Tectum Layer Research Articles

The methyl-CpG-binding domain family member PEM1 is essential for Ubisch body formation and pollen exine development in rice.

Pollen exine is composed of finely-organized nexine, bacula and tectum, and is crucial for pollen viability and function. Pollen exine development involves a complicated molecular network that coordinates the interaction between pollen and tapetal cells, as well as the biosynthesis, transport and assembly of sporopollenin precursors; however, our understanding of this network is very limited. Here, we report the roles of PEM1, a member of methyl-CpG-binding domain family, in rice pollen development. PEM1 expressed constitutively and, in anthers, its expression was detectable in tapetal cells and pollen. This predicted PEM1 protein of 240 kDa had multiple epigenetic-related domains. pem1 mutants exhibited abnormal Ubisch bodies, delayed exine occurrence and, finally, defective exine, including invisible bacula, amorphous and thickened nexine and tectum layer structures, and also had the phenotype of increased anther cuticle. The mutation in PEM1 did not affect the timely degradation of tapetum. Lipidomics revealed much higher wax and cutin contents in mutant anthers than in wild-type. Accordingly, this mutation up-regulated the expression of a set of genes implicated in transcriptional repression, signaling and diverse metabolic pathways. These results indicate that PEM1 mediates Ubisch body formation and pollen exine development mainly by negatively modulating the expression of genes. Thus, the PEM1-mediated molecular network represents a route for insights into mechanisms underlying pollen development. PEM1 may be a master regulator of pollen exine development.

Development of the permanent tetrad wall in Juncus L. (Juncaceae, Poales).

Juncaceae, a cosmopolitan family, belong to the cyperid clade of Poales together with Cyperaceae and Thurniaceae. Pollen grain of Juncaceae, as in Thurniaceae, is dispersed in a permanent tetrad, and knowledge about the ontogeny of its wall is still incipient, based on data from only one species. This study aims to analyze the formation of the pollen wall of seven Juncus species in order to characterize the timing and the ontogenetic events that lead to the cohesion of the four pollen grains in a permanent tetrad. Anthers at different developmental stages were submitted to techniques of light microscopy and transmission electron microscopy; dehiscent anthers with mature pollens were also analyzed in scanning electron microscopy. In all the species here studied, callose deposits around each microsporocyte, with dissolution prior to meiosis. Microspore wall starts at the end of the second meiotic division with formation of primexine. Exine comprises tectum, columellae, and foot layer. During cytokinesis, cell plates form the internal wall of the pollen tetrad. Mature permanent tetrad is enveloped externally by both the exine and intine and internally by the intine and the foot layer, which forms the continuous internal wall. Callose was detected in the early stages of microsporocytes, although reported to be absent in Juncaceae. Our data confirm the variation in Juncaceae cytokinesis and the occurrence of simple cohesion due to the presence of a continuous tectum along the pollen tetrad.

Morphology, ultrastructure and pollen germination in Ibicella lutea (Martyniaceae)

Pollen morphology, ultrastructure and germination mode of Ibicella lutea was investigated and documented using light microscopy (LM), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Ibicella pollen is clypeate, spheroidal, apolar, inaperturate, insulated, large, with an average diameter of 80 μm. The pollen wall is formed in the insula by two distinct layers; the outer layer, the exine, which is semitectate, with ectexine constituted by columellae, tectum and foot layer and the granular endexine, being the only layer between the insulae and the inner layer, the thick intine. The generative nucleus is of irregular shape and the vegetative nucleus is spheroidal. This is the first report of the presence of microbodies within the vegetative cell in pollen of Martyniaceae. The probable function of the microbodies in the lipid metabolism of the vegetative cell is discussed. The pollen tube emerges through the thinner areas of the wall between the insulae. In these areas the cytoplasm shows vacuoles and secretory vesicles produced by dictyosomes.

Nicotinic potentiation of frog retinotectal transmission in tectum layer F by α3β2, α4β2, α2β4, α6β2, or α7 acetylcholine receptor subtypes

