Expression of proteins supporting visual function in heterobranch gastropods.

To sense light, animals often utilize mechanisms that rely on visual pigments composed of opsin and retinal. The photon-induced isomerization of 11-cis-retinal to the all-trans configuration triggers phototransduction cascades, resulting in a change in the membrane potential of the photoreceptor. In mollusks, the most abundant opsin in the eye is Gq-coupled rhodopsin (Gq-rhodopsin). The Gq-rhodopsin-based visual pigment is bistable, with the regeneration of 11-cis-retinal occurring in a light-dependent manner without leaving the opsin moiety. 11-cis-retinal is also regenerated by the action of retinochrome in the cell bodies. Retinal binding protein (RALBP) mediates retinal transport between Gq-rhodopsin and retinochrome in the cytoplasm. However, recent studies have identified additional bistable opsins in mollusks, including Opn5 and xenopsin. It is unknown whether these bistable opsins require RALBP and retinochrome for the continuous regeneration of 11-cis-retinal. In the present study, we examined the expression of RALBP and retinochrome in the photoreceptors expressing Opn5 or Xenopsin in the heterobranch gastropods Limax and Peronia. Our findings revealed that retinochrome, but not RALBP, was present in some of the Opn5A-positive brain photosensory neurons of Limax. The ciliary cells in the dorsal eye of Peronia, which express Xenopsin2, lacked both retinochrome and RALBP. Therefore, bistable opsins do not necessarily depend on the RALBP-retinochrome system in a cell. We also examined the expression of other proteins that support visual function, such as ß-arrestin, Gq, and Go, in all types of photoreceptors in these animals, and uncovered differences in the molecular composition among the photoreceptors.

Functional characterization of four opsins and two G alpha subtypes co-expressed in the molluscan rhabdomeric photoreceptor.

BACKGROUND: Rhabdomeric photoreceptors of eyes in the terrestrial slug Limax are the typical invertebrate-type but unique in that three visual opsins (Gq-coupled rhodopsin, xenopsin, Opn5A) and one retinochrome, all belonging to different groups, are co-expressed. However, molecular properties including spectral sensitivity and G protein selectivity of any of them are not determined, which prevents us from understanding an advantage of multiplicity of opsin properties in a single rhabdomeric photoreceptor. To gain insight into the functional role of the co-expression of multiple opsin species in a photoreceptor, we investigated the molecular properties of the visual opsins in the present study. RESULTS: First, we found that the fourth member of visual opsins, Opn5B, is also co-expressed in the rhabdomere of the photoreceptor together with previously identified three opsins. The photoreceptors were also demonstrated to express Gq and Go alpha subunits. We then determined the spectral sensitivity of the four visual opsins using biochemical and spectroscopic methods. Gq-coupled rhodopsin and xenopsin exhibit maximum sensitivity at ~ 456 and 475 nm, respectively, and Opn5A and Opn5B exhibit maximum sensitivity at ~ 500 and 470 nm, respectively, with significant UV sensitivity. Notably, in vitro experiments revealed that Go alpha was activated by all four visual opsins, in contrast to the specific activation of Gq alpha by Gq-coupled rhodopsin, suggesting that the eye photoreceptor of Limax uses complex G protein signaling pathways. CONCLUSIONS: The eye photoreceptor in Limax expresses as many as four different visual opsin species belonging to three distinct classes. The combination of opsins with different spectral sensitivities and G protein selectivities may underlie physiological properties of the ocular photoreception, such as a shift in spectral sensitivity between dark- and light-adapted states. This may be allowed by adjustment of the relative contribution of the four opsins without neural networks, enabling a simple strategy for fine-tuning of vision.

Compensation mechanism for membrane potential against hypoosmotic stress in the Onchidium neuron.

