SUMMARY: Leptin is a 16 kilodaltons hormone secreted by adipose tissue and in the past few years it has been related to the reproductive system regulation. Leptin and its receptor (OBR) have been described in several reproductive organs and in different species but, in epididymis, there is still a lack of information. The aim of this work is to establish if leptin and its receptor are present on epididymis and where the production is occurring. At mRNA level the cauda portion showed a high expression of leptin (p<0.025) and OBRa (p<0.002) while at protein level the OBR expression was lower in cauda region (p<0.025) and leptin was not detected. The ratio between OBRa and OBRb was higher in both regions despite its total amount. By immunohistochemistry leptin and OBR were detected on epididymis epithelia, restricted to clear cells (CC). After efferent duct ligation (EDL) a decrease on leptin staining on CC was observed, suggesting that despite of epididymis production, most of leptin source may probably come from testis. Our results show that leptin and OBR, both mRNA and protein, are present on epididymis and exclusively in CC, suggesting that this tissue is responsive to the hormone and may have an important role on CC regulation.

RESUMEN: La leptina es una hormona de 16 kilodaltons secretada por el tejido adiposo y se ha relacionado en los últimos años con la regulación del sistema reproductivo. La leptina y su receptor (OBR) se han reportado en varios órganos reproductores y en diferentes especies sin embargo, en el epidídimo aún falta información. El objetivo de este trabajo fue establecer si la leptina y su receptor están presentes en el epidídimo y donde se produce. A nivel de ARNm la porción de cauda mostró una alta expresión de leptina (p <0,025) y OBRa (p <0,002) mientras que a nivel de proteína la expresión de OBR fue menor en la región de la cauda epididimaria (p <0,025) y no se detectó leptina. La relación entre OBRa y OBRb fue mayor en ambas regiones a pesar de su cantidad total. Por inmunohistoquímica se detectaron leptina y OBR en el epitelio del epidídimo restringido a células claras (CC). Después de la ligadura del conducto deferente (EDL) se observó una disminución en la tinción de leptina en CC, lo que sugiere que a pesar de la producción del epidídimo, la mayor parte de la fuente de leptina puede provenir probablemente del testículo. Nuestros resultados mostraron que la leptina y OBR, mRNA y proteína, están presentes en el epidídimo y exclusivamente en CC, lo que sugiere que este tejido es sensible a la hormona y puede tener un papel importante en la regulación CC.

PURPOSE: To investigate the effects of the maternal caffeine consumption during pregnancy to adult male testis mice offspring.METHODS: Twenty pregnant mice were divided into control group (c) and caffeine group (cf). dams received daily saline or 20 mg/kg of caffeine subcutaneously. Male offspring were monitored daily until 13th week. The testis were used to evaluate both the proliferation (pcna) and apoptosis (bax); leptin receptor (ob-r); aromatase; follicle stimulating hormone (fshr), luteinizing hormone (lhr) and androgen receptors (ar); steroidogenic acute regulatory protein (star); vascular endothelial growth factor (vegf) and estrogen receptors (erα and erβ) by western blotting. Serum concentrations of testosterone, estradiol and leptin were measured.RESULTS: There was a significant reduction in food intake and the body mass gain (p<0.05) in the cf ; pcna (p=0.01), fshr (p=0.02), star (p=0.0007), vegf (p=0.009), ar (p=0.03) in the cf. while an increase were note in bax (p=0.01), ob-r (p=0.02), lhr (p=0.04) and in the aromatase (p=0.03) in the cf. only erα and erβ were not changed by maternal caffeine. The serum testosterone levels in the cf offspring were 90% lower than in the c offspring (p=0.04).CONCLUSION: Maternal caffeine consumption has a role and alters the testis of the offspring in adulthood.

