RESUMEN

Present study describes the spawning induction of striped Snakehead, Channa striata using carp pituitary extract (CPE) and LH-RH agonist i.e. Buserelin (Glp-His-Trp-Ser-Tyr-Ser-tBu-Leu-Arg-Pro-NHEt). Total four treatments were designed under both hormones trail and treated as control group, T1, T2, and T3 with three replications of each treatment. While breeders under all hormone treatments showed spawning performances, no spawning performance was observed in control group. Latency time after hormonal treatment was lowest (20-24 hrs) in case of CPE than Buserelin (25-30 hrs). Regarding to CPE, spawning, fertilization and hatching rate were higher with the increasing doses of CPE in different treatments. The highest mean ± Standard deviation spawning, fertilization and hatching rate were 85.60±8.58 %, 79.38±4.89 % and 64.33±6.60 % respectively in T3 where dose of CPE was 80 mg kg-1. Similarly, in case of Buserelin hormone highest spawning rate was found in T3 (80.61±5.59) where dose of Buserelin was 0.80 µg kg-1 body weight. Fertilization rate was on the level 48.57±5.99, 70.62±5.33 and 90.32±4.79 respectively for T1, T2, and T3.Whilst, hatching rate was found 20.81±4.91, 37.11±4.50 and 61.33±6.61 in T1, T2, and T3 treatments respectively. However, T3 exhibited best performance regarding spawning, fertilization and hatching rate which were significantly higher than other two treatments.The current study revealed that spawning induction using carp pituitary extract and Buserelin is effective and might be useful for artificial breeding of Channa striata. Regarding to dose application i.e. 80 mg kg-1 of CPE and 0.80 µg kg-1 of Buserelin may be successfully applied to ovulation stimulation of Channa striata.

Asunto(s)

Buserelina , Peces , Conducta Sexual Animal , Carpas , Embrión no Mamífero , Masculino , Femenino , Animales , Hormonas Hipofisarias/metabolismo , Gonadotropinas/metabolismo , Cruzamiento , Peces/crecimiento & desarrollo , Fertilización

RESUMEN

BACKGROUND: The pituitary glands normally produce and stores gonadotropic hormones (GnH) that are responsible for ovulation and spermiation in animals. However, whether fish pituitary extracts can elicit same effects in treated animals need elucidation as a prelude to their practical usage in animals. OBJECTIVES: The aim of this study was to investigate the oestrus-inducing potential of the pituitary gland extract of the Africa Catfish (Clarias gariepinus) in immature Wistar rats. METHODS: The experiment involved the use of 18 immature female Wistar rats and 10 male catfish brood stocks with the use of six Wistar rats per groups as follows: Group A had human chorionic gonadotropin (hCG) treatment. Group B had only normal saline treatment as the control whereas Group C had the C. gariepinus pituitary extract administration to induce oestrus with treatments occurring twice six hours apart in each group. RESULTS: There was an obvious expression of visible signs of heat and the presence of uterine horn oedema with significant (p < 0.05) increase in reproductive tract weight and uterine width and length. However, only progesterone levels increased significantly (p < 0.05) in the hCG and the C. gariepinus pituitary extract treated groups compared to other assayed hormones. CONCLUSION: These results showed that C. gariepinus pituitary extract has the capacity to induce oestrus in Wistar rats because of its gonadotropic effects, which needs further investigations at higher doses and for longer exposure periods for possible oestrus induction and synchronization in higher mammals. Further favourable results could herald the possible patent of the catfish pituitary extract for either experimental or commercial use in mammals.

Asunto(s)

Bagres , Ratas , Animales , Humanos , Femenino , Masculino , Ratas Wistar , Patentes como Asunto , Hipófisis , Mamíferos , Hormonas

RESUMEN

Tumour development and progression is dependent upon tumour cell interaction with the tissue stroma. Bioengineering the tumour-stroma microenvironment (TME) into 3D biomimetic models is crucial to gain insight into tumour cell development and progression pathways and identify therapeutic targets. Ameloblastoma is a benign but locally aggressive epithelial odontogenic neoplasm that mainly occurs in the jawbone and can cause significant morbidity and sometimes death. The molecular mechanisms for ameloblastoma progression are poorly understood. A spatial model recapitulating the tumour and stroma was engineered to show that without a relevant stromal population, tumour invasion is quantitatively decreased. Where a relevant stroma was engineered in dense collagen populated by gingival fibroblasts, enhanced receptor activator of nuclear factor kappa-B ligand (RANKL) expression was observed and histopathological properties, including ameloblastoma tumour islands, developed and were quantified. Using human osteoblasts (bone stroma) further enhanced the biomimicry of ameloblastoma histopathological phenotypes. This work demonstrates the importance of the two key stromal populations, osteoblasts, and gingival fibroblasts, for accurate 3D biomimetic ameloblastoma modelling.