The effect of acute hypoosmotic stress on the neural response was investigated using the neurons identified in the abdominal ganglion of the amphibious mollusk Onchidium. The membrane potential of an identified neuron (Ip-1/2) was not significantly altered in 50% hypoosmotic artificial sea water. In isotonic 50% artificial seawater (ASW) with osmolarity that was compensated for using glycerol or urea, the membrane potentials of Ip-1/2 were also not altered compared to those in 50% hypoosmotic ASW. However, hyperpolarization was induced in isotonic 50% ASW when osmolarity was compensated for using sucrose or mannose. In the presence of volume-regulated anion channel (VRAC) inhibitors (niflumic acid and glibenclamide), the Ip-1/2 membrane potentials were hyperpolarized in 50% hypoosmotic ASW. These results suggest that there is a compensatory mechanism involving aquaglyceroporin and VRAC-like channels that maintains membrane potential under hypoosmotic conditions. Here, we detected the expression of aquaglyceroporin mRNA in neural tissues of Onchidium.

Integration of ocular and non-ocular photosensory information in the brain of the terrestrial slug Limax.

In the terrestrial slugs Limax, most of the photosensory information is thought to be acquired by an eye located on the superior tentacles, by which the slugs avoid light. Recent studies, however, suggested that the brain also plays a role as a photosensor in their negative phototaxis behavior. In the present study, we investigated how the photosensory information acquired by the eye and brain is integrated. The visual pathway in the brain was traced by incorporating tracer molecules from the cut end of an optic nerve, and commissural interactions were found in optic neuropiles located in the lateral regions of the cerebral ganglia. A cluster of neuronal cell bodies located near the dorsal surface of the cerebral ganglion had connections with the contralateral optic neuropile via gap junctions. Some of these neuronal cell bodies were Opn5A-immunoreactive, and contained numerous photic vesicle-like structures. Light-induced spikes were recorded extracellularly from the dorsal surface of these neuronal clusters, and they were synchronous with the spikes recorded from the cut end of the cerebral commissure. This study suggests that both the light information from the eye and the contralateral cerebral ganglion are integrated in the optic neuropile.

Distribution and physiological effect of enterin neuropeptides in the olfactory centers of the terrestrial slug Limax.

In gastropods, the function of neuropeptides has been studied primarily in the peripheral motor systems. Their functional roles in the central nervous system have received less attention. The procerebrum, the secondary olfactory center of the terrestrial slug Limax, consists of several hundred thousand interneurons, and plays a pivotal role in olfactory learning and memory. In the present study, we found that enterin, known as a myoactive peptide functioning in the enteric and vascular system of Aplysia, is expressed in the procerebrum of Limax. These enterin-expressing neurons primarily make projections within the cell mass layer of the procerebrum. The oscillatory frequency of the local field potential in the procerebrum was reduced by an exogenous application of enterin. The local field potential oscillation in the tentacular ganglion, the primary olfactory center, was also modulated by enterin. Whole-cell patch-clamp recordings revealed that the modulatory effect in the procerebrum was due to the inhibitory effect of enterin on the bursting neurons, which function as the kernels determining the oscillatory activity of the procerebrum. Our results revealed the novel role of the myoactive neuropeptide enterin in the higher olfactory function in terrestrial gastropods.

Light avoidance by a non-ocular photosensing system in the terrestrial slug Limax valentianus.

Although the eye is the best-studied photoreceptive organ in animals, the presence of non-ocular photosensing systems has been reported in numerous animal species. However, most of the roles that non-ocular photosensory systems play remain elusive. We found that the terrestrial slug Limax valentianus avoids light and escapes into dark areas even if it is blinded by the removal of the bilateral superior tentacle. The escape behaviour was more evident for short-wavelength light. Illumination to the head with blue but not red light elicited avoidance behaviour in the blinded slugs. Illumination to the tail was ineffective. The light-avoidance behaviour of the blinded slugs was not affected by the removal of the penis, which lies on the brain in the head, suggesting that the penis is dispensable for sensing light in the blinded slug. mRNA of Opn5A, xenopsin, retinochrome and, to a lesser extent, rhodopsin was expressed in the brain according to RT-PCR. Light-evoked neural responses were recorded from the left cerebro-pleural connective of the isolated suboesophageal ganglia of the brain, revealing that the brain is sensitive to short wavelengths of light (400-480 nm). This result is largely consistent with the wavelength dependency of the light-avoidance behaviour of the blinded slugs that we observed in the present study. Our results strongly support that the terrestrial slug L. valentianus detects and avoids light by using its brain as a light-sensing organ in the absence of eyes.