OBJETIVO: Investigar os efeitos da desnutrição proteica materna durante a lactação sobre as fibras elásticas da traqueia de filhotes de ratos Wistar. MÉTODOS: Ao nascimento, 12 filhotes machos de duas ratas Wistar foram igualmente divididos em dois grupos: grupo controle, cuja mãe recebeu água e dieta padrão de laboratório ad libitum durante a lactação, e grupo restrição proteica (RP), cuja mãe recebeu água ad libitum e dieta isoenergética com RP (8% de proteína). Aos 21 dias de vida, os filhotes foram sacrificados, e suas traqueias foram ressecadas. As fibras elásticas foram coradas pelo método de resorcina-fucsina de Weigert (precedido de oxidação) e avaliadas sob microscopia óptica. As determinações morfométricas foram feitas por estereologia, utilizando o método de contagem de pontos, e expressas em densidade volumétrica. RESULTADOS: As fibras elásticas foram identificadas abaixo da mucosa traqueal, sendo a maioria em distribuição longitudinal. Além disso, camadas circulares bem definidas de fibras elásticas envolviam as superfícies interna e externa do anel cartilaginoso. Não houve diferenças entre os grupos quanto à organização e distribuição das fibras elásticas. A densidade volumétrica das fibras elásticas dos filhotes nos grupos controle e RP foi de, respectivamente, 2,46 ± 0,99% e 3,25 ± 1,13% (p < 0,01). CONCLUSÕES: Nossos resultados sugerem que a densidade volumétrica de fibras elásticas é maior em filhotes de ratos alimentados por fêmeas submetidas a dieta com RP do que naqueles de mães recebendo dieta normal.

OBJECTIVE: To investigate the effects of maternal protein malnutrition during lactation on the elastic fibers in the tracheas of Wistar rat pups. METHODS: At delivery, 12 male pups of two Wistar rat dams were equally divided into two groups: control, in which the dam received water and standard rat chow ad libitum during lactation; and protein-restricted (PR), in which the dam received water ad libitum and an isoenergetic PR diet (8% protein). At 21 days of age, the pups were killed and their tracheas were excised. The elastic fibers were stained with Weigert's resorcin-fuchsin (after oxidation) and evaluated under light microscopy. Morphometric determinations were performed by stereology, with the point-counting method, and expressed as volumetric densities. RESULTS: Elastic fibers, most having a longitudinal distribution, were identified beneath the tracheal mucosa. In addition, well-defined circular layers of elastic fibers were found around the inner and outer surfaces of the cartilaginous ring. There were no differences between the groups regarding the organization and distribution of the elastic fibers. The volumetric density of the elastic fibers of the pups in the control and PR groups was 2.46 ± 0.99% and 3.25 ± 1.13%, respectively (p < 0.01). CONCLUSIONS: The volumetric density of elastic fibers appears to be greater in rat pups breastfed by dams receiving a PR diet than in those breastfed by dams receiving a normal diet.

OBJETIVO: Avaliar se o tratamento neonatal com leptina durante os primeiros dias de vida poderia programar o peso dos orgãos do sistema reprodutor masculino e o perfil lipídico. MÉTODOS: Ao nascimento seis ratas-mãe foram distribuídas em dois grupos: Leptina - cada filhote recebeu 50μL de leptina recombinante (80ng/gPC, SC) nos primeiros 10 dias de lactação; Controle - cada filhote recebeu o mesmo volume de salina. Após o desmame, todos os filhotes tiveram acesso ilimitado a ração até 190 dias de idade quando foram mortos. Os dados são expressos como média ± erro padrão e foram analisados pelo teste t Student. RESULTADOS: O tratamento com leptina resultou em aumento significativo no peso corporal (Control= 411.8±16.31; Leptin= 481.8±11.29, p=0.005) e consumo alimentar (Control= 25.32±0.09; Leptin= 32.42±0.15, p=0.0001) e redução significativa nos níveis séricos de triglicerídeos (Control= 540.0±117.9; Leptin= 93.25±15.21, p=0.006), no peso do hipotálamo (Control= 0.234±0.016; Leptin= 0.154±0.015, p=0.007), hipófise (Control= 0.104±0.0120; Leptin= 0.033±0.012, p=0.003), testículo (Control= 3.75±0.055; Leptin= 3.19±0.10, p=0.002) e próstata (Control=1.641±0.1389; Leptin= 0.91±0.07, p=0.001). CONCLUSÃO: O tratamento com leptina nos primeiros dias de vida pode programar o peso dos órgãos do sistema reprodutor e o perfil lipídico da prole.