RESUMEN

The European eel (Anguilla anguilla) is a commercially valued species for aquaculture. Over the past decades, it has experienced a drastic reduction in its natural stocks. Thus, breeding in captivity is considered essential, nowadays, to guarantee the eel aquaculture and to reduce pressure on natural populations. Traditionally, the European eel has been sexually matured by means of weekly hormonal injections, which cause stress to the fish. The purpose of this research study was to assess the use of osmotic pumps as a new method to induce sexual maturation in male and female European eels, without the weekly injection. The control groups were treated with weekly hormone injections (recombinant human chorionic gonadotropin for males and carp pituitary extract for females), and the implanted groups were treated with osmotic pumps (ALZET® osmotic pumps) loaded with the respective hormones. Regarding male European eels, this study shows that the use of controlled release systems was able to induce the maturation and spermiation, but without the necessary capacity to produce enough gametes with acceptable quality parameters that could meet the needs of a commercial eel hatchery. Concerning female European eels, the study demonstrates that the use of osmotic pumps loaded with CPE became an effective method, generating early maturations (4 to 10 weeks) in 50% of the females, so this method could become a viable alternative for eel hatchery procedures.

RESUMEN

Egg biochemical composition is among the main factors affecting offspring quality and survival during the yolk-sac stage, when larvae depend exclusively on yolk nutrients. These nutrients are primarily embedded in the developing oocytes during vitellogenesis. In aquaculture, assisted reproduction procedures may be applied enabling gamete production. For the European eel (Anguilla anguilla), reproductive treatment involves administration of pituitary extracts from carp (CPE) or salmon (SPE) to induce and sustain vitellogenesis. In the present study, we compared the influence of CPE and SPE treatments on offspring quality and composition as well as nutrient utilization during the yolk-sac stage. Thus, dry weight, proximal composition (total lipid, total protein), free amino acids, and fatty acids were assessed in eggs and larvae throughout the yolk-sac stage, where body and oil-droplet area were measured to estimate growth rate, oil-droplet utilization, and oil-droplet utilization efficiency. The results showed that CPE females spawned eggs with higher lipid and free amino acid contents. However, SPE females produced more buoyant eggs with higher fertilization rate as well as larger larvae with more energy reserves (estimated as oil-droplet area). Overall, general patterns of nutrient utilization were detected, such as the amount of total lipid and monounsaturated fatty acids decreasing from the egg stage and throughout the yolk-sac larval stage. On the contrary, essential fatty acids and free amino acids were retained. Notably, towards the end of the yolk-sac stage, the proximal composition and biometry of surviving larvae, from both treatments, were similar.

Asunto(s)

Anguilla , Hormonas/farmacología , Óvulo/química , Vitelogénesis , Saco Vitelino/química , Aminoácidos/química , Animales , Extractos Celulares , Ácidos Grasos/química , Femenino , Larva , Hipófisis , Vitelogénesis/efectos de los fármacos

RESUMEN

In carp aquaculture, hormonal manipulation with an analog of GnRH (Ovopel) and carp pituitary extract (CPE), which act at different levels of the hypothalamic-pituitary-gonadal axis, is a routine practice to enhance sperm production. Our recent studies revealed that hormonal stimulation of male carp was associated with changes in the seminal plasma proteome, including blood origin proteins. Here, we explored whether Ovopel and CPE could affect the blood proteome of male carp. Both preparations induced increases in semen volume, total number of sperm, and testosterone level. However, hormonal stimulation did not affect the plasma cortisol and glucose levels. A comparative proteomic analysis of carp blood plasma between the control (PBS) and the hormonally treated males revealed significant changes (>1.2 <-1.2-fold change, P < 0.05) in the abundance of 30 spots (14 up- and 16 downregulated) and 44 spots (28 up- and 16 downregulated) upon CPE and Ovopel treatment, respectively. The most significantly affected pathways were acute phase response signaling, the coagulation system, LXR/RXR and FXR/RXR activation; however, there were different sets of proteins in Ovopel- and CPE-treated males. The majority of differentially abundant proteins were involved in the regulation of the immune defense response, the response to stress, and complement activation. Moreover hormonal stimulation with CPE markedly increased the bactericidal activity of blood and both preparations caused profound changes in gene expression in hematopoietic organs. This work is important in understanding the biological processes behind the protein-based response to hormonal stimulation of sperm production in fish.

