Anatomical and developmental patterns of interleukin-2 gene expression in the mouse: analysis of IL-2 expressing cells and partial characterization of interactions involved in mediating IL-2 gene expression in vivo

Interleukin-2 (IL-2) is an important mediator in the vertebrate immune system. IL-2 is a potent growth factor that mature T lymphocytes use as a proliferation signal and the production of IL-2 is crucial for the clonal expansion of antigen-specific T cells in the primary immune response. IL-2 driven proliferation is dependent on the interaction of the lymphokine with its cognate multichain receptor. IL-2 expression is induced only upon stimulation and transcriptional activation of the IL-2 gene relies extensively on the coordinate interaction of numerous inducible and constitutive trans-acting factors. Over the past several years, thousands of papers have been published regarding molecular and cellular aspects of IL-2 gene expression and IL-2 function. The vast majority of these reports describe work that has been carried out in vitro. However, considerably less is known about control of IL-2 gene expression and IL-2 function in vivo.

To gain new insight into the regulation of IL-2 gene expression in vivo, anatomical and developmental patterns of IL-2 gene expression in the mouse were established by employing in situ hybridization and immunohistochemical staining methodologies to tissue sections generated from normal mice and mutant animals in which T -cell development was perturbed. Results from these studies revealed several interesting aspects of IL-2 gene expression, such as (1) induction of IL-2 gene expression and protein synthesis in the thymus, the primary site of T-cell development in the body, (2) cell-type specificity of IL-2 gene expression in vivo, (3) participation of IL-2 in the extrathymic expansion of mature T cells in particular tissues, independent of an acute immune response to foreign antigen, (4) involvement of IL-2 in maintaining immunologic balance in the mucosal immune system, and (5) potential function of IL-2 in early events associated with hematopoiesis.

Extensive analysis of IL-2 mRNA accumulation and protein production in the murine thymus at various stages of development established the existence of two classes of intrathymic IL-2 producing cells. One class of intrathymic IL-2 producers was found exclusively in the fetal thymus. Cells belonging to this subset were restricted to the outermost region of the thymus. IL-2 expression in the fetal thymus was highly transient; a dramatic peak ofiL-2 mRNA accumulation was identified at day 14.5 of gestation and maximal IL-2 protein production was observed 12 hours later, after which both IL-2 mRNA and protein levels rapidly decreased. Significantly, the presence of IL-2 expressing cells in the day 14-15 fetal thymus was not contingent on the generation of T-cell receptor (TcR) positive cells. The second class of IL-2 producing cells was also detectable in the fetal thymus (cells found in this class represented a minority subset of IL-2 producers in the fetal thymus) but persist in the thymus during later stages of development and after birth. Intrathymic IL-2 producers in postnatal animals were located in the subcapsular region and cortex, indicating that these cells reside in the same areas where immature T cells are consigned. The frequency of IL-2 expressing cells in the postnatal thymus was extremely low, indicating that induction of IL-2 expression and protein synthesis are indicative of a rare activation event. Unlike the fetal class of intrathymic IL-2 producers, the presence of IL-2 producing cells in the postnatal thymus was dependent on to the generation of TcR+ cells. Subsequent examination of intrathymic IL-2 production in mutant postnatal mice unable to produce either αβ or γδ T cells showed that postnatal IL-2 producers in the thymus belong to both αβ and γδ lineages. Additionally, further studies indicated that IL-2 synthesis by immature αβ -T cells depends on the expression of bonafide TcR αβ-heterodimers. Taken altogether, IL-2 production in the postnatal thymus relies on the generation of αβ or γδ-TcR^+ cells and induction of IL-2 protein synthesis can be linked to an activation event mediated via the TcR.

