Efeitos da hipóxia-isquemia pré-natal durante o desenvolvimento: receptores e transportadores glutamatérgicos e comunicação celular in vitro

O cérebro infantil humano submetido à hipóxia-isquemia (HI) apresenta perda de oligodendrócitos, hipomielinização, astrogliose, alterações no desenvolvimento cortical e no comportamento motor, incluindo a paralisia cerebral. O cerebelo desempenha um importante papel no controle motor e diversos danos vêm sendo observados em humanos e animais que sofreram HI. A excitotoxicidade glutamatérgica é frequentemente associada à HI e junções celulares podem ser responsáveis pela transferência de moléculas capazes de modular os danos decorrentes. Dados prévios de nosso grupo utilizando um modelo de HI pré-natal em ratos demonstraram danos permanentes na estrutura cerebelar, indicando que os efeitos deletérios da HI pré-natal podem ser mantidos até a vida adulta. O objetivo deste trabalho foi caracterizar os níveis de conexinas, receptores e transportadores de glutamato ao longo do desenvolvimento do cerebelo HI, e avaliar a configuração das junções celulares em culturas de astrócitos derivadas do cerebelo de ratos submetidos a esse modelo. Ratas no 18 dia de gestação, após anestesia, tiveram as quatro artérias uterinas obstruídas por 45 minutos (Grupo HI). Animais controle tiveram os úteros expostos sem sofrer a obstrução (Grupo SH). A gestação prosseguiu e apenas filhotes nascidos a termo foram utilizados. Os animais foram decapitados aos 2 (P2), 9 (P9), 16 (P16),23 (P23), 30 (P30), 45 (P45) e 90 (P90) dias pós-natal. Os cerebelos foram submetidos à técnica de Western blotting utilizando os anticorpos anti-NR2B, anti-GluR3, anti-EAAT1, anti-GFAP e anti-Cx43. Para a cultura de astrócitos foram utilizados cerebelos de animais P2. Após terem atingido confluência, as células foram fixadas e imunomarcadas com os anticorpos anti-Cx43, anti-GFAP, anti-nestina e anti-A2B5. Nossos resultados demonstram diferenças nos níveis de GluR3 durante o desenvolvimento do cerebelo SH e HI, havendo uma redução significativa da expressão desta subunidade no grupo HI em P9. Por outro lado, não foram verificadas alterações nos níveis de NR2B e de GFAP entre os grupos nas diferentes idades. Observou-se redução significativa de Cx43 em animais HI em P2 bem como nos astrócitos HI em cultura, os quais também apresentaram alterações morfológicas e diferenças na expressão do marcador A2B5. A alteração referente a GluR3 no grupo HI pode ser causada pela redução da arborização das células de Purkinje e pela redução no número de precursores de oligodendrócitos no cerebelo de animais HI em P9, já observadas em nosso laboratório. A diminuição de Cx43 indica que a passagem de substâncias por canais astrocitários pode estar reduzida e contribuir para a expansão dos danos persistentes descritos em HI. Alterações morfológicas e na expressão de marcadores da diferenciação de astrócitos podem refletir os potenciais efeitos de HI sobre a maturação destas células a longo-prazo. Nossos resultados apontam que a HI sistêmica pré-natal pode ser responsável por alterações que caracterizam a excitotoxicidade glutamatérgica. Ressaltamos também a importância da comunicação entre astrócitos como estratégia neuroprotetora nesta lesão.

