Testosterone prevents synaptic loss in the perineal motoneuron pool in the spinal cord in male rats exposed to chronic stress

Chronic stress is known to induce disorders of reproductive neuroendocrine functions. Motoneurons of the spinal nucleus of the bulbocavernosus (SNB) in male rats play an important role in copulatory behavior. In the present study, it was examined whether chronic stress would alter synaptic organization of the SNB motoneurons and whether androgen would modify the changes under chronic stress. Five male rats were under restraint stress for 5 days per week for 3 weeks, and five males implanted subcutaneously with Silastic capsules containing testosterone were also exposed to stress. Five males served as unstressed controls. After 3 weeks of restraint stress, cholera toxin-horseradish peroxidase (CT-HRP) was injected into the bulbocavernosus muscles and animals were killed 2 days later. The spinal cords containing the SNB were dissected, processed with a modified tetramethylbenzidine (TMB) method for visualization of retrogradely transported CT-HRP, and examined ultrastructurally. Neuronal structures apposing the membranes of 150 SNB motoneurons (total for three groups) were analyzed by measuring the percentage of somatic membranes covered by synaptic contacts. The mean percentage of somatic membranes covered by synapses in males exposed to chronic stress was significantly less than that in controls or stressed males treated with testosterone. Size and number of synaptic contacts per unit length of somatic membranes in males exposed to stress were also significantly less than those in controls or stressed males treated with testosterone. There was no significant difference in any of the parameters between controls and stressed males treated with testosterone. Changes in plasma levels of testosterone showed the same profile as changes in the synaptic contacts. These results suggest that the SNB motoneurons of male rats exposed to chronic stress retain a considerable synaptic plasticity in response to androgen, and that androgen treatment can rescue the SNB system in male rats when under chronic restraint stress.     

Adrenal mediation of intermale aggression maintained by aromatized and reduced metabolites of testosterone

Individually housed CD-1 mice were either sham castrated or castrated and treated with testosterone (T), estradiol benzoate (EB), dihydrotestosterone (DHT), a combination of EB and DHT (EB+DHT), or the injection vehicle. Following 16 days of isolation and subcutaneous injections, animals were tested repeatedly for fighting behavior in paired encounters with nonaggressive stimulus males. Results indicated that the T and EB+DHT groups fought to the same extent as the gonadally intact group. Both the EB and DHT groups fought more than the vehicle-treated group but less than the T, EB+DHT and sham castrated groups. A similar study was subsequently performed with adrenalectomized animsls. Adrenalectomy eliminated agonistic responses in animals receiving metabolites of testosterone (EB, DHT, EB+DHT) but had only slight effects in gonadally intact and T-treated, castrated mice. The results suggested that a) EB and DHT, either singly or in combination, maintain aggression through a synergism with adrenal steroids; b) the combined effects of EB and DHT reflect an additive action rather than synergistic interaction, notwithstanding the synergism with adrenal steroids; c) metabolism of testosterone to estrogen and dihydrotestosterone does not sufficiently account for the action of testosterone.     

Ablation of cerebellar nuclei prevents H-reflex down-conditioning in rats

While studies of cerebellar involvement in learning and memory have described plasticity within the cerebellum, its role in acquisition of plasticity elsewhere in the CNS is largely unexplored. This study set out to determine whether the cerebellum is needed for acquisition of the spinal cord plasticity that underlies operantly conditioned decrease in the H-reflex, the electrical analog of the spinal stretch reflex. Rats in which the cerebellar output nuclei dentate and interpositus (DIN) had been ablated were exposed for 50 d to the H-reflex down-conditioning protocol. DIN ablation, which in itself had no significant long-term effect on H-reflex size, entirely prevented acquisition of a smaller H-reflex. Since previous studies show that corticospinal tract (CST) transection also prevents down-conditioning while transection of the rubrospinal tract and other major descending tracts does not, this result implies that DIN output that affects cortex is essential for generation of the CST activity that induces the spinal cord plasticity, which is, in turn, directly responsible for the smaller H-reflex. The result extends the role of the cerebellum in learning and memory to include participation in induction of plasticity elsewhere in the CNS, specifically in the spinal cord. The cerebellum might simply support processes in sensorimotor cortex or elsewhere that change the spinal cord, or the cerebellum itself might undergo plasticity similar to that occurring with vestibulo-ocular reflex (VOR) or eyeblink conditioning.     

