Physiological cell death is a widespread phenomenon in the development of both vertebrates and invertebrates. This review concentrates on an aspect of developmental cell death that has tended to be neglected, the manner in which the cells are dismantled. It is emphasized that the dying cells may adopt one of at least three different morphological types: "apoptotic", "autophagic", and "non-lysosomal vesiculate". These probably reflect a corresponding multiplicity of intracellular events. In particular, the destruction of the cytoplasm in these three types appears to be achieved primarily by heterophagy, by autophagy and by non-lysosomal degradation, respectively. The various mechanisms underlying both nuclear and cytoplasmic destruction are reviewed in detail. The multiplicity of destructive mechanisms needs to be born in mind in studies of other aspects of cell death such as the signals which trigger it, since different signals probably trigger different types of cell death.

Developmental cell death: morphological diversity and multiple mechanisms.

The early ontogeny of the monoamine neuron systems in the rat brain has been analysed using Falck-Hillarp fluorescence histochemistry. Serial sagittal sections of embryos with a crown rump length between 7 and 13 mm, approximately corresponding to gestational days 12 to 15 were obtained from mothers treated with a monoamine oxidase inhibitor given in order to increase the monoamine levels of the embryos. 5-hydroxytryptamine (5-HT)-neurons made their first appearance in the 8 mm embryo, dopamine (DA)-neurons in the 9 mm embryo, and noradrenaline (NA)-neurons in the 11 mm embryo. Small, rounded, weakly fluorescent cell bodies forming sparse aggregations appeared first. Fluorescent processes of two types soon appeared. Short processes from the cell bodies were running perpendicular to the long axis of the brain stem within the cell groups, while long slender axon bundles could be traced ascending through the met- and mesencephalon and into the prosencephalon as well as descending in the myelencephalon andspinal cord. In the 12 mm embryo the primordial DA cell formation of the substantia nigra with its striatal projections, the 5-HT neuron formations of the caudal mesencephalon, met- and myelencephalon as well as the NA neurons of the met- and myelencephalon are relatively well developed. It is concluded that the monoamine-neurons develop mechanisms for synthesis and storage of amines at a very early stage during ontogeny, thus recapitulating the phylogeny of these old systems. Likewise, monoamine oxidase is present early. The presence of neurotransmitters specifying the different developing neurons long before development of their nerve terminal areas and therefore before the establishment of normal synaptic function may indicate a role of these substances during ontogeny other than transmission of nerve impulses.

Late prenatal ontogeny of central monoamine neurons in the rat: Fluorescence histochemical observations

Gap junctions in the rat cochlea were investigated using immunostaining for connexin26 and transmission electron microscopy. Electron microscopy of normal and pre-embedded immunostained material showed that there were gap junctions between and among all cells that light microscopy showed to have immunostained appositions. Light microscopy showed immunostaining between and among all cell types that electron microscopy showed to be joined by gap junctions. Immunostaining for connexin26 was therefore taken as providing a reasonable approximation of the locations of gap junctions throughout the cochlea and was used to provide an overview of the extent of those locations. Cells interconnected via gap junctions fell into one of two groups. The first group consists of nonsensory epithelial cells and includes interdental cells of the spiral limbus, inner sulcus cells, organ of Corti supporting cells, outer sulcus cells, and cells within the root processes of the spiral ligament. The second group consists of connective tissue cells and includes various fibrocyte types of the spiral limbus and spiral ligament, basal and intermediate cells of the stria vascularis, and mesenchymal cells which line the scala vestibuli. The present work represents a first attempt towards a description of how serial gap junctions among cochlear cells reflect a level of organization of the tissue. The organization described here, together with a great deal of information from previous investigators, suggest that serially arranged gap junctions of both epithelial and connective tissue cells serve as the strucural basis for recycling endolymphatic potassium ions that pass through the sensory cells during the transduction process.

Gap junctions in the rat cochlea: immunohistochemical and ultrastructural analysis.

K+ cycling and the endocochlear potential.

Outer hair cells (OHC) from the organ of Corti are capable of fast voltage-induced length changes (Santos-Sacchi and Dilger, 1988), suggesting that an associated voltage sensor should reside in the OHC plasma membrane. Voltage-dependent mechanical responses and nonlinear charge movement of isolated OHCs from the guinea pig were analyzed using the whole-cell voltage-clamp technique. Ionic currents in the cells were blocked. Nonlinear voltage-dependent charge movement or, correspondingly, voltage-dependent capacitance was measured with step or AC analysis. OHC movements were measured either from video or using a differential photodiode technique. Maximum charge movements up to 2.5 pC were measured in OHCs from the low-frequency region of the cochlea. Both AC and step analyses indicated a peak nonlinear capacitance of 16- 17 pF. The voltage dependence was fit to a Boltzmann relation with the step analysis indicating a maximum nonlinear capacitance at -23 mV step potential from a holding potential of about -120 mV, and AC analysis indicating a maximum at a holding potential near -40 mV. AC analysis probably provides a more accurate evaluation of voltage dependence. Measures of OHC motility magnitude versus voltage follow the nonlinear capacitance-voltage function obtained from AC measures. Treatment of the cells with gadolinium ions (0.5-1 mM) blocked OHC motility. This treatment also produced a shift of the nonlinear capacitance function along the voltage axis in the depolarizing direction, which can be explained by membrane surface charge screening. However, maximum capacitance was reduced as well and may correspond to the reduction or abolition of OHC motility in response to gadolinium treatment. Gadolinium effects were reversible. Nonlinear capacitance is not a function of membrane deformation due to length changes, since removal of OHC cytosol via the patch pipette abolished longitudinal movement but did not reduce nonlinear charge movement. It is interesting to note that the nonlinear capacitance will dynamically influence the time constant of the OHC during acoustically evoked receptor potential generation.

https://www.jneurosci.org/content/jneuro/11/10/3096.full.pdf

Reversible Inhibition of Voltage-dependent Outer Hair Cell Motility and Capacitance

Intercellular junctions in the vascular stria and spiral ligament.

The junctions between the cells of the organ of Corti have been investigated with the freeze-fracture method in the chinchilla inner ear. Numerous gap junctions have been found among all the supporting cells, particularly at the level of their basal part. This means that the supporting cells have their cytoplasm electro-tonically and metabolically coupled. Gap junctions were not observed between supporting and sensory cells nor between sensory cells and nerve endings. At this level the transmission should be chemically mediated.

L. Luciano

Papers

Intercellular junctions in the vascular stria and spiral ligament.

Intercellular Junctions in the Organ of Corti as Revealed by Freeze Fracturing

Intercellular junctions and other membrane specializations in developing spinal ganglia: a freeze-fracture study.

Histochemical localization of acetylcholinesterase (AChE) activity in the inner ear.

Cholinesterase activity in spinal ganglia neuroblasts: A histochemical study at the electron microscope