Distribuição da caspase-3 clivada no cerebelo de ratos durante o desenvolvimento em um modelo de hipóxia-isquemia sistêmica pré-natal

A Hipóxia-isquemia (HI) perinatal é um problema de saúde pública, e ocorrem aproximadamente 1,5 casos de encefalopatias por HI por 1000 nascidos vivos. Dos que sobrevivem 25-60% sofrem de deficiências permanentes do desenvolvimento neurológico, incluindo paralisia cerebral, convulsões, retardo mental, e dificuldade de aprender. Neurônios e oligodendrócitos, especialmente os progenitores, são os mais afetados pela HI. Existem vários modelos de HI, no entanto, poucos levam em consideração as intercorrências maternas, a importância da atividade placentária, e as trocas entre mãe-filho, que são clinicamente observadas em humanos. Robinson estabeleceu um modelo de HI sistêmica pré-natal transitório, onde o fluxo das artérias uterinas da rata grávida era obstruído por 45 minutos no décimo oitavo dia (E18) de gestação. Neste modelo foram observadas alterações que são similares às observadas em cérebros humanos que passaram por hipóxia perinatal, dentre as quais foram relatados aumento no nível de apoptose. Caspase-3 é descrita como uma enzima que atua na apoptose, e é amplamente utilizada como marcador para células apoptóticas. Vários autores vêm mostrando, entretanto, que a enzima caspase-3 pode estar ativada para fins não apoptóticos. No modelo de HI sistêmica pré-natal, foram observados astrogliose na substância branca, morte de oligodendrócitos, lesão em axônios tanto na substância branca como no córtex cerebral, e danos motores. Pouco se sabe da influencia do insulto HI no desenvolvimento do cerebelo, considerando que o cerebelo junto com o córtex motor, contribui para o controle motor. O objetivo desse trabalho foi avaliar a distribuição da caspase-3 clivada durante o desenvolvimento do cerebelo em um modelo de HI pré-natal. Os resultados deste trabalho demonstram que as células caspase-3 clivadas apresentaram duas morfologias distintas em ambos os grupos. Uma onde a caspase-3 foi observada apenas no núcleo, oscilando entre células com imunorreatividade fraca a intensa, e de células com a presença da caspase-3 no corpo celular, nos prolongamentos condensados e presença de fragmentos ao redor do soma, morfologia típica de célula em apoptose. A HI pré-natal, assim como nos hemisférios cerebrais, levou ao aumento de células caspase-3 clivadas com morfologia de progenitores de oligodendrócitos no cerebelo do grupo HI em P2, mas não em P9 e P23. Também foi demonstrado que a HI pré-natal não levou a uma ativação da apoptose em oligodendrócitos, neurônios e microglia (identificados por seus respectivos marcadores, CNPase, NeuN e ED1) apresentando marcação no núcleos de células GFAP+, na substância branca, camada granular e nas células da glia de Bergmann, em P9 e P23 no cerebelo. Podemos concluir que a HI pré-natal aumentou o número de células imunorreativas para a caspase-3 em um período crítico do desenvolvimento da oligodendroglia no cerebelo, e que a diminuição de progenitores de oligodendrócitos no cerebelo decorrente do insulto pré-natal visto em trabalhos anteriores, pode estar relacionada a morte celular por apoptose, embora não se possa descartar a hipótese da participação dessas células que apresentam caspase-3 clivada em outros eventos não apoptóticos desencadeados pela hipóxia-isquemia.

Internal models in the cerebellum.

This review will focus on the possibility that the cerebellum contains an internal model or models of the motor apparatus. Inverse internal models can provide the neural command necessary to achieve some desired trajectory. First, we review the necessity of such a model and the evidence, based on the ocular following response, that inverse models are found within the cerebellar circuitry. Forward internal models predict the consequences of actions and can be used to overcome time delays associated with feedback control. Secondly, we review the evidence that the cerebellum generates predictions using such a forward model. Finally, we review a computational model that includes multiple paired forward and inverse models and show how such an arrangement can be advantageous for motor learning and control.

Nueva especie del género Mugil (Pisces: Mugilidae) de aguas cubanas.

A new species of mullet (Mugil) from Nipe Bay. North coast of Oriente Province, Cuba. The absence of axillary scale of pectoral fins, a very slender body and a notably elongated caudal peduncle are among the most diagnostic features of a new species.