C-Arm Cone Beam Computed Tomography Guidance for Radiofrequency Ablation in Hepatocellular Carcinoma.

OBJECTIVE: To assess the usefulness of C-arm cone beam computed tomography (CBCT) combined with ultrasound for the treatment of hepatocellular carcinoma (HCC) by radiofrequency ablation (RFA). METHODS: Patients underwent RFA following transcatheter arterial chemoembolization (TACE) or RFA alone under ultrasound or CBCT guidance combined with ultrasound-based techniques. They were divided into 2 groups based on the use (C group) and nonuse (NC group) of CBCT guidance. The technical success of RFA and local tumor progression after the first RFA session were evaluated by dynamic contrast-enhanced imaging methods. Between-group differences were assessed retrospectively. RESULTS: We enrolled 198 patients with 260 HCC nodules. The complete ablation rates were 63.0 and 89.4% in the NC and C groups, respectively. In log-rank testing, local tumor progression occurred significantly more often in the NC group when RFA was used without TACE, in males when des-gamma-carboxy prothrombin was ≥29 mAU/mL, and when the diameter of a nodule was ≥18 mm. On Cox proportional-hazards regression analysis, the NC group, RFA alone without TACE, and male gender were significant independent variables. CONCLUSION: TACE followed by RFA under CBCT and ultrasound guidance improves the reliability of ablation of target HCC nodules, reduces the need for additional treatment sessions, and prevents local tumor progression.

Expression and light-dependent translocation of ß-arrestin in the visual system of the terrestrial slug Limax valentianus.

Vertebrates, cephalopods and arthropods are equipped with eyes that have the highest spatiotemporal resolution among the animal phyla. In parallel, only animals in these three phyla have visual arrestin specialized for the termination of visual signaling triggered by opsin, in addition to ubiquitously expressed ß-arrestin that serves in terminating general G protein-coupled receptor signaling. Indeed, visual arrestin in Drosophila and rodents translocates to the opsin-rich subcellular region in response to light to reduce the overall sensitivity of photoreceptors in an illuminated environment (i.e. light adaptation). We thus hypothesized that, during evolution, visual arrestin has taken over the role of ß-arrestin in those animals with eyes of high spatiotemporal resolution. If this is true, it is expected that ß-arrestin plays a role similar to visual arrestin in those animals with low-resolution eyes. In the present study, we focused on the terrestrial mollusk Limax valentianus, a species related to cephalopods but that has only ß-arrestin, and generated antibodies against ß-arrestin. We found that ß-arrestin is highly expressed in photosensory neurons, and translocates into the microvilli of the rhabdomere within 30 min in response to short wavelength light (400 nm), to which the Limax eye exhibits a robust response. These observations suggest that ß-arrestin functions in the visual system of those animals that do not have visual arrestin. We also exploited anti-ß-arrestin antibody to visualize the optic nerve projecting to the brain, and demonstrated its usefulness for tracing a visual ascending pathway.

Context-Dependent Passive Avoidance Learning in the Terrestrial Slug Limax.

