Immunocytochemical localization of corticotropin-releasing factor (CRF) in the rat brain

The immunocytochemical localization of neurons containing the 41 amino acid peptide corticotropin-releasing factor (CRF) in the rat brain is described. The detection of CRF-like immunoreactivity in neurons was facilitated by colchicine pretreatment of the rats and by silver intensification of the diaminobenzidine end-product. The presence of immunoreactive CRF in perikarya, neuronal processes, and terminals in all major subdivisions of the rat brain is demonstrated. Aggregates of CRF-immunoreactive perikarya are found in the paraventricular, supraoptic, medial and periventricular preoptic, and premammillary nuclei of the hypothalamus, the bed nuclei of the stria terminalis and of the anterior commissure, the medial septal nucleus, the nucleus accumbens, the central amygdaloid nucleus, the olfactory bulb, the locus ceruleus, the parabrachial nucleus, the superior and inferior colliculus, and the medial vestibular nucleus. A few scattered perikarya with CRF-like immunoreactivity are present along the paraventriculo-infundibular pathway, in the anterior hypothalamus, the cerebral cortex, the hippocampus, and the periaqueductal gray of the mesencephalon and pons. Processes with CRF-like immunoreactivity are present in all of the above areas as well as in the cerebellum. The densest accumulation of CRF-immunoreactive terminals is seen in the external zone of the median eminence, with some immunoreactive CRF also present in the internal zone. The widespread but selective distribution of neurons containing CRF-like immunoreactivity supports the neuroendocrine role of this peptide and suggests that CRF, similarly to other neuropeptides, may also function as a neuromodulator throughout the brain.     

Immunocytochemical ultrastructural study of hypothalamic neurons containing corticotropin-releasing factor in normal and adrenalectomized rats

The neurons of the rat hypothalamus which secrete corticotropin-releasing factor were studied by using a pre-embedding immunocytochemical staining technique that improves both the penetration of immunoreagents within the tissue and the preservation of the ultrastructural morphology of labeled structures. Comparison was made between the subcellular location of corticotropin-releasing factor-41 in perikarya of the paraventricular nucleus and axons of the median eminence, both in intact and adrenalectomized animals either untreated or 24 h after the intracerebral injection of colchicine. Morphometric analysis of the numerical density and of the diameter of corticotropin-releasing factor immunoreactive neurosecretory granules in axons of the median eminence of rats not treated with colchicine, indicated that the main modifications induced by adrenalectomy concerned (1) the differential repartition of labeled granules within the preterminal and terminal axonal portions of the median eminence, and (2) the enlargement of the diameter of labeled granules contained in these axons (from 98 nm to 165 nm). In the hypothalamus of intact and adrenalectomized rats, colchicine treatment increased the number of corticotropin-releasing factor-immunoreactive granules in the neuronal perikarya and reduced their number in the axons, but both these variations were much more marked in adrenalectomized rats. Although the corticotropin-releasing factor immunoreactive granules that accumulated in the perikarya after colchicine treatment were slightly smaller than those in the corresponding axons, the diameter of perikaryal-labeled granules was larger in adrenalectomized than in intact animals (129 nm vs 93 nm). These findings fit the idea that adrenalectomy markedly stimulates both the synthesis and axonal excretion of secretory granules in the hypothalamic neurons secreting corticotropin-releasing factor. They also indicate that suppression of circulating corticosteroids induces qualitative modifications in these neurons leading to the visualization of larger neurosecretory granules, which may reflect differential synthesis and granular packing of synergistic peptides other than corticotropin-releasing factor and/or changes in the process of intragranular maturation of hormonal material.     

Immunohistochemical localization of corticotropin-releasing factor (CRF)-like immunoreactivity in the hypothalamus of the newt, Triturus cristatus

The distribution of corticotropin-releasing factor (CRF) immunoreactivity was investigated in the hypothalamus and preoptic area of the newt by single and double immunocytochemical procedures. CRF immunopositive cell bodies were seen in the preoptic area (from the anterior wall of the preoptic recess to the dorsal parts of the preoptic nucleus) and in the tuberal portions of the posterior hypothalamus. Abundant nerve fibres are seen in the outer zone of the median eminence, while the pars nervosa lacks CRF-immunoreactivity. CRF immunoreactive material is seemingly separated from neurophysins- and somatostatin immunoreactive cell bodies and fibres.     