Asunto(s)

Carpas , Proteoma , Animales , Proteínas Sanguíneas , Carpas/microbiología , Carpas/fisiología , Proteínas de Choque Térmico , Masculino , Plasma , Proteómica , Electroforesis Bidimensional Diferencial en Gel

RESUMEN

The aim of this study was to evaluate induced reproduction in tambaqui females using buserelin acetate as compared with the traditional treatment regimen with carp pituitary extract (CPE). Reproductive traits of females with a body weight (BW) of 8.47 ±â€¯1.52 kg were evaluated in ten females treated with buserelin acetate at the dose of 0.5 mL/kg BW, in a single application, and in ten females treated with CPE at the dose of 5.5 mg/kg BW, in two applications (10 % and 90 %, with a 12-h interval between applications). Spawning rate did not differ between the females treated with buserelin acetate (40 %) and CPE (40 %). Weight, fertilization rate and hatching rate did not differ between the two treatment groups. Degree-hours (determined as the average temperature multiplied by time, in hours, for spawning after the treatment with the second dose of CPE and after the single treatment with buserelin acetate) for spawning and number of oocytes per gram of gametes collected were greater (P < 0.05) in the females treated with buserelin acetate than in the females treated with CPE. Production index, absolute fecundity and relative fecundity were greater (P < 0.05) in the females treated with CPE. The hormone buserelin acetate promotes reproduction in tambaqui females with there being a similar spawning rate and oocyte quality, however, lesser production indices and fecundity than when there is the conventional treatment regimen imposed with carp pituitary extract.

Asunto(s)

Buserelina/farmacología , Characiformes/fisiología , Fármacos para la Fertilidad Femenina/farmacología , Reproducción/efectos de los fármacos , Animales , Femenino , Reproducción/fisiología

RESUMEN

As the male reproductive organ, the main task of the testis is the production of fertile, haploid spermatozoa. This process, named spermatogenesis, starts with spermatogonial stem cells, which undergo a species-specific number of mitotic divisions until starting meiosis and further morphological maturation. The pituitary gonadotropins, luteinizing hormone, and follicle stimulating hormone, are indispensable for vertebrate spermatogenesis, but we are still far from fully understanding the complex regulatory networks involved in this process. Therefore, we developed an ex vivo testis cultivation system which allows evaluating the occurring changes in histology and gene expression. The experimental circulatory flow-through setup described in this work provides the possibility to study the function of the male tilapia gonads on a cellular and transcriptional level for at least 7 days. After 1 week of culture, tilapia testis slices kept their structure and all stages of spermatogenesis could be detected histologically. Without pituitary extract (tilPE) however, fibrotic structures appeared, whereas addition of tilPE preserved spermatogenic cysts and somatic interstitium completely. We could show that tilPE has a stimulatory effect on spermatogonia proliferation in our culture system. In the presence of tilPE or hCG, the gene expression of steroidogenesis related genes (cyp11b2 and stAR2) were notably increased. Other testicular genes like piwil1, amh, or dmrt1 were not expressed differentially in the presence or absence of gonadotropins or gonadotropin containing tilPE. We established a suitable system for studying tilapia spermatogenesis ex vivo with promise for future applications.

Asunto(s)

Cíclidos/fisiología , Proteínas de Peces/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Hormonas Esteroides Gonadales/farmacología , Espermatogénesis , Espermatogonias/metabolismo , Animales , Proteínas de Peces/genética , Perfilación de la Expresión Génica , Masculino , Espermatogonias/efectos de los fármacos

RESUMEN

This study was conducted to determine the efficacy of preserved and fresh pituitary extract of Clarias gariepinus (Burchell, 1822) to induced spawning in the same species. Growth performance of fry was also monitored for 15 days to determine the possible effect of natural hormone treatment on the fry. Pituitary glands (PG) were obtained from 12 male broodstocks, six extracted PG were preserved in 96% ethanol 24 h before injection, while the other were extracted and used in their fresh state. The PG solutions (1 g ml-1 of saline water) were injected at 1 ml kg-1 of the female broodstock. Ovaprim® (a synthetic hormone) was used as the control and administered at a rate of 0.5 ml kg -1 of fish. After a uniform latency period of 9 h 30 min, fish were striped, fertilized, incubated and the performance evaluated. Results obtained revealed better hatching performance using Ovaprim® synthetic hormone (64.52%), however, preserved PG gave better hatchability (59.74%) than fresh PG extract (51.39%). After 15 days of feeding ad libitum with shell-free Artemia cysts, the growth of fry spawn with preserved PG was comparable with that of the control, while least performance was observed using the fresh PG. It was concluded that preservation of PG in ethanol 24 h before injection had a positive effect on breeding performance and could be exploited in the commercial production of C. gariepinus fingerlings.

