The role of target muscles in the expression of calcitonin gene-related peptide mRNA in the spinal nucleus of the bulbocavernosus.

Previous studies have shown that calcitonin gene-related peptide (CGRP) mRNA steady-state levels and CGRP immunoreactivity in motoneurons of the spinal nucleus of the bulbocavernosus (SNB) are inversely related to the gonadal steroid environment in male rats. Since both the SNB motoneurons and their target muscles, the bulbocavernosus and the levator ani, are steroid sensitive, gonadal steroids may act at either site to regulate CGRP expression. In the present study, we tested the hypothesis that gonadal steroids influence CGRP expression in SNB motoneurons through their effects on the bulbocavernosus and levator ani muscles. We determined the levels of alpha-CGRP mRNA and immunoreactive CGRP in SNB motoneurons of adult male rats following injection of the bulbocavernosus with muscle extracts from bulbocavernosus/levator ani of castrated rats, paralysis of the bulbocavernosus or pudendal nerve cuts. Following injection of the bulbocavernosus/levator ani with extracts from castrated rats, the level of CGRP expression and the number of SNB motoneurons with alpha-CGRP message were increased. These studies suggest that the bulbocavernosus/levator ani muscles from castrated rats produce a factor that increases levels of CGRP. Injections of extract prepared from the bulbocavernosus and levator ani muscles of gonadally intact rats did not change the expression of alpha-CGRP mRNA in the SNB. Paralysis of the bulbocavernosus/levator ani with a local anesthetic increased the number of SNB motoneurons expressing alpha-CGRP mRNA and CGRP immunoreactive neurons. To determine whether nerve damage accounted for the observed effects following injection of anesthetic, the pudendal nerves were cut bilaterally.(ABSTRACT TRUNCATED AT 250 WORDS)     

Neonatal androgen maintains sexually dimorphic muscles in the absence of innervation

We examined the site of androgen action in maintaining the sexually dimorphic spinal nucleus of the bulbocavernosus (SNB) and its target perineal muscles, the bulbocavernosus (BC) and levator ani (LA), in rats. To determine whether androgen action on SNB motoneurons is crucial for BC/LA survival, we removed SNB cells in newborn female rats by lumbosacral spinalectomy, administered testosterone propionate (TP) on days 1 and 3 of life, and examined for the presence of BC/LA muscles in adulthood. BC/LA muscles were present in all TP-treated spinalectomized females, and staining of these muscles with alpha-bungarotoxin or for acetylcholinesterase showed no evidence of cholinergic innervation. Thus, complete neonatal denervation of the BC/LA does not prevent TP from sparing these muscles, suggesting that androgen acts directly upon BC/LA muscles to maintain them during development. This androgenic maintenance of the BC/LA may be crucial for the survival of SNB motoneurons.     

Increase in motoneurons in the spinal nucleus of the bulbocavernosus of prepubertally castrated male Mongolian gerbils following delayed treatment with testosterone

Sexual dimorphism in the spinal nucleus of the bulbocavernosus (SNB) of the Mongolian gerbil is achieved by two periods of postnatal increase, one in the first month after birth and one at puberty. The pubertal increase in motoneuron number is of particular interest because it occurs in a nearly adult animal. The purpose of this research was threefold. The first was to determine the response of the SNB in prepubertally castrated male gerbils receiving delayed hormone replacement as adults. Testosterone propionate (TP) treatment resulted in numbers of SNB motoneurons comparable to those seen in intact males, whereas androgen metabolites were less effective. The second purpose was to determine the latency of motoneurons to appear in response to TP. New SNB motoneurons appeared within 2 days of delayed TP replacement in prepubertally castrated males, and 16 days of treatment did not further increase SNB motoneuron numbers. The response of the motoneurons to TP appeared more rapid than the response of the bulbocavernosus (BC) muscle, scent gland, and seminal vesicles. The third purpose was to determine whether the new cells were connected to a target muscle. After 16 days of TP treatment, more motoneurons were labeled in the SNB following injection of a retrograde tract tracer into the BC muscle compared with the number seen in control animals. Thus, new motoneurons appeared in the SNB of prepubertally castrated male Mongolian gerbils within 2 days of the start of delayed TP treatment and were connected to a target within 16 days of TP treatment.     