Vaginal bleeding and spotting in transgender men after initiation of testosterone therapy: A prospective cohort study (ENIGI)

Abstract

Background: Previous studies have cross-sectionally described amenorrhea in cohorts of transgender men on intramuscular or subcutaneous testosterone injections. It remains uncertain which testosterone preparations most effectively suppress vaginal bleeding and when amenorrhea occurs after testosterone initiation.

Aim:To investigate the clinical effects of various testosterone preparations on vaginal bleeding and spotting in transgender men.

Methods: This prospective cohort study was part of the European Network for the Investigation of Gender Incongruence (ENIGI). Data on the persistence and intensity of vaginal bleeding and spotting, serum sex steroid levels and body composition were prospectively and cross-sectionally assessed in 267 transgender men during a three-year follow-up period, starting at the initiation of various testosterone preparations.

Results: After three months of testosterone, 17.9% of transgender men reported persistent vaginal bleeding and 26.8% reported spotting. The percentages reporting vaginal bleeding and spotting decreased over the first year of testosterone (bleeding 4.7% and spotting 6.9% at 12?months, respectively), with no participants reporting vaginal bleeding or spotting after 18?months of testosterone. Factors associated with vaginal bleeding or spotting included lower serum testosterone levels and being on testosterone gel as compared to injections (e.g., esters or undecanoate preparations). If vaginal bleeding persisted, starting progestogens at three months resulted in a decrease in the intensity of vaginal bleeding and spotting.

Discussion: Transgender men and hormone-prescribing providers can be reassured that vaginal bleeding and spotting usually stop within three months after testosterone initiation. If not, serum testosterone levels should be measured and testosterone dose adjusted to achieve serum testosterone levels in the physiologic male range. Adding a progestin can be considered after three to six months if bleeding persists. Providers should be aware that cessation of bleeding can be more difficult to achieve in transgender men with lower serum testosterone levels or those on testosterone gel.     

Tolerance to the antinociceptive effect of intrathecal morphine in intact and chronic spinal rats

The antinociceptive effect of daily acute intrathecal morphine injections on the tail-flick withdrawal response was compared in Intact rats and rats that were spinally transected 3 to 4 weeks prior to morphine administration (Spinal rats). Spinal rats became tolerant to repeated intrathecal injections of 5, 15, or 30 micrograms of morphine within 3 days. Intact rats were not tolerant to these doses after 4 daily injections. Spinal rats, made tolerant to repeated intrathecal morphine injections, were not tolerant to a subcutaneous injection of 6.0 mg/kg of morphine. These data suggest that, in intact animals, tolerance to the antinociceptive effect of spinal morphine is modulated by descending, supraspinal input.     

Retrieval-induced forgetting of arithmetic facts across cultures

Retrieving a single-digit multiplication fact (3×4 =12) can slow response time (RT) for the corresponding addition fact (3+4=7). The present experiment investigated effects of problem type (i.e., tie addition problems such as 3+3 vs. non-ties such as 3+4) and cultural background on this retrieval-induced forgetting (RIF) phenomenon in young adults. Canadians answering in English (n=36), Chinese adults answering in English (n=36), and Chinese answering in Chinese (n=36) received four blocks of multiplication practice and then two blocks of the addition counterparts and control additions. Tie addition problems presented a robust RIF effect that did not differ between groups, but only the Canadian group showed RIF for non-ties and only for small non-ties with sum≤10 (3+4). The Chinese groups' RIF effect for addition ties, but not small non-ties, converges with recent evidence that ties are solved by direct memory retrieval whereas small non-ties may be solved by highly efficient procedural processes in skilled performers.     

