performed and the results were applied to our system. As a demonstration, a laser-printed output was scanned by the microscope. Line bars with a line width of 100 μm and a spacing between lines of 200 μm were clearly imaged. This paper was presented at the 8th International Superconductive Electronics Conference, Osaka, Japan, 19–22 June 2001.

A new type of superconducting interference device (SQUID) microscope was designed and fabricated. A direct-coupled SQUID magnetometer with a high µ-metal needle was used and the substrate was machined to create a dimple for the needle at the center of the pick-up loop. One end of the needle penetrated through the superconducting pick-up loop in a vacuum; the needle was fixed in the vacuum window with the other end at room temperature in the outside atmosphere. As a demonstration, a laser printed output was scanned by the microscope. Line bars with a line width of 100 µm and a spacing between lines of 200 µm were clearly imaged.