Lymphatic vascular endothelial hyaluronan receptor (LYVE)-1- and CCL21-positive lymphatic compartments in the diabetic thymus.

To explore the biological significance of the lymphatics in the autoimmune process, the thymus from non-obese diabetic (NOD) mice was evaluated by histochemistry and western blot analysis. Thymic lymphatic endothelial cells showed suggestive expression patterns of the functional molecules lymphatic vascular endothelial hyaluronan receptor (LYVE)-1, CCL21, CD31 and podoplanin. With increasing age, the expression of CCL21 was reduced in the medullary epithelial cells and lymphatics. Of note, LYVE-1-expressing lymphatics, filled with a cluster of thymocytes, increased in number and size and extended from the corticomedullary boundary into the medulla as the insulitis progressed. The development of lymphatic compartments was occasionally accompanied by a regional disappearance between the cortex and medulla. The CD4- and CD8-positive T cells frequently penetrated through the slender lymphatic walls. The epithelial reticular cell layer lining the perivascular spaces was extensively stained with cytokeratin, but the expression of cytokeratin showed an age-dependent decrease. These findings indicate that the occurrence of LYVE-1-expressing lymphatic compartments and the alteration of CCL21 expression in the lymphatics may be involved in defective thymocyte differentiation and migration, and play a significant role in insulitic and diabetic processes.

Transmission of Homoiogenetic Induction in Presumptive Ectoderm of Newt Embryo.

Using newt, Cynops pyrrhogaster, embryos, the production and/or transmission of a homoiogenetic effect of the induction within a presumptive ectoderm was investigated. The primary inductor was the swimbladder of a crucian carp, Caracius auratus. In the first experiment, a piece of presumptive ectoderm was isolated from an early gastrula and one-third of its inner surface was placed in contact with the swimbladder for 30 min. After removal of the inductor, the ectodermal piece was allowed to stand by itself for several hours (pre-cultivation), and then it was divided into three parts of equal size, i.e. the end that had been placed in contact with the swimbladder (P1), the next middle (P2) and the end part (P3). The tissues produced in each part were examined after 10 days' cultivation in Holtfreter's solution. The induction was evoked not only in P1, but also in P2 and P3 which had been free from the inductor. The incidences of tissue differentiation in P2 and P3 increased with the lapse of the pre-cultivation time, and the rise in P3 came after P2. These results suggested that the mesodermal tissues in the parts not placed in contact with the swimbladder (P2 and P3) were evoked by the homoiogenetic stimulus which came from P1 and P2, respectively. In the next experiment, P2 of the ectodermal piece was substituted by an aged ectoderm which had lost its primary competence. In this system, the mesodermal induction was not evoked in P3. This suggested that the production of the homoiogenetic activity of the ectoderm might be associated with its competenece.

DIFFERENTIATION OF PARTIALLY MESODERMALIZED ECTODERM; HOMOIOGENETIC AND HETEROGENETIC INDUCTION BY PRIMARILY INDUCED PART OF THE ECTODERM.

Using embryos of the Japanese newt, Cynops pyrrhogaster, homoiogenetic and heterogenetic induction were investigated in the partially mesodermaelzed presumptive ectoderm. Half of the isolated presumptive ectoderm was placed in contact with the swimbladder of the crucian carp, Carasius auratus, for 15 or 60 min, while the other half was stained with Nile blue sulfate at the same time. The distribution of the stained cells in the tissues evoked in the explants was examined after cultivation for 10 days. Some mesodermal tissues were composed of both stained and unstained cells. This indicates homoiogenetic induction by the primarily induced part of the ectoderm on the other half. The neural and epidermal tissues in the explants were composed of stained cells only, except in one case. We conclude that the neural tissues are derived from cells not placed in contact with the swimbladder and that they are induced by the primarily induced part of the ectoderm.

Vegetalising factor extracted from the fish swimbladder and tested on presumptive ectoderm ofTriturus embryos.

Vegetalising factor was isolated from swimbladder of crusian carp (Carassius auratus) by solubilishing with 8 M urea the precipitate obtained after digesting the swimbladder with collagenase. The urea-soluble fraction vegetalised isolated presumptive ectoderm ofTriturus gastrula and produced both undifferentiated mesodermal and endodermal cells. Brief heating of the fraction changed its capacity to produce organised mesodermal tissues, such as notochord and somite, and the frequency of induction of undifferentiated cells was reduced. By inserting the urea-soluble fraction into the blastocoel of an early gastrula, embryos without epidermis were obtained. Some of the embryos consisted of undifferentiated mesodermal and endodermal cells, but in the remaining ones small fragments of notochord, small numbers of somites and pronephros developed, enclosed by endodermal cells.