The terrestrial slug Limax has been used as a model animal for studying the neural mechanisms underlying associative olfactory learning. The slug also innately exhibits negative phototactic behavior using its eyes. In the present study, we developed an experimental paradigm for quantification of slug's negative phototaxis behavior, and investigated whether the nature of the negative phototaxis can be modified by learning experience. The experimental set-up consists of light and dark compartments, between which the slug can move freely. During conditioning, the slug was placed in the light compartment, and an aversive stimulus (quinidine sulfate solution) was applied when it reached the dark compartment. After a single conditioning session, the time to reach the dark compartment significantly increased when it was tested following 24 hr or one week. Protein synthesis inhibition immediately following the conditioning impaired the memory retention at one week but not at 24 hr. The retrieval of the memory was context-dependent, as the time to reach the dark compartment did not significantly increase if the slug was placed on a floor with a different texture in the memory retention test. If the aversive stimulus was applied when the slug was in the light compartment, the time to reach the dark compartment did not increase after 24 hr. This is the first report demonstrating the capability of the slug to form context-dependent passive avoidance memory that can be established in a single conditioning session.

Olfactory Memory Storage and/or Retrieval Requires the Presence of the Exact Tentacle Used During Memory Acquisition in the Terrestrial Slug Limax.

Terrestrial pulmonates can form odor-aversion memories once a food odor is presented in combination with an aversive stimulus. Most of the olfactory information ascends via a tentacular ganglion located in the tip of the two pairs of tentacles, and is then transmitted to the higher olfactory center, the procerebrum. The procerebrum is the locus of memory storage and has been shown to be necessary for odor-aversion learning. However, it is unknown whether the procerebrum is the sole locus in which the memory engram resides. By exploiting the regenerative ability of tentacles, here we investigated whether tentacles function merely in transmitting olfactory information to the procerebrum, or constitute a part of the memory engram. We showed that after removal of the tentacles used during memory acquisition, slugs were unable to retrieve the memory, even if these tentacles were regenerated sufficiently to subserve memory function. Our results support the view that tentacles are more than conduits of odor information; they also participate in the formation of the memory engram.

Photo-tropotaxis based on projection through the cerebral commissure in the terrestrial slug Limax.

In the terrestrial slug, Limax, eyes are located at the tip of the superior tentacles. This animal has long been believed to show negative phototaxis through tropotaxis, i.e., it compares the two light intensities detected by bilateral eyes to move away from a light source. As one of the possible manifestations of such negative phototaxis, a circling movement has been observed: if one of the superior tentacles is removed, the slugs continuously move in the direction of the removed side. However, there has been no evidence demonstrating that this behavior is actually based on negative phototropotaxis. In this study, we showed that the slugs do not exhibit the circling behavior in the absence of light, and that amputation of the cerebral commissure also diminishes the circling behavior under light. We could detect light-evoked responses during electrical recording from the cut edge of the cerebral commissure. Labeling of the optic nerve with neurobiotin also revealed the presence of the commissural fibers that potentially transmit the light information to the contralateral cerebral ganglion. Our study suggests that the slug's circling behavior is based on phototropotaxis in which the light intensities detected by the bilateral eyes are compared through the cerebral commissure.

Visual afferents from an eye in the terrestrial slug Limax valentianus.

Terrestrial gastropods have a lens-bearing eye on the tip of their tentacles. There are two morphologically distinct photoreceptors, called Type-I and Type-II photoreceptors, in the retina. Type-I photoreceptors are equipped with highly developed photoreceptive microvilli in their outer rhabdomeric segment, whereas Type-II photoreceptors have short and fewer microvilli. Although both types of photoreceptors send afferent projections directly to the brain, their destinations in the brain, called optic neuropiles, have not been sufficiently investigated. Our recent studies revealed that there are commissural fibers in the cerebral ganglia that transmit photic information acquired by bilateral eyes. Moreover, some of the retinal photoreceptors are connected by gap junctions to the photosensitive brain neurons, suggesting the functional interaction of the photic information between the eye and brain photoreceptors, as well as between bilateral eyes. However, it has not been clarified which type of retinal photoreceptors send commissural projections to the contralateral hemiganglion nor interact with the brain photoreceptors. In the present study, we demonstrated by molecular histological analyses and tracer injections that (1) Type-I and Type-II photoreceptors send glutamatergic afferent projections to the medial and lateral lobes of the ipsilateral optic neuropile, respectively, (2) direct synaptic interaction between bilateral optic nerves occurs in the medial lobe of the optic neuropile, and (3) brain photosensory neurons form gap junctions with the medial lobe of the contralateral optic neuropile. These results reveal an ordered pattern of afferent projections from the retina and provide insight into the different functional roles of retinal photoreceptors.

