Quadrupedal Walking with the Skin: The Ambulatory Flaps in "Walking" Cuttlefish (Paintpot Cuttlefish, Metasepia tullbergi).

AbstractThe locomotion strategy of cephalopods is an important factor that influences their ability to exploit various oceanic environments. Particularly, Metasepia cuttlefish have a unique locomotion strategy; they prefer slow walking (ambling) on the seafloor over swimming. For this locomotion, they use their ventral arms as forelimbs and ambulatory flaps as hindlimbs. This locomotion is similar to the gait of quadruped vertebrates, where the forelimbs and hindlimbs on the left and right move alternately. The original description and some textbooks have considered these flaps to be muscular; however, this has not been proven. Here, we report the histological morphology of the ambulatory flaps of Metasepia tullbergi and their ambling locomotion. Histological observations indicated that the ambulatory flaps had a papillae structure comprising papillae musculature (dermal erector or retractor muscles) and connective tissue in the skin. Behavioral observations indicated that the ambulatory flaps changed their shape during ambling, which could explain the existence of the skin papillae. Our results suggest that ambulatory flaps are skin papillae, which can change shape by using their papillae musculature and connective tissue. This is a unique feature of Metasepia species that use the skin papillae for locomotion.

Suppression of colonic oxidative stress caused by chronic ethanol administration and attenuation of ethanol-induced colitis and gut leakiness by oral administration of sesaminol in mice.

Chronic consumption of excess ethanol is one of the major risk factors for colorectal cancer (CRC), and the pathogenesis of ethanol-related CRC (ER-CRC) involves ethanol-induced oxidative-stress and inflammation in the colon and rectum, as well as gut leakiness. In this study, we hypothesised that oral administration of sesaminol, a sesame lignan, lowers the risk of ER-CRC because we found that it is a strong antioxidant with very low prooxidant activity. This hypothesis was examined using a mouse model, in which 2.0% v/v ethanol was administered ad libitum for 2 weeks with or without oral gavage with sesaminol (2.5 mg per day). Oral sesaminol administration suppressed the ethanol-induced colonic lesions and the ethanol-induced elevation of the colonic levels of oxidative stress markers (8-hydroxy-2'-deoxyguanosine, malondialdehyde, and 4-hydroxyalkenals). It consistently suppressed the chronic ethanol-induced expressions of cytochrome P450-2E1 and inducible nitric oxide synthase and upregulated heme oxygenase-1 expression, probably via the nuclear factor erythroid-derived 2-like 2 pathway in the mouse colon. Oral sesaminol administration also suppressed the chronic ethanol-induced elevation of colonic inflammation marker levels, such as those of tumour necrosis factor-α, interleukin-6, and monocyte chemoattractant protein-1, probably via the nuclear factor-kappa B pathway. Moreover, it prevented the chronic ethanol-induced gut leakiness by restoring tight junction proteins, giving rise to lower plasma endotoxin levels compared with those of ethanol-administered mice. All of these results suggest that dietary supplementation of sesaminol may lower the risk of ER-CRC by suppressing each of the above-mentioned steps in ER-CRC pathogenesis.

Sesaminol prevents Parkinson's disease by activating the Nrf2-ARE signaling pathway.

Parkinson's disease (PD) is a neurodegenerative disease caused by the degeneration of substantia nigra neurons due to oxidative stress. Sesaminol has strong antioxidant and anti-cancer effects. We investigated the preventive effect on PD as a new physiological action of sesaminol produced from sesaminol glycoside using in vitro and in vivo PD models. To prepare an in vitro PD model, 6-hydroxydopamine (6-OHDA) was added to human neuroblastoma (SH-SY5Y cells). The viability of SH-SY5Y cells decreased dose-dependently following 6-OHDA treatment, but the addition of sesaminol restored viability to the control level. 6-OHDA increased intracellular reactive oxygen species production, and the addition of sesaminol significantly suppressed this increase. No Nrf2 expression in the nucleus was observed in the control group, but a slight increase was observed in the 6-OHDA group. The sesaminol group showed strong expression of Nrf2 in the cytoplasm and nucleus. NAD(P)H: quinone oxidoreductase (NQO1) activity was enhanced in the 6-OHDA group and further enhanced in the sesaminol group. Furthermore, the neurotoxine rotenone was orally administrated to mice to prepare an in vivo PD model. The motor function of rotenone-treated mice was shorter than that of the control group, but a small amount of sesaminol restored it to the control level. The intestinal motility in the rotenone group was significantly lower than that in the control group, but it remained at the control level in the sesaminol group. The expression of α-synuclein in the substantia nigra increased in the rotenone group but decreased in the sesaminol group. The rotenone group exhibited shortening and damage to the colonic mucosa, but these abnormalities of the colonic mucosa were scarcely observed in the sesaminol group. These results suggest that sesaminol has a preventative effect on PD.

Comparative Anatomy of Trunk Musculature in Salamandridae with Different Habitats.