Androgenic regulation of expression of androgen receptor protein in the perineal motoneurons of aged male rats.

Androgen might regulate expression of androgen receptors (AR) in AR-containing motoneurons in young animals. In the present study, it was examined whether expression of AR was also regulated by androgen in aged animals. Twelve male rats were castrated at 26 months of age. Five days following castration, the animals were treated with testosterone propionate (TP; six males) or vehicle (six males) and killed 2 hours later. Six sham-castrated rats served as controls. AR immunoreactivity was examined in motoneurons of the spinal nucleus of the bulbocavernosus (SNB) in these animals by immunohistochemistry using the polyclonal antibody PG21. In control animals, slightly intense AR immunoreactivity was confined to the nuclei of the motoneurons. AR immunoreactivity was completely eliminated in the motoneurons of castrated rats. In castrated, aged animals treated with TP, the intensity of AR immunoreactivity in the nuclei of SNB motoneurons was increased. Plasma levels of testosterone in castrated, aged animals 2 hours following treatment with TP were significantly greater than those in controls. These results suggest that expression of AR in motoneurons of the SNB in aged male rats is up-regulated in response to androgen and that androgen may be, at least in part, involved in the process of aging of the SNB in male rats.     

Ontogeny of steroid accumulation in spinal lumbar motoneurons of the rat: implications for androgen's site of action during synapse elimination.

Androgens influence the postnatal development of motoneurons in the spinal nucleus of the bulbocavernosus (SNB) by regulating neuromuscular synapse elimination, the process through which multiple axonal inputs are retracted from each muscle fiber until single innervation is established. In the rat levator ani (LA), one of the target muscles for SNB motoneurons, much of this loss of multiple innervation can be prevented by prepubertal androgen treatment. We used steroid autoradiography to measure the ontogeny of steroid accumulation in the SNB and the retrodorsolateral nucleus (RDLN), two motoneuronal groups thought to differ in their sensitivity to androgens. Spinal cord tissue was analyzed from castrated male rats at 7, 14, 21, and 60 days of age after injection of radiolabelled testosterone. SNB and RDLN motoneurons differ in the ontogeny of androgen accumulation. Over 80% of SNB motoneurons develop the capacity to accumulate androgen during the second week after birth, during the period when androgen regulates synapse elimination in the LA. In contrast, androgen accumulation in RDLN motoneurons develops much later (after 21 days). These data suggest that androgen may act directly on SNB motoneurons to influence synapse elimination.     

Direct androgenic regulation of calcitonin gene-related peptide expression in motoneurons of rats with mosaic androgen insensitivity.

The spinal nucleus of the bulbocavernosus (SNB) and its target muscles, bulbocavernosus and levator ani (BC/LA), form a sexually dimorphic neuromuscular circuit whose development and maintenance are androgen-dependent. The mechanisms whereby androgen regulates gene expression in the SNB of adult rats are largely unknown, although a retrograde influence from the BC/LA muscles has been suggested to underlie the suppression of calcitonin gene-related peptide (CGRP) expression observed in SNB motoneurons after systemic androgen treatment. A mosaic paradigm was used to determine the site of action of androgen in the regulation of CGRP expression in SNB motoneurons. As a consequence of random X chromosome inactivation, androgenized female rats heterozygous for the tfm androgen receptor (AR) mutation (XwtXtfm-mosaics) express a mosaic of androgen-sensitive and androgen-insensitive motoneurons in the SNB, whereas the BC/LA target musculature appears to be uniformly sensitive to androgens. In adult mosaics, testosterone administration resulted in a reduction in the proportion of androgen-sensitive cells expressing CGRP, whereas no such reduction was observed in the androgen-insensitive population, indicating that neuronal AR plays an essential role in the neuromuscular regulation of CGRP expression in these motoneurons. This provides the first in vivo demonstration of AR regulation of gene expression unambiguously localized to a neuronal population.     