Larval ontogeny of a percichthyid fish, Synagrops philippinensis (Günther) in Kagoshima Bay, southern Japan

The larval ontogeny of a developmental series (1.2-8.3mm body length, BL) of Synagrops philippinensis from Kagoshima Bay, southern Japan is described and illustrated. The yolk was completely absorbed in larva of ≥1.5 mm BL. Notochord flexion commenced at about 3.5mm BL and was completed by about 4.0-4.5mm BL. S. philippinensis larvae were distinguished from their congeners based on melanophore patterns, head spination and fin spines and rays. Larvae of 7.5-8.3 mm BL were characterized by anteriorly serrated pelvic spine, two anal spines, nine inner preopercular spines and no melanophore on lateral side of the caudal peduncle; 7.0 to 7.5mm BL larvae by the above characters except serration on pelvic spine; and yolk-sac, pre-flexion, flexing and post-flexion larvae up to 7.0mm BL by unique melanophores on lower lobe of pectoral finfold/fin.

Fat and water distribution patterns in the flesh of the common cat-fish Wallago attu (Bl. & Schn.)

The quantitative distributional pattern of fat and water has been studied in the flesh of the various zones corresponding to the dorsal and ventral regions of the body of a common cat-fish, Wallago attu. The ventral aspect of the body showed more accumulation of fat than the dorsal aspect. The fat content in the ventral region also exhibited an increase from the anterior to the posterior zones. In the dorsal region, however, the back portion of the body, behind the dorsal fin, generally possessed low fat content. On an average, the highest fat accumulation was observed in the caudal peduncle region. The distribution of water followed an almost opposite pattern of distribution, indicating an inverse relationship with fat.

Identification of genes differentially expressed in wild type and Purkinje cell degeneration mice

Purkinje cell degeneration (pcd) mice are characterized by death of virtually all cerebellar Purkinje cells by postnatal day 30. In this study, we used DNA microarray analysis to investigate differences in gene expression between the brains of wild type and pcd mice on postnatal day 20, before the appearance of clear-cut phenotypic abnormalities. We identified 300 differentially expressed genes, most of which were involved in metabolic and physiological processes. Among the differentially expressed genes were several calcium binding proteins including calbindin -28k, paravalbumin, matrix gamma-carboxygluta mate protein and synaptotagamins 1 and 13, suggesting the involvement of abnormal Ca2+ signaling in the pcd phenotype.

Central representation of dynamics when manipulating handheld objects.

To explore the neural mechanisms related to representation of the manipulation dynamics of objects, we performed whole-brain fMRI while subjects balanced an object in stable and highly unstable states and while they balanced a rigid object and a flexible object in the same unstable state, in all cases without vision. In this way, we varied the extent to which an internal model of the manipulation dynamics was required in the moment-to-moment control of the object's orientation. We hypothesized that activity in primary motor cortex would reflect the amount of muscle activation under each condition. In contrast, we hypothesized that cerebellar activity would be more strongly related to the stability and complexity of the manipulation dynamics because the cerebellum has been implicated in internal model-based control. As hypothesized, the dynamics-related activation of the cerebellum was quite different from that of the primary motor cortex. Changes in cerebellar activity were much greater than would have been predicted from differences in muscle activation when the stability and complexity of the manipulation dynamics were contrasted. On the other hand, the activity of the primary motor cortex more closely resembled the mean motor output necessary to execute the task. We also discovered a small region near the anterior edge of the ipsilateral (right) inferior parietal lobule where activity was modulated with the complexity of the manipulation dynamics. We suggest that this is related to imagining the location and motion of an object with complex manipulation dynamics.

