To localize the different domains of the laminin-1 molecule in tissues and gain insight into their in vivo relevance, we raised rat anti-mouse monoclonal antibodies (MAbs) against the entire molecule. Then we tested eight of the 20 clones producing anti-laminin-1 MAbs to specify their reactivity towards the al-, PI-, and yl-chains and the elastase-cleaved fragments of the laminin-l molecule. We found three MAbs with high titers in ELISA that showed good reactivity in embedded tissue. One of these reacted specifically against the El fragment, one against the ES fragment, and one MAb detected the al-chain of laminin-1 but not the PI-or ylchain. AU three MAbs are useful for light immunohistochemical investigations on a y d o n s and on paraffra-embedded material, and for ultrastructural localization of laminin-l in LR Gold-embedded mouse tissue. Antibody staining of the El and ES domains of laminin-1 revealed distinct localization of the molecule in the proximal tubule basement mem-
Differentiated C1300 mouse neuroblastoma cells were treated with 10(-4)-10(6) M gamma-aminobutyric acid (GABA) and/or sodium bromide (NaBr) for 2 days and then fixed. Quantitative studies revealed an increase in the length and branching of the processes, as well as an increase in the number of cells when compared to the controls. It is suggested that the above changes contribute to the augmentation of specialized contacts between cells and processes as well as the further maturation of the primitive stages of synaptogenesis as discussed.
We wish to report the isolation in pure form of a porcine calcitonin (PC-1) and the elucidation of its structure.1(1) We also wish to acknowledge the valuable assistance of C. Pidacks for chromatography development work, as well as E. Lindemann and H. Falk for biological assay data.
An in situ embedding technique for monolayer cultures for subsequent ultra-structural studies is described. Dehydration is carried out in graded ethanol series without propylene oxide, and Epon is used as a final embedding medium. The advantage of this method is that the plastic flask with the embedded material can be stripped off easily without affecting the monolayer culture and that the plastic does not dissolve in Epon.
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