Asunto(s)

Bagres/fisiología , Hipófisis/química , Extractos de Tejidos/farmacología , Conservación de Tejido/métodos , Animales , Acuicultura/métodos , Domperidona/farmacología , Combinación de Medicamentos , Etanol/química , Femenino , Hormona Liberadora de Gonadotropina/farmacología , Masculino , Técnicas Reproductivas Asistidas

RESUMEN

Reproduction in captivity is a key study issue in Anguilla anguilla as a possible solution for its dwindling population. Understanding the mechanisms controlling the production of ribosomal building blocks during artificially induced oocyte maturation could be particularly interesting. Transcription levels of ribosomal biogenesis associated genes could be used as markers to monitor oogenesis. Eels from the Albufera Lagoon were injected with carp pituitary extract for 15weeks and ovaries in previtellogenic (PV) stage (non-injected), in early-, mid-, late-vitellogenesis (EV, MV, LV), as well as in migratory nucleus stage (MN) were analysed. 5S rRNA and related genes were highly transcribed in ovaries with PV oocytes. As oocytes developed, transcriptional levels of genes related to 5S rRNA production (gtf3a), accumulation (gtf3a, 42sp43) and nucleocytoplasmic transport (rpl5, rpl11) and the 5S/18S rRNA index decreased (PV>EV>MV>LV>MN). On the contrary, 18S rRNA was at its highest at MN stage while ubtf1 in charge of activating RNA-polymerase I and synthesising 18S rRNA behaved as 5S related genes. Individuals that did not respond (NR) to the treatment showed 5S/18S index values similar to PV females, while studied genes showed EV/LV-like transcription levels. Therefore, NR females fail to express the largest rRNAs, which could thus be taken as markers of successful vitellogenesis progression. In conclusion, we have proved that the transcriptional dynamics of ribosomal genes provides useful tools to characterize induced ovarian development in European eels. In the future, such markers should be studied as putative indicators of response to hormonal treatments and of the quality of obtained eel oocytes.

Asunto(s)

Diferenciación Celular/genética , Anguilas/genética , Oocitos/citología , Oogénesis/fisiología , Hormonas Hipofisarias/administración & dosificación , Animales , Femenino

RESUMEN

Resumen El yaque Leiarius marmoratus, es una especie de siluriforme nativo con gran potencial para la diversificación de la acuicultura colombiana; sin embargo, la espermiación en cautiverio aún presenta dificultades debido al bajo volumen seminal liberado con los protocolos de inducción tradicionales. En consecuencia, el objetivo fue evaluar los efectos de cinco protocolos de inducción hormonal, basados en Extracto de Hipófisis de Carpa (EHC), sGnRHa + domperidona (OVAPRIM®) y Gonadotropina Coriónica Humana (GHC) (FERTIVET®), sobre la calidad y volumen seminal de la especie. Fueron seleccionados 18 machos sexualmente maduros de 54,4 ± 2,2 cm de longitud total y 1,6 ± 0,1 kg peso corporal. Se evaluaron 5 tratamientos, así: T1: 0,25 ml/kg (0 h) y 1 ml/kg de OVAPRIM® (12 h); T2: 1 mg/kg de EHC (0 h) y 3 mg/kg de EHC (12 h); T3: dosis única de 200 UI/kg de GCH; T4: 3 mg/kg de EHC (0 h) y 200 UI/Kg de GHC (8h); T5: 1 ml/kg de OVAPRIM® (0h) y 200 UI/kg de GHC (8h); Control: 1 ml de suero fisiológico. El semen se colecto 6 h después de aplicada la última dosis hormonal. El T1 y T5 mostraron un volumen mayor (p<0,05) (5,4 ± 1,1; 4,8 ± 1,2 ml) cuando comparados con T2 (3,1 ± 1ml), T3 (2,3 ml) T4 (2,4 ± 1,1 ml) y el control (0,2 ml). En cuanto a la movilidad y tiempo de activación, T1 y T5 presentaron los mejores resultados (90 ± 0% y 54,2 ± 7 s; 90 ± 0% y 52,9 ± 3,8 s, respectivamente). La concentración espermática fue mayor (p<0,05) en T2 y T5 (1,275 ± 322; y 1,261 ± 225 sptz x 103/µl) comparadas con los otros tratamientos. Se concluye que la inducción hormonal con OVAPRIM® combinada con HCG, son efectivas para la producción seminal en la especie.

