Effects of tamoxifen on neuronal morphology, connectivity and biochemistry of hypothalamic ventromedial neurons: Impact on the modulators of sexual behavior.

Tamoxifen (TAM) is a selective estrogen receptor modulator, widely used in the treatment and prevention of estrogen-dependent breast cancer. Although with great clinical results, women on TAM therapy still report several side effects, such as sexual dysfunction, which impairs quality of life. The anatomo-functional substrates of the human sexual behavior are still unknown; however, these same substrates are very well characterized in the rodent female sexual behavior, which has advantage of being a very simple reflexive response, dependent on the activation of estrogen receptors (ERs) in the ventrolateral division of the hypothalamic ventromedial nucleus (VMNvl). In fact, in the female rodent, the sexual behavior is triggered by increasing circulation levels of estradiol that changes the nucleus neurochemistry and modulates its intricate neuronal network. Therefore, we considered of notice the examination of the possible neurochemical alterations and the synaptic plasticity impairment in VMNvl neurons of estradiol-primed female rats treated with TAM that may be in the basis of this neurological disorder. Accordingly, we used stereological and biochemical methods to study the action of TAM in axospinous and axodendritic synaptic plasticity and on ER expression. The administration of TAM changed the VMNvl neurochemistry by reducing ERα mRNA and increasing ERß mRNA expression. Furthermore, present results show that TAM induced neuronal atrophy and reduced synaptic connectivity, favoring electrical inactivity. These data suggest that these cellular and molecular changes may be a possible neuronal mechanism of TAM action in the disruption of the VMNvl network, leading to the development of behavioral disorders.

Autophagy plays beneficial effect on diabetic encephalopathy in type 2 diabetes: studies in vivo and in vitro.

OBJECTIVES: The hypothalamus regulates metabolism and feeding behavior by perceiving the levels of peripheral insulin. However, little is known about the hypothalamic changes after aberrant metabolism. In this study, we investigated the changes of insulin and autophagy relevant signals of hypothalamus under diabetes mellitus. METHODS: C57B/L mice were injected with low-dose streptozotocin (STZ) and fed with high-fat diet to induce type 2 diabetes mellitus. In vitro, PC12 cells were treated with oleic acid to mimic lipotoxicity. RESULTS: Results showed that the cholesterol level in the hypothalamus of the diabetic mice was higher than that of the normal mice. The expression of insulin receptors and insulin receptor substrate-1 were downregulated and the number of Fluoro-Jade C positive cells significantly increased in the hypothalamic arcuate nucleus of the diabetic mice. Furthermore, Upregulation of mammalian target of rapamycin (mTOR) and downregulation of LC 3II were obvious in the hypothalamus of the diabetic mice. In vitro, results showed that high-lipid caused PC12 cell damage and upregulated LC3 II expression. Pretreatment of cells with 3-methyladenine evidently downregulated LC3 II expression and aggravated PC12 cell death under high lipid conditions. By contrast, pretreatment of cells with rapamycin upregulated LC3 II expression and ameliorated PC12 cell death caused by lipotoxicity. CONCLUSION: These results demonstrate that autophagy activation confers protection to neurons under aberrant metabolism and that autophagy dysfunction in the hypothalamus occurs in the chronic metabolic disorder such as T2DM.

Subcellular localization of NAPE-PLD and DAGL-α in the ventromedial nucleus of the hypothalamus by a preembedding immunogold method.

The hypothalamus and the endocannabinoid system are important players in the regulation of energy homeostasis. In a previous study, we described the ultrastructural distribution of CB1 receptors in GABAergic and glutamatergic synaptic terminals of the dorsomedial region of the ventromedial nucleus of the hypothalamus (VMH). However, the specific localization of the enzymes responsible for the synthesis of the two main endocannabinoids in the hypothalamus is not known. The objective of this study was to investigate the precise subcellular distribution of N-arachidonoylphospatidylethanolamine phospholipase D (NAPE-PLD) and diacylglycerol lipase α (DAGL-α) in the dorsomedial VMH of wild-type mice by a high resolution immunogold electron microscopy technique. Knock-out mice for each enzyme were used to validate the specificity of the antibodies. NAPE-PLD was localized presynaptically and postsynaptically but showed a preferential distribution in dendrites. DAGL-α was mostly postsynaptic in dendrites and dendritic spines. These anatomical results contribute to a better understanding of the endocannabinoid modulation in the VMH nucleus. Furthermore, they support the idea that the dorsomedial VMH displays the necessary machinery for the endocannabinoid-mediated modulation of synaptic transmission of brain circuitries that regulate important hypothalamic functions such as feeding behaviors.