With regard to tissue specificity of IL-2 gene expression in vivo, analysis of whole body sections obtained from normal neonatal mouse pups by in situ hybridization demonstrated that IL-2 mRNA^+ cells were found in both lymphoid and nonlymphoid tissues with which T cells are associated, such as the thymus (as described above), dermis and gut. Tissues devoid of IL-2 mRNA^+ cells included brain, heart, lung, liver, stomach, spine, spinal cord, kidney, and bladder. Additional analysis of isolated tissues taken from older animals revealed that IL-2 expression was undetectable in bone marrow and in nonactivated spleen and lymph nodes. Thus, it appears that extrathymic IL-2 expressing cells in nonimmunologically challenged animals are relegated to particular epidermal and epithelial tissues in which characterized subsets of T cells reside and thatinduction of IL-2 gene expression associated with these tissues may be a result of T-cell activation therein.

Based on the neonatal in situ hybridization results, a detailed investigation into possible induction of IL-2 expression resulting in IL-2 protein synthesis in the skin and gut revealed that IL-2 expression is induced in the epidermis and intestine and IL-2 protein is available to drive cell proliferation of resident cells and/or participate in immune function in these tissues. Pertaining to IL-2 expression in the skin, maximal IL-2 mRNA accumulation and protein production were observed when resident Vγ_3^+ T-cell populations were expanding. At this age, both IL-2 mRNA^+ cells and IL-2 protein production were intimately associated with hair follicles. Likewise, at this age a significant number of CD3ε^+ cells were also found in association with follicles. The colocalization of IL-2 expression and CD3ε^+ cells suggests that IL-2 expression is induced when T cells are in contact with hair follicles. In contrast, neither IL-2 mRNA nor IL-2 protein were readily detected once T-cell density in the skin reached steady-state proportions. At this point, T cells were no longer found associated with hair follicles but were evenly distributed throughout the epidermis. In addition, IL-2 expression in the skin was contingent upon the presence of mature T cells therein and induction of IL-2 protein synthesis in the skin did not depend on the expression of a specific TcR on resident T cells. These newly disclosed properties of IL-2 expression in the skin indicate that IL-2 may play an additional role in controlling mature T-cell proliferation by participating in the extrathymic expansion of T cells, particularly those associated with the epidermis.

Finally, regarding IL-2 expression and protein synthesis in the gut, IL-2 producing cells were found associated with the lamina propria of neonatal animals and gut-associated IL-2 production persisted throughout life. In older animals, the frequency of IL-2 producing cells in the small intestine was not identical to that in the large intestine and this difference may reflect regional specialization of the mucosal immune system in response to enteric antigen. Similar to other instances of IL-2 gene expression in vivo, a failure to generate mature T cells also led to an abrogation of IL-2 protein production in the gut. The presence of IL-2 producing cells in the neonatal gut suggested that these cells may be generated during fetal development. Examination of the fetal gut to determine the distribution of IL-2 producing cells therein indicated that there was a tenfold increase in the number of gut-associated IL-2 producers at day 20 of gestation compared to that observed four days earlier and there was little difference between the frequency of IL-2 producing cells in prenatal versus neonatal gut. The origin of these fetally-derived IL-2 producing cells is unclear. Prior to the immigration of IL-2 inducible cells to the fetal gut and/or induction of IL-2 expression therein, IL-2 protein was observed in the fetal liver and fetal omentum, as well as the fetal thymus. Considering that induction of IL-2 protein synthesis may be an indication of future functional capability, detection of IL-2 producing cells in the fetal liver and fetal omentum raises the possibility that IL-2 producing cells in the fetal gut may be extrathymic in origin and IL-2 producing cells in these fetal tissues may not belong solely to the T lineage. Overall, these results provide increased understanding of the nature of IL-2 producing cells in the gut and how the absence of IL-2 production therein and in fetal hematopoietic tissues can result in the acute pathology observed in IL-2 deficient animals.

