Androgenic, but not estrogenic, protection of motoneurons from somal and dendritic atrophy induced by the death of neighboring motoneurons

Motoneuron loss is a significant medical problem, capable of causing severe movement disorders or even death. We have been investigating the effects of motoneuron loss on surviving motoneurons in a lumbar motor nucleus, the spinal nucleus of the bulbocavernosus (SNB). SNB motoneurons undergo marked dendritic and somal atrophy following the experimentally induced death of other nearby SNB motoneurons. However, treatment with testosterone at the time of lesioning attenuates this atrophy. Because testosterone can be metabolized into the estrogen estradiol (as well as other physiologically active steroid hormones), it was unknown whether the protective effect of testosterone was an androgen effect, an estrogen effect, or both. In the present experiment, we used a retrogradely transported neurotoxin to kill the majority of SNB motoneurons on one side of the spinal cord only in adult male rats. Some animals were also treated with either testosterone, the androgen dihydrotestosterone (which cannot be converted into estradiol), or the estrogen estradiol. As seen previously, partial motoneuron loss led to reductions in soma area and in dendritic length and extent in surviving motoneurons. Testosterone and dihydrotestosterone attenuated these reductions, but estradiol had no protective effect. These results indicate that the neuroprotective effect of testosterone on the morphology of SNB motoneurons following partial motoneuron depletion is an androgen effect rather than an estrogen effect.     

知母中甾体皂苷类成分的研究(英文)

从中药知母(Anemarrhena asphodeloides Bge.)的70%乙醇提取物中分离得到6个化合物,经波谱学方法和与已知样品对照鉴定为:3-O-β-D-glucopyranosyl-(1→2)-β-D-galactopyranosyl-(3β,5β)-pregn-16(17)-ene-20-one(1)、timosaponin AIII(2)、timosaponin BIII(3)、22-hydroxy-5β-furost-3β,15α-diol-3-O-β-D-glucopyranosyl(1→2)-β-D-galactopyranoside(4)、timosaponin G(5)、β-daucosterol(6)。其中化合物1为新天然产物,命名为知母孕甾A,是知母中首次发现的C21甾体类化合物,经HR ESI-MS、1D NMR、2D NMR确定其结构,并首次对其核磁数据进行归属。     

角果藜的化学成分研究(英文)

从新疆特色植物角果藜(Ceratocarpus arenarius L.)地上部分乙醇提取物的乙酸乙酯和氯仿萃取部分共分离得到11个化合物,通过波谱数据结合理化性质分别鉴定为7-酮基-β-谷甾醇(1)、β-谷甾醇(2)、豆甾-3-O-β-D-吡喃葡萄糖苷(3)、正十六碳醇(4)、[24S]豆甾-4-烯-3-酮(5)、173-脱镁叶绿素乙酯(6)、麦黄酮(7)、穆坪马兜铃酰胺(8)、邻苯二甲酸二(2-乙基己基)酯(9)、β-谷甾醇亚油酸酯(10)、丁香树脂酚葡萄糖苷(11)。所有化合物均为首次从该植物中分离得到。     

Testosterone action on erythropoiesis does not require its aromatization to estrogen: Insights from the testosterone and estrogen treatment of two aromatase-deficient men

Androgens act on erythropoiesis, but the relative role of testosterone (T) and estradiol (E₂) on erythropoietic parameters in men is a poorly investigated issue. In order to evaluate separately the effects on erythropoiesis of high-dose T administration alone and of physiological dose of E₂ administration alone two adult men with aromatase deficiency were assessed before and during each treatment. Blood cell count, hemoglobin (Hb), hematocrit (Hct), erythrocyte mean cell volume (MCV), erythrocyte mean corpuscular hemoglobin (MCH), erythrocyte mean corpuscular hemoglobin concentration (MCHC), serum ferritin, iron and total iron-binding capacity (TIBC), serum erythropoietin, serum total testosterone and estradiol were evaluated. Hb, Hct and red cell count rose during testosterone treatment, consistently with the increase in circulating testosterone, but failed to increase during estradiol treatment. A decrease in Hb, Hct and red cell count was recorded in one of the two subjects during estradiol treatment, with a concomitant decrease in serum testosterone. Circulating T alone is capable of and sufficient to influence erythropoiesis, especially at supraphysiological dosage, while circulating E₂ have not the same effect on erythropoietic parameters, suggesting the hypothesis that the erythropoietic changes induced by androgens are not mediated via its aromatization to estrogens.     

Positive Darwinian selection results in resistance to cardioactive toxins in true toads (Anura: Bufonidae)

Members of the Family Bufonidae, true toads, are famous for their endogenously synthesized cardioactive steroids that serve as defensive toxins. Evolution of resistance to these toxins is not understood. We sequenced a key region of the toxin''s binding site in the Na⁺/K⁺ ATPase for relevant taxa representing Hyloidea (including bufonids), Ranoidea and Archaeobatrachia and tested for positive selection in a phylogenetic context. Bufonidae were distinct from other Hyloidea at 4–6 of 12 sites and, with one exception, had a homologous amino acid sequence. Melanophryniscus stelzneri had a distinct sequence, consistent with other independent evidence for a differentiated toxin. Tests within Bufonidae detected positive selection within the binding region, providing, to our knowledge, the first evidence of this type for positive selection within Amphibia. There was no evidence for positive selection on Bufonidae or M. stelzneri lineages. Sequence change in Leptodactylus ocellatus, a leptodactylid predator of Bufonidae, provides a molecular basis for predator resistance possibly associated with gene duplication.     

