RESUMO
Mammary ductal morphogenesis during prepuberty occurs mainly in response to insulin-like growth factor-1 (IGF-1) and estradiol stimulation. Dairy heifers infected with gastrointestinal nematodes have reduced IGF-1 levels, accompanied by reduced growth rate, delayed puberty onset, and lower parenchyma-stroma relationship in their mammary glands. Immunohistochemical studies were undertaken to determine variations in cell division rate, IGF-1 system components, and estradiol receptors (ESR) during peripubertal development in the mammary glands of antiparasitic-treated and untreated Holstein heifers naturally infected with gastrointestinal nematodes. Mammary biopsies were taken at 20, 30, 40, and 70 wk of age. Proliferating cell nuclear antigen immunolabeling, evident in nuclei, tended to be higher in the parenchyma of the glands from treated heifers than in those from untreated. Insulin-like growth factor binding proteins (IGFBP) type 2 and type 3 immunolabeling was cytoplasmic and was evident in stroma and parenchyma. The IGFBP2-labeled area was lower in treated than in untreated heifers. In the treated group, a maximal expression of this protein was seen at 40 wk of age, whereas in the untreated group the labeling remained constant. No differences were observed for IGFBP3 between treatment groups or during development. Immunolabeling for α ESR (ESR1) was evident in parenchymal nuclei and was higher in treated than in untreated heifers. In the treated group, ESR1 peaked at 30 wk of age and then decreased. These results demonstrate that the parasite burden in young heifers negatively influence mammary gland development, affecting cell division rate and parameters related to estradiol and IGF-1 signaling in the gland.
Assuntos
Proliferação de Células , Receptor alfa de Estrogênio/metabolismo , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Glândulas Mamárias Animais/metabolismo , Infecções por Nematoides/veterinária , Animais , Anti-Helmínticos/farmacologia , Bovinos/parasitologia , Estradiol/metabolismo , Receptor alfa de Estrogênio/genética , Feminino , Fenbendazol/farmacologia , Trato Gastrointestinal/parasitologia , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Fator de Crescimento Insulin-Like I/genética , Fator de Crescimento Insulin-Like I/metabolismo , Ivermectina/farmacologia , Levamisol/farmacologia , Glândulas Mamárias Animais/citologia , Nematoides , Transdução de SinaisRESUMO
The insulin-like growth factor (IGF) system is related to quality of oocytes and embryos. The aim of this study was to investigate the mRNA levels of IGF1 and IGF2 and their receptors, IGFR1 and IGFR2, as well as IGFBP2, IGFBP4, and PAPP-A in oocytes from Nelore compared to Holstein cows. Pools of oocytes (20 oocytes/pool) from Nelore (n=8 pools) and Holstein (n=4 pools) were obtained via ovum pick-up (OPU, 10 sessions) and cumulus cells and zona pellucida were removed. The pools were submitted to total RNA extraction. Expression of members of the IGF system was assessed by real time RT-PCR. The mRNA expression of IGF1 and IGF2, IGFR1 and IGFR2, IGFBP2 and IGFBP4 was significantly higher (P<0.01) in oocytes from Holstein whereas the expression of PAPP-A was significantly higher (P<0.05) in oocytes from Nelore cows. The high PAPP-A expression and the low expression of IGFBP2 and IGFBP4 are associated with more efficient degradation of IGFBPs, which results in greater bioavailability of IGF in Nelore oocytes when compared to the Holstein.