Study on the effects of three fishing baits on the catch composition of malian traps in Lake Kainji

The studies on the effects of three fishing baits on the catch composition of Malian traps in Lake Kainji were investigated. The traps were set between Monai and Taafa fishing villages in the Southern basin of the lake, baited with their respective treatment and were inspected daily for twelve days. A total of 218 fish were caught, of which the highest (54.59%) was caught by corn bran, while the lowest (11.01%) was caught by stomach content and rice bran caught 34.4%. The fish caught comprised of 15 species belonging to 8 families. There was no significant different (P>0.05) in the catch of the various baits. The weight also followed the same trend as the number of fish caught. However, both baits showed better efficiency for Alestes baremose. Tilapia zilli, S. galilaeus, Oreochromis niloticus, Labeo coubie and Distichodus rostratus than other species caught. There was a wide range between the inimum and maximum size of species caught, which showed the efficiency of the traps in capturing small size, juveniles and the adult of large fish species due to small mesh size (1") net-cover of the trap. Recommendations were made on the use of corn and rice bran as baits enhancing catch efficiency for fishes such as O.niloticus, T. zilli, T. galilaeus and D. rostratus

Fischerei und fischereibiologische Untersuchungen im Nordost-Atlantik im 1. Halbjahr 1996

Biological investigations were carried out onboard the commercial freezing trawler "KIEL" off the Norwegian coast in February/March and in the Bear Island area in May 1996. Data will be used as German contribution to the assessments of the ICES "Arctie Fisheries Working Group". Informations about the fishery and the concentrations of cod, haddock, saithe and redfish in the areas between Röst-, Nordvest-, Fuglöy-Bank and Bear Island are given. Length - and age distributions as well as preliminary results of stomach- and gonad investigations are represented.

Gute Kabeljau- und Schellfischvorkommen bei der Bären Insel

Biological investigations were carried out onboard the commercial freezing trawler "Kiel" in the Bear Island area in May/June 1995. Data will be used as German contribution to the assessments of the ICES "Artic Fisheries Working Group". Concentrations of cod were found southwest of Bear Island. Catches ammounted to 5-60 t/haul. Relatively good catches (5-25 t) with high proportions of haddock (40-80 %) were observed to the northwest of the island. Length- and age distributions as well as preliminary results of stomach- and gonad- investigations are presented.

Feeding selectivity of brown trout (Salmo trutta) in Loch Ness, Scotland

The aim of this study was to compare statistically the zooplankton assemblage ingested by brown trout (Salmo trutta) in Loch Ness with that of the zooplankton in the water column. This would allow the examination of the apparent paradox that very few copepods appear to be consumed by trout at a time of year when they are numerous and readily available as food. The investigation was limited to the crustacean zooplankters, since the Rotifera are generally so small that they are only of interest to fish in the first few days of life. 25 trout were obtained from anglers, and the stomach contents of non-"ferox" animals analysed. Samples of pelagic zooplankton were obtained approximately monthly from 30-m vertical net-hauls (mesh size 100 km). It is concluded that the variation in dietary composition with trout wet weight indicates an ontogenetic habitat shift producing spatial separation of young and older individuals.

Ecologia de Astyanax taeniatus na Bacia do Rio Mato Grosso, Saquarema - RJ: Dieta, uso do micro-hábitat e uma breve análise comportamental

