Postnatal testicular development in Vietnamese pot-bellied pigs.

The Vietnamese pot-bellied pig is a breed with high investigation potential. However, at the reproductive level, its testicular characteristics are still unknown, as well as the different stages of its development. Therefore, the objective of this work is to describe the postnatal testicular development of Vietnamese pot-bellied pigs. In this study, we used pigs grouped into the neonatal stage, animals at zero weeks; prepubertal stage, animals at three and eight weeks; pubertal stage, animals at twelve and sixteen weeks; and postpubertal stage animals at twenty, twenty-four, twenty-eight and thirty-two weeks of age. The neonatal stage is characterized by gonocytes at different migration phases. In the prepubertal stage, gonocytes were differentiated into spermatogonia at 3 weeks of age, and the first spermatocytes were observed at 7 weeks of age. Puberty was determined to start at 12 weeks because seminiferous tubules are found with complete spermatogenesis and the highest peaks in positive cell counting of androgen receptors (AR) and proliferating cell nuclear antigen (PCNA) factor that later decreased and further stabilized in the following weeks. In the postpubertal stage, an increase in seminiferous tubule areas was observed. The number of apoptotic cells ranged from low to null at all ages. Testosterone (T) and gonadotropin levels had two important peaks at 3 and 24 weeks. The seminiferous epithelium cycle was found to have 11 stages according to acrosome development. These characteristics of Vietnamese pot-bellied pig testes, which are different from rat testes and more similar to human testicles, make them a viable model to study human male reproductive biology. The postnatal testicular development of pot-bellied pigs is different from the postnatal testicular development of other breeds. Therefore, due to this difference in size and easy manipulation, the Vietnamese pig is an alternative for investigation compared to other pig breeds.

Inhibition of the nitric oxide/cyclic guanosine monophosphate pathway limited the cardioprotective effect of post-conditioning in hearts with apical myocardial infarction.

Reperfusion damage involves opening of the mitochondrial permeability transition pore (mPTP) and loss of ATP synthesis. Several cardioprotective pathways are activated by ischemic or pharmacological post-conditioning (PC). The mechanisms that are activated by PC in no co-morbidity murine models include: activation of rescue kinases, oxidative stress reduction, glycolytic flux regulation and preservation of ATP synthesis. However, relatively scarce efforts have been made to define whether the efficacy of PC signaling is blunted by risk factors or systemic diseases associated with ischemic heart pathology. Experimental evidence has shown that the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) signaling is a main mechanism activated by PC in hearts without pathological history. In this work we evaluated the participation of the NO pathway, through downstream kinase activation and inhibition of mPTP in hearts with previous infarct. Myocardial infarction was induced with a single dose of isoproterenol (85 mg/kg i.p.) to male Wistar rats. After 24 h, the hearts were mounted into the Langendorff system and subjected to 30 min of ischemia and 60 min of reperfusion. PC consisted of 5 cycles of 30 s of reperfusion/30 s of ischemia, then the hearts were reperfused with or without inhibitors of the NO/cGMP pathway. PC activates the NO/cGMP pathway, as increased cGMP and NO levels were detected in isoproterenol-treated hearts. The cardioprotective effect of PC was abolished with both L-NAME (inhibitor of constitutive NO synthase) and ODQ (inhibitor of soluble guanylate cyclase), whereas the NO donor (DETA-NO) restored cardioprotection even in the presence of L-NAME or ODQ. We also found that mitochondrial structure and function was preserved in PC hearts. We conclude that PC exerts cardioprotection in hearts with previous infarct by maintaining mitochondrial structure and function through NO-dependent pathway.

Prenatal diagnosis in a cystic fibrosis family: a combined molecular strategy for a precise diagnosis.

INTRODUCTION: The high genetic heterogeneity in populations with a wide spectrum of mutations in the CF transmembrane conductance regulator gene (CFTR), makes the detection of mutations a very hard and difficult task, thereby limiting the accurate diagnosis of the disease, mainly in patients with uncharacterized mutations. MATERIAL AND METHODS: Molecular strategies, like targeted identification of the most frequent CFTR mutations in Mexican population combined with linkage analysis using markers, is very useful for carrier detection and for prenatal diagnosis in affected families with CF. In this paper we show that the combination of methodologies was a crucial alternative to reach a precise prenatal CF diagnosis. We documented CF diagnosis in a 14th-week fetus combining the screening of the most common mutations in Mexican population with linkage analysis of two extragenic polymorphisms (XV2C/TaqI and KM19/PstI). RESULTS: We determined that the fetus inherited the PG542X mutation from its mother and an unknown mutation from its father through the chromosomal phases analysis.

