Taurine: a promising nutraceutic in the prevention of retinal degeneration

Taurine is considered a non-essential amino acid because it is synthesized by most mammals.However,dietary intake of taurine may be necessary to achieve the physiological levels required for the development,maintenance,and function of certain tissues.Taurine may be especially important for the retina.The concentration of taurine in the retina is higher than that in any other tissue in the body and taurine deficiency causes retinal oxidative stress,apoptosis,and degeneration of photoreceptors and retinal ganglion cells.Low plasma taurine levels may also underlie retinal degeneration in humans and therefore,taurine administration could exert retinal neuroprotective effects.Taurine has antioxidant,anti-apoptotic,immunomodulatory,and calcium homeostasis-regulatory properties.This review summarizes the role of taurine in retinal health and disease,where it appears that taurine may be a promising nutraceutical.

Increased retinal venule diameter as a prognostic indicator for recurrent cerebrovascular events:a prospective observational study

Microvasculature of the retina is considered an alternative marker of cerebral vascular risk in healthy populations.However,the ability of retinal vasculature changes,specifically focusing on retinal vessel diameter,to predict the recurrence of cerebrovascular events in patients with ischemic stroke has not been determined comprehensively.While previous studies have shown a link between retinal vessel diameter and recurrent cerebrovascular events,they have not incorporated this information into a predictive model.Therefore,this study aimed to investigate the relationship between retinal vessel diameter and subsequent cerebrovascular events in patients with acute ischemic stroke.Additionally,we sought to establish a predictive model by combining retinal veessel diameter with traditional risk factors.We performed a prospective observational study of 141 patients with acute ischemic stroke who were admitted to the First Affiliated Hospital of Jinan University.All of these patients underwent digital retinal imaging within 72 hours of admission and were followed up for 3 years.We found that,after adjusting for related risk factors,patients with acute ischemic stroke with mean arteriolar diameter within 0.5-1.0 disc diameters of the disc margin(MAD_(0.5-1.0DD))of≥74.14μm and mean venular diameter within 0.5-1.0 disc diameters of the disc margin(MVD_(0.5-1.0DD))of≥83.91μm tended to experience recurrent cerebrovascular events.We established three multivariate Cox proportional hazard regression models:model 1 included traditional risk factors,model 2 added MAD_(0.5-1.0DD)to model 1,and model 3 added MVD0.5-1.0DD to model 1.Model 3 had the greatest potential to predict subsequent cerebrovascular events,followed by model 2,and finally model 1.These findings indicate that combining retinal venular or arteriolar diameter with traditional risk factors could improve the prediction of recurrent cerebrovascular events in patients with acute ischemic stroke,and that retinal imaging could be a useful and non-invasive method for identifying high-risk patients who require closer monitoring and more aggressive management.

Interplay between mesenchymal stromal cells and the immune system after transplantation: implications for advanced cell therapy in the retina

Advanced mesenchymal stromal cell-based therapies for neurodegenerative diseases are widely investigated in preclinical models.Mesenchymal stromal cells are well positioned as therapeutics because they address the underlying mechanisms of neurodegeneration,namely trophic factor deprivation and neuroinflammation.Most studies have focused on the beneficial effects of mesenchymal stromal cell transplantation on neuronal survival or functional improvement.However,little attention has been paid to the interaction between mesenchymal stromal cells and the host immune system due to the immunomodulatory properties of mesenchymal stromal cells and the long-held belief of the immunoprivileged status of the central nervous system.Here,we review the crosstalk between mesenchymal stromal cells and the immune system in general and in the context of the central nervous system,focusing on recent work in the retina and the importance of the type of transplantation.

P-aminobenzoic acid promotes retinal regeneration through activation of Ascl1a in zebrafish

The retina of zebrafish can regenerate completely after injury.M ultiple studies have demonstrated that metabolic alte rations occur during retinal damage;however to date no study has identified a link between metabolites and retinal regeneration of zebrafish.Here,we performed an unbiased metabolome sequencing in the N-methyl-D-aspartic acid-damaged retinas of zebrafish to demonstrate the metabolomic mechanism of retinal regeneration.Among the differentially-ex pressed metabolites,we found a significant decrease in p-aminobenzoic acid in the N-methyl-D-aspartic acid-damaged retinas of zebrafish.Then,we investigated the role of p-aminobenzoic acid in retinal regeneration in adult zebrafish.Impo rtantly,p-aminobenzoic acid activated Achaetescute complex-like 1a expression,thereby promoting Müller glia reprogramming and division,as well as Müller glia-derived progenitor cell proliferation.Finally,we eliminated folic acid and inflammation as downstream effectors of PABA and demonstrated that PABA had little effect on Müller glia distribution.Taken together,these findings show that PABA contributes to retinal regeneration through activation of Achaetescute complex-like 1a expression in the N-methyl-Daspartic acid-damaged retinas of zebrafish.