ObjectiveThe aim of the study was to explore the effect of semi-specific antagonists and agonists of the nicotinic acetylcholine receptors on the paired-pulse facilitation and nicotinic tonic and phasic potentiation of the frog retinotectal synaptic transmission. Materials and methodsThe experiments were performed in vivo on adult frogs, Rana temporaria. An individual retina ganglion cell (or its retinotectal fiber) was stimulated by current pulses delivered through multichannel stimulating electrode positioned on the retina. Responses to a discharge of a single retinal ganglion cell were recorded in the tectum by an extracellular carbon-fiber microelectrode positioned in the terminal arborization of the retinotectal fiber in the tectum layer F. The effect of the antagonists and agonists of the nicotinic acetylcholine receptors on the tectal responses has been tested. ResultsWe found that the antagonists, MLA and DHβE, and agonists, RJR-2403 and choline, of the nicotinic acetylcholine receptors of the α3β2, α4β2, α2β4, α6β2 or α7 subtypes have had no effect on the phasic and tonic potentiation of the retinotectal transmission. The paired-pulse facilitation of the retinotectal transmission was not appreciably affected by the antagonists, but the choline, agonist of the α7 subtype receptor, has significantly decreased the paired-pulse facilitation. ConclusionsThe tonic and phasic potentiation of the retinotectal transmission in the tectum layer F were not mediated by the receptors of α3β2, α4β2, α2β4, α6β2 or α7 subtype. The results suggest that presynaptic nicotinic acetylcholine receptors of the frog optic fibers are different from those of the mammalian optic fibers.

Inhibition of Dendritic L-Type Calcium Current by Memantine in Frog Tectum

The aim of the study was to explore the effects of memantine on responses elicited in the frog tectum by the bursts of spikes of moderate strength of a single retina ganglion cell and to gain an insight about the effect of memantine on the L-type Ca(2+) current. The experiments were performed in vivo on adult frogs (Rana temporaria). An individual retina ganglion cell (or its retinotectal fiber) was stimulated by current pulses delivered through a multichannel stimulating electrode positioned on the retina. Responses to the discharge of a single retinal ganglion cell were recorded in the tectum by an extracellular carbon-fiber microelectrode positioned in the terminal arborization of the retinotectal fiber in the tectum layer F. The solution of memantine (1-amino-3,5-dimethyladamantane) hydrochloride (30 or 45 μM) was applied onto the surface of the tectum by perfusion at a rate of 0.4 mL/min. Memantine (30-45 μM) largely inhibited the L-type Ca(2+) channel-mediated slow negative wave and late discharges seen in the tectum responses without any effect on fast synaptic retinotectal transmission. Our results suggest that the neuroprotective effect of memantine could arise not only through the inhibition of the NMDA receptor current but also through the suppression of the L-type Ca(2+) current.

Phasic nicotinic potentiation of frog retinotectal transmission facilitates eliciting of higher activity level of the tectum column

Nicotinic acetylcholine receptors contribute to the mediation of cholinergic role in attention, vigilance, orienting and detection of behavioral significant stimuli. We have recently demonstrated an increase of the intrinsic recurrent excitatory activity of the tectum column caused by the phasic (after-burst) nicotinic potentiation of a frog single axon retinotectal transmission to the tectum layer F. We have shown in the present study that the phasic nicotinic potentiation facilitates eliciting of higher activity level of the tectum column featured by generation of output signals from the tectum column. Since these signals can lead to an escape from danger reactions, a functional significance of nicotinic modulation of the neural network has been demonstrated. The phasic nicotinic potentiation that facilitates eliciting of higher activity level of the tectum column can be considered as a mechanism of vigilance and cue detection at the level of small neural network.

Frog retinal ganglion cells projecting to the tectum layer F release acetylcholine as co-mediator

Neurons may release more than one classical neurotransmitter (co-mediator). It has been demonstrated in a recent study that a burst of action potentials in frog retina ganglion cells induces an after-burst increase (phasic potentiation) of the retinotectal transmission that lasts tens of seconds. This increase is mediated by presynaptic nicotinic acetylcholine receptors that are activated by the endogenous acetylcholine released during the burst of action potentials of the retinotectal fiber. The objective of the present study was to find out the origin of acetylcholine release. We show that reduction of the retinotectal transmission to the subthreshold level by application of moderate concentrations of kynurenic acid or CNQX had no effect on the phasic nicotinic potentiation of the retinotectal transmission. This demonstrates that the retinotectal terminals are the source of acetylcholine – responsible for the phasic potentiation of retinotectal transmission. The acetylcholine is thus co-released with glutamate.