Neste trabalho tivemos como objetivo caracterizar a dieta, uso do habitat e padrões comportamentais de Astyanax taeniatus da bacia do Rio Mato Grosso, que encontra-se na porção leste do Estado do Rio de Janeiro (22 52 S; 42 40 W e 22 53 S; 42 34 W). Para a análise da dieta, os exemplares foram coletados bimestralmente entre março de 2006 e janeiro de 2007 em três localidades que diferiram pelas variáveis físicas. As observações de uso dos recursos do habitat foram realizadas por observação subaquática, na posição focal dos exemplares avistados, enquanto a quantificação da disponibilidade foi realizada em 50 quadrats de 20x20cm (400cm2) ao longo dos mesmos 50m onde foi realizada a observação sub-aquática. A análise do conteúdo estomacal de 651 exemplares foi realizada sob microscópio estereoscópico de acordo com métodos qualitativos e quantitativos (Freqüência de Ocorrência e Volumétrica). A participação relativa de cada item registrado nos estômagos em relação à totalidade da dieta foi analisada através do Índice Alimentar (IAi). Para verificar possíveis diferenças entre as proporções dos itens de origem animal e vegetal, autóctone e alóctone, os valores proporcionais foram testados pelo 2 de contingência. A partir dos dados de comprimento padrão e comprimento do intestino, foi calculado o valor do quociente intestinal. Os itens de origem vegetal tiveram maior contribuição na dieta da espécie para as localidades com maior altitude, enquanto os itens animais tiveram maior contribuição na localidade baixa. A diferença na contribuição dos itens de origem autóctone e alóctone também foi significativa. Na dieta de jovens e adultos, houve diferença significativa na contribuição de itens de origem vegetal e animal somente na localidade mais alta, onde os adultos consumiram maior quantidade de matéria vegetal. Os valores médios de quociente intestinal em jovens e adultos foram significativamente diferentes nas localidades de maior altitude, com valores maiores para indivíduos adultos. Observamos 52% dos indivíduos em profundidades entre 30 e 45 cm, 72% em áreas de rápido, 72% em velocidades entre 0 e 0,5km/h, 66% encontravam-se distantes da margem entre 40 e 120 cm, 37,6% em substrato do tipo areia e 34,4% em substrato do tipo pedra. De todos os padrões comportamentais observados, aquele que mais se destacou foi o forrageamento, onde 70,91% dos indivíduos estavam forrageando no meio da coluna dágua. Os resultados da dieta reforçam a idéia de as espécies de Astyanax têm hábito alimentar onívoro e oportunista, onde a espécie alimentou-se dos recursos disponíveis no ambiente evidenciando sua alta plasticidade alimentar ao longo do riacho. Espécies do gênero Astyanax são consideradas generalistas em relação ao uso do habitat e altamente ativas, corroborando com os resultados do presente estudo.

Comparative drift studies on streams in lower Austria (Flysch, gneiss and chalk formations) [Translation from: Schweizerische Zeitschrift fur Hydrobiologie 30 138-185, 1968.]

Seasonal changes and flooding have an extraordinarily great influence on the drift of organisms. The free water space plays the main part in the provision of food for some fish (Salmo trutta - trout): drift and content of the stomach are balanced here (Simuliidae): whereas others (Thymallus vulgaris) only selectively chose certain animals living at the bottom (molluscs). The total drift, drift of organisms and drift of organic material and minerals, plays a main role in the rate of production in streams. Besides the biology of the organisms living on the river bed, also the geological and hydrographical situation of the area plays a very important role for the composition of the drift. During the years 1964-1966 three streams in the characteristical geological formations flysch, gneiss and chalk of lower Austria were studied in regard to their drift. The Tulln (above St. Christopen), the Krems (above Senftenberg) and the Schwarza (above Hirschwang) seemed to be ideal for this comparative study because they are easy to reach. After summarising the hydrography and chemistry of examined rivers, the author examines the relationship between water level and total drift and the stratification of the total drift before analysing the drift of living organisms. Also considered are seasonal changes of drift of organisms and drift of exuviae.

Zooplankton - fish interaction in the littoral zone of Nyanza Gulf, Lake Victoria

The zooplankton community of the littoral zone of Nyanza Gulf, Lake Victoria, was studied between June 1998 and June 1999 to identify and quantify various zooplankton groups, and investigate the interactions that occur between them and the littoral fish through the food chain. Zooplankton samples were collected from five stations using a 83 micro-m mesh size plankton net hauled vertically through the water column. Fish samples were obtained by beach seine, except at Gingra (May 1999), where trawl samples were used. Gut/stomach analysis was carried out on the three major commercial species, Lates niloticus (L.), Oreochromis niloticus (L.) and Rastrineobola argentea (Pellegrin).