DNA endoreplication in the brain neurons during body growth of an adult slug.

Endoreplication is DNA synthesis without cell division. Giant neurons observed in the brains of mollusks are thought to be generated as a result of DNA endoreplication. It has been hypothesized that neuronal size becomes larger in parallel with an increase in body size and that DNA endoreplication is involved in this process to meet the increasing demand for macromolecules in neurons. There is, however, no experimental evidence for this hypothesis to date. In the present study, we investigated the following quantitatively: (1) the size of the brain and each ganglion, (2) the size of identified neurons, (3) the total number of neurons undergoing DNA endoreplication, (4) the total number of the neurons containing a cardioexcitatory peptide, and (5) the gene expression level per neuron, using terrestrial slugs whose body growth was regulated through the amount of food supplied in the laboratory. The body growth was accompanied by increases in the sizes of both neurons and ganglia and triggered more frequent DNA endoreplication events in each ganglion of the growth-promoted slugs, without increasing the total number of neurons. Increase in the neuronal size also involved the increase in the amount of transcripts expressed in a single neuron. This is the first quantitative evidence showing that the DNA endoreplication, neuronal size, and gene expression are increased concomitantly with body growth in adult mollusks.

Excitatory effects of GABA on procerebrum neurons in a slug.

Classical neurotransmitters, such as glutamate and γ-aminobutyric acid (GABA), often have different actions on invertebrate neurons from those reported for vertebrate neurons. In the terrestrial mollusk Limax, glutamate was found to function as an inhibitory transmitter in the procerebrum (PC), but it has not yet been clarified how GABA acts in the PC. We thus examined what effects GABA exerts on PC neurons in the present study. For this purpose, we first applied GABA to isolated PC preparations and recorded postsynaptic currents and potentials in PC neurons. The GABA application reduced the amplitude of inhibitory postsynaptic currents and depolarization-induced outward currents recorded in nonbursting neurons and increased the number of spontaneous spikes of nonbursting neurons. However, direct GABA-induced currents were not observed in either bursting or nonbursting neurons. These results suggest a potential direct effect of GABA on outward currents resulting in enhanced excitability of PC neurons. Next, we measured the change in [Ca(2+)](i) in cultured PC neurons by application of GABA. The GABA application increased spontaneous Ca(2+) events in cultured neurons. These Ca(2+) events were ascribable to the influx of extracellular Ca(2+). We then confirmed the presence of GABA and GABA receptors in the PC. The GABA-like immunoreactivity was observed in the neuropil layers of the PC, and the mRNAs for both GABA(A) and GABA(B) receptors were expressed in the PC. In particular, GABA(B) receptor mRNA, rather than GABA(A), was found to be more abundantly expressed in the PC. These results suggest that GABA functions as an excitatory modulator for PC neurons via mainly GABA(B) receptors.

Whole genome amplification in large neurons of the terrestrial slug Limax.

The brain of gastropod mollusks contains giant neurons whose nuclei are enlarged with a large amount of genomic DNA. Such DNA is produced by repeated endoreplication. We have previously demonstrated that the frequency of the neuronal DNA endoreplication is correlative to the body growth of the adult land slug and to the increase in the amount of transcripts within the neuron. However, it has long been controversial whether the neuronal DNA endoreplication entails whole genome amplification (polyploidy), or whether only the necessary genomic loci are amplified (polyteny, polysomy, or cis-amplification by unequal recombination). In the present study, we adopted two modern techniques - quantitative genomic PCR and 5'-bromodeoxyuridine labeling - to distinguish between these two possibilities. Our results demonstrated that multiple genomic loci were amplified to the same extent irrespective of the transcriptional activities at these loci. Moreover, the visceral giant cell, the biggest neuron in the slug's brain, was estimated to contain approximately 10 000-times as much genomic DNA as the haploid amount. The 5'-bromodeoxyuridine-labeling experiments also revealed a uniform DNA synthesis within the nucleus. These results strongly support the idea that the giant neurons contain a polyploid genome rather than a locus-specific amplified genome.