Homer1a reduces inflammatory response after retinal ischemia/reperfusion injury

Elevated intraocular pressure(IOP)is one of the causes of retinal ischemia/reperfusion injury,which results in NRP3 inflammasome activation and leads to visual damage.Homerla is repo rted to play a protective role in neuroinflammation in the cerebrum.However,the effects of Homerla on NLRP3inflammasomes in retinal ischemia/reperfusion injury caused by elevated IOP remain unknown.In our study,animal models we re constructed using C57BL/6J and Homer1^(flox/-)/Homerla^(+/-)/Nestin-Cre^(+/-)mice with elevated IOP-induced retinal ischemia/repe rfusion injury.For in vitro expe riments,the oxygen-glucose deprivation/repe rfusion injury model was constructed with M uller cells.We found that Homerla ove rexpression amelio rated the decreases in retinal thickness and Muller cell viability after ischemia/reperfusion injury.Furthermore,Homerla knockdown promoted NF-κB P65^(Ser536)activation via caspase-8,NF-κB P65 nuclear translocation,NLRP3 inflammasome formation,and the production and processing of interleukin-1βand inte rleukin-18.The opposite results we re observed with Homerla ove rexpression.Finally,the combined administration of Homerla protein and JSH-23 significantly inhibited the reduction in retinal thickness in Homer1^(flox/-)Homer1a^(+/-)/Nestin-Cre^(+/-)mice and apoptosis in M uller cells after ischemia/reperfusion injury.Taken together,these studies demonstrate that Homer1a exerts protective effects on retinal tissue and M uller cells via the caspase-8/NF-KB P65/NLRP3 pathway after I/R injury.

Circadian rhythm disruption and retinal dysfunction:a bidirectional link in Alzheimer’s disease?

Dysfunction in circadian rhythms is a common occurrence in patients with Alzheimer’s disease.A predominant function of the retina is circadian synchronization,carrying information to the brain through the retinohypothalamic tract,which projects to the suprachiasmatic nucleus.Notably,Alzheimer’s disease hallmarks,including amyloid-β,are present in the retinas of Alzheimer’s disease patients,followed/associated by structural and functional disturbances.However,the mechanistic link between circadian dysfunction and the pathological changes affecting the retina in Alzheimer’s disease is not fully understood,although some studies point to the possibility that retinal dysfunction could be considered an early pathological process that directly modulates the circadian rhythm.

Retinal displacement after surgery for idiopathic macular hole

AIM:To review and summarize the mechanism hypothesis,influencing factors and possible consequences of macular retinal displacement after idiopathic macular hole(IMH)surgery.METHODS:PubMed and Web of Science database was searched for studies published before April 2023 on“Retinal displacement”,“Idiopathic macular holes”,and“Macular displacement”.RESULTS:Recently,more academics have begun to focus on retinal displacement following idiopathic macular holes.They found that internal limiting membrane(ILM)peeling was the main cause of inducing postoperative position shift in the macular region.Moreover,several studies have revealed that the macular hole itself,as well as ILM peeling method,will have an impact on the result.In addition,this phenomenon is related to postoperative changes in macular retinal thickness,cone outer segment tips line recovery,the occurrence of dissociated optic nerve fiber layer(DONFL)and the degree of metamorphopsia.CONCLUSION:As a subclinical phenomenon,the clinical significance of postoperative macular displacement cannot be underestimated as it may affect the recovery of anatomy and function.