Pollen development of Cardiocrinum giganteum (Wall.) Makina in China

In this article, we studied the pollen morphology and wall development, microsporogenesis, male gametophyte development, and anther wall structure changes during pollen development of Cardiocrinum giganteum (Wall.) Makina from the genus Cardiocrinum (Endl.) Lindl. (Liliaceae) using paraffin sections, scanning and transmission electron microscopy, and fluorescence microscopy. The results showed that C. giganteum has oval-shaped pollen with a single sulcus and reticulate ornamentation. The exine is of the semi-tectum type and can be divided into the tectum layer, columellate layer and basal layer. Meiosis in the microsporocyte is accompanied by successive cytokinesis. The mature pollen is three-celled. The anther wall prior to maturity is built by one layer of epidermis, 1–2 layers of endothecium cells, 4–5 middle layers and 2 layers of tapetum, while upon maturity it is only built by one layer of epidermis, one layer of endothecium cells and one middle layer. The tapetal cells are secretory, with two or more nuclei. Ubisch bodies originate from rough endoplasmic reticulum except a few from mitochondria.

Presynaptic nicotinic potentiation of a frog retinotectal transmission evoked by discharge of a single retina ganglion cell

It was demonstrated in our previous studies of the frog retinotectal transmission that retinotectal synaptic potentials are enhanced by a factor of 1.5 due to the tonic presynaptic nicotinic potentiation, caused by the ambient level of the acetylcholine in the frog tectum. Furthermore, the results of those studies have indicated that the mechanism of the nicotinic potentiation is only partially exploited, because the application of the cholinergic agonist had increased the retinotectal transmission more than 2 times above the level of the tonic potentiation. The purpose of the present study was to explore this additional potentiation. We have shown that: (1) Bursts of 4–10 action potentials of a frog retina ganglion cell gave rise to an increase (phasic potentiation) of the retinotectal transmission 1.4–2.2 times, depending on the burst strength, that lasted tens of seconds. (2) This increase has been mediated through the presynaptic nicotinic acetylcholine receptors activated by the endogenous acetylcholine released into the tectum during relatively strong bursts of the retina ganglion cell. (3) Two types of the nicotinic acetylcholine receptors are co-localized in the presynaptic terminals of the individual retinotectal input to the tectum layer F – high-affinity (tonic) and low-affinity (phasic) nicotinic receptors.

Components of the pigeon tectothalamic visual pathway, revealed with aid of study of cytochrome oxidase activity and immunoreactivity to calcium-binding proteins

Distribution of activity of cytochrome oxidase (CO) and immunoreactivity to parvalbumin (Pv) and calbindin (Cb) was studied in the optic tectum of the pigeon (Columba livia). In the first link of the tectofugal pathway in the central gray layer (SGC = layer 13), small amounts of the CO-active and Pv-immunoreactive (Pv-ir) cellular bodies were revealed in its internal part (sublayer 13b). Some of these neurons located along the SGC lower boundary had long dendritic processes ascending into the superficial tectum layer (SGF). In the retinorecipient SGF sublayers and particularly in neuropil of the sublayers 4 and 7, the high CO activity correlating with Pv-immunoreactivity was found. It is suggested that a great contribution to metabolic activity of these sublayers is made by the largely branching dendritic processes of Pv-ir neurons of sublayer 13b. The projectional neurons SGC located in its external part (sublayer 13a) were CO-inactive and contained Cb. They sent long dendrites into sublayer 5b; in its neuropil, the high density of Cb-immunoreactivity and a moderate CO activity were detected. Thus, the tectal link of the pigeon tectofugal visual channel consists of two components--the Pv-specific, highly metabolically active and the Cb-specific, metabolically less active ones that transduce visual information from different retinorecipient SGF sublayers. The absence of the significant amount of CO-positive bodies of projectional neurons in SGC can be due to that metabolically more active are their dendritic arborizations in the SGF sublayers.

Muscarinic inhibition of recurrent glutamatergic excitation in frog tectum column prevents NMDA receptor activation on efferent neuron

It is widely recognized that neuronal network activity can be modulated via activation of nicotinic and muscarinic acetylcholine receptors located pre- and postsynaptically. It was established in our earlier study that the activation of presynaptic nicotinic receptors greatly facilitates the retinotectal glutamatergic transmission. In the present study, we have determined a transmitter of tectal recurrent excitation and explored the effects of muscarinic acetylcholine receptor activation on the recurrent excitation and the activity of frog tectum column in vivo. Discharge of a single retinal ganglion cell was elicited by a minimal electrical stimulation of the retina. Evoked activity of the tectum column was recorded using the carbon-fiber microelectrode inserted into the tectum layer F. We found the following: 1. The recurrent excitation in the tectum column was not affected by d-tubocurarine (10μM) and was greatly depressed by the kynurenic acid (500μM), demonstrating glutamatergic nature of the recurrent excitation. 2. The glutamatergic recurrent excitation was largely reduced by carbamylcholine (100μM) and oxotremorine-M (10μM), demonstrating that the activation of muscarinic receptors, located, presumably, on the presynaptic terminals of recurrent pear-shaped neurons, inhibits the recurrent excitation in the tectum column. 3. The muscarinic inhibition of glutamatergic recurrent transmission had critical influence on the activity of the tectum column, preventing the generation of an output signal through suppression of the NMDA receptor activation and establishing necessary conditions for returning of the network to its resting state.