DNA methylation and behavioral changes induced by neonatal spinal transection

Although the importance of epigenetic mechanisms in behavioral development has been gaining attention in recent years, research has largely focused on the brain. To our knowledge, no studies to date have investigated epigenetic changes in the developing spinal cord to determine the dynamic manner in which the spinal epigenome may respond to environmental input during behavioral development. Animal studies demonstrate that spinal cord plasticity is heightened during early development, is somewhat preserved following neonatal transection, and that spinal injured animals are responsive to sensory feedback. Because epigenetic alterations have been implicated in brain plasticity and are highly responsive to experience, these alterations are promising candidates for molecular substrates of spinal plasticity as well. Thus, the current study investigated behavioral changes in the development of weight-bearing locomotion and epigenetic modifications in the spinal cord of infant rats following a neonatal low-thoracic spinal transection or sham surgery on postnatal day (P)1. Specifically, global levels of methylation and methylation status of the brain-derived neurotrophic factor (Bdnf) gene, a neurotrophin heavily involved in both CNS and behavioral plasticity, particularly in development, were examined in lumbar tissue harvested on P10 from sham and spinal-transected subjects. Behavioral results demonstrate that compared to shams, spinal-transected subjects exhibit significantly reduced partial-weight bearing hindlimb activity. Molecular data demonstrate group differences in global lumbar methylation levels as well as exon-specific group differences in Bdnf methylation. This study represents an initial step toward understanding the relationship between epigenetic mechanisms and plasticity associated with spinal cord and locomotor development.     

Limb Segment Load Inhibits the Recovery of Soleus H-Reflex After Segmental Vibration in Humans

We investigated the effects of vertical vibration and compressive load on soleus H-reflex amplitude and postactivation depression. We hypothesized that, in the presence of a compressive load, limb vibration induces a longer suppression of soleus H-reflex. Eleven healthy adults received vibratory stimulation at a fixed frequency (30 Hz) over two loading conditions (0% and 50% of individual's body weight). H-reflex amplitude was depressed ～88% in both conditions during vibration. Cyclic application of compression after cessation of the vibration caused a persistent reduction in H-reflex excitability and postactivation depression for > 2.5 min. A combination of limb segment vibration and compression may offer a nonpharmacologic method to modulate spinal reflex excitability in people after CNS injury.     

Modulation of afterhyperpolarization evoked by doublets and increasing number of stimuli in rat motoneurons

The authors studied the influence of variable stimulation patterns on parameters of afterhyperpolarization (AHP) in rat spinal motoneurons using intracellular recording of antidromic action potentials. The action potentials analyzed were evoked by either (a) a single stimulus or pair of stimuli with gradually increasing interpulse intervals (IPI) of 5-20 ms or (b) an increasing number of stimulus pulses, from 1 to 4, at a constant IPI. It was demonstrated that modulations of AHP parameters after 2 pulses depended on the IPI, whereas the most significant changes in these parameters were found after the application of 2 or 3 stimuli in a series. The authors propose that changes in the AHP parameters have functional significance for motoneuronal firing pattern and therefore for motor unit force development during tetanic contractions.     

Is a lower dose of cyproterone acetate as effective at testosterone suppression in transgender women as higher doses?

Objective: The recommended dose of cyproterone acetate (CPA), an anti-androgen that is commonly used in the hormonal treatment of transgender women, is 50–100 mg daily. Our objective was to determine whether CPA at 25 mg daily would suppress total testosterone as effectively as 50 mg daily in transgender women.

Methods: We conducted a retrospective cohort analysis of transgender women attending an endocrinology clinic between April 1, 2009, and June 30, 2015. We used a generalized linear mixed model to compare total testosterone between patients on CPA 25 mg versus CPA 50 mg or higher. In a subgroup of patients for which the CPA dose was decreased from 50 mg to 25 mg, we compared total testosterone levels before and after the decrease.

