Afferent connections of the prelunate visual association cortex (areas V4 and DP).

The afferent and efferent connections of the prelunate visual association area V4 of macaque monkeys were investigated by means of the horseradish peroxidase (HRP) method. The specific thalamic afferents from the dorsolateral segment of the medial pulvinar and the lateral segment of the inferior pulvinar were topographically organized. A band of cells was labelled in the intralaminar nuclei (nucl. centr. med. and lat., reaching into LD and the most dorsal part of VL), and a few cells in the interlaminar layers of the lateral geniculate body. Other diencephalic afferents included the claustrum, the nucleus basalis Meynert and the pars compacta of the substantia nigra. Ipsilateral cortical areas which projected into V4 included area 18 (V2), the inferior parietal cortex, the anterior and posterior parts of the superior temporal sulcus, the frontal eye fields and the temporo-basal association cortex on the lateral half of the parahippocampal gyrus and around the occipito-temporal sulcus. In the contralateral cortex, discontinuous regions in areas V4 and V5 on the prelunate gyrus and some cells at the 17/18-border were labelled. All regions in which labelled cells were found and, in addition a restricted region in the dorsal cap of the head and the tail of the caudate nucleus showed fibre and terminal labelling. In addition mesencephalic afferents and efferents were identified but not investigated in detail. An attempt to estimate the quantitative contribution of the various afferent systems to the prelunate cortex was made by counting the labelled cells in the different areas. The afferent and efferent organization of the prelunate visual association area indicates that it is incorporated in a network of cortical and subcortical regions involved in various aspects of visual behavior.

An improved silver stain for developing nervous tissue.

A reduced silver technique using physical development to stain embryonic nervous tissue is described. Brains are fixed in Bodian's fixative. Paraffin sections are pretreated with 1% chromic acid or 5% formol. They are impregnated with 0.01% silver nitrate dissolved in 0.1 M boric acid/sodium tetraborate buffer of pH 8 or with silver proteinate. Finally they are developed in a special physical developer which contains 0.1% silver nitrate, 0.01-0.1% formol as reducing agent, 2.5% sodium carbonate to buffer the solution at pH 10.3, 0.1% ammonium nitrate to prevent precipitation of silver hydroxide, and 5% tungstosilicic acid as a protective colloid. The development takes several minutes in this solution, thus the intensity of staining can be controlled easily. The method yields uniform, complete and reproducible staining of axons at all developmental stages of the nervous tissue and is easy to handle.

mu1A-adaptin-deficient mice: lethality, loss of AP-1 binding and rerouting of mannose 6-phosphate receptors.

The heterotetrameric AP-1 complex is involved in the formation of clathrin-coated vesicles at the trans-Golgi network (TGN) and interacts with sorting signals in the cytoplasmic tails of cargo molecules. Targeted disruption of the mouse mu1A-adaptin gene causes embryonic lethality at day 13.5. In cells deficient in micro1A-adaptin the remaining AP-1 adaptins do not bind to the TGN. Polarized epithelial cells are the only cells of micro1A-adaptin-deficient embryos that show gamma-adaptin binding to membranes, indicating the formation of an epithelial specific AP-1B complex and demonstrating the absence of additional mu1A homologs. Mannose 6-phosphate receptors are cargo molecules that exit the TGN via AP-1-clathrin-coated vesicles. The steady-state distribution of the mannose 6-phosphate receptors MPR46 and MPR300 in mu1A-deficient cells is shifted to endosomes at the expense of the TGN. MPR46 fails to recycle back from the endosome to the TGN, indicating that AP-1 is required for retrograde endosome to TGN transport of the receptor.