Faunistic studies

An update on the work undertaken by the authors, Macan and Hynes, over the past year. Macan provides details of additional work undertaken since his publication of faunistic work on adult animals, including details on additional field data, further developments of the conclusions intially drawn and further comparisons with fauna of the highlands of Scotland. Two histograms are provided showing succession of Corixidae in two separate waterbodies. Habitat conditions are also discussed for this species and a number of locations are mentioned including, Blelham Tarn, Derwentwater and Rusland Moss. An update on stonefly work undertaken by Hynes is provided, specifically stomach contents of nymphs and habitat preference. Details are also given of aquatic insects being breed by the staff for research.

Effects of Cow Green Reservoir upon downstream fish populations

Post impoundment observations on the fish populations of the River Tees, downstream from the Cow Green Reservoir were made between 1971 and 1980. Both bullhead (Cottus gobio L.) and brown trout (Salmo trutta L.) population densities increased in the river after impoundment but changes in growth rate were small. Investigations into the stomach contents of the two species reflects the results of other work on increases of benthos in the river. Following regulation there was an increase in the quantity of Ephemerella ignita found in trout stomachs while in the bullhead, regulation caused an increase in the importance of Mollusca and a decrease in importance of Ephemeroptera and Plecoptera.

The food of coarse fish

Remarkably little has been published on the feeding habits of the non-salmonid fishes of British fresh waters. The following report briefly summarizes the results obtained from the examination of the stomach contents of some 2,700 fish, belonging to 19 species, which were obtained during 1939. The results of all examinations of gut contents were analysed, species by species, upon a simple basis of the presence of different types of food. Foodstuffs were divided up into six main categories— fish, molluscs, insects, crustaceans, higher plants together with filamentous algae, and diatoms—and the occurrence of members of any of these categories was recorded for each fish.

Dieta do boto-cinza (Sotalia guianensis Van Benédén, 1864) (Cetacea: Delphinidae) na Baía de Sepetiba (RJ)

O boto-cinza (Sotalia guianensis Van Benédén, 1864) é um pequeno cetáceo da família Delphinidae. Ocorre em águas costeiras da América do Sul e Central, associado à ambientes estuarinos, às baías e áreas protegidas. Estudos sobre hábitos alimentares são importantes para avaliar os padrões de relação entre presa e predador. Desta forma, o conhecimento da composição da dieta do predador pode fornecer informações a respeito de sua distribuição, padrões de migração e de seu comportamento, além de contribuir com informações sobre a biologia e comportamento de suas presas. Neste trabalho, são abordados aspectos da ecologia alimentar do boto-cinza da Baía de Sepetiba, com o objetivo de caracterizar a dieta e compará-la em relação ao sexo, classe etária e estações do ano, a partir da análise do conteúdo estomacal de 76 botos-cinza encalhados entre 2005 e 2011. Os itens alimentares recuperados dos estômagos como otólitos e ossos de peixes, bicos de cefalópodes, carapaças de crustáceos, foram identificados baseando-se em trabalhos de identificação e em uma coleção de referência. O comprimento e biomassa das presas foram estimados com o uso de equações de regressão encontradas na literatura para estas espécies. As amostras foram separadas em machos adultos, fêmeas adultas e juvenis, e em estações quente/chuvosa (out-abr) e fria/seca (mai-set), em que os estômagos foram recuperados. Um total de 1800 presas foi identificado, relativo a 23 espécies de teleósteo, quatro espécies de cefalópode e três de crustáceo. O boto-cinza da Baía de Sepetiba apresentou uma dieta tipicamente piscívora, com um hábito alimentar no qual, poucas espécies foram consumidas em alta frequencia. Todas as presas identificadas têm distribuição costeira sendo a maioria estuarina de pequeno porte ou juvenil. As presas mais importantes na dieta do boto-cinza, segundo o índice de importância relativa (IIR), foram Cetengraulis edentulus; Micropogonias furnieri; Mugil spp.; Chloroscombrus chrysurus; Cynoscion jamaicensis; Stellifer sp.; e Sciadeichthys luniscutis. Dentre os cefalópodes, a lula Doryteuthis plei foi à presa mais importante. Este estudo indica que o boto-cinza apresenta variações intra-específicas no seu hábito alimentar entre fêmeas adultas, machos adultos e juvenis, além de variações sazonais na composição de sua dieta. A partir do conhecimento do comportamento e hábito de suas presas, pode-se concluir que, Sotalia guianensis da Baía de Sepetiba se alimenta ao longo de toda a Baía, além de utilizar áreas costeiras próximas para atividades de alimentação e forrageio.