Results: We divided the sixty-eight patients included in the study into 4 groups: group 1, CPA 25 mg (N =31); group 2, CPA 50 mg or higher (N = 19); group 3, CPA dose lowered from 50 mg to 25 mg (N = 15); group 4, CPA dose increased from 25 mg to 50 mg (N = 3). The mean total testosterone on treatment was 0.9 nmol/L (95% CI 0.7 to 1.1) in group 1 and 1.2 nmol/L (95% CI 0.9–1.5) in group 2 and were not significantly different (p = 0.087). In group 3, there was no significant difference between total testosterone levels before and after decreasing the dose of CPA from 50 mg to 25 mg, p = 0.86. Group 4 was excluded from analysis.

Conclusions: We found that 25 mg of CPA daily was effective at suppressing testosterone levels to within normal female range when used in combination with recommended estrogen therapy in transgender women. Clinicians should consider using a lower dose of CPA in order to minimize potential adverse effects.     

Cross over of forebrain and brainstem neuronal projections to spinal cord sympathetic preganglionic neurons in the rat

Neuronal inputs from the forebrain and the brainstem to sympathetic preganglionic neurons in the spinal cord were investigated by the transneuronal retrograde tracing technique using pseudorabies virus in intact and brainstem-lesioned rats. After unilateral subcutaneous viral inoculations into the hind limb of intact rats, infected neurons were then visualized by immunostaining. At 3.5 days after inoculation, infected neurons appeared in the thoracic (T10) intermediolateral (IML) cell column. On the 4th day, infected neurons were present in the C1, A5, A6, A7 catecholamine cell groups and the rostral ventromedial medulla (RVMM). On the 5th day, viral labeling was seen in the hypothalamic paraventricular and arcuate nuclei and the lateral hypothalamic area. In all of these nuclei, the infected cells appeared bilaterally. However, the appearance of virus-labeled cells in these nuclei was unilateral following unilateral coronal sections between the medulla and the spinal cord (depending on the side of hemisection, but not on the site of virus inoculation). Midsagittal sections throughout the entire medulla oblongata did not alter the topographical pattern of virus-infected neurons in the forebrain or the brainstem. These findings indicate that descending fibers to the spinal neurons may not cross over in the lower brainstem but that they decussate within the spinal cord.     

Exogenous testosterone and social experience each enhance the development of aggressive behavior in Cyprinodon variegatus

The combined effects of testosterone and social experience from the time of hatching on development of aggressive behaviors in Cyprinodon variegatus were investigated. Social experience was defined as a rearing condition allowing visual, tactile, chemosensory, and auditory contact with zero, three, or 15 additional conspecifics. Behaviors were videotaped once weekly from 3 weeks to 4 months post-hatching, using the focal animal method, and an ethogram was constructed. At 3 months post-hatching, half the fish were injected once with testosterone propionate (2 μg/g b. w., i. p., vehicle-Ringer's solution) and the other half were injected with Ringer's isotonic saline solution. Testosteronetreated subjects showed significantly greater frequencies and durations of behaviors that may be related to aggression than did saline controls. Two weeks after the treatments, a round-robin tournament was conducted between testosterone- and saline-treated fish from each rearing condition. Testosterone-treated fish won significantly more encounters than did saline-treated subjects over all rearing conditions. Furthermore, significantly more aggressive acts were displayed by fish raised in the groups of 16 and the groups of four than did the Isolates. Although the number of aggressive acts exhibited by the group of 16 was greater than the group of four, the difference was not statistically significant.     

Neural pathways mediating vaginal function: the vagus nerves and spinal cord oxytocin

The initial observations, made in our laboratory with Knut Larsson, of the ability of vaginocervical stimulation (VCS) to block withdrawal responses to foot pinch in rats has led to findings of multiple behavioral, autonomic, and neuroendocrine effects of this potent stimulus in rats and also in women. It has led to an understanding of: (1) the neuroanatomical and neurochemical basis of a novel and potent pain-blocking mechanism; (2) likely neuroanatomical pathways mediating both the Ferguson reflex and a specific autonomic response - the pupil-dilating effect of VCS; (3) a role for oxytocin as a putative central nervous system neurotransmitter that stimulates autonomic sympathetic preganglionic neurons within the spinal cord; and (4) a novel pathway that can convey sensory activity from the cervix, adequate to induce orgasm, via the vagus nerves. This latter pathway bypasses the spinal cord and projects directly to the medulla oblongata, and thus can convey genital afferent activity despite complete spinal cord injury at any level.     

