Immature rat intestinal stem cells (IEC-6) given the ability to express the transcription factor, pancreatic duodenal homeobox 1 (Pdx-1), yielded YK cells. Although these cells produced multiple enteroendocrine hormones, they did not produce insulin. Exposure of YK cells to 2 nmol/l betacellulin yielded BYK cells that showed the presence of insulin expression in cytoplasm and that secreted insulin into culture media. By examining the mechanism of differentiation in BYK cells, we found that another transcription factor, islet factor 1 (Isl-1) was newly expressed with the disappearance of Pax-6 expression in those cells after exposure to betacellulin. These results indicated that combined expression of Pdx-1 and Isl-1 in IEC-6 cells was required for the production of insulin. In fact, overexpression of both Pdx-1 and Isl-1 in IEC-6 cells (Isl-YK-12, -14, and -15 cells) gave them the ability to express insulin without exposure to betacellulin. Furthermore, implantation of the Isl-YK-14 cells into diabetic rats reduced the animals' plasma glucose levels; glucose levels dropped from 19.4 to 16.9 mmol/l 1 day after the injection of cells. As expected, the plasma insulin concentrations were 2.7 times higher in the diabetic rats injected with Isl-YK-14 cells compared to in controls. In summary, our results indicated that immature intestinal stem cells can differentiate into insulin-producing cells given the ability to express the transcription factors Pdx-1 and Isl-1.

The ascending projections of the inferior colliculus (IC) in the cat were traced by the autoradiographic method, with special reference in the differenial projections of each subnucleus of IC. The laminated ventrolateral part of the central nucleus of IC (CNv) projects to the ventral and medial divisions of the ipsilateral medial geniculate body (MGB). The projections to the ventral division are topographically organized in the mediolateral direction, the terminals being arranged in the form of lamina, while those to the medial division are diffuse. The unlaminated dorsomedial part of the central nucleus of IC (CNd) sends fibers to every division of the ipsilateral MGB, particularly to the dorsal division and the ventromedial portion of the ventral division. It is noteworthy that the external nucleus of IC (EN) projects to the superior colliculus, part of the pretectum, and the anterior extremity of MGB ipsilaterally, in addition to the ventral and medial divisions of MGB. The posterior cap of IC, regarded as the pericentral nucleus of IC (PC), projects ipsilaterally to the ventral part of the caudal tip of MGB and the posterior part of the suprapeduncular nucleus. In addition of these projections, the parabrachial region and interstitial nucleus of the brachium of IC (BIC) are identified as common targets of projections of each nucleus of IC on the ipsilateral side. Contralaterally, every subnucleus of IC except for PC projects via the commissure of IC to areas corresponding to the targets of the ipsilateral projections, such as the ventral and medial divisions of MGB and the parabrachial region and the interstitial nucleus of BIC, although these contraleral projections are in general much sparser than those ipsilateral. Intrinsic and commissural connections within IC are also revealed in this study, providing characteristic configurations of each subnucleus of IC. It is concluded that the ascending projections of IC in the cat are highly differentially organized according to its subnucleus.

The morphology of electrophysiologically identified neurons was examined in the primary auditory cortex (AI) of the cat. After stimulation of the medial geniculate nucleus (MG), second auditory cortex, posterior ectosylvian gyrus, contralateral AI, or corpus callosum, intracellular potentials were recorded from AI neurons, which were then injected intracellularly with horseradish peroxidase and recovered. Layer IV neurons, which receive MG fibers monosynaptically, are spiny and nonspiny stellate cells, small and medium-sized nonspiny tufted cells, and fusiform cells. They send their axons to layer III of the AI. Corticocortical AI neurons are medium-sized pyramidal cells in layer III. They receive axons from layer IV neurons of the AI and send their axons to layers I, II, IV, and V of the AI. Horizontal cells in layer I receive slow-conducting MG fibers monosynaptically, and send their axons to layer II of the AI. Stellate cells and small pyramidal cells in layer II receive afferent inputs polysynaptically from the MG. Layer II pyramidal cells receive afferent inputs from the MG via AI neurons in layers I and III, and send their axons to layers V and VI. The axons of layer II stellate cells were distributed within layer II. Pyramidal cells which send their axons to the MG are located in layers V and VI, distributing their axon collaterals to layers III-VI of the AI.

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Premotor neurons sending their axons to the trigeminal motor nucleus were observed in the cat by light and electron microscopy after labeling the neurons retrogradely or anterogradely with horseradish peroxidase (HRP). After HRP injection into the trigeminal motor nucleus, retrogradely labeled neurons were seen most frequently in the parvocellular reticular formation bilaterally. Many labeled neurons were also seen contralaterally in the intermediate zone at the rostralmost levels of the cervical cord and its rostral extension into the caudalmost levels of the medulla oblongata. Additionally, some neurons were labeled ipsilaterally in the mesencephalic trigeminal nucleus, contralaterally in the main sensory trigeminal nucleus and the trigeminal motor nucleus, and bilaterally in the oral and interpolar subnuclei of the spinal trigeminal nucleus. Only a few labeled neurons were seen in the confines of the gigantocellular reticular formation. All labeled neurons were small or of medium size; no large neurons were labeled. After HRP injection into the regions around the trigeminal motor nucleus or the parvocellular reticular formation, axodendritic terminals containing HRP granules were found contralaterally within the trigeminal motor nucleus. Some of these labeled terminals were filled with round synaptic vesicles and others contained pleomorphic synaptic vesicles. The varied morphology of labeled axon terminals was considered to reflect the functional heterogeneity of the premotor neurons for the trigeminal motor nucleus.