Effects of ethanol on hypothalamic luteinizing hormone-releasing hormone (LHRH) in the male rat an immunocytochemical study

Summary These studies were designed to determine if the acute alcohol-induced decreases in luteinizing hormone (LH) seen in previous studies using rats could be due to an inhibitory effect of ethanol (ETOH) on hypothalamic LHRH release. Thus, effects of multiple injections of ETOH on the relative amount of immunoreactive LHRH fibers in the hypothalamus and median eminence (ME) of castrate and intact male rats were determined immunocytochemically. Brains were removed following cardiac perfusion of 10% phosphate-buffered formalin. A block containing the hypothalamus with the ME was isolated from each brain, then postfixed in Bouin's solution. Paraffin sections were rehydrated and stained for LHRH with the peroxidaseantiperoxidase technique using an antiserum to synthetic LHRH conjugated to bovine serum albumen. Differences visualized immunocytochemically between saline-treated intact and castrate rats indicated that the LHRH content of the ME was markedly depleted after castration. Conversely, castrate rats treated with ETOH showed only a slight reduction in immunoreactive LHRH fibers. In ETOH-treated intact animals, the LHRH fiber content of both the hypothalamus and ME appeared to be slightly greater than the saline-treated intact controls. Thus, these data support the hypothesis that ETOH diminishes LHRH release, and hence provides an explanation for the depressed plasma LH levels observed in ETOH-treated intact and castrate rats.This work was supported by Grants NIH-BRSG-3-81 and TAMU-ORR-8-82     

生长抑素样神经元及其纤维在人胎下丘脑的分布与发育研究

本文用免疫细胞化学技术观察了人胎下丘脑内含生长抑素样神经元及其纤维的分布与发育。胎龄14周,见浅染的生长抑素免疫反应阳性神经元散布于下丘脑前部、室周带、第三脑室壁室管膜上皮深面。15～16周胎龄,含生长抑素样神经元除见于上述部位外,还见于腹内侧基底部、室旁核、视上核和室管膜上皮内,阳性纤维散布于正中隆起,室管膜上皮。17—18周胎龄,含生长抑素样结构在上述部位变化明显,神经元更加密集,分布范围扩大,反应染色加深,阳性突起延长。21周胎龄至足月胎儿,含生长抑素神经元及其纤维在下丘脑的分布变化不明显。生长抑素样神经元以中、小型细胞为主,室旁核内见有少量大细胞。电镜观察21周和29周胎龄正中隆起发现:毛细血管为窗孔型,基底膜外有生长抑素样阳性终末靠近。在阳性终末及非阳性终末内可见分泌囊泡。本结果提示人下丘脑内生长抑素系统在出生前已经发育并可能参与内分泌活动的调节。     

Light- and electron-microscopic immunocytochemistry of glutamic acid decarboxylase (GAD) in the basal hypothalamus: Morphological evidence for neuroendocrine γ -aminobutyrate (GABA)

GABAergic cells and axon terminals were localized in the basal hypothalamus of different species (rat, mouse and cat), by means of an immunocytochemical approach using a specific and well-characterized antiserum to the GABA biosynthetic enzyme, glutamate decarboxylase. Lightmicroscopic visualization was performed with an indirect immunofluorescence method and electron-microscopic observations were made on material with pre-embedding staining and use of the peroxidase-antiperoxidase procedure.At the light-microscopic level, a dense immunofluorescent plexus was observed over both the medial and lateral parts of the external layer of the median eminence. The labelling extended from the rostral part of the median eminence up to the pituitary stalk. Over the subependymal and internal layers only a few immunoreactive dots were visible, except around the blood vessels where they appeared more concentrated. Immunoreactive varicosities could be found following the outlines of the capillary loops and lining tanycyte processes, especially in the median eminance midportion.At the electron-microscopic level, the immunolabelling was exclusively found over neuronal profiles in the median eminence. The latter represented a small fraction of the total number of varicosities visible on the same section. Labelled profiles typically contained numerous small clear synaptic vesicles and only a few or no dense-core vesicles. In the subependymal and internal layers, rare labelled endings were found close to ependymal cells or among transversally cut fibers, respectively. In the palisadic zone, elongated positive boutons were visible intermingled with bundles of unlabelled axons and glial or ependymal processes. In the neurohemal contact zone, immunoreactive endings were observed among unlabelled neurosecretory endings in close vicinity to fenestrated capillary perivascular space.Small moderately intense immunofluorescent varicosities were observed all over the hypothalamus. The density of the glutamate decarboxylase-positive network was higher than in most diencephalic regions. Intraventricular or topical injection of colchicine allowed the visualization of small lightly immunoreactive cells in the diffusion area of colchicine. In the arcuate nucleus labelled axonal endings containing small pleomorphic synaptic vesicles and sometimes a few dense-core vesicles were observed at the electron-microscopic level. Typical synaptic junctions were commonly found between positive endings and unlabelled perikarya, or more frequently, unlabelled dendrites.These findings show that glutamate decarboxylase-containing endings are localized in several strategic sites for potential GABAergic neuroendocrine regulations. The GABAergic endings found among neurosecretory endings in the neurohemal contact zone may provide the morphological support for the release of γ -aminobutyrate into the portal blood flow as an hypothalamic hypophysiotropic hormone. Alternatively, neurosecretory cells might be under GABAergic control expressed either at their terminal level within the median eminence or the cell body level within the parvicellular hypothalamic nuclei.     