Microvascular alterations of the ocular surface and retina in connective tissue disease-related interstitial lung disease

AIM:To examine the disparities in macular retinal vascular density between individuals with connective tissue disease-related interstitial lung disease(CTD-ILD)and healthy controls(HCs)by optical coherence tomography angiography(OCTA)and to investigate the changes in microvascular density in abnormal eyes.METHODS:For a retrospective case-control study,a total of 16 patients(32 eyes)diagnosed with CTD-ILD were selected as the ILD group.The 16 healthy volunteers with 32 eyes,matched in terms of age and sex with the patients,were recruited as control group.The macular retina’s superficial retinal layer(SRL)and deep retinal layer(DRL)were examined and scanned using OCTA in each individual eye.The densities of retinal microvascular(MIR),macrovascular(MAR),and total microvascular(TMI)were calculated and compared.Changes in retinal vascular density in the macular region were analyzed using three different segmentation methods:central annuli segmentation method(C1-C6),hemispheric segmentation method[uperior right(SR),superior left(SL),inferior left(IL),and inferior right(IR)],and Early Treatment Diabetic Retinopathy Study(ETDRS)methods[superior(S),inferior(I),left(L),and right(R)].The data were analyzed using Version 9.0 of GraphPad prism and Pearson analysis.RESULTS:The OCTA data demonstrated a statistically significant difference(P<0.05)in macular retinal microvessel density between the two groups.Specifically,in the SRL and DRL analyses,the ILD group exhibited significantly lower surface density of MIR and TMI compared to the HCs group(P<0.05).Furthermore,using the hemispheric segmentation method,the ILD group showed notable reductions in SL,SR,and IL in the superficial retina(P<0.05),as well as marked decreases in SL and IR in the deep retina(P<0.05).Similarly,when employing the ETDRS method,the ILD group displayed substantial drops in superficial retinal S and I(P<0.05),along with notable reductions in deep retinal L,I,and R(P<0.05).In the central annuli segmentation method,the ILD group exhibited a significant decrease in the superficial retinal C2-4 region(P<0.05),whereas the deep retina showed a notable reduction in the C3-5 region(P<0.05).Additionally,there was an observed higher positive likelihood ratio in the superficial SR region and deep MIR.Furthermore,there was a negative correlation between conjunctival vascular density and both deep and superficial retinal TMI(P<0.001).CONCLUSION:Patients with CTD-ILD exhibits a significantly higher conjunctival vascular density compared to the HCs group.Conversely,their fundus retinal microvascular density is significantly lower.Furthermore,CTD-ILD patients display notably lower superficial and deep retinal vascular density in comparison to the HCs group.The inverse correlation between conjunctival vascular density and both superficial and deep retinal TMI suggests that detecting subtle changes in ocular microcirculation could potentially serve as an early diagnostic indicator for connective tissue diseases,thereby enhancing disease management.

Single-cell RNA sequencing analysis of the retina under acute high intraocular pressure

High intraocular pressure causes retinal ganglion cell injury in primary and secondary glaucoma diseases,yet the molecular landscape characteristics of retinal cells under high intraocular pressure remain unknown.Rat models of acute hypertension ocular pressure were established by injection of cross-linked hyaluronic acid hydrogel(Healaflow■).Single-cell RNA sequencing was then used to describe the cellular composition and molecular profile of the retina following high intraocular pressure.Our results identified a total of 12 cell types,namely retinal pigment epithelial cells,rod-photoreceptor cells,bipolar cells,Müller cells,microglia,cone-photoreceptor cells,retinal ganglion cells,endothelial cells,retinal progenitor cells,oligodendrocytes,pericytes,and fibroblasts.The single-cell RNA sequencing analysis of the retina under acute high intraocular pressure revealed obvious changes in the proportions of various retinal cells,with ganglion cells decreased by 23%.Hematoxylin and eosin staining and TUNEL staining confirmed the damage to retinal ganglion cells under high intraocular pressure.We extracted data from retinal ganglion cells and analyzed the retinal ganglion cell cluster with the most distinct expression.We found upregulation of the B3gat2 gene,which is associated with neuronal migration and adhesion,and downregulation of the Tsc22d gene,which participates in inhibition of inflammation.This study is the first to reveal molecular changes and intercellular interactions in the retina under high intraocular pressure.These data contribute to understanding of the molecular mechanism of retinal injury induced by high intraocular pressure and will benefit the development of novel therapies.

Postnatal development of rat retina:a continuous observation and comparison between the organotypic retinal explant model and in vivo development

