Spermatogenic cycle length and spermatogenic efficiency in the gerbil (Meriones unguiculatus)

The gerbil (Meriones unguiculatus) is a rodent native of the and regions of Mongolia and China. Because the gerbil can be easily bred in laboratory conditions, this species has been largely used as an experimental model in biomedical research. However, there is still little information concerning the testis structure and function in the gerbil. In this regard, we performed a detailed morphofunctional analysis of the gerbil testis and estimated the spermatogenic cycle length utilizing H-3-thymidine as a marker for germ cell progression during their evolution through the spermatogenic process. The stage frequencies of the XII stages characterized according to the acrosome formation and development were (I-XII) 13.8, 10.1, 8.1, 7.8, 4.0, 11.2, 7.5, 7.1, 5.9, 7.6, 8.1, and 8.9. The mean duration of each seminiferous epithelium cycle was determined to be 10.6 +/- 1.0 days and the total duration of spermatogenesis, based on 4.5 cycles, was approximately 47.5 days. The volume density of tubular and interstitial compartments was approximately 92% and 8%, respectively. Based on the volume occupied by seminiferous tubules in the testis and the tubular diameter, about 9 and 18 m of seminiferous tubules were found per testis and per gram of testis, respectively. Twelve primary spermatocytes were formed from each type A1 spermatogonia. The meiotic index was 2.8, indicating that 30% of cell loss occurs during meiosis. The number of Leydig and Sertoli cells per gram of the testis was 28 million and each Sertoli cell was able to support approximately 13 spermatids. The daily sperm production per gram of testis (spermatogenic efficiency) was 33 million. Taken together, these data indicate that, mainly due to the high seminiferous tubule volume density and Sertoli cell support capacity for germ cells, the gerbil presents high spermatogenic efficiency compared with other mammalian species already investigated. The data obtained in the present study might provide the basis for future research involving the reproductive biology in this species.

Prevalência de patologias no trato genital de machos bubalinos (Bubalus bubalis) nos estados do Pará, Amapá e baixo Amazonas, Brasil

Distúrbios reprodutivos estão associados a problemas adquiridos (estresse, manejo inadequado e de origem infecciosa), e genéticos (alta consangüinidade existente em alguns rebanhos do Brasil). Embora mostre o grau do potencial de fertilidade do macho, o exame andrológico e, em particular, o teste de funcionalidade, tem sido pouco utilizado, principalmente na seleção dos touros, embora, estes mesmos estudos, mencionem a importância do sêmen para avaliar a eficiência reprodutiva do macho. O objetivo do estudo foi determinar a prevalência de patologias clinicamente notáveis, através de palpação e inspeção e caracterizar o perfil espermático de búfalos criados em sistemas de produção, intensivo e extensivo. O estudo foi realizado em propriedades do Estado do Pará, nos Municípios de Belém, Ipixuna, Mojú, Nova Timbotéua, Paragominas, Santarém Novo e São Caetano de Odivelas, Muaná, Soure, Chaves e Ponta de Pedras. No estado do Amapá, nos Municípios de Cutias, Itaubal, Tartarugalzinho e no Arquipélago do Bailique. Os animais eram criados no sistema intensivo e extensivo, respectivamente. O período do estudo foi de março de 2008 a março de 2009. Foram utilizados 305 touros da raça Murrah, Mediterrânea e mestiços, com idade variando entre 2 a 15 anos, sendo que 160 no Estado do Pará, e 145 no Estado do Amapá. Os dados dos exames clínicos - andrológicos, assim como o espermiograma foram documentados em fichas especiais. A obtenção de sêmen foi realizada pela massagem das ampolas a avaliação física do sêmen contou da cor, aspecto, volume, motilidade, vigor, concentração e a avaliação morfológica e pH. A concentração foi aferida por espectrofotômetro da Central de Biotecnologia Animal (CEBRAN). Para a morfologia espermática foi utilizado o método de Cerovsky. Foram analisadas 200 espermatozóides em microscópio óptico em objetiva de imersão. Para análise estatística, foi empregado o programa estatístico BIOESTAT 5.0, estatísticas descritivas para todas as variáveis estudadas, biometrias testiculares, patologias do sistema reprodutor e características físicas e morfológicas do sêmen, registrando-se as médias, os desvios padrões e a distribuição de freqüências para as características de classes andrológicas. Os dados quantitativos referentes às idades dos touros, biometrias testiculares, avaliações físicas e morfológicas do sêmen foram submetidos à ANOVA, e quando houve efeito significativo pelo teste F, às médias foram comparadas pelo teste de Tukey a 5 % de probabilidade de erro. Dos 305 búfalos, nos animais do Estado do Amapá a media e o desvio padrão da circunferência escrotal foi superior ao do Estado do Pará independente do tipo de manejo. Indiferente de idade e sistemas de criação não foi encontrado nenhuma alteração no pênis. A bolsa escrotal bífida (fenda escrotal) foi a alteração mais prevalente. A maior freqüência de alterações no epidídimo foi epididimite granulomatosa tipo tuberculóide, observada nos animais de criação extensiva no estado do Amapá, com uma tendência aos touros de mais de quatro anos. Entre as alterações do testículo a mais prevalente foi à torção testicular e de causas infecciosa destacamos a ocorrência de casos de orquite interticial crônico ativa, orquite granulomatosa tipo tuberculóide, hipoplasia testicular grau I, hipoplasia testicular grau III associada à orquite tuberculóide e hipoplasia grau II (parcial). Outra observação bem evidente é com relação às percentagens de espermatozóide com defeitos maiores e menores que se apresentam muito altas as recomendadas pelo CBRA. Deste estudo pode-se concluir que os reprodutores criados de forma extensiva apresentam patologias de origem genética e infecciosa muito prevalente.