大鼠下丘脑生长抑素神经元性差别的观察

本实验用PAP免疫组织化学方法对比观察了雄性与雌性大鼠下丘脑生长抑素(SRIF)神经元胞体、纤维在正常和改变性激素代谢条件下数量或染色程度的差别。结果表明,在室周核(PV)、室旁核(PVN)和视交叉上核(SCN)内,雄性大鼠的阳性细胞和纤维较雌性多。此外,在视前内侧区(MPO)雄性有大量细丝样阳性纤维网络,而雌性只有较稀疏的阳性纤维。切除睾丸后大鼠PV、PVN、SCN阳性细胞和纤维减少,MPO和正中隆起阳性纤维密度降低。而切除卵巢的大鼠在上述区域的阳性胞体和纤维密度增强,当给去性腺的雌、雄性大鼠以同性激素替代后,染色结果接近正常。     

下丘脑生长抑素神经元的发生

实验用免疫组化ＰＡＰ漂浮法研究了下丘脑生长抑素免疫反应阳性神经元的发生发育规律。结果表明生长抑素免疫反应阳性神经元的发生发育规律可归为三种模式；（１）生长抑素阳性神经元或纤维逐渐增至成年大鼠水平、代表核区为室周核和正中隆起；（２）阳性神经元或纤维数目很快增至峰值，然后又降至成年大鼠水平，该模式最普遍，大多数核区阳性神经元或纤维的发生都属于该模式。     

Immunoreactivity of hypothalamo-neurohypophysial neurons which secrete corticotropin-releasing hormone (CRH) and vasopressin (Vp): immunocytochemical evidence for a correlation with their functional state in colchicine-treated rats

Summary The specific immunoreactivity of neurons containing corticotropin-releasing hormone (CRH) or vasopressin (Vp) was studied both centrally, in the parvocellular division of the paraventricular nucleus, and distally, in the external median eminence. Control rats were compared with adrenalectomized rats and with animals supplemented with corticosterone or dexamethasone, either without additional treatment, or 24, and 48 h after an intraventricular injection of colchicine. In all groups of animals, colchicine induced a progressive and parallel decrease in both CRH and Vp immunoreactivity within the axons of the external median eminence. A semi-quantitative estimation of this axonal immunostaining showed that the decrease was clearly correlated with the axons' releasing activity according to the different functional states of the adrenocorticotropic system. Increased rates of hormonal release induced by adrenalectomy could be seen in the accelerated depletion of axonal immunoreactivity whereas corticosteroid supplementation had the opposite effect. Correspondingly, the progressive intensification of the CRH and Vp immunoreactivity within the perikarya following colchicine treatment was further markedly enhanced in adrenalectomized rats and diminished after corticosteroid supplementation. Taken together, these data suggest that in these neurons, perikaryal hormone synthesis may be closely related to the releasing activity of the axon terminals. They further point to appropriate colchicine treatment as useful tool for evaluating the functional state of CRH and Vp neurons of the parvocellular paraventricular nucleus under various experimental conditions.     