Single retinal ganglion cell evokes the activation of L-type Ca 2+-mediated slow inward current in frog tectal pear-shaped neurons

The dendrites of neurons from many regions of the nervous system contain voltage-sensitive channels that generate persistent inward currents. We have recently suggested that a slow negative wave (sNW), extracellularly observed in the frog tectum during the burst discharge of a single retinal ganglion cell, can be generated as a result of the persistent inward current in dendrites of tectal pear-shaped neurons. The aim of this study is to substantiate this hypothesis by simulation using a quasi-reconstructed pear-shaped neuron with bistable dendrites and experimental investigation of the sNW. In the experiments, the discharge of a single retinal ganglion cell was elicited by an electrical stimulation of the retina. The evoked electrical activity of the tectum was recorded using a carbon-fiber microelectrode inserted into tectum layer F. We found the following: (1) Slow inward current or plateau potential in bistable dendrites is reflected in the extracellular space as a sNW. (2) The sNW evoked by the burst discharge of a single retinal ganglion cell projecting to frog tectum layer F is generated by the activation of L-type calcium channels in the dendrites of pear-shaped neurons. (3) A few pear-shaped neurons may be suprathresholdly excited during the development of the sNW.

Pollen Morphology and Ultrastructure of Calyceraceae

Pollen morphology of 13 species from all six genera of Calyceraceae (Acicarpha, Boopis, Calycera, Gamocarpha, Moschopsis, and Nastanthus) and representatives of the Campanulaceae and Goodeniaceae is examined with light (LM), scanning (SEM), and transmission (TEM) electron microscopy. Acicarpha, Calycera, and Nastanthus pollen grains are distinguished by angulaperturate apertures, colpar ledges and surface depressions between colpi known as intercolpar concavities (IC). Pollen of Gamocarpha and Moschopsis is tricolporate rather than angulaperturate and without an IC. Some species of Boopis are similar to the preceding genera (e.g., B. graminea), while others (e.g., B. gracilis) are angulaperturate with ICs. Structural features derived from fractured pollen in SEM and sections in TEM show pollen walls composed of prominent columellae ca. 0.55–1.1 µm high and <0.25 µm wide. The columellae terminate distally into a complex of shortened columellae ca. 1.5 µm in length and are separated by an illdefined irregular internal tectum layer. This structural complex is well known in several tribes of the Asteraceae and referred to as the Anthemoid type. In those grains with an IC, the structure consists of essentially short (ca. 1 µm), unbranched columellae, similar to those found within the Asteraceae subfamily Barnadesioideae (Dasyphyllum and Schlechtendalia). Goodeniaceae (including Brunonia) pollen has angulaperturate apertures, spinules (i.e., minute spines), problematic IC and some structural similarity to Calyceraceae pollen. The tendency within Calyceraceae to develop colpar ledges, ektexine bridges, and ICs may be a synapomorphy uniting the family with Goodeniaceae. If the ancestral pollen type for the Calyceraceae, Asteraceae, and Goodeniaceae clade is the Gamocarpha type (convex intercolpar regions; no colpar ledges and no ektexine bridges), then the appearance of these structures within each family may be a synapomorphy supporting their close phylogenetic relationship suggested by molecular analyses.

Suprathreshold excitation of frog tectal neurons by short spike trains of single retinal ganglion cell.