The organotypic retinal explant culture has been established for more than a decade and offers a range of unique advantages compared with in vivo experiments and cell cultures.However,the lack of systematic and continuous comparison between in vivo retinal development and the organotypic retinal explant culture makes this model controversial in postnatal retinal development studies.Thus,we aimed to verify the feasibility of using this model for postnatal retinal development studies by comparing it with the in vivo retina.In this study,we showed that postnatal retinal explants undergo normal development,and exhibit a consistent structure and timeline with retinas in vivo.Initially,we used SOX2 and PAX6 immunostaining to identify retinal progenitor cells.We then examined cell proliferation and migration by immunostaining with Ki-67 and doublecortin,respectively.Ki-67-and doublecortin-positive cells decreased in both in vivo and explants during postnatal retinogenesis,and exhibited a high degree of similarity in abundance and distribution between groups.Additionally,we used Ceh-10 homeodomain-containing homolog,glutamate-ammonia ligase(glutamine synthetase),neuronal nuclei,and ionized calcium-binding adapter molecule 1 immunostaining to examine the emergence of bipolar cells,Müller glia,mature neurons,and microglia,respectively.The timing and spatial patterns of the emergence of these cell types were remarkably consistent between in vivo and explant retinas.Our study showed that the organotypic retinal explant culture model had a high degree of consistency with the progression of in vivo early postnatal retina development.The findings confirm the accuracy and credibility of this model and support its use for long-term,systematic,and continuous observation.

Experiment Study of Retinal Ultrastructure after Intravitreal FK506

Purpose:To investigate the retinal toxicity of FK506 by intravitreal administration.Methods:Twenty-two eyes of 14 New Zealand rabbits were investigated. FK506 at concentrations of 5,25 and 50μg/eye was injected into the vitreous cavities respectively. The control eyes were received mixed solution of balanced salt and ethanol. All eyes were examined by tonometry, slit lamp and indirect ophthalmoscopy preoperatively and postoperatively at the 1st, 3rd, 7th,and 14th day respectively. In the final examination, all eyes were enucleated and processed for light and electron microscopy. Results:No evidence of toxic reaction was seen in the eyes received 25μg FK506 or less of FK506. Several eyes received 50μg FK506 and control eyes developed conjunctival congestion and slightly bloody exudates in anterior chamber which may be related to irritation of ethanol. Two of five eyes received 50μg developed transient vitreous opacities. Electron microscopically, the mitochondria of the photoreceptor cells were swelled in the eyes treated with 50μg FK506.Conclusion:It is safety with intravitreal FK506. There are no irritation and toxicity to the rabbits eyes with the intravitreal doses of 25μg FK506 or less. The doses of 50μg FK506 are proved to be toxic to the retina. Eye Science 2004;20:34-38.

Physiology and Ultrastructure of Azolla imbricata as Affected by Hg 2+ and Cd 2+ Toxicity

The toxic effects of different gradient concentrations of Hg2+ and Cd2+ on chlorophyll content, chlorophyll a/b value, photosynthetic O-2 evolution, respiration rate, anti-oxidase system (superoxide dismulase (SOD), catalase (CAT), peroxidase (POD)) and ultrastructure of the cells of Azolla imbricata (Roxb.) Nakai were studied. The results showed that with Hg2+ and Cd2+ increase, chlorophyll content and chlorophyll a/b value, photosynthetic O-2 evolution decreased drastically; respiration rate peaked at 2 mg/L pollutant and declined afterwards. The activities of SOD, CAT and POD increased first and decreased afterwards except the activity of POD, which decreased with the increasing of Cd2+ concentration. Ultrastructural observation showed that the extent of ultrastructural damage was much more serious with higher pollutant concentration and longer time of stress. This resulted in swelling of chloroplast, disruption and disappearance of chloroplast membrane and disintegration of chloroplasts; swelling of cristae of mitochondria, deformation and vacuolization of mitochondria; condensation of chromatin in nucleus, dispersion of nucleolus and disruption of nuclear membrane. The experimental results showed: (1) Hg2+ and Cd2+ pollution not only destroyed physiological activities, but also caused irreversible damage to its ultrastructure, thus leading the cells to death; (2) With increase in the stress of Hg2+ and Cd2+, ultrastructural damage was related to the changes of plant physiology; (3) The toxic symptoms of plant showed an evident correlation between dose and effect; (4) The toxicity of Cd2+ on A. imbricata is heavier than that of Hg2+ under the same treatment time and concentration. The lethal concentration of Hg2+ to A. imbricata ranged from 3.5 to 4 mg/L, and that of Cd2+ ranged from 3 to 3.5 mg/L. The damage of cell ultrastructure on Anabaena azollae Strasburger was observed. The results indicated that tolerance of Azolla imbricata for Hg2+ and Cd2+ was higher than that of A. imbricata.