A new species of the genus Tor from Yunnan, China (Teleostei : Cyprinidae)

A new species of the subfamily Barbinae, Tor yingjiangensis, is recognized from Yunnan province, China. It can be distinguished from other Tor species by the following combination of characters: last simple dorsal-fin ray osseous and non-serrated; no forward directed predorsal procumbent spine; head length considerably longer than body depth; no tubercles on the snout or sides of the face; 18-20 gill rakers on the outside of first gill arch; 24-26 lateral line scales; median lobe of lower lip short, its posterior margin truncate, not extending to the vertical across the inner corners of the mouth; and the condition of the lower lip consistent in individuals of different sizes. The new species has been misidentified previously as Tor putitora (Hamilton, F. 1822. An account of the fishes found in the River Ganges and its Branches. Edinburgh & London. 405 pp), which occurs in the Ganges and Indus River basins. The new species can easily be distinguished from T. putitora by having 3-3.5 (vs. 2.5) scales from lateral line to pelvic-fin origin, shorter caudal peduncle length (13.0% vs. 17.2% of standard length), lesser body depth (26.4% vs. 24.0% of standard length) and longer caudal peduncle depth (12.0% vs. 10.9% of standard length), no longitudinal stripe present along side of body, and eyes visible in ventral view of head.

Balitora nantingensis (Teleostei : Balitoridae), a new hillstream loach from Salween drainage in Yunnan, Southwestern China

A new species of hillstream loach genus Balitora Gray is discovered from Nanting River drainage, which drains to Salween, Yongde County, Yunnan, China. Balitora nantingensis can be distinguished from B. lancangjiangensis by a triangle-like blotch in nape absent vs. present; area from halfway of pelvicfin ray to anus covered by scales vs. no scales in this area; the first pectoral ray thickened vs. not; posterior end of pelvic-fin surpasses middle point from posterior end of pelvic-fin base to anal-fin origin vs. the end from posterior extremity of pelvic-fins to anus; fewer lateral line scales, 59-64 vs. 66-70; slightly longer head, SL 4.2-5.1 (4.6) times of HL vs. 5.0-5.9 (5.4); slightly larger eyes, HL 4.0-6.4 (5. 1) times of eye diameter vs. 5.5-7.5 (6.3). Balitora nantingensis can be distinguished from B. burmanica by body rounded vs. slender in dorsal view; pointed snout vs. blunt in dorsal view; a deeper body, body depth 12.5-16.9% of SL vs. 10-12%; a deeper caudal-peduncle, length of caudal-peduncle 2.0-2.8 times of its depth vs. 3.0-4.2; a shorter caudal-peduncle, 14.3-15.6% of SL vs. 16-21%; the average ratio of body width at pelvic-fin origin and distance between inner pelvic-fin base 2.0 vs. 3.4; body width 1.1-1.5 times of body depth vs. 1.5-1.6; posterior end of pelvic-fin almost one or slight more than one eye diameter from anus vs. closet to anus; outside rays of pelvic-fin extend more posteriorly than inside rays and make posterior outline of fin acclivitous and straight vs. the outside ones almost equal to the inside ones and make the fin slightly convex.

Schistura cryptofasciata, a new loach (Cypriniformes : Balitoridae) from Salween drainage in Yunnan, Southwestern China

Schistura cryptofasciata is distinguished from its congeners by following combination characters: lateral line complete; 8(1)/(2) branched dorsal-fin rays; 9+8 branched caudal-fin rays; no sexual dimorphism; 8-11 indistinct bars, wider than interspaces; caudal-peduncle length 44.1-16.5% standard length, 68.1-90.4% dorsal head length; anterior portion of body scaleless; air bladder entirely embedded in bony capsule; lateral head length 104-122% dorsal head length; lower jaw with a notch at symphysis; lower lip completely interrupted; interorbital width 1.9-2.2 times eye diameter; pelvic-fin origin ventral to dorsal-fin origin; anus 1.0-1.5 eye diameters in front of anal-fin origin; the widest part of black basal caudal bar is the midlateral, and then the upper and lower ends.