Assuntos
Bovinos , Recuperação de Oócitos/veterinária , Oócitos/metabolismo , Transdução de Sinais/genética , Somatomedinas/genética , Somatomedinas/metabolismo , Animais , Bovinos/genética , Bovinos/metabolismo , Feminino , Regulação da Expressão Gênica , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Proteína 4 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Proteína 4 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Proteína Plasmática A Associada à Gravidez/genética , Proteína Plasmática A Associada à Gravidez/metabolismo , Receptor IGF Tipo 1/genética , Receptor IGF Tipo 1/metabolismo , Receptor IGF Tipo 2/genética , Receptor IGF Tipo 2/metabolismo , Transdução de Sinais/fisiologia , Especificidade da EspécieRESUMO
Ovarian cysts are one of the major causes of infertility in dairy cows. The development is associated with an endocrine imbalance in the hypothalamo-hypophyseal-gonadal axis in which endocrine factors participate in follicular growth and differentiation and in the secretion of ovarian hormones. Insulin-like growth factor family are essential local regulators of ovarian follicle development and functionality and actions are mediated by binding protein activity. The aim of the present study was to analyze the expression of IGFBP-2 and IGFBP-3 in developing follicles of normal estrous cycling animals and with spontaneous and induced cystic ovarian disease (COD) to determine IGF bioavailability. The mRNA of IGFBP-2 and IGFBP-3 in follicular walls was quantified by reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization. Protein expression was analyzed by immunohistochemistry. The results demonstrated reduced amounts of mRNA of both IGFBPs in the granulosa cells of ovarian follicles of animals with COD (P<0.05). The present study suggests that the IGF system or imbalances between IGFs and IGFBPs may be involved in COD of cattle.
Assuntos
Doenças dos Bovinos/metabolismo , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Cistos Ovarianos/veterinária , Animais , Bovinos , Feminino , Regulação da Expressão Gênica/fisiologia , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Proteína 3 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Cistos Ovarianos/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Overexpression of the EGFR, IGFBP-2 and HIF-2A genes has been observed in high-grade astrocytomas and these genes seem to be functionally related to one another. This study aimed to define the profile of their expressions, interactions and correlation with clinical features and prognostic significance in microdissected tumor samples from 84 patients with astrocytomas of different grades and from 6 white matter non-neoplasic brain tissue sample. EGFR, IGFBP-2 and HIF-2A gene expression levels were analyzed by quantitative real-time PCR and differed significantly between grades I-IV astrocytic tumors (P < 0.0001, P < 0.0001 and P: 0.0013, respectively) when analyzed by the Kruskal-Wallis test. Grade I astrocytomas presented gene expression levels similar to those encountered in samples of microdissected white matter of non-neoplastic brain tissue Overexpression of the EGFR, IGFBP-2 and HIF-2A genes was significantly associated with lower 2-year survival (P: 0.009, P: 0.0002 and P: 0.008, respectively). Co-overexpression of these genes was strongly associated with high-grade gliomas and lower survival in univariate (P < 0.0001) and multivariate (P: 0.009) analysis, suggesting that the co-expression of the EGFR/IGFBP-2/HIF-2A pathway genes may have a more important clinical and biological impact than the expression of each individual gene alone. These data support the existence of a common pathway involving these genes that could contribute to the design of new target treatments.
Assuntos
Astrocitoma/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Neoplasias Encefálicas/genética , Receptores ErbB/genética , Regulação Neoplásica da Expressão Gênica , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Adulto , Astrocitoma/metabolismo , Astrocitoma/patologia , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Criança , Receptores ErbB/metabolismo , Feminino , Humanos , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Masculino , Microdissecção , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Prognóstico , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Neoplásico/genética , RNA Neoplásico/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Protein intake is a critical regulatory factor of the GH/IGF-I axis. Recently, it has been shown that splenic GH/IGF-I may respond to nutritional stress by preserving tissue homeostasis. To study the effects of exogenous administration of rhIGF-I on the splenic GH/IGF-I axis in protein malnourished rats, six-week-old male rats were assigned to one of four isocaloric diets differing in the protein content (0%, 4%, 12% and 20%) for a period of 12 days. Animals in the same dietary group on day 5 were randomly divided into two groups and during 7 days received a continuous subcutaneous infusion of either vehicle or rhIGF-I (300 microg/day). A low protein intake decreased the circulating levels of IGF-I, IGFBP-3, GH and insulin whereas the serum levels of IGFBP-1 were increased. Splenic IGFBP-3, -4 and -6 mRNA expression were up-regulated by protein malnutrition. Similarly, IGF-IR and GHR mRNA expression were significantly increased by the lack of dietary protein, whereas the levels of IGF-I mRNA remained unchanged. Exogenous rhIGF-I administration increased the circulating levels of IGFBP-1 and -3 in protein malnourished rats and reduced significantly the GH and insulin levels in well-fed rats. Similarly, rhIGF-I increased significantly the expression of the GHR in the spleen and splenic weight in all dietary groups, whereas nitrogen balance was enhanced only in the high-protein diet group. Among the cell subpopulations, B lymphocytes showed the highest GHR expression. These results suggest that in catabolic stress, induced by protein malnutrition the splenic GH/IGF-I axis is an important modulator and contributes to the maintenance of the homeostasis of the immune system.
