Male hamster sociosexual behaviors: effects of testosterone and its metabolites

Male hamsters were tested for copulatory behavior (CB) with receptive females, for investigatory responses to the females' ano-genital region (A/G), and for attraction to female hamster vaginal secretion (FHVS). After castration, the males received Silastic capsules containing one of two doses of testosterone (T), 5 alpha-dihydrotestosterone (DHT), estradiol (E2) or DHT + E2, and the maintenance of their copulatory and chemoinvestigatory responsiveness was assessed during weekly tests for the next month. The major findings were: (1) T thresholds for the maintenance of CB were lower than they were for the maintenance of A/G behavior and FHVS attraction; (2) DHT + E2 or DHT alone were more effective in maintaining A/G and FHVS attraction than was E2 alone; (3) DHT + E2 or DHT alone maintained ejaculatory behavior in some animals but E2 did not; (4) the posttreatment maintenance of normal ejaculation latencies and intromissions to ejaculation shown by intact and T-treated males was not demonstrated by males receiving DHT or DHT + E2. These results are consistent with the hypothesis that copulatory and chemoinvestigatory behaviors may be subserved by distinct neuroendocrine mechanisms in male hamsters.     

Androgen-mediated sex differences of cardiovascular responses in rats

Sex differences in the systemic depressor response to arachidonic acid (50 or 150 microgram/kg iv) were observed in intact and castrated anesthetized Sprague-Dawley rats. The rank order of responsiveness was: castrate males, castrate females, females, males; all four groups were significantly different (P less than 0.05) at the higher dose. Castrated males pretreated with testosterone (1 mg/kg sc) 5 or 7 days previously gave a response at the higher arachidonate dose levels that was of the same order as that obtained with intact males. Similar treatment of castrate males with androgen potentiated (P less than 0.05) the vasopressor action of norepinephrine (0.25 microgram/kg) on day 7 after the testosterone pretreatment. In contrast, treatment with depot estradiol (100 microgram/kg sc) in castrate males produced no significant change in the response to either of the vasoactive compounds on both days 5 and 7 after pretreatment. These data suggest that testosterone may be a significant factor in the development of sex differences in the cardiovascular systems of rats.     

Vomeronasal organ removal before sexual experience impairs male hamster mating behavior

Removal of vomeronasal chemoreceptors before sexual experience in male hamsters resulted in complete failure to mate in some animals but removal of these receptors after sexual experience had no effect. Animals were tested for mating behavior with intact behaviorally receptive females and also with anesthetized males scented with vaginal fluid. The two tests produced essentially the same result. Histological analysis of the lesions and radioimmunoassay of androgen levels showed no group differences, other than vomeronasal organ removal, that could account for the results. The behavioral data suggest that the vomeronasal system may be concerned with the production of preprogrammed behavior.     

Effects of volatile and nonvolatile chemical signals on male sex behaviors mediated by the main and accessory olfactory systems

Estrous hamster vaginal discharge (HVD) contains both volatile and nonvolatile chemical signals which collectively elicit both male attraction to females and male mating behavior. These two aspects of normal sexual behavior are differentially affected by lesions involving afferents of the main and the accessory olfactory systems. The results of lesions that involve the main olfactory system suggest that it provides information primarily concerning those volatile components of HVD that normally signal the presence of a female. Lesions restricted to the accessory olfactory system do not impair a male's interest in or relative preference for HVD but do significantly interfere with subsequent stages of sexual behavior by reducing the amount of mating obtained upon exposure to HVD. Stimulation of the accessory olfactory system by nonvolatile components of HVD has thus been implicated in the production of mating behavior. In these experiments, male hamsters were attracted to female odor and engaged in significant amounts of mating behavior with surrogate females when only the volatile components of HVD were available to them. These behaviors were further enhanced when both volatile and nonvolatile components of HVD were provided. Lesion studies of the afferents involved reinforce the hypothesis that the main olfactory system is preferentially involved with processing those volatile chemical signals in HVD that denote female attractiveness whereas the accessory olfactory system is preferentially involved with processing those chemical signals, both volatile and nonvolatile, that evoke subsequent steps in male sexual behavior.     