Development and Ultrastructure of the Phloem Ganglion in Bamboo Node

The development, cytological characters and ultrastructure of phloem ganglion in the nodal region of Phyllostachys edulis (Carr.) H. de Lehaie, a most economically important bamboo, were investigated and the possible physiological function of this special structure was proposed. The phloem ganglion derived directly from procambium is situated at the sites where the vascular bundle forks and is present in pairs. The phloem ganglion is spindle_like in appearance and usually consists of 4 to 6 layers. Two kinds of cells in the ganglion could be distinguished. In the middle, there are two layers of filiform cells with pointed ends so that there are no normal sieve plates. Nevertheless, there are many pits on the lateral wall of the filiform cells. The other type of cells located at both ends of the spindle which possess an intermediate form between the filiform cell and the normal sieve tube. The walls of these cells towards the filiform cells are strongly convex forming a special sieve plate. Ultrastructure study showed that cells in the ganglion are connected by enriched plasmodesma. During early differentiation, the paramural body and the ingrowth of cell wall could be observed. It indicates that the cells of phloem ganglion have the character of transfer cells. The organelles in the mature cells are mainly plastids with abundant accumulation of proteins of crystalline structure. The above_mentioned results suggest that the physiological function of the phloem ganglion is closely related with substance transport.

Ultrastructure of Epidermis and Flesh of the Developing Apple Fruit

The ultrastructure of the epidermis and flesh of apple ( Malus domestica Borkh cv. Red Fuji) fruit was systematically observed during the fruit development via transmission electron microscopy. The results showed that, in spite of the ultrastructural changes in many aspects of the developing fruit epidermal cells, it remained almost unchanged throughout the whole developmental process that the cytoplasm was filled with numerous endoplasmic reticula (ER). Most of these endoplasmic reticula were tube_like and rough_ER with enlarged cisterna from which many vesicles were produced. Some of the vesicles were shown to merge into vacuole. Some dynamic Golgi bodies were also found. All the ultrastructural characteristics showed that the epidermal cells have the features of excretory cells. The ultrastructure of the fruit flesh cells at the young fruit stage were shown to be metabolically active, characterized by the presence of numerous clustered plasmodesmata, cisterna enlarged_ and rough_ER filling the cytoplasm, plenty of vesicles and Golgi bodies, indicating their dynamic cellular transport function. Some giant_circular rough_ERs were found. All the ultrastructural features at this early developmental stage should be closely associated with the enlargement of the young fruit. At the rapid growing phase of the fruit the main changes were characterized by: the starch grain_filled amyloplasts, furcating of the single orifice of plasmodesmata, and the cytoplasm enrichment of both the Golgi body_formed vesicles and other vesicles. These features correspond well with those of a photoassimilate sink_cell. An ultrastructural degeneration phenomenon was observed at the fruit ripening stage, but the mitochondria and plasmalemma still remained intact, which might be related to the continuous development of fruit quality during the fruit ripening.

Ultrastructure of the Multicellular Nodules in Hypericum perforatum Leaves

With the development of the multicellular nodules in the leaves of Hypericum per plastids increased both in number and volume, and some plastids degenerated and were surrounded by dark tubular elements and vesicles. Some vesicles fused with vacuoles and secreted substances into vacuoles. There are many multivesicles, multimembrane structures and osmiophilic droplets in vacuoles of nodule cells. Meanwhile, dictyosome secreted vesicles into the vacuoles. However, there is a large central vacuole completely filled with secretory materials (hypericin) in matured multicellular nodules. This suggests that the osmiophilic droplets may be a precursor of hypericin originated from the degenerated plastids. There were abundant endoplasmic reticulums and dictyosomes between the plastids and osmiophilic droplets, suggesting that they may be involved in the synthesis and/or transport of the precursor of hypericin.

Ultrastructure of Oogenesis in Osmunda cinnamomea var. asiatica

The ultrastructure of oogenesis in the fern Osmunda cinnamomea L. var. asiatica Fernald has been studied by electron microscopy. During oogenesis, numerous vesicles not only moved towards the periphery, but also were arranged in line along the inside of plasmalemma, and in addition aggregated outside the plasmalemma by exocytosis. They released or excreted osmiophilic material. It was observed that a few vesicles containing lamellar osmiophilic material situated closely along the plasmalemma, seemed to break open. Simultaneously, a separation cavity between egg cell and archegonium wall formed. Its width was broader than the other advanced ferns reported previously, and an extra egg membrane occurred outside around the plasmalemma of the egg, its thickness being greater than in Pteridium and Dryopteris. Amyloplasts around the nucleus were filled with large triangular semicircular or subelliptical starch grains, but as the egg matured they progressively decreased. Nucleus was large and flattened, and paired membrane in two to three couples laid within the nucleus, close and parallel to the nuclear membrane. No nuclear evagination was observed. Mitochondria seemed to have been undeveloped, but finally recovered normally.