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Abstract The yaque Leiarius marmoratus is specie of native silurid, with great potential for the diversification of the colombian aquaculture; however, the spermiation in captive yet presents difficulties due to the low seminal volume released with traditional induction protocols. In consequence, the aim of this study was to evaluate the effects of five protocols of hormonal induction based on carp pituitary extract (EHC), sGnRHa + domperidone (OVAPRIM®) and human chorionic gonadotropin (GHC) (FERTIVET®) on seminal quality and volume of the specie. Were selected 18 sexually mature males of 54,4 ± 2,2 cm in total length and 1,6 ± 0,1 kg body weight. 5 treatments were evaluated so: T1 : 0,25 ml/kg (0 h) and 1 ml/kg of OVAPRIM® (12 h ); T2: 1 mg/kg of EHC (0 h) and 3 mg/kg of EHC (12 h) ; T3 : single dose of 200 UI/kg HCG; T4: 3 mg/kg of EHC (0 h) and 200 UI /kg GHC (8h) ; T5: 1 ml/kg of OVAPRIM® (0h) and 200 U.I / kg of GHC (8h) ; Control: 1 ml of saline solution. Semen was obtained 6 hours after the last dose hormone. T1 and T5 showed a larger volume (p<0.05) (5,4 ± 1,1, 4,8 ± 1,2 ml) compared to T2 (3,1 ± 1ml), T3 (2,3 ml) T4 (2,4 ± 1.1 ml) and control (0.2 ml). Regarding motility and activation time, T1 and T5 had the best results (90 ± 0 % and 54,2 ± 7 s; 90 ± 0 % and 52,9 ± 3,8 s, respectively). The sperm concentration was higher in (p<0,05) T2 and T5 (1,275 ± 322, ± 225 and 1,261 sptz x 103/ul) compared with the other treatments. In conclusion, hormonal induction with OVAPRIM® combined with HCG, are effective for the seminal production in the specie.

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Resumo O yaque Leiarius marmoratus, é uma espécie siluriforme nativa com grande potencial para a diversificação da aquicultura colombiana; No entanto, a spermiação em cativeiro ainda apresenta dificuldades devido ao baixo volume seminal liberado com protocolos de indução tradicionais. Por conseguinte, o objetivo foi avaliar os efeitos de protocolos de indução de cinco hormonais base de extrato de hipófise de carpa (EHC), sGnRHa + domperidona (Ovaprim®) e Gonadotropina coriónica humana (GHC) (FERTIVET®) sobre a qualidade e volume seminal das espécies. Foram Selecionados 18 machos sexualmente maduros de 54,4 ± 2,2 cm de comprimento total e 1,6 ± 0,1 kg de peso corporal. Foram avaliados cinco tratamentos: T1: 0,25 ml / kg (0 h) e 1 ml / kg de OVAPRIM® (12 h); T2: 1 mg / kg de EHC (0 h) e 3 mg / kg de EHC (12 h); T3: dose única de 200 UI / kg de HCG; T4: 3 mg / kg de EHC (0 h) e 200 UI / kg de GHC (8h); T5: 1 ml / kg de OVAPRIM® (0h) e 200 UI / kg de GHC (8h); Controle: 1 ml de solução salina fisiológica. O sêmen foi coletado 6 h após a última dose hormonal foi aplicada. T1 e T5 mostraram um volume maior (p <0,05) (5,4 ± 1,1, 4,8 ± 1,2 ml) quando comparado com T2 (3,1 ± 1 ml), T3 (2,3 ml) T4 (2,4 ± 1,1 ml) e controle (0,2 ml). Em relação à mobilidade e ao tempo de ativação, T1 e T5 apresentaram os melhores resultados (90 ± 0% e 54,2 ± 7 s, 90 ± 0% e 52,9 ± 3,8 s, respectivamente). A concentração de esperma foi maior (p <0,05) em T2 e T5 (1275 ± 322 e 1261 ± 225 sptz x 103 / µl) em comparação com os outros tratamentos. Conclui-se que a indução hormonal com OVAPRIM® combinada com HCG, é efetiva para a produção seminal nas espécies.