[Changes of hypothalamic ventromedial nucleus structure in rats of different age with experimental diabetes mellitus].

This work describes morpho-functional organization of the hypothalamic ventromedial nucleus (VMN) in rats of different age with streptozotocin-induced diabetes mellitus (DM). On day 7 of experimental DM, the development of a stress reaction was observed in 3-month-old animals, which was characterized by the ultrastructural signs of increased functional activity of light neurosecretory cells (NSC) of the VMN. Meanwhile, NSC of the VMN of 24-month-old animals remained intact. These changes in the VMN of 3-month-old animals with DM mellitus are probably related to much higher levels of glucose. On day 28 of experimental DM, the decrease in NSC numbers in the VMN was observed in the animals of both age groups because of apoptosis and hydropic degeneration. Such changes were accompanied by the deterioration in NSC trophism due to the development of the initial stages of diabetic microangiopathy and destructive changes in glial cells.

Importance of mitochondrial dynamin-related protein 1 in hypothalamic glucose sensitivity in rats.

AIMS: Hypothalamic mitochondrial reactive oxygen species (mROS)-mediated signaling has been recently shown to be involved in the regulation of energy homeostasis. However, the upstream signals that control this mechanism have not yet been determined. Here, we hypothesize that glucose-induced mitochondrial fission plays a significant role in mROS-dependent hypothalamic glucose sensing. RESULTS: Glucose-triggered translocation of the fission protein dynamin-related protein 1 (DRP1) to mitochondria was first investigated in vivo in hypothalamus. Thus, we show that intracarotid glucose injection induces the recruitment of DRP1 to VMH mitochondria in vivo. Then, expression was transiently knocked down by intra-ventromedial hypothalamus (VMH) DRP1 siRNA (siDRP1) injection. 72 h post siRNA injection, brain intracarotid glucose induced insulin secretion, and VMH glucose infusion-induced refeeding decrease were measured, as well as mROS production. The SiDRP1 rats decreased mROS and impaired intracarotid glucose injection-induced insulin secretion. In addition, the VMH glucose infusion-induced refeeding decrease was lost in siDRP1 rats. Finally, mitochondrial function was evaluated by oxygen consumption measurements after DRP1 knock down. Although hypothalamic mitochondrial respiration was not modified in the resting state, substrate-driven respiration was impaired in siDRP1 rats and associated with an alteration of the coupling mechanism. INNOVATION AND CONCLUSION: Collectively, our results suggest that glucose-induced DRP1-dependent mitochondrial fission is an upstream regulator for mROS signaling, and consequently, a key mechanism in hypothalamic glucose sensing. Thus, for the first time, we demonstrate the involvement of DRP1 in physiological regulation of brain glucose-induced insulin secretion and food intake inhibition. Such involvement implies DRP1-dependent mROS production.

GABAergic and cortical and subcortical glutamatergic axon terminals contain CB1 cannabinoid receptors in the ventromedial nucleus of the hypothalamus.