The diversity of trunk muscle morphology of Salamandridae occupying different habitats (aquatic: Pachytriton labiatus; terrestrial: Tylototriton kweichowensis and Salamandra salamandra salamandra) was examined. Trunk muscles were dissected, and muscle weight ratios were quantified. The terrestrial species have larger abdominal trunk muscles than the aquatic species do. In contrast, the lateral hypaxial muscles are larger in the aquatic species. The M. rectus abdominis profundus is located subjacent to the M. rectus abdominis in the terrestrial species. In the aquatic species, the ventral muscle is composed of the M. rectus abdominis alone. The lateral hypaxial muscles in the aquatic species are suited to lateral bending during underwater locomotion in the denser aquatic medium. Larger abdominal muscles may be used in supporting body weight against gravity in the terrestrial species. The function of the M. rectus abdominis profundus may be to support the M. rectus abdominis in the terrestrial species. These findings suggest a divergent evolution of trunk muscle characteristics within the Salamandridae, which correlate with both habitats and modes of locomotion.

The functional-morphological adaptive strategy of digestive organs of decapodiform cephalopods.

The digestive organs in decapodiform cephalopod species morphologically vary by individual lifestyle. We examined the following six species of adult decapodiformes cephalopods representing different habitats: Todarodes pacificus, Loligo bleekeri, Loligo edulis, Watasenia scintillans (pelagic), Sepia lycidas and Euprymna morsei (benthic). L. bleekeri and L. edulis possess a bursiform cecal sac connected to the cecum. Pelagic species have a single digestive gland smaller than in benthic species. T. pacificus has an oval digestive gland larger than that of L. bleekeri and L. edulis, which possess withered-looking and smaller digestive glands. In contrast, the digestive glands in benthic species are paired. S. lycidas and E. morsei have well-developed and larger digestive glands than those of the pelagic species. Well-developed digestive duct appendages are found in benthic species. In qualification of the mass of digestive organs, pelagic species have smaller stomachs, digestive glands and digestive ducts' appendages than benthic species. Because pelagic species need to swim, they may possess smaller stomachs and larger cecums for more rapid digestion. A smaller digestive gland may have the advantage of reducing the body weight in pelagic species for rapid swimming. In contrast, since benthic species require a longer time for digestion than pelagic species, they compact more food in their stomachs and absorb nutrients via more organs, such as the digestive grand and digestive duct appendages, in addition to cecum.

Positional strategy of trunk muscles among aquatic, semi-aquatic and terrestrial species in Urodela.

Clarification of the trunk structure in Urodela is important in understanding the locomotive evolution of basal tetrapods. The components of the muscular trunk wall among Urodela using different modes of locomotion were compared. Since the whole trunk may be used for swimming and the effect of limbs may be small in the more aquatic species, they showed smaller differences in the trunk muscles among anterior, middle and posterior sections of the trunk. By contrast, in the more terrestrial species, the dorsal and abdominal muscles are larger in the middle section than those in the anterior and posterior sections. High compressive stresses occur in the supporting limbs and their insertion at the trunk on the ventral side, and spread from the forelimbs along the back to the supporting hindlimbs on the dorsal side. Tensile stresses occur in the middle ventral part. The components of the trunk muscles among the three sections may reflect differences in stresses occurring in the trunk of the more terrestrial species. The findings also suggest that in the middle section, larger dorsal muscles for stiffening the back to maintain posture and larger abdominal muscles are responsible for balancing the body weight while it is supported by the limbs in the more terrestrial species.

Locomotion pattern and trunk musculoskeletal architecture among Urodela.

We comparatively examined the trunk musculature and prezygapophyseal angle of mid-trunk vertebra in eight urodele species with different locomotive modes (aquatic Siren intermedia, Amphiuma tridactylum, Necturus maculosus and Andrias japonicus; semi-aquatic Cynops pyrrhogaster, Cynops ensicauda; and terrestrial Hynobius nigrescens, Hynobius lichenatus and Ambystoma tigrinum). We found that the more terrestrial species were characterized by larger dorsal and abdominal muscle weight ratios compared with those of the more aquatic species, whereas muscle ratios of the lateral hypaxial musculature were larger in the more aquatic species. The lateral hypaxial muscles were thicker in the more aquatic species, whereas the M. rectus abdominis was more differentiated in the more terrestrial species. Our results suggest that larger lateral hypaxial muscles function for lateral bending during underwater locomotion in aquatic species. Larger dorsalis and abdominal muscles facilitate resistance against sagittal extension of the trunk, stabilization and support of the ventral contour line against gravity in terrestrial species. The more aquatic species possessed a more horizontal prezygapophyseal angle for more flexible lateral locomotion. In contrast, the more terrestrial species have an increasingly vertical prezygapophyseal angle to provide stronger column support against gravity. Thus, we conclude trunk structure in urodeles differs clearly according to their locomotive modes.

Ontogenetic changes of trunk muscle structure in the Japanese black salamander (Hynobius nigrescens).