Human lumbar cord circuitries can be activated by extrinsic tonic input to generate locomotor-like activity

We have demonstrated that non-patterned electrical stimulation of the lumbar cord can induce stepping-like activity in the lower limbs of complete spinal cord injured individuals. This result suggested the existence of a human lumbar locomotor pattern generator, which can convert a tonic input to a rhythmic motor output. We have studied the human lumbar cord in isolation from supraspinal input but under extrinsic tonic input delivered by spinal cord stimulation. Large-diameter afferents within the posterior roots are directly depolarized by the electrical stimulation. These afferents project to motoneurons as well as to lumbar interneurons involved in the motor control of lower limbs. Stimulation at 25-50 Hz can elicit rhythmic alternating flexion/extension movements of the lower limbs in supine individuals. Reducing the tonic input frequency to 5-15 Hz initiates lower limb extension. Epidural stimulation applied during manually assisted treadmill stepping in complete spinal cord injured persons immediately increases the central state of excitability of lumbar cord networks and enhances stepping-like functional motor outputs. Sustained, non-patterned tonic input via the posterior roots can activate human lumbar cord networks. Pattern generating configurations of these multifunctional circuitries can be set-up depending on the stimulation parameters and particularly on the input frequency.     

The relationship between testosterone and aggression: a meta-analysis

In non-human animals, the relationship between testosterone and aggression is well established. In humans, the relationship is more controversial. To clarify the relationship, Archer conducted three meta-analyses and found a weak, positive relationship between testosterone and aggression. Unfortunately, each of the analyses included only five to six studies. The aim of the present study was to re-examine the relationship between testosterone and aggression with a larger sample of studies. The present analyses are based on 45 independent studies (N=9760) with 54 independent effect sizes. Only studies that reported a p-value or effect size were included in the analyses and the sample may underestimate the proportion of non-significant findings in the population. Correlations ranged from −0.28 to 0.71. The mean weighted correlation (r=0.14) corroborates Archer's finding of a weak positive relationship.     

Effects of chronic administration with high doses of testosterone propionate on behavioral and physiological parameters in mice with differing basal aggressiveness

The effects of testosterone propionate, an anabolic‐androgenic steroid, on the behavior displayed during a social encounter by gonadally intact male mice were investigated. Animals were distributed into three groups according to their attack latency in a pre‐screening test (high‐, moderate‐, and low‐ attacking mice) and each group received weekly injections of 60 or 120 mg/kg of testosterone or sesame oil for 10 weeks. Behavioral tests were then carried out. Afterwards, organs were weighed and blood samples collected in order to obtain hormonal data. Treatment had a differential impact on attack in the three groups of animals. Only the high‐attacking testosterone‐treated mice showed lower total duration of attack than their controls. Those that received 60 mg/kg spent more time exhibiting exploratory behaviors. As an index of the anabolic activity of the drug, all testosterone‐treated mice had heavier kidneys and, as an index of the androgenic activity of testosterone propionate, they had heavier seminal vesicles, lighter testes, and showed higher testosterone levels in a dose‐dependent way than their controls. Hence, the effect of treatment on peripheral physiological parameters was similar in all three groups whereas behavioral effects differed depending on basal aggressiveness, considered a characteristic of coping style. Aggr. Behav. 29:173–189, 2003. © 2003 Wiley‐Liss, Inc.     