BACKGROUND: Type-1 cannabinoid receptors (CB(1)R) are enriched in the hypothalamus, particularly in the ventromedial hypothalamic nucleus (VMH) that participates in homeostatic and behavioral functions including food intake. Although CB(1)R activation modulates excitatory and inhibitory synaptic transmission in the brain, CB(1)R contribution to the molecular architecture of the excitatory and inhibitory synaptic terminals in the VMH is not known. Therefore, the aim of this study was to investigate the precise subcellular distribution of CB(1)R in the VMH to better understand the modulation exerted by the endocannabinoid system on the complex brain circuitries converging into this nucleus. METHODOLOGY/PRINCIPAL FINDINGS: Light and electron microscopy techniques were used to analyze CB(1)R distribution in the VMH of CB(1)R-WT, CB(1)R-KO and conditional mutant mice bearing a selective deletion of CB(1)R in cortical glutamatergic (Glu-CB(1)R-KO) or GABAergic neurons (GABA-CB(1)R-KO). At light microscopy, CB(1)R immunolabeling was observed in the VMH of CB(1)R-WT and Glu-CB(1)R-KO animals, being remarkably reduced in GABA-CB(1)R-KO mice. In the electron microscope, CB(1)R appeared in membranes of both glutamatergic and GABAergic terminals/preterminals. There was no significant difference in the percentage of CB(1)R immunopositive profiles and CB(1)R density in terminals making asymmetric or symmetric synapses in CB(1)R-WT mice. Furthermore, the proportion of CB(1)R immunopositive terminals/preterminals in CB(1)R-WT and Glu-CB(1)R-KO mice was reduced in GABA-CB(1)R-KO mutants. CB(1)R density was similar in all animal conditions. Finally, the percentage of CB(1)R labeled boutons making asymmetric synapses slightly decreased in Glu-CB(1)R-KO mutants relative to CB(1)R-WT mice, indicating that CB(1)R was distributed in cortical and subcortical excitatory synaptic terminals. CONCLUSIONS/SIGNIFICANCE: Our anatomical results support the idea that the VMH is a relevant hub candidate in the endocannabinoid-mediated modulation of the excitatory and inhibitory neurotransmission of cortical and subcortical pathways regulating essential hypothalamic functions for the individual's survival such as the feeding behavior.

Ventromedian forebrain dysgenesis follows early prenatal ethanol exposure in mice.

Ethanol exposure on gestational day (GD) 7 in the mouse has previously been shown to result in ventromedian forebrain deficits along with facial anomalies characteristic of fetal alcohol syndrome (FAS). To further explore ethanol's teratogenic effect on the ventromedian forebrain in this mouse model, scanning electron microscopic and histological analyses were conducted. For this, time mated C57Bl/6J mice were injected with 2.9g/kg ethanol or saline twice, at a 4h interval, on their 7th day of pregnancy. On GD 12.5, 13 and 17, control and ethanol-exposed specimens were collected and processed for light and scanning electron microscopic analyses. Gross morphological changes present in the forebrains of ethanol-exposed embryos included cerebral hemispheres that were too close in proximity or rostrally united, enlarged foramina of Monro, enlarged or united lateral ventricles, and varying degrees of hippocampal and ventromedian forebrain deficiency. In GD 12.5 control and ethanol-exposed embryos, in situ hybridization employing probes for Nkx2.1 or Fzd8 to distinguish the preoptic area and medial ganglionic eminences (MGEs) from the lateral ganglionic eminences, respectively, confirmed the selective loss of ventromedian tissues. Immunohistochemical labeling of oligodendrocyte progenitors with Olig2, a transcription factor necessary for their specification, and of GABA, an inhibitory neurotransmitter, showed ethanol-induced reductions in both. To investigate later consequences of ventromedian forebrain loss, MGE-derived somatostatin-expressing interneurons in the subpallial region of GD 17 fetal mice were examined, with results showing that the somatostatin-expressing interneurons that were present were dysmorphic in the ethanol-exposed fetuses. The potential functional consequences of this insult are discussed.

Effects of estrogens and progesterone on the synaptic organization of the hypothalamic ventromedial nucleus.