Androgen stimulates neuronal plasticity in the perineal motoneurons of aged male rats

Ten aged male rats (24 months of age) were castrated and implanted subcutaneously with Silastic capsules containing testosterone (T)(5 males) or nothing (5 males). Five sham-castrated males (25 months of age) served as controls. Four weeks after castration, cholera toxin-horseradish peroxidase (CT-HRP) was injected into the bulbocavernosus muscles and animals were killed 2 days later. The spinal cords containing the spinal nucleus of the bulbocavernosus (SNB) were dissected, processed with a modified tetramethylbenzidine method for visualization of retrogradely transported CT-HRP, and examined ultrastructurally. Neuronal structures apposing the membranes of 150 CT-HRP-labeled SNB motoneurons were analyzed by measuring the percentage of somatic membranes covered by synaptic contacts, synaptoid contacts, and neuron-neuron contacts. Most of the neuronal structures in the control and experimental SNB motoneurons consisted of synaptic contacts. The mean percentage of somatic membranes covered by synapses in castrated, aged males treated with T was significantly greater than that in control or castrated animals. Size and number of synaptic contacts per unit length of somatic membranes in castrated, aged males treated with T were also significantly greater than those in control or castrated animals. Plasma levels of T in castrated, aged males treated with T were significantly greater than that in controls. These results suggest that the SNB motoneurons of aged male rats retain a considerable synaptic plasticity in response to androgen, and that androgen may be, at least in part, involved in the process of aging of the SNB system in male rats.     

Axotomy transiently down-regulates androgen receptors in motoneurons of the spinal nucleus of the bulbocavernosus

Testosterone is an important trophic factor for motoneurons in the spinal nucleus of the bulbocavernosus (SNB), and SNB motoneurons are more responsive to testosterone than are other motoneurons. Axonal injury during early postnatal life prevents the normal development of steroid-sensitivity by adult SNB motoneurons. Axonal injury also causes changes in the expression by motoneurons of a wide range of proteins, including the up-regulation of trophic factor receptors. We have used a polyclonal antibody (PG-21; G.S. Prins) to study the expression of androgen receptors in SNB motoneurons after axonal injury. PG-21 labeled motoneuronal nuclei in the lower lumbar spinal cord of rats in a pattern that matched autoradiograpic reports of androgen accumulation in this region of the nervous system. A population of numerous, small cells located dorsal to the central canal also showed evidence of androgen receptor expression. Cutting the axons of SNB motoneurons in adulthood or in development caused a decrease in androgen receptor immunoreactivity in SNB motoneurons. This is the first report that a trophic factor receptor in motoneurons is down-regulated after axonal injury, and is interesting in light of reports that testosterone treatment can facilitate motoneuronal regeneration after nerve cut. Androgen receptor levels subsequently returned to normal, regardless of the age at axotomy, providing no evidence for a lasting effect of developmental axotomy on androgen receptor levels in SNB motoneurons. Thus, axotomy-induced down-regulation of androgen receptors does not underlie the inability of SNB motoneurons to respond to androgen treatment several months after pudendal nerve cut in development.     

Morphology of rat spinal motoneurons with normal and hormonally altered specificity

Potential determinants of motoneuronal morphology were examined by using a sexually dimorphic, steroid-sensitive neuromuscular system in the rat spinal cord. In males, the spinal nucleus of the bulbocavernosus (SNB) innervates the perineal muscles bulbocavernosus (BC) and levator ani (LA), and the dorsolateral nucleus (DLN) innervates the ischiocavernosus muscle (IC). Adult females normally lack these motoneurons and the peripheral targets. Prenatal exposure of females to the androgen dihydrotestosterone propionate (DHTP) partially masculinizes this neuromuscular system and alters moto-neuron-to-muscle specificity, resulting in retained SNB target muscles anomalously innervated by motoneurons in the DLN. Because the morphology of SNB and DLN motoneurons normally differs significantly, the influence of spinal cord location and peripheral target on motoneuron morphology can be directly compared. Injection of cholera toxin conjugated to horseradish peroxidase (CTHRP) into the LA of DHTP-treated females labeled motoneurons predominantly in the SNB. These (SNB-LA) motoneurons in DHTP females were identical in all morphological measures to those of normal males. CTHRP injection into the BC of DHTP females labeled motoneurons in both the SNB and the DLN. SNB-BC motoneurons in DHTP females resembled those of normal males in process number and orientation, but were significantly smaller in dendritic length per motoneuron and in soma size. The DLN motoneurons anomalously projecting to the BC in DHTP females differed significantly from SNB-BC motoneurons in soma size and number and orientation of primary processes. However, these motoneurons were identical in all respects to DLN-IC motoneurons in DHTP females; DLN-IC motoneurons were similar to those of normal males in the orientation of their dendritic arbor, but were significantly smaller in dendritic length, soma size, and number of primary processes. These comparisons make it clear that DHTP selectively affects motoneuronal specificity and morphology in specific motoneuron classes. Further, motoneuronal morphology in the SNB/DLN system appears to be influenced more by spinal cord location than by peripheral target.     