Ultrastructure of the Blepharoplast and the Multilayered Structure in Spermatogenesis in Osmunda cinnamomea var. asiatica

The ultrastructure of the blepharoplast and the multilayered structure (MLS) in the fern Osmunda cinnamomea var. asiatica Fernald have been studied by electron microscopy with respect to spermatogenesis. The blepharoplast appears in the young spermatid. The differentiating blepharoplast is approximately a spherical body, which is composed of densely stained granular material in the center and some cylinders outside of it. The differentiated blepharoplast is also a sphere, but without the densely stained material in the center, consisting of scattered or radially arranged cylinders. The MLS seen in the spermatid lies between the basal bodies and the giant mitochondrion. In the early developmental stage, the MLS only consists of lamellar layers, each of which runs parallel to one another and forms a strip. In the mid stage, the MLS is composed of the microtubular ribbon (MTr), the lamellar layers and a layer of plaque. In the late stage, the MLS forms accessory band, osmiophilic crest and a layer of osmiophilic material. The MTr grows out from the MLS and extends along the surface of the nucleus to unite with the nuclear envelope in a complex. The basal body coming from the cylinder produces the axoneme of the flagella in the distal end and the wedge-shaped structure in the proximal end, respectively. In the present study, the ultrastructural features of blepharoplast and the MLS of the protoleptosporangiopsida fern, O. cinnamomea var. asiatica, have been described and compared with those of other kinds of pteridophytes in detail. The lamellar layers appearing before the formation of the MTr was found and reported for the first time.

ULTRASTRUCTURE OF THE VEGETATIVE CELLS OF NOSTOC FLAGELLIFORME PREPARED WITH HIGH PRESSURE FREEZING AND FREEZE SUBSTITUTION TECHNIQUE

The ultrastructure of the vegetative cells of Nostoc flagelliforme Born. et Flah. was investigated with high pressure freezing and freeze substitution technique and compared with the results obtained by using conventional preparation methods. During the processes of chemical fixation, dehydration and embedding, the cell structures might be more artificially modified than that obtained from high pressure freezing and freeze substitution. With the present method, the sheath of N. flagelliforme could be well penetrated and no extra big space could exist between the cell and the sheath. The cell protoplasm rarely shrinked. Some fine structures of cell inclusions and unit membranes became visualized. Many bacteria were harbored in the sheath. In addition, the presence of big vacuoles in the cell of N. flagelliforme as well as the presence of bacteria in the sheath shown in the present preparation for cyanobacteria has not been described so far in the literature.

Effects of CO_2 Enrichment on Microstructure and Ultrastructure of Two Species of Freshwater Green Algae

order to investigate the morphological response of freshwater green algae to elevated CO2 concentration, Chlamydomonas reinhardtii Dang and Scenedesmus obliquus Kutz were cultured with enriched CO2, and their microstructure and ultrastructure were examined by microscopy and electron microscopy. The effect of CO2 enrichment to 186 mumol/L, was insignificant on the shape and size of C. reinhardtii, but significant in reducing the volume of S. obliquus. High-CO2 increased the amount of chloroplast. The pyrenoids occurred in low-CO2-grown cells but not in high-CO2-grown ones and more starch granules were observed in the former.

Evidence of Ultrastructure and Physiology of F-actin as Component of Plasmodesmata

The characters and ultrastructure of the intercellular connection were revealed in the outer epidermis of the garlic clove sheath by means of fluorescent probe TRITC_Phalloidin (TRITC_Ph) labeling combined with confocal laser scanning microscopy (CLSM), immuno_gold labeling and transmission electron microscopy. These results show that transcellular channel is a complex of rod_like cytoplasm channel and grouped plasmodesmata (PDs) in pit. The former remains a portion of the cell protoplast. The diameter of PD is normally 60-70 nm. The PDs are the real intercellular symplasmic connections of the cells. The transcellular fibers labeled with the TRITC_Ph obviously become narrow in the primary pit fields, which is the same as the characters observed under the electron microscope. The bright fluorescent spot in the primary wall reflects the grouped PDs in pit, and hence the presence of F_actin in the PDs can be confirmed. In immuno_gold labeling experiment, a lot of gold particles were massively distributed in the rod_like cytoplasm channel and grouped PDs. The result provides effective support that these fluorescent filaments possibly are the existing form of F_actin.