Immunocytochemical localization of Na+, K+-ATPase in primary cultures of rat retina

Conditions have been established which allow growth of embryonic rat retinal cells in dissociated cell culture for up to one month. Na⁺,K⁺-ATPase localization was stuied in both neuronal and mixed neuronal-glial (flat cell) cultures. High Na⁺,K⁺-ATPase-like-immunoreactivity was associated with plasma membranes of neuronal cell bodies and their processes. Markedly lower immunoreactivity was found in the underlying flat cells in mixed cultures. Staining was generally uniform over perikaryal plasma membranes and showed a bead-like appearance in neuronal processes, supporting previous studies in brain tissue which used histocytochemical procedures specific for the Na⁺,K⁺-ATPase. This system should be useful for examining distribution of the enzyme in developing nerve and glial cells and may help to resolve questions regarding Na⁺–K⁺ homeostasis by neurons and glia.Dedicated to Henry McIlwain.     

Root response to above-ground light quality – Differences between rhizomatous and non-rhizomatous clones of Festuca rubra

Root response to reduced red/far red ratio above ground was studied in an experiment with 12 vegetatively propagated clones of Festuca rubra, a rhizomatous grass species. A new method suitable for describing the node position within rhizome systems was developed. It uses a combination of developmental and functional directions of ordering, adopting methods of topology. At each node the root structure was defined as consisting of three root types: unbranched, long branched and fine branched. Root system size was expressed as the number and length of roots of these individual types. Rhizomatous and non-rhizomatous clones of Festuca rubra differed in the size and structure of their root systems and in the response to lowered red/far red light ratio. This response was caused more by the behaviour of the rhizomes than of the roots alone. In rhizomatous clones, the largest differences in root system structure were between the mother-nodes and the nodes on the rhizomes. The size of the root system also depended on the node position. Response to the red/far red ratio was clone specific only in some of the root parameters, especially in traits of unbranched roots at young nodes. The role of the three root morphotypes in the plant's ontogeny, nutrient uptake and ability to cope with environmental heterogeneity both above- and below ground, is discussed.     

Subcellular localization of the sites of conversion of 1-aminocyclopropane-1-carboxylic acid into ethylene in plant cells

The subcellular localization of the sites of 1-aminocyclopropane-1-carboxylic acid (ACC) conversion into ethylene was studied by comparing the specific radioactivity of ethylene evolved from the whole cells with that of intra- and extracellular pools of labelled ACC. We demonstrate that some cells cultured in vitro (Vitis vinifera L. cv. Muscat) or leaf tissues (Hordeum vulgare L. and Triticum aestivum L.) have two sites of ethylene production: (i) an external site, converting apoplastic ACC, located at the plasma membrane, and very sensitive to high osmotica and, (ii) an intracellular site, converting internal ACC and remaining unaffected even under severe plasmolysis. In other cells cultured in vitro (Vitis vinifera L. cv. Gamay) and pea leaves (Pisum sativum L.), only the intracellular site operates and ethylene production is almost unaffected by plasmolysis. Protoplasts obtained from plasmolysis-sensitive Muscat cells lose 97% of their capacity for ethylene production compared with the parent cell, while those from plasmolysisinsensitive Gamay cells retain up to 50%. Protoplasts from both Gamay and Muscat cells cultured for 8 d in vitro, recover the full capacity of ethylene production of the initial whole cells, whether or not they are allowed to reform their cell wall. Therefore, we exclude a cooperation between the cell wall and the plasma membrane in ethylene production.Abbreviations ACC 1-aminocyclopropane-1-carboxylic acid - EFE ethylene-forming enzyme We are grateful to Dr. Philip John (Reading, UK) for useful discus sions made possible by a North Atlantic Treaty Organization Colla borative Grant (No. 0383/88) and Dr. Yves Meyer (Perpignan, France) for his collaboration in culturing protoplasts.