Target-dependent hormonal control of neuron size in the rat spinal nucleus of the bulbocavernosus

The spinal nucleus of the bulbocavernosus (SNB) in the rat is a cluster of sexually dimorphic motoneurons that innervate perineal muscles. In adult male rats, the size of SNB neurons is reduced following castration, and this effect is reversed by treatment with testosterone. However, androgen receptors are present in the perineal muscles as well as in SNB neurons. Therefore, it is not clear whether the neuronal size is regulated by direct action of the hormone on SNB neurons or by a target-derived factor that may be controlled via hormonal action on the innervated muscle. To address this question, the peripheral (pudendal) nerve of SNB neurons on one side was cut and united to the grafted soleus muscle, which lacks androgen sensitivity. On the control side, the pudendal nerve was similarly cut but was allowed to reinnervate the perineal muscles. The size of SNB neurons was measured on both sides after a postoperative period of 10 weeks, during which the animal had been castrated or treated with testosterone after castration. The size of SNB neurons that had reinnervated perineal muscles was reduced following castration and enlarged by testosterone treatment. In contrast, the size of SNB neurons that had innervated the soleus muscle remained unaltered in response to testosterone manipulation. It is concluded that hormonal regulation of the size of SNB neurons in adult rats is mediated by their target muscles.     

Development of androgen receptor and p75(NTR) mRNAs and peptides in the lumbar spinal cord of the gerbil

Development of sex differences in the spinal cord appears to be largely under the control of androgen and although neurotrophins may also have a role. Spinal cords of male and female neonatal gerbils (postnatal days 1, 5, 7, 10, 23) and adult gerbils (postnatal day 150) were examined to determine the relative temporal expression of androgen receptor (AR) and the low-affinity neurotrophin receptor (p75) mRNAs within the spinal nucleus of the bulbocavernosus (SNB) and dorsolateral nucleus (DLN). Furthermore, prepubertal male gerbils were placed into one of six gonadal hormone treatment groups at weaning: Either sham castrate, castrated with gonadal hormone replacement, or castrated without gonadal hormone replacement. Ten weeks later gerbils were aldehyde-perfused, spinal cords removed and processed for presence of AR and p75 immunoreactivity (ir) in motoneurons of the SNB and DLN. During neonatal development, there were significant increases in androgen receptor mRNA within the SNB and DLN. In the SNB, the increase in androgen receptor mRNA preceded the increase in p75 mRNA. Peripubertally, significantly more SNB than DLN motoneurons contained AR- and p75-ir. These data demonstrate that AR expression occurs along the same developmental time frame as the development of the SNB and DLN and the organizational effects of androgens on their development continues through puberty in the male gerbil.     

Long-term castration effects motoneuron size but not number in the spinal nucleus of the bulbocavernosus in the adult male Mongolian gerbil

