Neurotoxic and cytoprotective mechanisms in the ischemic neocortex.

Neuronal damage in ischemic stroke occurs due to permanent imbalance between the metabolic needs of the brain and the ability of the blood-vascular system to maintain glucose delivery and adequate gas exchange. Oxidative stress and excitotoxicity trigger complex processes of neuroinflammation, necrosis, and apoptosis of both neurons and glial cells. This review summarizes data on the structural and chemical changes in the neocortex and main cytoprotective effects induced by focal ischemic stroke. We focus on the expression of neurotrophins (NT) and molecular and cellular changes in neurovascular units in ischemic brain. We also discuss how these factors affect the apoptosis of cortical cells. Ischemic damage involves close interaction of a wide range of signaling molecules, each acting as an efficient marker of cell state in both the ischemic core and penumbra. NTs play the main regulatory role in brain tissue recovery after ischemic injury. Heterogeneous distribution of the BDNF, NT-3, and GDNF immunoreactivity is concordant with the selective response of different types of cortical neurons and glia to ischemic injury and allows mapping the position of viable neurons. Astrocytes are the central link in neurovascular coupling in ischemic brain by providing other cells with a wide range of vasotropic factors. The NT expression coincides with the distribution of reactive astrocytes, marking the boundaries of the penumbra. The development of ischemic stroke is accompanied by a dramatic change in the distribution of GDNF reactivity. In early ischemic period, it is mainly observed in cortical neurons, while in late one, the bulk of GDNF-positive cells are various types of glia, in particular, astrocytes. The proportion of GDNF-positive astrocytes increases gradually throughout the ischemic period. Some factors that exert cytoprotective effects in early ischemic period may display neurotoxic and pro-apoptotic effects later on. The number of apoptotic cells in the ischemic brain tissue correlates with the BDNF levels, corroborating its protective effects. Cytoprotection and neuroplasticity are two lines of brain protection and recovery after ischemic stroke. NTs can be considered an important link in these processes. To develop efficient pharmacological therapy for ischemic brain injury, we have to deepen our understanding of neurochemical adaptation of brain tissue to acute stroke.

Topography of neurotrophins in the rat neocortex and their role in neuron apoptosis after experimental ischemic stroke.

Neuronal loss due to apoptosis after ischemic injury depends on the trophic support of neurons and cytoprotective effects of neurotrophins (NTs). Different NTs may activate both pro- and antiapoptotic factors. Their distribution in the ischemic core (IC) and penumbra (IP) has been poorly studied. The available data on the localization of NTs in the ischemic brain are contradictory and depend to a certain degree on the pathogenetic model used. The distribution of NTs in different layers of the ischemic cortex is also largely unknown hindering our understanding of their exact effects and targets in different zones of the ischemic brain. We examined the immunolocalization of brain-derived neurotrophic factor (BDNF), neurotrophin-3, and glial cell line-derived neurotrophic factor (GDNF) in the parietal cortex using a rat model of ischemic stroke due to permanent occlusion of the middle cerebral artery. The spatial density of immunoreactive (IR) cells varied across the cortical layers and changed with time after ischemic injury. Their distribution in the IC differed considerably from that in the IP. The immunolocalization of neurotrophins in the contralateral hemisphere was similar to that in IP. We also studied the distribution of pro- and anti-apoptotic factors in IC and IP with and without intravenous BDNF administration. In the model without BDNF administration, the proportions of Bcl-2-, p53-, caspase-3-, and Mdm2-IR cells showed different dynamics during the ischemic period. In the model with BDNF administration, Mdm2 immunoreactivity was mainly observed in pyramidal cells of layers V/VI, and Bcl-2, in interneurons of layers II and III. The dynamics of p53 immunoreactivity was opposite to that of caspase-3 throughout the ischemic period. The present results suggest that after ischemic injury, 1) the number of neurotrophin-positive cells increases in the early ischemic period and decreases afterwards; 2) there is a close metabolic relationship between astrocytes and neurons contributing to their adaptation to ischemic conditions; 3) the IP borders undergo constant changes; 4) in the IP, neuronal loss occurs mainly by apoptotic pathway throughout the ischemic period; 5) BDNF may enhance considerably antiapoptotic mechanisms with a predominance of Mdm-2 activity in pyramidal neurons.

Structural and chemical changes in glial cells in the rat neocortex induced by constant occlusion of the middle cerebral artery.