Grading ductal carcinoma in situ of the breast using an automated proliferation index

Tumor grade, size and margin status are the most significant factors in predicting the behavior of ductal carcinoma in-situ (DCIS). The inclusion of necrosis and nuclear grade in the grading of DCIS has demonstrated a fair but suboptimal agreement between pathologists. The grading of DCIS was studied and compared to the Van Nuys (VN) system, by using our newly proposed unifying "nuclear grade + proliferation index (N+P) grading system for invasive carcinomas. 162 DCIS tumors were studied including 49 VN I, 31 VN II, and 82 VN III cases. The VN and N+P systems were compared with each other and correlated with tumor size, ER, PR, p53, Her-2, EGFR, Bcl-2, p27 and p21 status. The two systems demonstrated similar frequencies for the different grades and an agreement with each other for all of the biomarkers studied. The greatest difference between the two systems was observed for those tumors initially classified as VN II (94% being down-graded to N+P I) and VN III (80% being down-graded to N+P II). These results suggest that the N+P system, combining nuclear grade with automated MIB-1 count, is a potentially valid and reproducible grading system for both non-invasive and invasive mammary carcinomas. It is automated, less subjective in assessing mitotic activity and necrosis and correlates with other prognostic biomarkers.     

Male hamster copulatory responses to a high molecular weight fraction of vaginal discharge: effects of vomeronasal organ removal

The importance of the vomeronasal (accessory olfactory) system for the copulatory responses of male hamsters to a high molecular weight fraction (HMF) of vaginal discharge was assessed in animals that had their vomeronasal organs (VNO) removed. These organs were extirpated bilaterally using an oral approach through the palate so as to eliminate the peripheral afferents to the accessory olfactory bulb (AOB) with minimal or no damage to the main olfactory system. The selective peripheral deafferentation procedure was verified by applying horseradish peroxidase intranasally following intraperitoneal injections of epinephrine to facilitate the vomeronasal pumping mechanism that draws fluids into the VNO. Heavy, bilateral anterograde labeling was evident in the olfactory nerve afferents within the main olfactory bulb of males that had their VNO removed and of animals that received sham surgery. Sham-operated males also had heavy, bilateral labeling in the vomeronasal nerve afferents within the AOB, whereas no such labeling occurred among animals with bilateral removal of the VNO. In sham-operated animals, both the HMF and the unfractionated discharge significantly increased the incidence of intromission attempts toward anesthetized males (surrogate females) whose hindquarters were scented with these stimuli. The unfractionated discharge also produced a significant elevation of overt copulatory behavior in males with selective peripheral deafferentation of the vomeronasal system, whereas the HMF did not facilitate copulatory behavior in these animals.(ABSTRACT TRUNCATED AT 250 WORDS)     

国内110株新生隐球菌临床株变种、基因型和交配型分析

目的 对国内部分地区的新生隐球菌(Cryptococcus neoformans)临床株进行分子流行病学调查,分析其变种、基因型和交配型的构成和分布.方法 (1)PCR指纹分型法:以野生型噬菌体M13中针对小卫星DNA的核心序列为单引物对模板进行PCR扩增,将所有受试菌株鉴定到8种主要基因型水平.(2)利用变种和交配型特异性引物扩增分型法,区分格鲁比变种(C.neoformans var.grubii)、新生变种(C.neoformans var.neoformans)和格特变种(C.neoformans var.gattii),同时鉴定α和a交配型.结果 110株临床株中,98株(89.1%)为格鲁比变种,均为VNI基因型和α交配型;9株(8.2%)格特变种,包括VGⅠ基因型、α交配型8株(7.3%)和VGⅡ基因型、α交配型1株(0.9%);2株(1.8%)为AD杂合体,VNⅢ基因型,-/α和α/-交配型各1株;1株(0.9%)为新生变种,VNⅣ基因型和a交配型.结论 我国新生隐球菌临床株包含3个变种和AD杂合体.与国外情况比较,相似的是国内临床株中绝大部分为α交配型菌株,且格鲁比变种中的VNⅠ基因型占了其中的大部分;但未发现VNⅡ、VGⅢ和VGⅣ基因型菌株.     

The perirhinal-entorhinal cortex, but not the hippocampus, is critical for expression of individual recognition in the context of the Coolidge effect