RESUMEN

The migratory species piabanha does not reproduce in lentic environments since it requires environmental stimuli for the maturation and extrusion of gametes, and therefore hormonal induction is mandatory. Current study compares the seminal characteristics of Brycon insignis without any hormonal induction (Control - Ctrl) and with two types of hormonal inductors, or rather, carp pituitary extract (T1 - 2.5 mg kg-1 body weight) and GnRH analogues, the latter applied in two different concentrations (T2 - 0.7 mg kg-1 body weight and T3 - 1.4 mg kg-1 body weight). Post-induction analyses showed that the hormones increased the motility rate - Ctrl (95%), T1 (100%), T2 (100%) and T3 (98%), although sperm concentration - Ctrl (11.52 x 109); T1 (4.37 x 109); T2 (4.34 x 109); T3 (4.01 x 109) decreased. Assessments for sperm vigor, motility time and spermatic morphology did not vary with hormonal induction. Hormonal inducer does not alter negatively the seminal characteristics of the piabanha, and the choice for the proper hormone depends on the preference of the dispenser.

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A espécie migradora piabanha não possui a capacidade de reproduzir em ambientes lênticos devido à necessidade de estímulos ambientais para a maturação e extrusão dos gametas, por isso a necessidade da indução hormonal. No presente estudo, as características seminais do Brycon insignis foram comparadas sem indução hormonal (Ctrl) e utilizando dois tipos de indutores hormonais - Extrato de Hipófise de Carpa (T1 - 2,5 mg kg-1 de peso vivo) e Análogos de GnRH, sendo este último aplicado em duas concentrações distintas (T2 - 0,7 mg kg -1 de peso vivo e T3 - 1,4 mg kg-1 de peso vivo). As análises realizadas após a indução mostraram que os hormônios utilizados produziram um aumento da taxa de motilidade - Ctrl (95%), T1 (100%), T2 (100%) e T3 (98%), porém houve uma diminuição na concentração espermática - Ctrl (11,52 x 109), T1 (4,37 x 109), T2 (4,34 x 109) e T3 (4,01 x 109). Os restantes das avaliações, vigor espermático, tempo de motilidade e morfologia espermática não apresentaram variações com a indução hormonal. Portanto, a utilização do indutor hormonal não altera negativamente as características seminais de piabanha, e a escolha do mesmo se deve à preferência do manipulador.

Asunto(s)

Characidae , Semen

RESUMEN

1α-Hydroxylation of 25-hydroxyvitamin D3 is believed to be essential for its biological effects. In this study, we evaluated the biological activity of 25(OH)D3 itself comparing with the effect of cell-derived 1α,25-dihydroxyvitamin D3 (1α,25(OH)2D3). First, we measured the cell-derived 1α,25(OH)2D3 level in immortalized human prostate cell (PZ-HPV-7) using [(3)H]-25(OH)D3. The effects of the cell-derived 1α,25(OH)2D3 on vitamin D3 24-hydroxylase (CYP24A1) mRNA level and the cell growth inhibition were significantly lower than the effects of 25(OH)D3 itself added to cell culture. 25-Hydroxyvitamin D3 1α-hydroxylase (CYP27B1) gene knockdown had no significant effects on the 25(OH)D3-dependent effects, whereas vitamin D receptor (VDR) gene knockdown resulted in a significant decrease in the 25(OH)D3-dependent effects. These results strongly suggest that 25(OH)D3 can directly bind to VDR and exerts its biological functions. DNA microarray and real-time RT-PCR analyses suggest that semaphorin 3B, cystatin E/M, and cystatin D may be involved in the antiproliferative effect of 25(OH)D3.

Asunto(s)

Calcifediol/farmacología , Próstata/efectos de los fármacos , ARN Mensajero/genética , Receptores de Calcitriol/genética , Esteroide Hidroxilasas/genética , Línea Celular Transformada , Proliferación Celular/efectos de los fármacos , Cromatografía Líquida de Alta Presión , Cistatina M/genética , Cistatina M/metabolismo , Cistatinas/genética , Cistatinas/metabolismo , Regulación de la Expresión Génica , Humanos , Masculino , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , Próstata/citología , Próstata/metabolismo , Unión Proteica , ARN Mensajero/metabolismo , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Receptores de Calcitriol/antagonistas & inhibidores , Receptores de Calcitriol/metabolismo , Semaforinas/genética , Semaforinas/metabolismo , Transducción de Señal , Esteroide Hidroxilasas/antagonistas & inhibidores , Esteroide Hidroxilasas/metabolismo , Tritio , Vitamina D3 24-Hidroxilasa