[Laparoscopy-assisted ventriculoperitoneal and lumboperitoneal shunt surgery].

Recently, laparoscopy (also referred to as minimally invasive surgery) has been used during peritoneal catheter implantation in shunt placement for hydrocephalus; however, the procedure and devices for this technique have not yet been well established. We adopted umbilical and paraumbilical laparoscopy for peritoneal catheter insertion. In this paper, we describe the technique we used and its clinical results and benefits. Ten consecutive patients with hydrocephalus who underwent laparoscopic shunt surgery (6 cases of ventriculoperitoneal shunt and 4 of lumboperitoneal shunt) were enrolled for this study. The follow-up period ranged from 21 to 434 days (mean, 263 days). After a standard cranial/spinal procedure, an approximately 5-mm incision was made in the lateral side of the umbilicus, where the abdominal catheter was introduced subcutaneously. Thereafter, we inserted a laparoscope into the peritoneal cavity via a small incision beneath or just on the umbilicus. A shunt catheter was laparoscopically inserted through a peel-off cannula and placed after taking note of the outflow of cerebrospinal fluid (CSF) from the catheter tip. In all patients, the shunt was inserted with no complications, and good patency was achieved. Laparoscopy allows implantation of the catheter into the peritoneal cavity, and the outflow of CSF can be confirmed intraoperatively. Furthermore, the abdominal surgical wounds are minimal, even for obese patients, and fascia/muscle incisions are not needed. Laparoscopy-assisted shunt surgery for hydrocephalus is effective and safe and also has cosmetic advantages.

Regional expression of neuropeptides in the retina of the terrestrial slug Limax valentianus (Gastropoda, Stylommatophora, Limacidae).

Gastropods use lens-bearing eyes to detect ambient light. The retina contains photoreceptors that directly project to the brain. Here, we identified the neurotransmitters that the retinal cells use for projection to the brain in the terrestrial slug Limax. We identified 12 genes encoding neuropeptides as well as a novel vesicular glutamate transporter, a marker of glutamatergic neuron, expressed in the retinal cells. Spatial expression profiles of the neuropeptide genes were determined by in situ hybridization. WWamide/MIP1/Pedal peptide2 were coexpressed in the neurons of the accessory retina. In the main retina, prohormone-4 was expressed in the ventrolateral region. Clionin was expressed in the ventromedial region. Pedal peptide was expressed in the anterior region of the main retina and in the accessory retina. Enterin was expressed in many neurons, including the accessory retina, but not in the dorsal region. FxRIamide1 and 2 were coexpressed in the posterior region. Prohormone-4 variant was uniformly expressed in many neurons but scarcely in the accessory retina. MIP2 was widely expressed throughout the dorsoventral axis in the posteriolateral region of the main retina. Myo1 was expressed in many neurons of the main retina but predominantly in the dorsal region. These expression patterns were confirmed by immunohistochemistry with specific antibodies against the neuropeptides. Projections of these peptidergic retinal neurons were confirmed by immunostaining of the optic nerve. Our present study revealed regional differentiation of the retina with respect to the neurotransmitters that the retinal cells use.

Deterioration of postural control due to the increase of similarity between center of pressure and smooth-pursuit eye movements during standing on one leg.