The Coolidge effect is a phenomenon in which males show renewed sexual interest in a novel female following copulation to satiety with another female. In golden hamsters, this phenomenon depends on the ability to recognize conspecifics using chemosensory cues processed through the main olfactory system. Here we tested whether olfactory targets in the hippocampal system support this natural form of recognition memory. Male hamsters received ibotenic acid lesions of the perirhinal-entorhinal cortex (PR-ENT) or hippocampus (H) and were allowed to copulate to satiety with a female conspecific, then were presented with two anesthetized females, the familiar mate and an unfamiliar female that copulated with another male. Sham-operated and H-lesioned subjects preferentially investigated the novel female, indicating intact recognition of individual identity. By contrast, PR-ENT-lesioned males failed to discriminate familiar and novel females, and this deficit could not be attributed to abnormal copulatory behavior during mating. All subjects were able to detect and discriminate between female odors when presented in isolation during a habituation-discrimination test, indicating that behavioral deficits shown by PR-ENT males were not due to anosmia or a general investigatory deficit. Thus, the perirhinal-entorhinal cortex, but not the hippocampus, is critical for the recognition of familiar conspecifics in this naturalistic situation. This study reveals an essential role for the perirhinal-entorhinal cortex, but not the hippocampus, in a natural form of recognition memory within the social behavior of hamsters. The findings show a strikingly similar pattern to the effects of selective damage to the same brain regions on performance in standard recognition memory tasks by rats and monkeys. Therefore, the present data extend our understanding of the differential role of structures of the hippocampal memory system, showing continuity across species and between formal laboratory tests and the function of memory in natural social behavior.     

Learned aversion to the odors of male mice: Effects on agonistic behavior

Intact and castrate male mice were subjected to avoidance training using the urine of intact and castrate donors as stimuli. Both intacts and castrates learned to discriminate between the odors, with such learning modifying their spontaneous responsiveness to such odors in an open field in the predicted direction. This result suggests that the previously reported lack of responsiveness in castrates [15] was not due to a simple sensory deficit. A second experiment determined that increasing the relative aversion exhibited by intact subjects to the odors of intact donors also increased the subject's aggression directed at a castrate opponent swabbed with such odors. Modification of castrates' responsivity had no such effect on their aggression. This indicated that replacing the responsivity to aggression-related odors which is lost following castration, is not equivalent to replacing the hormone. The results are discussed with regard to possible hormonal effects on the social significance of olfactory cues to the recipient animal.     

Individual recognition of female hamsters by males: role of chemical cues and of the olfactory and vomeronasal systems

Sexually satiated male hamsters preferred to investigate and to mount an anesthetized, estrous, novel female over a similarly presented female with which the male had become satiated (the Coolidge effect); likewise, such males preferred a novel female recently mated with another male over the familiar female but showed no preference between fresh and mated novel females. Thus the Coolidge effect is at least partly dependent on discrimination of a new female by chemical cues. Another experiment indicated that transfer of a male's own scent during mating is not involved in discrimination between familiar and novel females. Flank gland secretion of females were sufficient for individual discrimination by males, whereas head region scents and vaginal secretions were not sufficient. The presence of female's flank glands was not, however, necessary for such discrimination. Lesions of or removal of the vomeronasal organ did not disrupt the preferences of sexually satiated males for a novel female, but elimination of main olfactory system function by ZnSO4 treatment of the olfactory mucosa did abolish such preferences. Thus olfactory cues are sufficient for individual discrimination of novel females by sexually satiated male hamsters, and such recognition leads to increased sexual arousal. These processes are mediated by the main olfactory system but not the vomeronasal accessory-olfactory system.     

Changes in TNFα, NFκB and MnSOD protein in the vestibular nuclei after unilateral vestibular deafferentation

Background  

Unilateral vestibular deafferentation results in strong microglial and astroglial activation in the vestibular nuclei (VN) that could be due to an inflammatory response. This study was aimed at determining if markers of inflammation are upregulated in the VN after chemical unilateral labyrinthectomy (UL) in the rat, and if the inflammatory response, if any, induces the expression of neuroprotective factors that could promote the plasticity mechanisms involved in the vestibular compensation process. The expressions of inflammatory and neuroprotective factors after chemical or mechanical UL were also compared to verify that the inflammatory response was not due to the toxicity of sodium arsanilate.     

Hypothalamic cuts suggest a brain site for the antigonadotrophic action of melatonin in the syrian hamster

Thrice daily melatonin injections inhibit reproductive functions in pinealectomized male hamsters, as measured by sex organ weights and plasma testosterone levels. Total or anterior, but not posterior, hypothalamic deafferentation prevented the antigonadotrophic effect of melatonin. The findings suggest a site of action of melatonin within the central nervous system.     

Fos expression in female hamsters after various stimuli associated with mating

Detection of the expression of c-fos mRNA or its protein product, Fos, has been used to indicate differences in neuronal response to exogenous stimuli. Factors contributing to differences in Fos expression as a result of various stimuli associated with mating have been extensively studied in the female rat. Less is known about the factors that contribute to Fos expression in female hamsters. Female hamsters differ from female rats in several aspects of sexual behavior; therefore, it seems likely that Fos expression may also differ. The purpose of this study was to determine which factors associated with mating selectively affect Fos expression in the female hamster. Animals were ovariectomized, hormone treated, and then exposed to several behavioral conditions. Fos expression in several brain areas was then assessed via immunocytochemistry (ICC). As has been found by others, mating increases Fos immunoreactivity in a number of brain regions. Specifically, vaginal–cervical stimulation (VCS) was determined to be the salient factor contributing to Fos expression in the preoptic area (POA) and bed nucleus of the stria terminalis (BNST) of ovariectomized hormone primed female hamsters that received a mating interaction.     