Stroke-induced changes in neuroglia determine the basic conditions for the survival and damage of neurons in the ischemic core. Here, we studied the immunolocalization of glial cell line-derived neurotrophic factor (GDNF), glial fibrillary acidic protein (GFAP), ionized calcium-binding adaptor molecule 1 (Iba-1), and S-100ß in the rat parietal cortex after constant occlusion of the middle cerebral artery. These cytoplasmic proteins are specific for different glial cell types. They are used as indicators of activated microglia and astrocytes in immunocytochemical studies. The distribution pattern of all markers changed dramatically with time. GFAP- and S-100ß-positive astrocytes were observed in the penumbra zone and marked its boundaries. In days 1-8 after surgery, in the ischemic core, the number of S-100ß-immunoreactive astrocytes decreased, and individual pyramidal cells appeared. S-100ß-expressing pyramidal cells were localized in cortical layers III and V. They were only found in the ischemic core. Their proportion to the total number of cells was 37.3 ± 3.9 %, 22.2 ± 1.2 %, 16.3 ± 2.3 %, and 5.4 ± 0.3 % on days 1, 3, 8, and 14 after surgery. On day 21, no S-100ß-expressing pyramidal cells were observed. The spatial density of GFAP- and S-100ß-positive astrocytes increased in the penumbra region adjacent to the ischemic core and decreased in the penumbral periphery. As a result, the width of the perifocal penumbra zone decreased substantially at later stages of the stroke. In the penumbra, on days 1-3 after ischemic injury, GDNF immunoreactivity was mainly localized in neurons, while later on (days 8-21) it was mainly constrained to astrocyte glia. In intact rats, GDNF-positive neurons were situated in cortical layers II/III and V/VI and made up 52 ± 4.5 % of the total neuron population. Their proportion to the total number of neurons was 29 ± 2.1 %, 13.8 ± 0.6 %, and 3.1 ± 0.2 % on days 1, 8, and 21 after surgery. The number of GDNF-positive astrocytes, on the opposite, increased with time after ischemic injury. Iba-1-reactive microglia was mainly localized to the ischemic core. Microglial cells appeared activated as evidenced by their increased size and decreased number of processes and branching density. The spatial density of microglia reached a peak ​​on day 8 after ischemic injury both in the ischemic core and penumbra. An increase in the number of Iba-1-reactive microglia in the ischemic core correlated with a decrease of the number of GFAP-positive astrocytes. The results are discussed in the context of participation of neuroglia in regulation of various neuroprotective and destructive processes.

Gaseous transmitters in human retinogenesis.

Endogenous gaseous transmitters (nitric oxide, carbon monoxide, and hydrogen sulphide) form a special neuromodulation system mediating the development and modification of nerve centers. Here, we examined the localization of key gaseous transmitter enzymes: cystathionine ß-synthetase (CBS), cystathionine γ-lyase (CSE), heme oxygenase 2 (HO-2), and constitutive NO synthase (nNOS) in the fetal human retina at different stages of development. The number of CBS- and CSE-positive photoreceptors and intermediate retinal neurons was high in trimester I and gradually decreased to the end of trimester III. The number of HO-2-positive cells followed the same trend. The number of nNOS-positive intermediate retinal neurons and neurons within the ganglion cell layer showed the opposite dynamics with the peak in trimester III. The results are interpreted in terms of the role of gaseous transmitters in retinogenesis and cytoprotection.

The modular architecture and neurochemical patterns in the cerebellar cortex.

The review deals with topical issues of the neuronal arrangement underlying basic cerebellar functions. The cerebellum and its auxiliary structures contain several hundreds of modules (so called "microzones"). Each module receives the corticopetal input specific for the lobule it belongs to and forms the topographic projection. The precision of the major input-output signal flow in the cerebellar cortex is provided by a pronounced stratification of its synaptic zones of a various origin and regular topography of its afferent connections, interneurons, and efferent neurons. There is a nice match between the anatomical and functional coordinates of the modules, whose spatial boundaries are determined by the spread of afferent excitation and local interneuron connections. The dynamic characteristics of the modules are analyzed by the example of the formation of the nitrergic neuron ensembles and cerebellar projections of corticopetal fibers. The authors discuss the cerebellar blood flow and its relation to the activity of NO/GABAergic Lugaro cells and other interneurons in the cerebellar cortex. A generalized scheme of intra- and intermodular communication is proposed.

Retinal ganglion cell topography and spatial resolving power in the oriental fire-bellied toad Bombina orientalis.