Paracobitis nanpanjiangensis, a new loach (Balitoridae: Nemacheilinae) from Yunnan, China

A new species of the genus Paracobitis, Paracobitis nanpanjiangensis is described from tributaries of the Nanpanjiang River drainage in China. It is distinguished from its congeners, except P. oligolepis and P. wujiangensis, by body scaleless or with rudimentary scales (caudal peduncle with several deeply embedded scales). It can be differentiated from P. wujiangensis by the complete lateral line (vs. incomplete), lower dorsal crest reaching the vertical of origin of anal fin (vs. shorter and higher dorsal crest not reaching the base of anal fin). It is distinguished from P. oligolepis by the following characters: branched dorsal fin with 81/2 (a few 91/2) rays (vs. 91/2), interspaces between bars in front of dorsal fin conspicuously thinner than those behind (vs. vermiform markings), dorsal head without vermiform markings or obscure (vs. clearly vermiform markings on dorsal head), vertebrae 4 + 36-38 (vs. 4 + 39-41).

A new species of sisorid catfish genus Glyptothorax (Teleostei: Sisoridae) from Salween drainage of Yunnan, China

A new species of the genus Glyptothorax, Glyptothorax obliquimaculatus sp. nov. is described from the Xiaohei River, a tributary of the Nanting River, Salween drainage, in southwestern Yunnan province, China. This new species can be distinguished from its congeners by the following combination of characteristics: unculiferous ridges of the thoracic adhesive apparatus extending anteriorly onto the gular region; body with irregular dark blotches scattered along lateral surface (blotches mostly oblique); skin smooth on head and body; dorsal spine smooth without serrations on its posterior margin; lips smooth; posterior margin of pectoral spine with 7-8 serrations; dorsal-fin base 11.0-13.2% SL; pectoral-fin length 15.6-19.6% SL; depth of caudal peduncle 8.6-9.8% SL; head width 19.1-24.0% SL; nasal barbel length 23.3-33.3% HL.

Onychostoma virgulatum, a new species of cyprinid fish (Pisces: Teleostei) from southern Anhui Province, South China

Onychostoma virgulatum, new species, is described from the Qiupu River, a tributary on the Southern bank of the lower Yangtze River at Shitai County, southern Anhui Province, South China. It shares with O. fusiforme and O. meridionale the presence of a longitudinal dark brown stripe extending along the lateral line, a character separating them from all other congeners of the moderate-mouth group diagnosed by having a slightly arched or nearly transverse mouth opening (with the extremities slightly curved posteriorly), its width being equal to or slightly less than the width of head at this same point, and a short postlabial groove extending along half of the length of the lateral margin of the lower jaw. Onychostoma virgulatum differs from both in the presence of two pairs of barbels in adults, from O. fusiforme in the body depth, caudal-peduncle depth, and position of pelvic and anal fins, and from O. meridionale in the structure of the last simple dorsal-fin ray.

Triplophysa lixianensis, a new nemacheiline loach species (Pisces : Balitoridae) from the upper Yangtze River drainage in Sichuan Province, South China

Triplophysa lixianensis, a new nemacheiline loach species, is described from the Min Jiang of the upper Yangtze River drainage in Sichuan Province, South China. It can be separated from all other species of Triplophysa by having a unique combination of the following characters: posterior chamber of gas bladder greatly reduced or absent; caudal peduncle columnar with a roughly round cross- section at its beginning; anterior edge of lower jaw completely exposed or uncovered by lower lip; intestine short, forming a zigzag loop below stomach; dorsal- fin origin closer to caudal- fin base than to snout tip; pelvic fin inserted anterior to dorsal- fin origin; snout length 50.6 - 57.5 % of head length; eye diameter 12.3 15.4 % of head length; caudal peduncle length 25.1 - 27.1 % of standard length; anal fin with five branched rays; lower lip greatly furrowed with two thick lateral lobes; and body smooth or scaleless.