We investigated ontogenetic changes in the trunk muscles of the Japanese black salamander (Hynobius nigrescens) before, during and after metamorphosis. Given that amphibians change their locomotive patterns with metamorphosis, we hypothesized that they may also change the structure of their trunk muscles. The trunk muscles were macroscopically observed, and the weight ratios of each trunk muscle group were quantified at six different developmental stages. Immediately after hatching, we found that the lateral hypaxial muscle was composed of one thick M. ventralis, from ventral edge of which M. transversus abdominis arose later, followed by M. obliquus externus and M. rectus abdominis. The weight ratios of the dorsal and abdominal muscles to the trunk muscles increased with growth. We suggest that a single thick and large lateral hypaxial muscle facilitates swimming during early developmental stages. The increase in the weight ratios of the dorsal and abdominal muscles with growth possibly assists with gravity resistance necessary for terrestrial life.

Functional morphology and comparative anatomy of appendicular musculature in Cuban Anolis lizards with different locomotor habits.

We examined the diversity of the musculoskeletal morphology in the limbs of Anolis lizards with different habitats and identified variations in functional and morphological adaptations to different ecologies or behaviors. Dissection and isolation of 40 muscles from the fore- and hindlimbs of five species of Anolis were performed, and the muscle mass and length of the moment arm were compared after body size effects were removed. Ecologically and behaviorally characteristic morphological differences were observed in several muscles. Well-developed hindlimb extensors were observed in ground-dwelling species, A. sagrei and A. bremeri, and were considered advantageous for running, whereas adept climber species possessed expanded femoral retractors for weight-bearing during climbing. Moreover, morphological variations were observed among arboreal species. Wider excursions of the forelimb joint characterized A. porcatus, presumably enabling branch-to-branch locomotion, while A. equestris and A. angusticeps possessed highly developed adductor muscles for grasping thick branches or twigs. These findings suggest divergent evolution of musculoskeletal characteristic in the limbs within the genus Anolis, with correlations observed among morphological traits, locomotor performance, and habitat uses.

Functional and morphological variety in trunk muscles of Urodela.

Trunk musculature in Urodela species varies by habitat. In this study, trunk musculature was examined in five species of adult salamanders representing three different habitats: aquatic species, Amphiuma tridactylum and Necturus maculosus; semi-aquatic species, Cynops pyrrhogaster; terrestrial species, Hynobius nigrescens and Ambystoma tigrinum. More terrestrial species have heavier dorsal and ventral trunk muscles than more aquatic forms. By contrast, the lateral hypaxial musculature was stronger in more aquatic species. The number of layers of lateral hypaxial musculature varied among Urodela species and did not clearly correlate with their habitats. The M. rectus abdominis was separated from the lateral hypaxial musculature in both terrestrial and semi-aquatic species. In aquatic species, M. rectus abdominis was not separated from lateral hypaxial musculature. Lateral hypaxial musculature differed in thickness among species and was relatively thinner in terrestrial species. In more terrestrial species, dorsal muscles may be used for stabilization and ventral flexing against gravity. Ventral muscle may be used in preventing dorsally concave curvature of the trunk by dorsal muscles and by weight. The lengthy trunk supported by limbs needs muscular forces along the ventral contour line in more terrestrial species. And, the locomotion on well-developed limbs seems to lead to a decrease of the lateral hypaxial musculature.

Purification, gene cloning, and biochemical characterization of a ß-glucosidase capable of hydrolyzing sesaminol triglucoside from Paenibacillus sp. KB0549.

The triglucoside of sesaminol, i.e., 2,6-O-di(ß-D-glucopyranosyl)-ß-D- glucopyranosylsesaminol (STG), occurs abundantly in sesame seeds and sesame oil cake and serves as an inexpensive source for the industrial production of sesaminol, an anti-oxidant that displays a number of bioactivities beneficial to human health. However, STG has been shown to be highly resistant to the action of ß-glucosidases, in part due to its branched-chain glycon structure, and these circumstances hampered the efficient utilization of STG. We found that a strain (KB0549) of the genus Paenibacillus produced a novel enzyme capable of efficiently hydrolyzing STG. This enzyme, termed PSTG, was a tetrameric protein consisting of identical subunits with an approximate molecular mass of 80 kDa. The PSTG gene was cloned on the basis of the partial amino acid sequences of the purified enzyme. Sequence comparison showed that the enzyme belonged to the glycoside hydrolase family 3, with significant similarities to the Paenibacillus glucocerebrosidase (63% identity) and to Bgl3B of Thermotoga neapolitana (37% identity). The recombinant enzyme (rPSTG) was highly specific for ß-glucosidic linkage, and k cat and k cat/K m values for the rPSTG-catalyzed hydrolysis of p-nitrophenyl-ß-glucopyraniside at 37°C and pH 6.5 were 44 s(-1) and 426 s(-1) mM(-1), respectively. The specificity analyses also revealed that the enzyme acted more efficiently on sophorose than on cellobiose and gentiobiose. Thus, rPSTG is the first example of a ß-glucosidase with higher reactivity for ß-1,2-glucosidic linkage than for ß-1,4- and ß-1,6-glucosidic linkages, as far as could be ascertained. This unique specificity is, at least in part, responsible for the enzyme's ability to efficiently decompose STG.