Neurotransmitters,neuropeptides and binding sites in the rat mediobasal hypothalamus: effects of monosodium glutamate (MSG) lesions

Summary Indirect immunofluorescence histochemistry and receptor autoradiography were used to study the localization of transmitter-/peptidecontaining neurons and peptide binding sites in the mediobasal hypothalamus in normal rats and in rats treated neonatally with repeated doses of the neurotoxin monosodium-glutamate (MSG). In the arcuate nucleus, the results showed a virtually complete loss of cell bodies containing immunoreactivity for growth hormone-releasing factor (GRF), galanin (GAL), dynorphin (DYN), enkephalin (ENK), corticotropin-like intermediate peptide (CLIP), neuropeptide Y (NPY), and neuropeptide K (NPK). Tyrosine hydroxylase(TH)-, glutamic acid decarboxylase(GAD)-, neurotensin(NT)- and somatostatin(SOM)-immunoreactive (IR) cells were, however, always detected in the ventrally dislocated, dorsomedial division of the arcuate nucleus. In the median eminence, marked decreases in numbers of GAD-, NT-, GAL-, GRF-, DYN-and ENK-IR fibers were observed. The numbers of TH-, SOM-and NPY-IR fibers were in contrast not or only affected to a very small extent, as revealed with the immunofluorescence technique. Biochemical analysis showed a tendency for MSG to reduce dopamine levels in the median eminence of female rats, whereas no effect was observed in male rats. Autoradiographic studies showed high to moderate NT binding sites, including strong binding over presumably dorsomedial dopamine cells. In MSG-treated rats, there was a marked reduction in GAL binding in the ventromedial nucleus. The findings implicate that most neurons in the ventrolateral and ventromedial arcuate nucleus are sensitive to the toxic effects of MSG, whereas a subpopulation of cells in the dorsomedial division of the arcuate nucleus, including dopamine neurons, are not susceptible to MSG-neurotoxicity. The results indicate, moreover that the very dense TH-IR fiber network in the median eminence predominantly arises from the dorsomedial TH-IR arcuate cells, whereas the GAD-, NT-, GAL-, GRF-and DYN-IR fibers in the median eminence to a large extent arise from the ventrolateral arcuate nucleus. Some ENK-and NPK-positive cells in the arcuate nucleus seem to project to the lateral palisade zone of the median eminence, but most of the ENK-IR fibers in the median eminence, located in the medial palisade zone, seem to primarily originate from an area(s) located outside the arcuate nucleus, presumably the paraventricular nucleus. The NPY-positive fibers in the median eminence contain to a large extent immunoreactive dopamine -hydroxylase (DBH), and do not arise from the ventromedial arcuate nucleus. SOM-IR cells in the dorsal periventricular arcuate nucleus do not send major projections to the median eminence. The present findings thus show that MSG treatment represents a valuable tool to clarify the organization of chemically identified neuron populations in the arcuate nucleus-median eminence complex and provide further information for understanding the neuroendocrine effects of neonatal MSG treatment.     

Adrenalectomy increases sprouting in a peptidergic neurosecretory system

The neurons of the paraventricular nucleus of the hypothalamus have a substantial vasopressin-containing projection to the zona externa of the median eminence. Several parameters of neuronal function are increased in these cells following adrenalectomy; these increases can be prevented by glucocorticoid treatment. We have now used this system to study the effect of adrenal removal on the sprouting of axons in response to a unilateral lesion of the paraventricular nucleus. Results obtained from both immunocytochemical and anterograde tracing experiments indicate that the capacity of the paraventricular nucleus axons to sprout is enhanced in adrenalectomized animals. Anterograde tracing experiments indicate that the projection from the paraventricular nucleus to the zona externa is predominately ipsilateral confirming the findings of Alonso & Assenmacher¹ and that sprouting does not occur in intact animals with a unilateral paraventricular nucleus lesion. However, the zona externa of adrenalectomized animals with a unilateral paraventricular nucleus lesion is filled bilaterally with neurophysin terminals; the volume occupied by such fibers is the same as in non-lesioned, adrenalectomized animals. These findings are interpreted to indicate that sprouting can occur in this system if the animal is adrenalectomized. To estimate the time course of sprouting, animals were first adrenalectomized for 2 weeks and then a unilateral lesion placed in the paraventricular nucleus. Animals were killed 1–26 days following lesion placement and the brains processed for immunocytochemistry. Initial denervation in the zona externa due to the lesion can be detected by day 6 post-lesions while reinnervation (sprouting) is seen by day 26.The results suggest that the increase in metabolic activity induced by adrenalectomy in the vasopressin neurons under study may influence the ability of the axons of these cells to produce new terminals.     