The majority of the studies on the actions of estrogens in the ventrolateral part of the hypothalamic ventromedial nucleus (VMNvl) concern the factors that modulate the receptive component of the feminine sexual behavior and the expression of molecular markers of neuronal activation. To further our understanding of the factors that regulate synaptic plasticity in the female VMNvl, we have examined the effects of estradiol and progesterone, and of estrogen receptor (ER) subtype selective ligands on the number of dendritic and spine synapses established by individual VMNvl neurons and on sexual behavior. In contrast to earlier studies that analyzed synapse densities, our results show that exogenous estradiol increases the number of spine as well as of dendritic synapses, irrespective of the dose and regimen of administration. They also reveal that an effective dose of estradiol administered as one single pulse induces the formation of more synapses than the same dose administered as two pulses on consecutive days. Our results further show that both ER subtypes are involved in the mediation of the synaptogenic effects of estrogens on VMNvl neurons since the administration of the selective ERalpha, propyl-pyrazole-triol (PPT), and ERbeta, diarylpropionitrile (DPN), agonists induced a significant increase in the number of synapses that, however, was more exuberant for PPT. Despite its relevant role in feminine sexual behavior, progesterone had no synaptogenic effect in the VMNvl as no changes in synapse numbers were noticed in rats treated with progesterone alone, with estradiol followed by progesterone or with the antiprogestin mifepristone (RU486). Except for the sequential administration of estradiol and progesterone, none of the regimens was associated with lordosis response to vaginocervical stimulation. Therefore, from the sex steroids that undergo cyclic variations over the estrous cycle, only estrogens, acting through both ERalpha and ERbeta, play a key role in the activation of the neural circuits involving the ventromedial nucleus of the hypothalamus.

Ultrastructure of the ventromedial hypothalamic nucleus in fasted and refed young and old rats.

Many hypothalamic nuclei are involved in the regulation of food intake and energy homeostasis. An ultrastructural investigation of the hypothalamic ventromedial nucleus (VMN), a hypothetical "satiety centre" was performed to explore the morphological basis of altered feeding behaviour of old rats in an experimental model of fasting/refeeding. Young (5 months old, n=12) and old (24 months old, n=12) male Wistar rats were fasted for 48 hours, then refed for 24 hours and sampled thereafter. Brain tissue was fixed by perfusion, histological and ultrathin sections were obtained by routine methods. Although food intake was similar in control young and old rats, during refeeding old animals consumed less chow than young ones. The EM analysis of VMN neurones of old control rats revealed, besides typical age-related residual bodies, deep indentations of the nuclear envelope and the presence of long, undulating rough endoplasmic reticulum cisternae in the cell periphery. In both young and old rats fasting for 48 hours led to the expansion of Golgi complexes and increased folds of the nuclear envelope, which is suggestive of enhanced cellular activity of the VMN neurones. These fasting-induced alterations were sustained in the VMN neurones of refed rats in both age groups. The results showed that the VMN neurones of old control rats differ at the ultrastructural level from young ones. However, starvation and subsequent refeeding cause similar alterations in the hypothalamic neurones of "satiety centre" of both young and old rats.

Metabolic activity of the human ventromedial nucleus neurons in relation to sex and ageing.

The ventromedial nucleus (VMN) in animals is involved in a number of sexually dimorphic behaviors, including reproduction, and is a well-documented target for sex steroids. In rats and in lizards, it is also characterized by the presence of structural sexual dimorphisms. In the present study, we determined whether the metabolic activity of human ventromedial nucleus neurons was sex- or age-related. The size of the immunocytochemically defined Golgi apparatus (GA) and cell profiles were determined as measures for neuronal metabolic activity in 12 male and 16 female control brains sub-divided into four groups with the dividing line being the age of 50. It appeared that the size of the GA relative to cell size was 34% larger in young women (<50 years old) than in young men and was 25% larger in elderly men (> or = 50 years old) than in young men. In addition, the GA/cell size ratio correlated significantly with age in men and not in women. Our data suggest that androgens play an inhibitory role with respect to the metabolic activity of the human VMN neurons.

Ultrastructural evidence for enkephalin mediated disinhibition in the ventromedial nucleus of the hypothalamus.