The spinal nucleus of the bulbocavernosus (SNB) is a sexually dimorphic group of motoneurons in Rexed's Lamina X of the lumbosacral spinal cord of the Mongolian gerbil. The SNB innervates the perineal musculature, the bulbocavernosus (BC), levator ani (LA), and external anal sphincter (EAS). Recent studies demonstrated a peripubertal component to the masculinization of the gerbil SNB with an apparent increase in both motoneuron size and number after puberty as measured with a Nissl stain. However, these studies could not determine if the apparent change in motoneuron number were due to the long-term castration involved in the methodology or due to a loss of motoneuron size beyond the point of being recognizable as motoneurons. Therefore, the current study was undertaken to examine this possibility by repeating the experimental protocol from previous peripubertal studies, on adult male gerbils with the addition of retrograde tract-tracing. Adult male gerbils were castrated at postnatal day (PND) 150 and given subcutaneous implants of testosterone proprionate (TP), dihydrotestosterone (DHT), estradiol benzoate (EB), EB and DHT, no steroid, or left intact. At PND215, the animals were injected with fluorogold (FG), a retrograde tract-tracer. At PND220, the animals were aldehyde perfused transcardially. The spinal cords were sectioned and alternate sections processed for either thionin-stain or FG visualization. Results indicated that long-term castration had no effect on SNB motoneuron number but did decrease SNB motoneuron size. TP, and to a lesser extent DHT, treatment could prevent the reduction in motoneuron size, however, EB could not. This study indicates that the maintenance of the adult male gerbil SNB-BC system is androgen, not estrogen, dependent and that long-term castration does not reduce motoneuron size to the point where they can no longer be distinguished.     

Androgen regulates synaptic input to motoneurons of the adult rat spinal cord

Adult male rats (Sprague-Dawley) were castrated and implanted subcutaneously with Silastic capsules containing testosterone or nothing. Sham-castrated males served as controls. Four weeks following castration, cholera toxin-horseradish peroxidase (CT-HRP) was injected bilaterally into the bulbocavernosus muscles and animals were sacrificed 2 d later. The spinal cords containing the spinal nucleus of the bulbocavernosus (SNB) were dissected, processed with a modified tetramethylbenzidine (TMB) method for visualization of retrogradely transported CT-HRP, and examined at the ultrastructural level. Neuronal structures apposing the membranes of 150 TMB-labeled SNB neurons were analyzed by measuring the percentage of somatic and proximal dendritic membranes covered by synaptic contacts, synaptoid contacts, and neuron-neuron contacts. Most of the neuronal structures in the control and experimental SNB motoneurons consisted of synaptic contacts. The mean percentage of somatic and proximal dendritic membranes covered by synapses 4 weeks after castration was reduced to approximately 30% of those in control animals. However, treatment with testosterone for 4 weeks after castration prevented this decline. Castration and testosterone treatment also influenced the size and number of synaptic contacts per unit length of somatic and proximal dendritic membranes, and the incidence of neuron-neuron contacts and double synapses onto SNB motoneurons. These results indicate that androgen is critical for maintaining the organization of synaptic inputs to these spinal motoneurons in adult male rats.     

N-methyl-D-aspartate receptor blockade inhibits estrogenic support of dendritic growth in a sexually dimorphic rat spinal nucleus

The lumbar spinal cord of rats contains the sexually dimorphic, steroid-sensitive spinal nucleus of the bulbocavernosus (SNB). Dendritic development of SNB motoneurons requires the action of both androgens and estrogens. Estrogenic effects are limited to the initial growth of SNB dendrites through 4 weeks of age. During this postnatal period, dendritic growth in other spinal motoneurons is regulated by N-methyl-D-aspartate (NMDA) receptor activation. In this study, we tested whether NMDA receptor activation was involved in SNB dendritic growth and whether the estrogenic support of SNB dendritic growth was dependent on the activation of NMDA receptors. Motoneuron morphology was assessed in normal males, intact males treated daily with the NMDA receptor antagonist MK-801, castrated males treated with estradiol benzoate (EB), and castrated males treated with both EB and MK-801. SNB motoneurons were retrogradely labeled with cholera toxin-horseradish peroxidase at 4 weeks of age (when dendritic length is normally maximal) and reconstructed in three dimensions. Somal area and dendritic length of SNB motoneurons in MK-801-treated, intact males were below those of normal males. Dendritic growth was partially supported in EB-treated castrates, but this growth was blocked by MK-801 treatment. These results suggest that, as in other motoneurons, dendritic development in the SNB involves NMDA receptors and, furthermore, that the estrogen-sensitive component of SNB dendritic development requires their activation.     