It has been established that coincident inputs from multiple presynaptic axons are required to achieve a suprathreshold level of excitation for the most of central neurons. The present study, however, was designed to determine whether a train of spikes of an individual retinal ganglion cell (that is, input from a single presynaptic axon) targeting a frog tectum layer F could evoke suprathreshold excitation of tectal neurons. The lungs of immobilized frog were artificially ventilated during experiments. An individual ganglion cell was electrically stimulated in the retina through a multi-channel electrode. Responses evoked in the tectum by the stimulation were recorded extracellularly from a terminal arborization of the retinotectal fiber using the carbon-fiber microelectrode. Negative and negative-positive spikes (referred to as first type population responses) and polyphasic spikes followed by excitatory synaptic potentials (referred to as second type population responses) were observed in the recordings of retinotectal activity. Usually, the population responses have ensued after the frequency facilitated first and/or second testing individual retinotectal synaptic potential and disappeared in a threshold manner with a reduction of retinotectal transmission by an application of kynurenic acid. These observations have suggested that the population responses were a consequence of a suprathreshold excitation of tectal neurons and, therefore, could serve as the sign for such an excitation. Recordings have also demonstrated that sources of the first type population responses (likely, the hillocks of axons or somas of postsynaptic neurons) lie deeper than the optic fiber layer F of the tectum, whereas sources of the second type population responses (likely, axon terminal arborizations of these postsynaptic neurons) are scattered throughout the optic fiber layers. The findings have suggested: 1) a short train of action potentials of an individual retinal ganglion cell (likely darkness, also known as 5th, detector) can excite tectal neurons to suprathreshold level; 2) tectal and perhaps, nucleus isthmi neurons that make up recurrent connection circuits to the optic fiber layers of the tectum are also activated; 3) a suprathreshold level for an individual retinotectal input is achieved primarily due to the frequency facilitation of synaptic potentials; and 4) an artificial ventilation of the lungs of immobilized frog favors the eliciting of a suprathreshold excitation of tectal neurons, demonstrating that the ventilation certainly improves the physiological condition of a frog.

Pollen morphology in the ericoid clade of the order Ericales, with special emphasis on Cyrillaceae

Pollen morphology and pollen wall structure in 23 species representing all families of the ericoid clade of order Ericales, i.e. Actinidiaceae, Sarraceniaceae, Roridulaceae, Clethraceae, Cyrillaceae, and Ericaceae, was investigated by means of light microscopy and SEM. The pollen wall of all taxa was found to consist of two solid strata, a tectum and foot layer, separated by a more or less narrow infratectum with granular elements. The three genera of Cyrillaceae were found to differ from each other. In Cyrilla and Cliftonia the pollen is psilate, often more than 20 mum in diameter, spheroidal with an indistinct colpus rim; in contrast, pollen of Purdiaea are rugulate and slightly different between species, smaller and often less than 20 mum in diameter, oblate with a projecting rim reaching over the colpus. In several characteristic features, pollen of Purdiaea is more similar to pollen of Clethra (Clethraceae), than to pollen of Cyrilla and Cliftonia .

Pollen morphology in thePortulacaceae (Centrospermae)

Pollen of 110 species from 18 genera in thePortulacaceae has been examined by light and scanning electron microscopy, and a representative number by transmission electron microscopy. Three basic pollen types were found: 3-colpate with thick tectum and foot layer with prominent unbranched columellae and an extremely thin endexine; pantoporate with thick tectum and foot layer with branched columellae enclosing pores and an endexine that is one to two layers thick; pantocolpate with thin tectum and foot layer with broad, short unbranched columellae and an inconspicuous endexine. All pollen types, however, have a spinulose and tubuliferous/punctate ektexine. Also, all the genera except three,Calandrinia H.B.K.,Montia L. andTalinumAdanson are stenopalynous. There is, however, no absolute correlation between pollen morphology and geographical distribution, although both the major centre of palynological diversity and the majority of all species with tricolpate grains occur in South America.

Neurofibrillar changes in goldfish (Carassius auratus L.) brain in relation to environmental temperature

Neurofibrillar changes occur in the brains of goldfish, Carassius auratus L., maintained at 5 degrees C for 177 days or more under a 12-hour photoperiod. In paraffin sections impregnated by the silver method of Holmes, the light microscopic appearance of the neurofibrillary change was evidenced by black rings 1 micrometer to 2 micrometer in outside diameter. A quantitative study showed the mean number of rings to be: optic tectum layer 3, 1.34 X 10(2)/mm(3), layer 5, 1.14 X 10(2)/MM(3), nucleus prerotundus 4.63 X 10(2)/mm(3), olfactory bulbs, 6.2 X 10(2)/mm3. In the brains of fish kept at 15 degrees C a few rings are found only in some olfactory bulbs and not in any other region of the brain. The brain wet weight also changes and was found to be significantly (p less than 0.01) less in 5 degrees C than in 15 degrees C fish, the mean values being 0.1588 g and 0.2091 g respectively. The significance of the observed changes is discussed. It is suggested that the smaller brain wet weight may reflect a change in the vasculature of the brain and be related to the acclimation process. It is hypothesized that the rings are a morphological expression of a physiological change caused by prolonged exposure to low temperatures; a neuronal response reflecting either early degeneration or a functional adaptation which may involve an alteration in axoplasmic transport. It is suggested that this system is a useful model for the study of relationships between neurofibrillar disturbances and neuronal function.