The ventromedial nucleus of the hypothalamus (VMN) regulates the estrogen-dependent appearance of female mating behavior, lordosis. Accumulating evidence suggests that estrogen might exert its control over lordosis by acting, in part, on neurons that contain enkephalin in the VMN. The expression of the enkephalin precursor gene is robustly stimulated by estrogen and is correlated with the later appearance of lordosis. GABA has also been implicated as an important neurotransmitter for the appearance of lordosis. Because enkephalin is thought to act in several brain areas to modulate the activity of GABAergic neurons, we studied the ultrastructural morphology and relationship between neurons containing these neurochemicals using dual-labeling immunocytochemistry in ovariectornized rats, half of which received estrogen replacement. Immunolabeling for enkephalin was almost always detected within axon terminals (695 axonal profiles sampled), while GABA immunoreactivity was more often localized to cell bodies and dendrites (191 profiles), than to axons (63 profiles). Axon terminals containing enkephalin immunolabeling provided a major innervation to soma or dendrites containing GABA. That is, over one third (94/245) of the axon terminals in contact with GABA-immunoreactive dendrites contained enkephalin. Furthermore, these GABA-immunoreactive dendrites accounted for a fifth of the somatodendritic processes associated with enkephalin-containing axon terminals. These findings support the hypothesis that enkephalin may act in the VMN by inhibiting GABAergic neurons, which could result in the disinhibition of neural circuits relevant for lordosis.

Pineal influence on annual nuclear volume changes in ventromedial hypothalamic nucleus (VMH) neurons of the male Wistar rat.

The ventromedial hypothalamic nucleus (VMH) regulates various autonomic, endocrine, and behavioral activities. These activities show annual changes, and the pineal gland is involved in their adjustment to environmental cues. Therefore, this study investigated whether the VMH belongs to the effector structures of the pineal gland. To abolish the rhythmic melatonin release, male Wistar rats were subjected to pinealectomy (PX) or ganglionectomy (sympathetic denervation of the pineal gland, GX) regularly at the beginning of any of the four seasons. Brains from animals of PX-, GX-, and sham-operated control groups were prepared 3 months later for measurement of the nuclear volume, which changes according to the general gene activity. At each of the four seasons, 2000 nuclei of VMH neurons stemming from 18 animals per group were measured to obtain both seasonal daily mean values and annual mean values, respectively, as well as to calculate annual curves of the nuclear volume using empirical regression and locally adjusted polynomial approximation. The major findings are the following. First, inactivation of the pineal function influences the nuclear activity of VMH neurons, (2) PX and GX mainly depress the nuclear activity, indicating that the pineal influence on the VMH may predominantly be a stimulatory one. Third, size and direction of the changes caused by PX and GX vary in a seasonally dependent manner. Fourth, the annual rhythm of the nuclear activity of the VMH is modified by PX and GX. To explain how the pineal effects on the VMH may be mediated, a possible inhibitory influence of the suprachiasmatic nucleus (SCN), which has been activated in the same animals following both PX and GX, is discussed. In conclusion, the results confirm that the nuclear activity of VMH neurons underlies pineal influences. This also indicates an involvement of the pineal gland in many VMH-regulated functions.

A cellular stress model for the differential expression of glial lysosomal cathepsins in the aging nervous system.

Activation of the endosomal-lysosomal system and altered expression of various lysosomal hydrolases have been implicated in several senescence-dependent neurodegenerative disorders and occurs, to a lesser extent, in the course of normal brain aging. The progressive accumulation of autofluorescent, peroxidase-positive astrocytic granules represents a highly consistent biomarker of aging in the vertebrate CNS. The sulfhydryl agent cysteamine greatly accelerates the accumulation of these glial inclusions in situ and in primary brain cell cultures. We previously determined that these glial inclusions are derived from abnormal mitochondria which undergo fusion with lysosomal elements in a complex autophagic process. In the present study, we demonstrate that cysteamine suppresses cathepsin B mRNA levels and immunoreactive protein in cultured astroglia, whereas cathepsin D mRNA and protein levels are significantly augmented by CSH exposure in these cells. Moreover, cathepsin D (but not cathepsin B) exhibits robust colocalization to the red autofluorescent inclusions. Concordant with our in vitro observations, cathepsin B immunoreactivity is prominent in the hypothalamic ventromedial nucleus which accumulates few autofluorescent glial inclusions during aging and is relatively inapparent in the heavily granulated hypothalamic arcuate nucleus. Conversely, cathepsin D is prominent in the aging arcuate nucleus where it colocalizes to the autofluorescent inclusions and exhibits scant immunoreactivity in the adjacent ventromedial nuclear complex. In senescent astroglia, oxidative stress may down-regulate the cathepsin B gene as part of a concerted cellular stress (heat shock) response. Glial cathepsin D, on the other hand, resists stress-related inhibition and may play an important role in disposing of oxidatively modified mitochondria in the aging and degenerating nervous system.