Seasonal influence on fecal immunoreactive testosterone concentrations of male Formosan black bears (<Emphasis Type="Italic">Ursus thibetanus formosanus</Emphasis>)

We investigated the effect of seasonal influence and geographic location on the testosterone concentrations of three adult male Formosan black bears kept at a Low Altitude Experimental Station, simulating natural breeding conditions. Estimations of fecal testosterone for males 1, 2, and 3 were made during January to October 2000, April 2001 to December 2003, and April 2002 to December 2003, respectively, using non-invasive enzyme-linked immunosorbent assays. Results indicated that fecal immunoreactive testosterone recrudesces after winter and ceases after summer. Mating behaviors were observed both before and after the fecal immunoreactive testosterone peak of March–May. In addition, fecal immunoreactive testosterone concentrations were significantly (p < 0.01) higher during January to June than from July to December. As expected, levels corresponded to the longer and earlier breeding seasons of subtropical zones. Formosan black bears appear to be seasonal breeders who breed primarily in spring, synchronized with sunlight and ambient temperature. The aim of this study was to determine if testosterone concentrations respond to seasonality thereby reflecting bear mating demands. Baseline parameters established in this study will help to evaluate endocrine information applicable for guidelines in future management and preservation of Formosan black bears in the field. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Seasonality, sociality, and reproduction: Long-term stressors of ring-tailed lemurs (Lemur catta)

Fecal glucocorticoid (fGC) concentrations are reliable, non-invasive indices of physiological stress that provide insight into an animal's energetic and social demands. To better characterize the long-term stressors in adult members of a female-dominant, seasonally breeding species - the ring-tailed lemur (Lemur catta) - we first validated fecal samples against serum samples and then examined the relationship between fGC concentrations and seasonal, social, demographic, genetic, and reproductive variables. Between 1999 and 2006, we collected 1386 fecal samples from 32 adult, semi-free-ranging animals of both sexes. In males and non-pregnant, non-lactating females, fGC concentrations were significantly elevated during the breeding season, specifically during periods surrounding known conceptions. Moreover, group composition (e.g., multi-male versus one-male) significantly predicted the fGC concentrations of males and females in all reproductive states. In particular, the social instability introduced by intra-male competition likely created a stressor for all animals. We found no relationship, however, between fGC and the sex, age, or heterozygosity of animals. In reproducing females, fGC concentrations were significantly greater during lactation than during the pre-breeding period. During pregnancy, fGC concentrations were elevated in mid-ranking dams, relative to dominant or subordinate dams, and significantly greater during the third trimester than during the first or second trimesters. Thus, in the absence of nutritional stressors, social dominance was a relatively poor predictor of fGC in this female-dominant species. Instead, the animals were maximally challenged by their social circumstances and reproductive events—males by competition for mating opportunities and females by late-term gestation and lactation.     

Relationship between androgens, environmental factors and reproductive behavior in male white rhinoceros (Ceratotherium simum simum)

We conducted a longitudinal study of the endocrine activity of free-range male white rhinos. An enzyme immunoassay to measure androgens in the feces was developed and validated to show that it can be used to study testicular activity. We identified two fecal metabolites similar to testosterone and dihydrotestosterone. Several lines of evidence suggest that these metabolites clearly reflect testicular activity. Firstly, the stimulation of testicular activity with synthetic GnRH caused a 156% increase in androgen metabolite concentrations in the feces 1 day after treatment. Secondly, androgen metabolite concentrations increased with sexual maturity in rhinos, and finally there was a correlation between testosterone concentrations in plasma and androgen metabolite concentrations in feces. Using the method that we developed, it was possible to establish whether a relationship exists between androgen metabolite concentrations, the behavior and environmental factors. Adult territorial males (n = 5) had elevated androgen metabolite concentrations during months of high rainfall (September-February) compared to months of little or no rainfall (March-August). The increase in concentrations coincided with the beginning of the rainy season, suggesting a seasonal trend in reproduction. This trend was confirmed by behavior observations showing both a higher frequency of conceptions within the first 4 months of increased androgen metabolite concentrations, and a higher number of inter-sexual conflicts, reflecting the initial aggression between the sexes during the consort period. It was also evident that males accompanying a receptive female had higher fecal androgen metabolite concentrations compared to being alone. The elevated levels were likely induced by female presence.     

Glutamic acid decarboxylase (GAD65) immunoreactivity in brains of aggressive, adolescent anabolic steroid-treated hamsters

Chronic anabolic-androgenic steroid (AAS) treatment during adolescence facilitates offensive aggression in male Syrian hamsters (Mesocricetus auratus). The current study assessed whether adolescent AAS exposure influenced the immunohistochemical localization of glutamic acid decarboxylase (GAD65), the rate-limiting enzyme in the synthesis of gamma-aminobutyric acid (GABA), in areas of hamster brain implicated in aggressive behavior. Hamsters were administered high dose AAS throughout adolescence, scored for offensive aggression, and then examined for differences in GAD65 puncta to regions of the hamster brain important for aggression. When compared with control animals, aggressive AAS-treated hamsters showed significant increases in the area covered by GAD65 immunoreactive puncta in several of these aggression regions, including the anterior hypothalamus, ventrolateral hypothalamus, and medial amygdala. Conversely, aggressive AAS-treated hamsters showed a significant decrease in GAD65-ir puncta in the lateral septum when compared with oil-treated controls. However, no differences in GAD65 puncta were found in other aggression areas, such as the bed nucleus of the stria terminalis and central amygdala. Together, these results support a role for altered GAD65 synthesis and function in adolescent AAS-facilitated offensive aggression.