Assuntos
Proteínas Alimentares/farmacologia , Hormônio do Crescimento/metabolismo , Fator de Crescimento Insulin-Like I/metabolismo , Fator de Crescimento Insulin-Like I/farmacologia , Somatomedinas/metabolismo , Baço/metabolismo , Animais , Hormônio do Crescimento/sangue , Insulina/sangue , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Linfócitos/metabolismo , Masculino , RNA Mensageiro/genética , Ratos , Baço/efeitos dos fármacosRESUMO
Dehydroepiandrosterone (DHEA) and its sulfate ester (DHEA-S) exert multiple effects in rodent and human brain. Several findings suggest that insulin-like growth factor-1 (IGF-1) is involved in the actions of DHEA. In this study, we assessed whether systemic administration of DHEA regulates the IGF-1 system in the hypothalamus, hippocampus, cerebral cortex, and cerebellum of adult rats. DHEA resulted in a significant reduction in IGF-1 receptor protein levels. This effect was dose dependent and restricted to the hypothalamus. In contrast to IGF-1 receptor, IGF-1-binding protein 2 levels were unaffected by DHEA treatment. IGF-1 levels were significantly increased in the hypothalamus of the rats treated with DHEA, whereas IGF-1 serum levels were not affected by DHEA. The effects of DHEA on the hypothalamic IGF-1 system may be highly relevant to the control and maintenance of hypothalamic neuroendocrine function.
Assuntos
Desidroepiandrosterona/farmacologia , Hipotálamo/efeitos dos fármacos , Fator de Crescimento Insulin-Like I/metabolismo , Animais , Western Blotting , Cerebelo/efeitos dos fármacos , Cerebelo/metabolismo , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Hipotálamo/metabolismo , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/metabolismo , Masculino , Radioimunoensaio , Ratos , Ratos Wistar , Receptor IGF Tipo 1/metabolismoRESUMO
The presence of insulin-like growth factors (IGF), IGF binding proteins (IGFBP) and IGF receptor type 1 (IGF-IR) in the human corpus luteum was investigated by examining the expression and production of related proteins throughout the lifespan of the corpus luteum and the action of nitric oxide upon their production. The expression of proteins in corpora lutea from the early, mid-and late luteal phases was assessed by immunohisto-chemistry, evaluated by a semi-quantitative analysis and the functional study was performed in corpus luteum explants incubated with nitric oxide donors. IGF-I and -II and IGFBP-1 and -3 were measured in the culture media by specific immunoassays. The results showed that IGF-I and -II, IGFBP-1 to -6 and IGF-IR were detected in the human corpus luteum throughout the luteal phase. Moreover, the expression and production of IGF-I and IGFBP-1 increased progressively from corpora lutea from the early to late luteal phases (P < 0.05), whereas the expression and production of IGFBP-2, -4 and -5 were significantly higher in corpora lutea from the mid-luteal phase (P < 0.05). No differences were observed in the expression of IGF-II, IGFBP-3 and -6 and IGF-IR throughout the lifespan of the corpus luteum. However, functional studies showed that nitric oxide donors elicited a stimulatory action on production of IGF-I in corpora lutea from the early luteal phase (80%) and on production of IGFBP-1 in corpora lutea from the late luteal phase (50%) (P < 0.05), whereas production of IGF-II and IGFBP-3 was not affected by nitric oxide. In conclusion, the components of the IGF-IGFBP system are expressed in the human corpus luteum throughout its lifespan. Nitric oxide regulates IGF-I and IGFBP-1 production, indicating that the growth factors may serve, at least in part, as mediators of the action of nitric oxide in the human corpus luteum.