PURPOSE: To evaluate whether the neonatal leptin treatment during the first days of life can program the male reproductive organs weight and the lipid profile. METHODS: At birth 6 dams were divided into 2 groups: Leptin - each pup was injected with 50μL of recombinant rat leptin (80ng/g BW, sc), for the first 10 d of lactation; Control - each pup received the same volume of saline. After weaning, all pups received unlimited access to food until 190 days of age when they were killed. Values are given as mean ± SEM of 6 animals and Test t Student was used to analyze the results. RESULTS: The leptin treatment resulted in a significant increase in body weight (Control= 411.8±16.31; Leptin= 481.8±11.29, p=0.005) and food consumption (Control= 25.32±0.09; Leptin= 32.42±0.15, p=0.0001) and a significant reduction in triglycerides levels (Control= 540.0±117.9; Leptin= 93.25±15.21, p=0.006) and in the weight of hypothalamus (Control= 0.234±0.016; Leptin= 0.154±0.015, p=0.007), pituitary (Control= 0.104±0.0120; Leptin= 0.033±0.012, p=0.003), testis (Control= 3.75±0.055; Leptin= 3.19±0.10, p=0.002) and prostate (Control=1.641±0.1389; Leptin= 0.91±0.07, p=0.001). CONCLUSION: Leptin treatment on the first days of life can program the reproductive organs weight and the lipid profile of the progeny.

Foi avaliado o consumo alimentar de animais adultos submetidos à restrição protéica, cujas mães receberam dieta hipoprotéica ou hipocalórica na lactação: controle (C) ração normal com 23% de proteína; restrição protéica (RP) 8% de proteína; restrição energética (RE) 23% de proteína, em quantidade restrita à ingerida pelo grupo restrição protéica. Após o desmame todos os filhotes receberam ração normal até 60 dias e nesta época foram submetidos, por 21 dias, ao seguinte tratamento: (C/C)-filhotes de mães C recebendo ração normal; (restrição protéica/controle)-filhotes de mães Controle, recebendo ração hipoprotéica; (C/RP)-filhotes de mães RP recebendo ração normal; (RP/RP)-filhotes de mães RP recebendo ração hipoprotéica; (C/RE)-filhotes de mães RE recebendo ração normal; (RP/RE)-filhotes de mães RE recebendo ração hipoprotéica. Os filhotes de mães RP consumiram menos ração até 57 dias (p<0,01), enquanto os filhotes de mães RE normalizaram a ingestão aos 37 dias. Aos 81 dias, os animais submetidos à restrição protéica consumiram menos ração (p<0,01). A dieta materna na lactação modificou o consumo alimentar e o peso corporal da prole na vida adulta, estando, possivelmente, a restrição protéica associada a uma alteração permanente no controle hipotalâmico da seleção de nutrientes da prole.

This study evaluated food intake of adult animals whose mothers received protein- or energy-restricted diets during lactation as follows: control (C)- 23% of protein; protein-restricted (PR) - 8% of protein; energy-restricted (ER) - 23% of protein, restricted in quantity. After weaning all pups received the control diet until 60 days, when they were separated into groups, with free access to the following diets during 21 days: (C/C)- pups of control dams fed control diet; (PR/C)-pups of control dams fed protein-restricted diet; (C/PR)- pups of protein-restricted dams fed control diet; (PR/PR)-pups of protein-restricted dams fed protein-restricted diet; (C/ER)-pups of energy-restricted dams fed control diet; (PR/ER)-pups of energy-restricted dams fed protein-restricted diet. Pups of protein-restricted dams consumed less food until day 57 (p<0.01), while pups of energy-restricted dams normalized their ingestion at 37 days. At 81 days, all malnourished animals consumed less food than the control (p<0.01). The kind of diet consumed in the lactation period changed the food intake and the body weight of the offspring in the adulthood. So, the protein restriction can possibly be associated with a permanent alteration in the hypothalamic control of nutrients selection of the offspring.