Bilateral organization of unilaterally generated activity in lumbar spinal motoneurons of the rat

The spinal nucleus of the bulbocavernosus (SNB) is a medially located, bilaterally organized sexually dimorphic motor nucleus in the lumbar spinal cord of the male rat. To begin to assess the potential functional significance of this bilateral organization, we recorded ipsi- and contralateral SNB motor nerve activity following unilateral spinal stimulation and examined the timing, pattern, and recruitment of population motoneuron activity. A possible mechanism for bilateral communication, gap junctional intercellular communication, was also investigated because dye coupling experiments indicate an extensive syncytium in which SNB motoneurons are coupled with each other and neighboring interneurons. An in vivo peripheral nerve recording paradigm was used: a bipolar stimulating electrode was placed on dorsal root L6, and bipolar recording electrodes were placed bilaterally on the SNB motor nerves. All processes were severed distal to electrode placement to isolate the central preparation; recruitment curves of motoneuronal activity were then generated. Amplitude of peak to peak recruitment was greater in the contralateral motor nerve than in the ipsilateral nerve. Response latency, Fourier transform and spike counts showed no evidence of ipsi/contralateral asymmetry. Recruitment was attenuated both ipsi- and contralaterally after pharmacological gap junction blockade, but antidromic stimulation could not drive activity in contralateral motor axons. These results indicate that unilateral input to the SNB may be differentially modulated to produce functionally distinct output in the two separate halves of the nucleus. We also discuss the potential modulatory role of gap junctions in the activity of the SNB.     

Steroid regulation of calcitonin gene-related peptide mRNA expression in motoneurons of the spinal nucleus of the bulbocavernosus

Motoneurons express calcitonin gene-related peptide (CGRP). Previous studies have shown that CGRP immunoreactivity is regulated by testosterone in the androgen-sensitive motoneurons of the spinal nucleus of the bulbocavernosus (SNB). In this research the effect of plasma levels of testosterone on the expression of alpha CGRP mRNA in the SNB motoneurons of adult male rats was studied with in situ hybridization. The number of motoneurons expressing alpha CGRP mRNA and the level of alpha CGRP mRNA expression was significantly higher in the SNB of castrated male rats than in the SNB of gonadally intact rats. Using a 5x background labeling criterion in castrated rats 88.1 +/- 4.5% while in intact rats 75.3 +/- 6.4% of SNB motoneurons expressed alpha CGRP mRNA. Testosterone replacement at the time of castration prevented the effect of castration on the expression of alpha CGRP mRNA in SNB motoneurons. In castrated rats, the increase in the number of SNB cells expressing CGRP was the result of increased steady state levels of alpha CGRP mRNA in all SNB neurons.     

Calcitonin gene-related peptide-like immunoreactivity in spinal motoneurons of the male mouse is affected by castration and genotype

Calcitonin gene-related peptide (CGRP) is found in motoneurons of the mammalian spinal cord, including motoneurons of the androgen-dependent spinal nucleus of the bulbocavernosus (SNB) of the mouse. Motoneurons of the SNB innervate the bulbocavernosus (BC), a striated muscle involved in penile reflexes. CGRP is thought to be a trophic factor produced by motoneurons to regulate the expression of the acetylcholine receptor at the neuromuscular junction. In rats, the number of SNB motoneurons containing CGRP is increased by gonadal steroids. This regulation appears to rely on an activity-dependent factor produced by the BC muscle. The purpose of the present study was to examine, using immunohistochemistry, the steroid dependence of CGRP in the SNB of male house mice. Genotypic differences in the steroid regulation of CGRP immunoreactivity were examined in three strains of mice that differ in their behavioral sensitivity to castration. The results demonstrate that castration reduces the number of CGRP-positive SNB motoneurons in mice. The magnitude of the change in CGRP in response to castration and the length of time required following castration to alter CGRP were dependent on genotype. Interestingly, the effect of castration in mice, to reduce the number of CGRP-immunoreactive SNB motoneurons, is opposite in direction from the increase in CGRP SNB motoneurons observed in rats observed following castration. These experiments suggest that androgens may alter neuromuscular junction function of mouse SNB by regulating the production of CGRP in a species-specific, genotypically dependent fashion.     