RESUMEN

AIM: To investigate the effects of bovine pituitary extract on the proliferation of keratocytes and maintaining the keratocyte phenotype in vitro. METHODS: Single keratocytes were isolated by enzyme digestion for in vitro culture. Three groups were designed according to the different culture media: a bovine pituitary extract (BPE) group, a fetal bovine serum (FBS) group and the control group. The phenotypes and proliferation of cultured cells were evaluated by morphology, immunofluorescent staining and mRNA expression of CD34, Lumican, VSX1, α-SMA and proliferating cell nuclear antigen (PCNA). In the BPE group, cells underwent serial subcultivation, and their phenotypes were identified by immunofluorescent staining. To analyze the proliferation of keratocytes in different concentrations of BPE, six different concentrations were designed to ascertain the most appropriate amount. RESULTS: In the BPE group, the cells spread out and presented dendritic morphology, and their dendrites connected to one another to form networks. On the third passage, most cells maintained their phenotype. In the FBS group, the cells exhibited a dendritic appearance in early cultured stages, but their morphology subsequently changed into a fibroblast-like shape. The number of dendritic cells in BPE group was more than FBS and control groups. Immunofluorescent staining and real-time polymerase chain reaction (PCR) confirmed that few keratocytes underwent fibroblastic transformation in the BPE and control groups, and that proliferation was higher in the BPE group than in the control group. Although the proliferation was higher in the FBS group, many keratocytes underwent fibroblastic transformation. The analysis of cell morphology and mRNA expressions of CD34, PCNA and VSX1 in six group showed that different concentrations of BPE affected the proliferation obviously but didn't affect the keratocyte phenotype, and the concentration of 40µg/mL was the most appropriate one. CONCLUSION: BPE can improve the proliferation of keratocytes and maintain their phenotype in vitro. Many keratocytes can be harvested rapidly and provide seeds for the construction of corneal stroma.

RESUMEN

O extrato hipofisário é provavelmente o indutor hormonal para desova e espermiação mais utilizado em piscicultura. A possibilidade de o produtor ter uma nova fonte de renda motivou a execução deste trabalho. O objetivo foi verificar o rendimento da hipófise no jundiá (Rhamdia quelen), em função do peso, e analisar a viabilidade econômica de sua extração. Exemplares (n=116) com pesos entre 158-1.543g foram abatidos para extração da hipófise. O índice gonadossomático (IGS) também foi calculado para verificar sua relação com o peso da hipófise. A hipófise apresentou relação significativa com o peso dos jundiás, mas não com o IGS. A extração da hipófise é mais uma opção de renda para a cadeia produtiva dessa espécie.

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The pituitary extract is probably the most used hormonal inducer to spawning and spermiation in fish culture. The possibility of a new income source for the fish farmer was the reason of this study. The aim of the present study was to verify the yield of the pituitary in silver catfish (Rhamdia quelen) as a function of weight and to analyze the economical viability of its extraction. Specimens (n=116) with 158-1543g were sacrificed for pituitary extraction. The gonadosomatic index (GSI) was also calculated to verify its relationship with the pituitary weight. The pituitary showed a significant relationship with silver catfish weight, but not with the GSI. The extraction of the pituitary is another profitable option to silver catfish productive market.

RESUMEN

El estudio fue realizado en el Municipio de San Carlos (Antioquia, Colombia), por la Facultad de Ciencias Agrarias de la Universidad de Antioquia, en convenio con la Corporación CORNARE, ISAGEN,durante la temporada reproductiva Mayo-Julio de 2008. 12 hembras y 24 machos de sabaleta Brycon hennide aproximadamente 100 g de peso corporal y 20 cm de longitud estándar, provenientes de un lote de 320 reproductores capturados del ambiente natural en Agosto del 2007 y antenidos en cautiverio en un estanque de tierra de 200 m², fueron seleccionados por sus características externas de madurez sexual como vientre abultado, papila genital enrojecida y expulsión de semen después de leve presión sobre la cavidadcelómica. Además como carácter de dimorfismo sexual se consideró para las hembras la presencia de aleta anal lisa o con espinaciones en máximo el 50% de su longitud, mientras que en los machos es rugosa en toda su extensión. También se tuvieron en cuenta características internas, en las hembras se realizó biopsia ovárica por aspiración de una muestra de ovocitos para observar la posición de sus núcleos y medición delde diámetro.. Las hembras fueron seleccionadas con la condición que presentaran ovocitos con diámetrossuperiores a 900 μm. Los animales fueron distribuidos para recibir uno de tres tratamientos, T1 Extracto de Hipófisis de Carpa (Argent, USA) (EHC), n=4; T2 Ovopel (Sigma Co., St Louis, Missouri), n=4; T3 Ovaprim (Syndel Laboratories Ltd, Vancouver, BC Canadá), n=4, durante cuatro fechas de muestreo y fertilizadas cada una con el semen de dos machos, inyectados previamente con EHC. 11 de las 12 hembras respondieron a los tratamientos y presentaron diámetros ovocitarios promedio de 1428.8 y 1679.8 (T1), 1531.2 y 1879.2 (T2) y de 1712.8 a 1803.3 (T3), antes y después del tratamiento hormonal, respectivamente.

