Application of the Mirror Technique for Three-Dimensional Electron Microscopy of Neurochemically Identified GABA-ergic Dendrites

In the nervous system synaptic input arrives chiefly on dendrites and their type and distribution have been assumed pivotal in signal integration. We have developed an immunohistochemistry (IH)-correlated electron microscopy (EM) method – the “mirror” technique – by which synaptic input to entire dendrites of neurochemically identified interneurons (INs) can be mapped due preserving high-fidelity tissue ultrastructure. Hence, this approach allows quantitative assessment of morphometric parameters of synaptic inputs along the whole length of dendrites originating from the parent soma. The method exploits the fact that adjoining sections have truncated or cut cell bodies which appear on the common surfaces in a mirror fashion. In one of the sections the histochemical marker of the GABAergic subtype, calbindin was revealed in cell bodies whereas in the other section the remaining part of the very same cell bodies were subjected to serial section EM to trace and reconstruct the synaptology of entire dendrites. Here, we provide exemplary data on the synaptic coverage of two dendrites belonging to the same calbindin-D28K immunopositive IN and determine the spatial distribution of asymmetric and symmetric synapses, surface area and volume of the presynaptic boutons, morphometric parameters of synaptic vesicles, and area extent of the active zones.

The hidden coloniality at the parasitic stage in Peltogaster reticulatus (Crustacea: Rhizocephala)

Upon infesting the hermit crab Pagurus proximus the parasitic barnacle Peltogaster reticulatus typically forms a single externa on each host. In vivo, histological and histochemical study revealed that most P. reticulatus with a single mature externa also exhibited three or four primordial externae and several buds of earlier developmental stages. It is shown that stolon buds and primordial externae in P. reticulatus internae intensively express alkaline phosphatase activity, the classical histochemical marker for mammalian embryonic stem and primary germ cells. Ovaries of the most developed primordial externae were filled with growing oocytes, so the process of oogenesis begins in the externa rudiments of P. reticulatus before the externae appear on the surface of the host body. Since the interna, which is connected to the mature externa, also integrated three or four additional primordial externae and several earlier buds, P. reticulatus, a non-colonial species based on the common presence of a single externa in each host, appears to have a hidden colonial organization at the parasitic stage of its life cycle.