Zinc-induced survival of Leydig cells in Fischer rats (Rattus norvegicus) treated with cadmium chloride.

Zinc is known to prevent cadmium-induced carcinogenesis and Leydig cell destruction in rat testes; however, the mechanism of action is not known, although it has been suggested that pituitary feedback increases the production of luteinizing hormone (LH) in response to low circulating androgen. We therefore examined the biological role of zinc in reducing cadmium toxicity in the Leydig cells of Fischer rats. Two groups of eleven 6-month-old rats were injected subcutaneously with 20 micromol CdCl2/kg weekly for 5 weeks; one of these groups also received 1 mmol/kg zinc acetate weekly for the same 5 weeks. A third group of rats received 1 mmol/kg zinc acetate weekly, and a fourth group was injected with saline weekly for 5 weeks. After 8 months of study, the animals were euthanized by CO2 inhalation. The results indicated that the number of surviving Leydig cells was significantly lower in the cadmium group (7.34% = 0.095 x 10(9)/cm3) than in the cadmium-zinc group (20.85%) or control animals (91.2%). Moreover, the concentrations of serum testosterone and LH were significantly higher in the cadmium group than in any of the other groups. This difference probably was due to the testosterone produced by a small reservoir of surviving Leydig cells and to other endocrine factors. These findings suggest that Fischer rat testis may be a good model system for testing the effects of cadmium and zinc on the production of LH and testosterone and other androgens before spontaneous cancers develop.

Gubernacular fibroblasts express the androgen receptor during testis descent in cryptorchid rats treated with human chorionic gonadotrophin.

Cryptorchidism was provoked in 3 day old rats treated with 17-beta-estradiol over 30 days to identify the cells that express the androgen receptor (AR) during experimental testis descent in the gubernaculum. In one group of animals, testis descent was induced with human chorionic gonadotrophin (hCG) applied daily for 5 or 10 days. A correlative study using a testosterone radioimmunoassay with electron microscopy and immunocytochemical detection of AR was performed in gubernacula of hCG treated and untreated control animals. The gubernaculum of rats undergoing testes descent showed a dramatic increase in the number of AR-positive cells. These were located in the connective tissue among smooth muscle cells in the gubernacular cord and between striated muscle fibers in the bulb. In both regions, the AR-positive cells were identified as fibroblasts. Several clusters of amorphous material appeared in the extracellular matrix of the connective tissue in hCG treated rats. Our results suggest that testosterone induces the expression of AR in gubernacular fibroblasts which seem to degrade the extracellular matrix during gubernacular involution.

Androgen receptor and calcitonin gene-related peptide in neurons of the genitofemoral nerve during testicular descent induced with human chorionic gonadotropin.

BACKGROUND: Low levels of circulating testosterone during testis descent cause cryptorchidism in humans and rats. Treatment with human chorionic gonadotrophin (hCG) induces testis descent by stimulating production of testosterone (T). Neurons of genitofemoral nerve (GFN), which innervate testicular gubernaculum, may play a role in testis descent. METHODS: In the current study, putative correlations were made between T and GFN motor and sensory neuron activity during inguinoscrotal testis descent. Cryptorchidism was provoked in prepuberal rats with estradiol. Rats with testicular descent induced with hCG and cryptorchid controls were used. Cells of spinal cord and dorsal root ganglia were labeled by retrograde staining with fast-blue. Expression of androgen receptor (AR) and calcitonin gene-related peptide (CGRP) were detected with indirect immunofluorescence. RESULTS: Neurons labeled with fast-blue were found in the center of motor horn and dorsal root ganglia at levels L1 and L2. While number of motor neurons expressing AR was significantly higher in the group treated with hCG, number expressing CGRP was higher in controls. In dorsal root ganglion, number of cells immunostained with CGRP antibody was similar in both groups but AR was not detected. CONCLUSIONS: Present results support the hypothesis that motor nucleus of the GFN is a direct target of testosterone and that regulation of CGRP in sensory nucleus may be involved in testicular descent.