The main olfactory system mediates pheromone-induced fos expression in the extended amygdala and preoptic area of the male Syrian hamster

Copulation in male hamsters is stimulated by exposure to vaginal secretions of conspecifics. These pheromones also stimulate fos expression in neural areas that regulate copulation including: the medial nucleus of the amygdala, the bed nucleus of the stria terminalis, and the preoptic area. The pheromones in vaginal secretions are detected by both the main and accessory olfactory systems. However, the accessory system plays the greater role in the regulation of mating behavior and has direct connections with the medial nucleus of the amygdala and bed nucleus of the stria terminalis. The goal of the present study was to determine which system mediates the effect of pheromones on the stimulation of more central areas by deafferenting these systems in experienced male hamsters before exposure to vaginal secretions. Destruction of the receptors in the main olfactory system with zinc sulfate eliminated the increase in fos immunoreactivity in the amygdala, bed nucleus of the stria terminalis and preoptic area following exposure to sexually stimulating pheromones. Deafferentation of the accessory olfactory system by removing the vomeronasal organ had no effect on pheromone-induced fos expression in these areas.We conclude that neurons expressing fos following exposure to vaginal secretions are stimulated via the main olfactory system and are not associated with the expression of copulatory behavior.     

p53, ErbB2, and TAG-72 expression in the spectrum of ductal carcinoma in situ of the breast classified by the Van Nuys system

CONTEXT: The Van Nuys (VN) classification system for ductal carcinoma in situ (DCIS) of the breast is a simplified morphology-based system that uses the presence of nuclear pleomorphism and comedo-type necrosis to stratify DCIS lesions into 3 prognostic groups. OBJECTIVE: To determine if there is an underlying biological basis that correlates with the morphologic aspects of the VN classification system. DESIGN: We evaluated the expression of markers implicated in the development of breast cancer (p53, ErbB2, and TAG-72) in DCIS classified with the VN system. Forty-five cases of pure DCIS were classified as 8 cases of VN1, 7 cases of VN2, and 30 cases of VN3. p53, ErbB2, and TAG-72 antigen expression was measured by immunohistologic means in each of the cases. RESULTS: Nuclear accumulation of p53 was only observed in VN3 (30%). ErbB2 overexpression was found only in VN2 (14%) and VN3 (43%). TAG-72 expression was observed in all categories of lesions but was more frequent in VN2 (71%) and VN3 (70%) compared with VN1 (25%). It appears that overexpression of ErbB2 and p53 are features associated with the high-grade lesions. CONCLUSION: The simplified VN classification system for DCIS has a clear underlying biological basis as evidenced by differential expression of tumor-associated antigens in each of the 3 morphologic categories. These differences may contribute to the differential clinical behavior of the separate groups.     

Fractal properties of sympathetic nerve discharge

Fano factor analysis and dispersional analysis were used to characterize time series of single and multifiber spikes recorded from the preganglionic cervical sympathetic nerve and cardiac-related slow-wave activity of the whole postganglionic sympathetic vertebral nerve (VN) in anesthetized cats. Fluctuations in spike counts and interspike intervals for single preganglionic fibers proved to be fractal (i.e., time-scale invariant), as reflected by a power law relationship between indices of the variance of these properties and the window size used to make the measurements. Importantly, random shuffling of the data eliminated the power law relationships. Fluctuations in spike counts in preganglionic multifiber activity also were fractal, as were fluctuations in the height and of the area of cardiac-related slow waves recorded from the whole postganglionic VN. These fractal fluctuations were persistent (i.e., positively correlated), as reflected by a Hurst exponent significantly >0.5. Although fluctuations in the interval between cardiac-related VN slow waves were random, those in the interval between heart beats were fractal and persistent. These results demonstrate for the first time that apparently random fluctuations in sympathetic nerve discharge are, in fact, dictated by a complex deterministic process that imparts "long-term" memory to the system. Whether such time-scale invariant behavior plays a role in generating the fractal component of heart rate variability remains to be determined.     