Simulation of motor-neuronal networks

This research develops a theoretical connectionist-type model of the operation of the human motor system. The model includes the operation of the cerebral cortex, pons, spinal cord, and muscles. It emphasizes the parallel passage of neuronal signals, the integration of multiple signals within nuclei, and the modulation of signals as a result of sensory input. A redundant Fast Fourier Transform (FFT) procedure that analyzes EEG signals by a method similar to the one used by pontine nuclei is described.     

The cerebellum in maintenance of a motor skill: a hierarchy of brain and spinal cord plasticity underlies H-reflex conditioning

Operant conditioning of the H-reflex, the electrical analog of the spinal stretch reflex, is a simple model of skill acquisition and involves plasticity in the spinal cord. Previous work showed that the cerebellum is essential for down-conditioning the H-reflex. This study asks whether the cerebellum is also essential for maintaining down-conditioning. After rats decreased the soleus H-reflex over 50 d in response to the down-conditioning protocol, the cerebellar output nuclei dentate and interpositus (DIN) were ablated, and down-conditioning continued for 50-100 more days. In naive (i.e., unconditioned) rats, DIN ablation itself has no significant long-term effect on H-reflex size. During down-conditioning prior to DIN ablation, eight Sprague-Dawley rats decreased the H-reflex to 57% (+/-4 SEM) of control. It rose after ablation, stabilizing within 2 d at about 75% and remaining there until approximately 40 d after ablation. It then rose to approximately 130%, where it remained through the end of study 100 d after ablation. Thus, DIN ablation in down-conditioned rats caused an immediate increase and a delayed increase in the H-reflex. The final result was an H-reflex significantly larger than that prior to down-conditioning. Combined with previous work, these remarkable results suggest that the spinal cord plasticity directly responsible for down-conditioning, which survives only 5-10 d on its own, is maintained by supraspinal plasticity that survives approximately 40 d after loss of cerebellar output. Thus, H-reflex conditioning seems to depend on a hierarchy of brain and spinal cord plasticity to which the cerebellum makes an essential contribution.     

Testosterone intake and aggressiveness: Real effect or anticipation?

In a double-blind experiment, human males (n = 27) were given either testosterone (40 mg/day), placebo, or no treatment, over a one week period. Subjective and observer assessed mood estimations were conducted before and after treatment. Testosterone levels in saliva were measured with radioimmunoassay. The results revealed a significant placebo effect [c. f. Medicine and Science in Sports 4: 124–126]: After treatment, the placebo group scored higher than both the testosterone and the control group on self-estimated anger, irritation, impulsivity, and frustration. Observer-estimated mood yielded similar results. The lack of a placebo effect in the testosterone group is intriguing, and may be due to secondary effects caused by suppression of the body's own testosterone production, since recorded non-protein bound testosterone did not significantly rise due to treatment. The resultss suggest that androgen usage causes expectations, rather than an actual increase of aggressiveness. © 1994 Wiley-Liss, Inc.     

Testosterone,aggressiveness, and antisocial personality

Testosterone levels were examined in prisoners convicted of violent crimes (n = 13), in men previously convicted of violent crimes but currently not in prison (n = 15), in nonviolent alcoholics (n = 15), and in randomly selected control males (n = 16). Morning, afternoon, and evening testosterone levels were assessed after a minimum alcohol abstinence period of 24 hr. Violent and nonviolent men did not differ in plasma total testosterone level on any sampling occasion. In violent men, however, testosterone levels were significantly correlated with hostility, as measured by the Derogatis Symptom Check List. Most violent men were diagnosed with Antisocial Personality Disorder (ASP) [DSM‐III‐R; 301.70], and the unweighted ASP symptom count also correlated significantly with testosterone levels in these subjects. We suggest that individuals whose life histories involve numerous antisocial behaviors tend to have high testosterone levels even when interpersonal violence is excluded. This, however, does not eliminate the possibility that males who are characterized by high hostility may also have elevated testosterone levels. Violent predisposition and antisocial conduct beginning in early adolescence predict adult aggressive behaviors, which are augmented by power‐related alcohol expectancies and alcohol abuse. Aggr. Behav. 25:113–123, 1999. © 1999 Wiley‐Liss, Inc.