The vertebrate visual system is determined by two main factors, a species' lifestyle and phylogenetic legacy. Studying the visual system in outgroup lineages may shed some light on the balance of these factors within a certain radiation. We studied the topography of retinal ganglion cells (RGCs) in the retina of the oriental fire-bellied toad Bombina orientalis. These toads belong to the ancient superfamily Discoglossoidea, a sister group to all extant Anura except for two small families. RGCs were retrogradely labeled with tetramethylrhodamine- dextran amine (TMR-DA) and examined in retinal wholemounts. RGCs occurred all over the retina except for the far periphery. Their total number was [Formula: see text] ([Formula: see text], [Formula: see text]). They comprised 73-77% of all cells in the ganglion cell layer. The spatial density of GCs increased gradually from the dorsal and ventral retinal periphery toward the equator to form a weak visual streak and a moderately pronounced area centralis. The minimum density was [Formula: see text], and the maximum, [Formula: see text]. The maximum density gradient was [Formula: see text]. The spatial resolution was minimum in the dorsal and ventral periphery ([Formula: see text] and [Formula: see text] cycles per degree in water and air, respectively). Intermediate values of spatial resolving power were found within the visual streak ([Formula: see text] and [Formula: see text] cycles per degree) and reached a peak in area centralis ([Formula: see text] and [Formula: see text] cycles per degree). This is sufficient for efficient prey location and capture. The relatively high RGC density and the presence of specialized retinal regions in oriental fire-bellied toads are consistent with their highly visual behavior. A brief review comparing the phylogeny and ecology of this with other anuran species suggests that the main factor shaping the RGC distribution in Anura is phylogenetic legacy; the environmental pressure results mainly in adjusting the maximum spatial density of RGCs (and hence the visual acuity) to meet the species' needs.

Retinal ganglion cells of the accessory optic system: a review.

The review deals with the morphology, physiology, topography, and central projections of direction-selective cells of the accessory optic system in vertebrates.

Retinal ganglion cells in the eastern newt Notophthalmus viridescens: topography, morphology, and diversity.

The topography and morphology of retinal ganglion cells (RGCs) in the eastern newt were studied. Cells were retrogradely labeled with tetramethylrhodamine-conjugated dextran amines or horseradish peroxidase and examined in retinal wholemounts. Their total number was 18,025 +/- 3,602 (mean +/- SEM). The spatial density of RGCs varied from 2,100 cells/mm(2) in the retinal periphery to 4,500 cells/mm(2) in the dorsotemporal retina. No prominent retinal specializations were found. The spatial resolution estimated from the spatial density of RGCs varied from 1.4 cycles per degree in the periphery to 1.95 cycles per degree in the region of the peak RGC density. A sample of 68 cells was camera lucida drawn and subjected to quantitative analysis. A total of 21 parameters related to RGC morphology and stratification in the retina were estimated. Partitionings obtained by using different clustering algorithms combined with automatic variable weighting and dimensionality reduction techniques were compared, and an effective solution was found by using silhouette analysis. A total of seven clusters were identified and associated with potential cell types. Kruskal-Wallis ANOVA-on-Ranks with post hoc Mann-Whitney U tests showed significant pairwise between-cluster differences in one or more of the clustering variables. The average silhouette values of the clusters were reasonably high, ranging from 0.52 to 0.79. Cells assigned to the same cluster displayed similar morphology and stratification in the retina. The advantages and limitations of the methodology adopted are discussed. The present classification is compared with known morphological and physiological RGC classifications in other salamanders.

Calcium-binding proteins in the cerebellar cortex of the bottlenose dolphin and harbour porpoise.

Studying the distribution of Ca2+-binding proteins allows one to discover specific neuron chemotypes involved in the regulation of the activity of various neural elements. While extensive data exist on Ca2+-binding proteins in the nervous system, in particular, in the cerebellar cortex of terrestrial mammals, the localization of these proteins in the cerebellar cortex of marine mammals has not been studied. We studied the localization of calretinin, calbindin, and parvalbumin immunoreactivity in the cerebellar cortex of the bottlenose dolphin Tursiops truncates and harbour porpoise Phocoena phocoena. In both species, most Purkinje cells were calbindin-immunoreactive, while calretinin and parvalbumin were expressed in a small portion of Purkinje cells. In addition, calretinin-immunoreactive unipolar brush and granule cells and calbindin- and parvalbumin-immunoreactive basket, stellate, and Golgi cells were observed. Calretinin-immunoreactive corticopetal (mossy and climbing) fibers were found. Based on the length of the primary dendrite, short-, middle-, and long-dendrite unipolar brush cells could be distinguished. The validity of this classification was supported using cluster analysis suggesting the presence of several natural types of these cells. The distribution of Ca2+-binding proteins in the cerebellar cortex of the cetaceans studied was generally similar to that reported for terrestrial mammals, suggesting that this trait is evolutionarily conservative in mammals.

Effects of the Briarosaccus callosus infestation on the commercial golden king crab Lithodes aequispina.

Commercial crab populations off the Kamchatka coasts are infested to a considerable degree by the rhizocephalan parasite Briarosaccus callosus: of 769 Lithodes aequispina males examined, 43 (5.7%) were parasitized. Infestations result in the feminization of the crabs, a significant decrease in the cheliped length, and a significant decrease in the carapace length and width. We suggest that commercial selection of healthy males, and the returning of unsuitable crabs, including infested ones, back into the sea, results in an increase of the proportion of infested crabs in the population, their elimination from reproduction, and, eventually, the gradual degradation of a whole population. To minimize as far as possible the negative effects of commercial crab harvesting, all infested crab specimens caught must be destroyed, either aboard or elsewhere, instead of throwing them back into the sea.