BDNF regulation of androgen receptor expression in axotomized SNB motoneurons of adult male rats

Brain-derived neurotrophic factor (BDNF) prevents the axotomy-induced loss of androgen receptor-like immunoreactivity (AR-LI) in the spinal nucleus of the bulbocavernosus (SNB) motoneurons of adult male rats. In this report, we investigated the dose-response effect of BDNF on androgen receptor expression in axotomized SNB motoneurons, and examined whether delayed application of BDNF to the cut SNB axons can completely reverse the axotomy-induced loss of androgen receptor expression. We also used autoradiography to test whether axotomy decreases the ability of SNB motoneurons to accumulate androgens. SNB motoneurons were axotomized bilaterally and BDNF or PBS was applied to the proximal ends of the axons. The percentage of SNB motoneurons expressing medium or high AR-LI was the major measure of androgen receptor expression. AR-LI was significantly higher on the BDNF-treated side than on the contralateral side treated with phosphate-buffered saline (PBS) for all three doses of BDNF (1.45, 2.9, and 5.8 mg/ml) and was higher than in rats treated bilaterally with PBS. Moreover, AR-LI at the highest dose of BDNF was not different from that in intact SNB motoneurons. Delayed application of BDNF to the axotomized SNB motoneurons restored the AR-LI to the intact level. The AR-LI decreased by axotomy started to increase significantly 4 days after BDNF application and returned to the intact level by 10 days. Furthermore, axotomy significantly decreased the percentage of SNB motoneurons to accumulate tritiated testosterone or its metabolites. In conclusion, our data demonstrate that BDNF completely prevents and reverses the axotomy-induced loss of AR-LI. Moreover, decrease of AR-LI by axotomy reflects the decrease in the ability of SNB motoneurons to accumulate androgens.     

Neurophysin-containing pathway from the paraventricular nucleus of the hypothalamus to a sexually dimorphic motor nucleus in lumbar spinal cord

A model that has been widely used in the study of steroid sensitive neurons, the spinal nucleus of the bulbocavernosus (SNB) is a sexually dimorphic motor nucleus in the lower lumbar spinal cord that innervates the striated bulbocavernosus (BC) muscle. The BC is responsible for penile reflexes in the male rat, which are important in ensuring pregnancy in females. The characterization of afferents to the SNB aids in the understanding of the neural circuitry involved in reproductive behavior. We have recently identified the paraventricular nucleus (PVN) as a possible source of afferents to the SNB. Because the PVN is the major source of oxytocin/vasopressin within the central nervous system (CNS), the purpose of the present study was to examine and characterize a neurophysin (NP)-containing pathway from the PVN to the SNB. The results demonstrate that neurons of the lateral parvicellular subnucleus of PVN, which project to levels of spinal cord containing SNB motoneurons, contain NP, the coproduct of oxytocin and vasopressin. NP-containing fibers and putative terminals were found in the region of the SNB and appear to contact the soma and proximal dendrites of SNB motoneurons which were retrogradely identified as BC-innervating. Electrolytic lesions, which destroy the lateral parvicellular subnucleus of PVN, abolish NP-containing fibers in the region of the SNB, suggesting that the PVN is the source of these NP fibers. The results of this study indicate a NP-containing projection from the hypothalamus directly to SNB motoneurons. It is suggested that this pathway may play a role in the integration of penile reflexes with other aspects of male copulatory behavior that are under hypothalamic control. © 1993 Wiley-Liss, Inc.     

Androgenic Regulation of Expression of β-Tubulin Messenger Ribonucleic Acid in Motoneurons of the Spinal Nucleus of the Bulbocavernosus

Expression of β-tubulin mRNA was examined in androgen-sensitive motoneurons of the spinal nucleus of the bulbocavernosus (SNB) in adult male rats by in situ hybridization histochemistry using cDNA encoding mouse β-tubulin. Hybridizable β-tubulin mRNA was localized in the somata and proximal dendrites of SNB motoneurons. Removal of androgen by castration significantly reduced the expression level of β-tubulin mRNA in the SNB motoneurons, whereas the change was prevented by testosterone treatment. On the contrary, castration or testosterone treatment did not induce any changes in the expression level of β-tubulin mRNA in the androgen-insensitive motoneurons of the retrodorsolateral nucleus. These results suggest that androgen regulates the expression of β-tubulin gene in the SNB motoneurons and may provide evidence for the molecular mechanisms of hormonally-induced neuronal plasticity in the SNB motoneurons.