Impact of adrenalectomy and dexamethasone treatment on testicular morphology and sperm parameters in rats: insights into the adrenal control of male reproduction

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Características seminais e resfriamento de sêmen de tamanduá (Myrmecophaga tridactyla) de vida livre

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

High-Fat Diet Obesity Associated With Insulin Resistance Increases Cell Proliferation, Estrogen Receptor, and PI3K Proteins in Rat Ventral Prostate

In this study, we evaluated the effects of obesity and insulin resistance induced by a high-fat diet on prostate morphophysiology, focusing on cell proliferation, expression of androgen (AR) and estrogen receptors (ER) and proteins of the insulin signaling pathway. Adult male Wistar rats were fed a high-fat diet (20% fat) for 15 weeks, whereas control animals received a balanced diet (4% fat). Both groups were then divided and treated for 2 weeks with 1 mg/kg body weight/day of the aromatase inhibitor letrozole or vehicle only. The ventral prostate was analyzed with immunohistochemical, histopathological, stereological, and Western blotting methods. Obese rats showed insulin resistance, hyperinsulinemia, and reduced plasma testosterone levels. The incidence of prostatic intraepithelial neoplasia (PIN) was 2.7 times higher in obese rats and affected 0.4% of the gland compared with 0.1% PIN areas found in control rats. Obesity doubled cell proliferation in both prostate epithelium and stroma. AR content decreased in the prostate of obese rats and estrogen receptor beta (ER beta) increased in this group. Protein levels of insulin receptor substrate 1 and protein kinase B diminished in the obese group, whereas phosphatidylinositol 3-kinase (PI3K) increased significantly. Most structural changes observed in the prostate of obese rats normalized after letrozole treatment, except for increased stromal cell proliferation and ER beta expression, which might be associated with insulin resistance. This experimental model of obesity and insulin resistance induced by a high-fat diet increases cell proliferation in rat prostate. Such alterations are associated with decreased levels of AR and increased ER beta and PI3K proteins. This change can facilitate the establishment of proliferative lesions in rat prostate.

Effects of bovine sperm cryopreservation using different freezing techniques and cryoprotective agents on plasma, acrosomal and mitochondrial membranes

The success of semen cryopreservation is influenced by several factors, such as freezing curves and cryoprotectants. These two factors are of special interest once they may lead to many important physical-chemical changes resulting in different degrees of damage in spermatozoa structure. This experiment was designed to compare the effect of bull semen cryopreservation using two freezing techniques: conventional (CT cooling rate of -0.55 degrees C min-1 and freezing rate of -19.1 degrees C min-1) and automated (AT cooling rate of -0.23 degrees C min-1 and freezing rate of -15 degrees C min-1), performed with different curves, and with three cryoprotectants (glycerol, ethylene glycol and dimethyl formamide) on bovine sperm motility and integrity of plasma, acrosomal and mitochondrial membranes. These variables were simultaneously evaluated using the fluorescence probes propidium iodide, fluorescein-conjugated Pisum sativum agglutinin and MitoTracker Green FM. The effects of freezing techniques, as well as of different cryoprotectants were analysed by the analysis of variance. The means were compared by Fishers test. There were no significant differences between freezing techniques (P > 0.05). Glycerol showed higher percentages of motility, vigour and integrity of plasma, acrosomal and mitochondrial membranes than other two cryoprotectants (P < 0.05). Ethylene glycol preserved higher motility and integrity of plasma and mitochondrial membranes than dimethyl formamide (P < 0.05). Sperm motility with glycerol was 30.67 +/- 1.41% and 30.50 +/- 1.06%, with ethylene glycol was 21.17 +/- 1.66% and 21.67 +/- 1.13% and with dimethyl formamide was 8.33 +/- 0.65% and 9.17 +/- 0.72% to CT and AT curves, respectively. The percentage of spermatozoa with simultaneously intact plasma membrane, intact acrosome and mitochondrial function (IPIAH) was 14.82 +/- 1.49% (CT) and 15.83 +/- 1.26% (AT) to glycerol, 9.20 +/- 1.31% (CT) and 9.92 +/- 1.29% (AT) to ethylene glycol 4.65 +/- 0.93% (CT) and 5.17 +/- 0.87% (AT) to dimethyl formamide. Glycerol provided the best results, although nearly 85% of spermatozoa showed some degree of injury in their membranes, suggesting that further studies are required to improve the results of cryopreservation of bovine semen.