Upright postural control is regulated by afferent and efferent/reafferent visual mechanisms. There are two types of efferent and conjugate eye movements: saccades and smooth pursuits. Although postural control is improved by saccades, the effects of smooth pursuits on postural control are still debated, because the difficulties of postural and visual tasks differ in the previous research. Additionally, the mechanisms that interfere with postural control and smooth pursuit are not fully understood. To address these issues, we examined the effects of different patterns of smooth-pursuit eye movement on the path length of the center of pressure (COP) displacement under bipedal and unipedal standing conditions. The relative frequency and amplitude of the COP displacement were remarkably increased when uniform linear visual targets were presented during unipedal standing. In addition, dynamic time warping analysis demonstrated that the similarity between the displacement of the COP and eye movements was increased by the presentation of uniform linear visual targets with orientation selectivity during unipedal standing but not during bipedal standing. In contrast, the attenuation of similarity between the displacement of the COP and eye movements significantly decreased the path length, relative frequency, and amplitude of the COP displacement. Our results indicate that postural stability is deteriorated by the increase of similarity between the displacement of the COP and smooth-pursuit eye movements under unstable conditions.

Opsins in the Cephalic and Extracephalic Photoreceptors in the Marine Gastropod Onchidium verruculatum.

AbstractThe marine gastropod Onchidium verruculatum has a pair of ocular photoreceptors, the stalk eyes, on the tip of its stalk near the head, as well as several extracephalic photosensory organs. The retinas of the stalk eye consist of two morphologically distinct visual cells, namely, the type I cells equipped with well-developed microvilli and the type II cells with less developed microvilli. The extracephalic photosensors comprise the dorsal eye, dermal photoreceptor, and brain photosensitive neurons. The characteristics of these cephalic and extracephalic photosensory organs have been studied from morphological and electrophysiological perspectives. However, little is known about the visual pigment molecules responsible for light detection in these organs. In the present study, we searched for opsin molecules that are expressed in the neural tissues of Onchidium and identified six putative signaling-competent opsin species, including Xenopsin1, Xenopsin2, Gq-coupled rhodopsin1, Gq-coupled rhodopsin2, Opsin-5B, and Gq-coupled rhodopsin-like. Immunohistochemical staining of four of the six opsins revealed that Xenopsin1, Gq-coupled rhodopsin1, and Gq-coupled rhodopsin2 are expressed in the rhabdomere of the stalk eye and in the dermal photoreceptor. Xenopsin2 was expressed in the type II photoreceptors of the stalk eye and in the ciliary photoreceptors of the dorsal eye. These immunohistochemical data were consistent with the results of the expression analysis, revealed by quantitative reverse transcription polymerase chain reaction. This study clarified the identities of opsins expressed in the extracephalic photosensory organs of Onchidium and the distinct molecular compositions among the photoreceptors.

Platelet administration via the portal vein promotes liver regeneration in rats after 70% hepatectomy.

OBJECTIVE: This study examines the applicability of platelet infusion therapy for liver regeneration in vivo. BACKGROUND: We recently reported that platelets accumulate in the liver immediately after extended hepatectomy and promote residual liver regeneration. Liver regeneration depends on the number of accumulated platelets in the sinusoids. METHODS: Male Sprague-Dawley rats underwent 70% hepatectomy and were then assigned to groups that were infused with 1 mL of either platelet-rich plasma (PRP; 1 × 10(9) platelets/mL) in normal saline (NS) or NS via the portal vein. We then analyzed liver regeneration and the signaling pathways that are related to liver regeneration and function. The dynamics of platelets infused via the portal vein were visualized before and after hepatectomy. RESULTS: The liver/body weight ratio after 70% hepatectomy was significantly higher and the Ki-67 labeling index was higher in the PRP, than in the NS group. The Akt pathway was activated earlier in the PRP, than in the NS group with concurrent ERK1/2 pathway activation, but this was prolonged in the PRP group. Many more platelets infused via the portal vein accumulated in the sinusoid after 70% hepatectomy, and serum liver function tests and histological findings revealed that portal infusion did not cause liver damage. CONCLUSIONS: Platelets infused via the portal vein promoted liver regeneration after 70% hepatectomy in rats without liver damage. These findings indicate that PRP administration could be a useful part of liver regeneration therapy.