Sexually dimorphic contribution from the fornix to the ventromedial hypothalamic nucleus: a quantitative electron microscopic study.

Quantitative electron microscopy confirmed that the neuropil of the ventrolateral part of the ventromedial hypothalamic nucleus (VL-VMHN) of the rat is sexually dimorphic with respect to the density of shaft and axo-spinous synapses, both of which are more numerous in the male. In addition, adult rats with complete interruption of the fornix displayed a sexually dimorphic input in the density of fornical synapses in the neuropil of the VL-VMHN, in which degenerating terminals were more numerous in the male. Perinatal exposure of the female to exogenous testosterone or castration of the newborn male 'inverted' these sex differences, demonstrating their hormonal dependence. It is concluded that (1) the fornix provides synaptic input to the VL-VMHN as proven by orthograde degeneration; (2) the number of fornical endings synapsing in the VL-VMHN is greater in the male than in the female; (3) this dimorphism depends of the organizational effect of gonadal sex steroids.

Ultrastructural effects of estradiol and 5 alpha-androstane-3 alpha, 17 beta-diol on neurons within the ventromedial nucleus of the hypothalamus.

The metabolite of dihydrotestosterone, 5 alpha-androstane-3 alpha, 17 beta-diol (3 alpha-Diol), is a potent inhibitor of estrogen-induced gonadotropin and prolactin secretion and lordosis behavior in the female rat. This study examined whether 3 alpha-Diol can counteract the ultrastructural changes which are known to occur in the ventromedial nucleus (VMN) of the hypothalamus following estrogen treatment. Ovariectomized rats were treated with estradiol (E2; n = 7), 3 alpha-Diol (n = 5), E2 and 3 alpha-Diol (n = 6), or received control (n = 6) treatments. E2 was administered in subcutaneous capsules for two discontinuous 2-hour periods separated by 5 h, a 'pulsed' treatment regimen known to mimic the timing of endogenous E2 action and to influence neuronal ultrastructure in the VMN. Animals given 3 alpha-Diol received subcutaneous injections (6 mg/kg) 3 h prior to each implantation of E2 or empty capsules. Control animals received vehicle 3 h prior to implantation of blank capsules. Animals were perfused 24 h after initial hormone treatment and neurons from the ventrolateral portion of the VMN were examined using electron microscopy. Separately, both E2 and 3 alpha-Diol treatment increased somal and nuclear size, altered somal and nuclear shape, and increased the numbers of lysosomes present in the cytoplasm above control levels. E2 treatment resulted in increased stacking of the rough endoplasmic reticulum while 3 alpha-Diol treatment resulted in an unusual plexiform rough endoplasmic reticulum distribution. In contrast, combined treatment with E2 and 3 alpha-Diol resulted in cells which were similar to ovariectomized control cells on these measures. All steroid treatments decreased the amount of heterochromatin present within the nucleus compared to that seen in controls. Thus, 3 alpha-Diol influences the ultrastructural characteristics of neurons within the VMN in a manner somewhat though not altogether similar to E2. However, 3 alpha-Diol given in combination with E2 counteracts or prevents the actions of E2 within these same neurons.

Distribution of somatostatin immunoreactivity in sheep hypothalamus: a comparison with that of the rat.