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The evaluation of three hormone treatments on the diameter size of ovocytes in Sabaleta wasconducted during the breeding season from May to July 2008. Twelve Brycon henni females and 24males, 100 g and a standard length of 24 cm, were selected according to their sexual maturity andexternal features such as bulging belly, red genital papilla and semen expulsion after softly pressingits coelomic cavity. The fish were selected from a 320 brood fish batch caught in the wild in August2007 and kept in captivity in a 200 m² earth pond. The presence of an anal fin with or without spines ofa maximum length of 50% was considered as sexual dimorphism in females. Internal characteristicswere also considered; e.g., an ovarian aspiration biopsy was performed in females in order to observe itsnuclei position and diameter. Ovocytes from selected females were at least 900 μm in diameter. Animalswere distributed into three treatment groups for four sampling dates: T1 carp pituitary extract (CPE)(Argent Chemical Laboratories Inc., USA), n = 4; T2 Ovopel (Sigma Co. USA), n = 4; and T3 Ovaprim(Syndel Laboratories Ltd., Canada), n = 4. Each female was fertilized with the semen of two malespreviously injected with CPE. 11 out of 12 females had a treatment response, and resulting ovocyteaverage diameters were 1428.8 and 1679.8 (T1), 1531.2 and 1879.2 (T2), and 1712.8 to 1803.3 (T3),before and after hormone treatment, respectively. Significant differences were found within T1 andT2 (p <0.05); however, this was not the case for T3. For T1, frequency distribution of the diameterof ovocytes before the hormonal treatment was bimodal. T2 had two modes before and one after thetreatment (bimodal and unimodal), and T3 had two before and two after (bimodal). Fertilization rateswere low in T1; nevertheless, T2 and T3 had high rates until 12 h eggs incubation.

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O estudo foi realizado no Município de San Carlos (Antioquia, Colombia), pela Facultad de CienciasAgrarias de la Universidad de Antioquia, conjuntamente com a corporação CORNARE, ISAGEN, entreos meses de maio e julho de 2008. Foram utilizadas 12 fêmeas e 24 machos de sabaleta Brycon henni deaproximadamente 100 g de peso corporal e 20 cm de comprimento, provenientes de um lote de reprodutorescapturados em ambiente natural em agosto de 2007. Os animais receberam um dos três tratamentos: T1extrato de hipófise de Hipófise de Carpa (Argent, USA) (EHC), n=4; T2 Ovopel (Sigma Co., St Louis,Missouri), n=4; T3 Ovaprim (Syndel Laboratories Ltd, Vancouver, BC Canadá), n=4, durante quatro datasde amostragem e fertilizadas cada uma com sêmen de dois machos, injetados previamente com EHC. 11das 12 fêmeas responderam aos tratamentos e apresentaram ovócitos entre 1428.8 e 1679.8 (T1), 1531.2 e1879.2 (T2) e de 1712.8 a 1803.3 (T3) de diâmetro, antes e depois do tratamento hormonal, respectivamente.Observaram-se diferencias significativas ao interior do T1 e T2 (p<0.05), mas não para o T3. A distribuiçãode freqüências do diâmetro do ovócito apresentou no T1, duas modas antes do tratamento hormonal e doisdepois do tratamento hormonal (bimodal); T2, duas modas antes e 1 depois (bimodal e unimodal) e T3,duas antes e duas depois (bimodal). As porcentagens de fertilização foram baixas em T1, e foram altas emT2 e T3 até as 12 h de incubação dos ovos.

Asunto(s)

Animales , Hormonas/administración & dosificación , Oocitos/crecimiento & desarrollo