Facilitation of sexual receptivity by hypothalamic and midbrain implants of progesterone in female hamsters

In the first experiment, ovariectomized female hamsters were stereotaxically implanted with bilateral guide cannulae aimed at the medial preoptic area (POA), ventromedial hypothalamus (VMH), or ventral tegmentum (VTA). The following week these females were injected SC with 10 micrograms estradiol benzoate (EB) and then had 27-gauge cannulae containing crystalline progesterone inserted through the guide tubes. Sexual receptivity was observed in 3 of 11 animals with VMH implants of progesterone, in 2 of 10 with VTA progesterone, but in none with POA implants. In the second experiment, the amount of intracranial progesterone was increased by mechanically expelling a 1.5 micrograms progesterone pellet from the tip of each cannula insert. This treatment facilitated receptivity in 10 of 20 hamsters with VTA implants and in 9 of 32 VMH-implanted animals. This induction of receptivity required approximately 2 hr. Progesterone pellets in the POA, mammillary region, and lateral mesencephalon were generally ineffective. In hamsters, progesterone into either the VMH or the VTA is sufficient to facilitate receptivity, although neither site is highly sensitive to progesterone. These results differ from those in recent studies in rats and this difference may reflect important species differences in the control of lordosis.     

Reliable confirmation of antibodies to bovine respiratory syncytial virus (BRSV) by enzyme-linked immunosorbent assay using BRSV nucleocapsid protein expressed in insect cells.

The nucleocapsid (N) protein of bovine respiratory syncytial virus (BRSV) in the baculovirus expression system was evaluated as a source of antigen in an enzyme-linked immunosorbent assay (ELISA) for the detection of respiratory syncytial virus (RSV) antibodies. The recombinant N protein was purified from infected-cell extracts by sucrose gradient centrifugation and used in the ELISA for the detection of antibodies to various RSV strains. The ELISA was compared with the virus neutralization (VN) test for determining BRSV antibodies in 10 consecutive serum samples from four calves vaccinated with a live modified BRSV vaccine and from two nonvaccinated control calves. The ELISA compared favorably with the VN test for detecting serological responses. All serum samples which were positive in the VN test were also positive in the ELISA. None of the serum samples collected from the two nonvaccinated calves reacted in the ELISA. To determine the usefulness of the ELISA for epidemiological studies, 58 cattle serum samples were tested in the ELISA and the VN test. Approximately 94% (42 of 45) of field serum samples which were positive in the ELISA were also positive in the VN test. No case was found in which the ELISA result was negative and the VN test result was positive. Thirteen of the serum samples were negative in both methods. Our results indicate that the ELISA with the baculovirus-expressed N protein as an antigen is an efficient, sensitive, and specific method for detecting serum antibodies to RSV.     

Effects of aging on sleep in the golden hamster

The golden hamster (Mesocricetus auratus) has been a model organism for the study of circadian rhythmicity and, in particular, the effects of age on the circadian system. Surprisingly, nothing is known about the effects of advanced age on sleep in this species. As a first step in determining the effects of aging on sleep in the golden hamster, we recorded sleep for 24 hours in 12 young (3 months) and 18 old (17-18 months) golden hamsters entrained to a 14:10 light:dark (LD) cycle. Aged hamsters exhibited small but significant increases in overall NREM sleep time, primarily due to an increase in time the old animals spent in the NREM sleep state during the dark period relative to the young hamsters. There were no significant differences in REM sleep, median sleep episode length, or the number of arousals. The most striking differences between the sleep of young and old hamsters was in NREM delta (0.5-4 Hz) power per epoch. Old hamsters showed approximately 27% less (p=0.0004) delta power per NREM epoch than young hamsters. It is possible that increased NREM sleep time in the old hamsters may be a failed attempt to maintain cumulative delta power; ie, old hamsters may have more NREM sleep in order to make up for the lower intensity of their sleep. This decline in delta power with age parallels earlier findings in cats and humans, although has it not been previously reported in rodents.     

A new enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against foot-and-mouth disease virus. I. Development and method of ELISA

A liquid-phase blocking sandwich ELISA has been developed for the quantification of antibodies against foot-and-mouth disease virus which may replace the virus neutralisation (VN) test. This test employs the incubation of a constant amount of antigen with a range of test serum dilutions in the liquid-phase before being assayed using a trapping ELISA. Thus it does not rely on the availability or growth of tissue culture cells. The assay is rapid and relatively simple to perform, reagents are used economically and results may be recorded within 24 h. The ELISA is sensitive and results are more specific and more reproducible than those obtained by VN. Results are expressed as reciprocal antibody titres which are analogous and of a similar order to those recorded by VN. Individual titres, therefore, may be easily assessed by workers in the field who are already familiar with VN.