The distribution of somatostatin immunoreactivity was determined throughout the hypothalamus of the sheep and comparisons were made with the known distribution of somatostatin immunoreactivity in the rat. Immunopositive perikarya were present in the sheep periventricular region from as far rostral as the supraoptic recess of the third ventricle to the posterior optic chiasm. In the basal hypothalamus, a thick shell of immunopositive neurons surrounded the ventromedial nucleus (VMH), and there were also neurons in the caudal arcuate nucleus. Somatostatin immunoreactive fibres were concentrated in the dorsal VMH and arcuate nucleus as well as in the median eminence. The distribution in sheep was similar to that in rats, but immunoreactive neurons around sheep VMH were distinctive, a characteristic that might relate to differences in growth hormone physiology in this species.

Postnatal development of the hypothalamic ventromedial nucleus: neurons and synapses.

1. In this report the postnatal differentiation of the hypothalamic ventromedial nucleus (VMN) was studied. The main maturational changes detected at the fine structural level occurred between 10 and 20 days of postnatal life. 2. In 5-day-old rats the majority of neurons was undifferentiated, with rudimentary cytoplasmic organelles. Dendritic profiles presented an empty appearance due to an electron-lucent matrix and scarce content of organelles. 3. At 10 days there was a significant proliferation of cytoplasmic organelles in the perikaryon, mainly of those involved in protein biosynthesis as the rough endoplasmic reticulum (RER) and the Golgi complex. 4. After 20 days of age the VMN neurons acquired the cytological appearance of adult neurons, with well-organized RER, Golgi complexes, and pleomorphic mitochondria. Concurrent with these changes, there was a marked development of other organelles in the neuropil, which was accompanied by an increase in synaptic density and differentiation of their subsynaptic structures.

5,7-Dihydroxytryptamine and gonadal steroid manipulation alter spine density in ventromedial hypothalamic neurons.

Golgi impregnation was used to examine changes in dendritic spine density in the ventromedial hypothalmic nucleus (VMN) of intact and gonadectomized (GDX) rats treated with 5,7-dihydroxytryptamine (5,7-DHT) and/or estrogen. In intact rats of both sexes 5,7-DHT injection resulted in a large increase in dendritic spine density in the VMN. The combination of 5,7-DHT and estrogen treatment in GDX females resulted in a further increase in dendritic spine density. In male rats gonadectomy increased dendritic spine density on VMN neurons, and estrogen treatment partially reversed this effect. When GDX males were treated with 5,7-DHT and estrogen, there was no effect on dendritic spine density. These results complement a previous study in which it was demonstrated that dendritic spine density in the VMN was greatest during proestrus and after estrogen administration to ovariectomized females. The relevance of these findings to gonadal-steroid-mediated lordosis is discussed.

Stereological analysis of the hypothalamic ventromedial nucleus. II. Hormone-induced changes in the synaptogenic pattern.

Stereological electron microscopic analysis of the rat hypothalamic ventromedial nucleus (VMN) throughout postnatal development revealed that synaptogenesis takes place up to day 45. Our results disclosed a sexual dimorphism in the synaptic organization of the neuropil of the ventrolateral (VL) subdivision of the VMN. The numerical densities of spine and shaft synapses in the adult male were higher than in the female. A dimorphic pattern in the numerical density of spine synapses occurred as early as day 5, and was present throughout postnatal life, even though in the adult rats both spine and shaft synapses were sexually dimorphic. Neonatal treatment of female rats with testosterone increased the numerical density of axodendritic synapses, inducing a pattern similar to the adult male. On the other hand, administration of tamoxifen to newborn male rats significantly reduced the numerical density of spine synapses to levels comparable to normal female rats.

Estrogen increases axodendritic synapses in the VMN of rats after ovariectomy.

Low doses of estrogen (E) were given to ovariectomized (OVX) rats and the ventromedial hypothalamic nucleus (VMN) was examined at the ultrastructural level. Administration of 10 micrograms of E for two consecutive days resulted in an increase in the number of axodendritic synapses in the VMN in comparison to OVX rats treated with oil. The percentage of axospine and multiple synapses and the length of the postsynaptic density were unchanged by E treatment. These results provide further support for the ability of low doses of E to alter neuronal morphology.