Hábitos alimentares e análise de sobreposição trófica de elasmobrânquios no litoral sul do Rio de Janeiro, Brasil

O presente trabalho analisou o conteúdo estomacal de cinco espécies de elasmobrânquios, sendo 9 estômagos de Psammobatis rutrum, 83 de Psammobatis extenta, 59 de Atlantoraja cyclophora, 112 de Rioraja agassizi e 43 de Rhizoprionodon lalandii, com o objetivo de obter informações sobre os hábitos alimentares dessas espécies, além de verificar a formação de guildas tróficas entre as espécies estudadas. As coletas ocorreram entre janeiro de 2006 e agosto de 2007, no litoral sul do Rio de Janeiro, ao largo da Ilha Grande. Os itens alimentares encontrados foram analisados quanto a Frequência de Ocorrência, Peso Percentual e Índice Alimentar, sendo a dieta de cada espécie avaliada considerando toda amostra, de acordo com o sexo dos indivíduos, estádios de maturidade e épocas do ano. Para investigar a similaridade entre as dietas e verificar a formação de guildas tróficas foi realizada uma Análise de Escalonamento Multidimensional (MDS). As dietas de Psammobatis rutrum e Psammobatis extenta foram consideradas carcino-bentófagas, com grande importância de camarões. O principal item para Atlantoraja cyclophora foi o siri Portunus spinicarpus, sendo a dieta considerada principalmente carcino-bentófaga, mas também com grande importância de teleósteos. Em Rioraja agassizi, a dieta foi considerada principalmente carcino-bentófaga, mas também com forte influência ictiófaga. Já o comportamento alimentar de Rhizoprionodon lalandii foi considerado ictio-bentófago. A dieta entre os sexos apresentou uma alta sobreposição em P. extenta, A. cyclophora e R. lalandii, mas para R. agassizi essa sobreposição foi considerada baixa. Entre os estádios de maturidade a sobreposição foi pequena para P. extenta e R. agassizi, entretanto para A. cyclophora e R. lalandii essa sobreposição foi elevada. Os exemplares maiores, aparentemente apresentaram preferência a se alimentarem de peixes, enquanto que predadores de menor porte se restringem aos invertebrados, basicamente crustáceos. Para todas as espécies foram identificadas alterações sazonais na importância dos itens alimentares, que pode ser explicado pelas oscilações naturais na composição e abundancia das presas potenciais em consequência das variações das massas de água na plataforma continental do sul fluminense. Os resultados indicaram a formação de 2 guildas, sendo a guilda 1 composta por P. rutrum e P extenta e com uma dieta principalmente de crustáceos (camarões) e a guilda 2 composta por A. cyclophora, R. agassizi e R. lalandii apresentando uma alimentação em Crustáceos e em Teleósteos.

Le régime alimentaire de Dentex angolensis, Poll et Maul, 1953 (Sparidae)

The food of Dentex angolensis, studied from 386 stomachs, consists essentially of small epibenthic crustaceans and, with less importance, of fishes. A systematic list with abundance and occurrence is given here. It is noted that the composition of the stomach content can differ with depth and season.

Note sur le régime alimentaire de quelques poissons démérsaux de Côte d'Ivoire

Stomach contents from 21 species of demersal fish caught on Grand Bassam (Côte d'Ivoire) fishing grounds at 20 and 35 m depth are analyzed. The results are compared with those of Longhurst (1957, 1960). The great importance of Crustacea in the food of those species appears clearly. A representative scheme of the diet is suggested.