Occurrence of galanin-like immunoreactivity in growth hormone-releasing factor (GRF)-containing neurons of the monkey (Macaca fascicularis) infundibular nucleus and median eminence.

The distribution of growth hormone-releasing factor (GRF)- and galanin (GAL)-immunoreactive (-IR) neurons in the mediobasal hypothalamus of the monkey (Macaca fascicularis) was studied with immunohistochemistry using a direct double-labelling method. GRF- and GAL-IR cell bodies were demonstrated in the ventral part of the infundibular nucleus and dense aggregations of GRF- and GAL-IR fibers were seen in the external layer of the median eminence, closely surrounding portal vessels. Double-staining revealed that GRF and GAL were colocalized in cell bodies of the infundibular nucleus and in nerve fiber varicosities in the external layer of the median eminence. GAL has been reported to stimulate the secretion of growth hormone in both rats and humans, most likely via hypothalamic mechanism(s).     

大鼠新生期皮下注射谷氨酸单钠对成年后下丘脑ACTH神经元和正中隆起CRF神经纤维免疫反应性的影响

本文采用过氧化酶—抗过氧化酶(PAP)的免疫组织化学和荧光免疫组织化学的方法,研究了新生期大鼠皮下注射谷氨酸单纳(MSG)对成年后下丘脑弓状核促肾上腺皮质激素(ACTH)神经元和正中隆起内促肾上腺皮质激素释放因子(CRF)神经纤维免疫反应的影响。新生期MSG处理后,弓状核中ACTH神经元的数目明显减少,正中隆起CRF样免疫反应末稍也大大减少,减少程度随MSG剂量增大而增加。提示MSG通过损伤前阿黑皮素(POMC)衍生的神经多肽,从而影响下丘脑—垂体—肾上腺皮质轴的功能。     

Autoradiographic localization of glucocorticosteroid binding sites in rat brain after in vivo injection of [3H]RU 28362

The autoradiographic distribution of glucocorticosteroid binding sites in the brain of adrenalectomized rats was studied following in vivo injection of a potent synthetic glucocorticosteroid agonist [3H]RU 28362. Analysis of the autoradiograms revealed a specific and dense labeling in the pyramidal cell layer of the Ammon's horn and in the granular cell layer of the dentate gyrus of the hippocampus. In the hypothalamus, the labeling was particularly high in the paraventricular nucleus (site of CRF synthesis), the arcuate, periventricular and the supraoptic nuclei as well as in the median eminence. Autoradiograms also revealed the presence of [3H]RU 28362 binding sites in several brain regions including the amygdala, the pineal gland, the entorhinal cortex, the interpeduncular, interfascicular and dorsal raphe nuclei, the central grey and the substantia nigra suggesting possible effects of glucocorticosteroids in these structures.     

Age-related changes in dopamine, LHRH and somatostatin in the rat hypothalamus

The distribution of dopamine, somatostatin and LHRH was examined in young and aged male rats of the Fisher 344 strain. Dopamine histofluorescence and peptide immunocytochemical staining were performed together in each animal by the use of a stimultaneous visualization technique. Comparative analysis or rats at 3, 12, 20, and 30 months of age revealed a general decrease in somatostatin and LHRH in the median eminence; dopamine fluorescence intensity also was depressed in the median eminence although dopaminergic perikarya of the arcuate nucleus of the hypothalamus appeared to increase in intensity with age. The age-related decline in median eminence LHRH may point to a central locus of reproductive senescence in the rat.     

后叶加压素在下丘脑的分布和释放途径及失水对正中隆起中后叶加压素的影响

我们用免疫细胞化学方法(ICC)对大白鼠下丘脑中加压素(VP)免疫反应阳性的神经元作了较详细研究。观察到VP阳性神经元存在于下丘脑室旁核(PVN)各亚核、视上核(SON)、交叉上核(SCN)、室周核(PN)及一些附属核团,包括环状核(CN)、交叉后核(RCN)、下丘脑外侧核(HLN)、穹窿周围核(PFN)。且发现一特殊的VP阳性胞体聚集区—很可能是多巴胺神经元聚集区A_(14)中的细胞,位于第三脑室侧壁中1/3段两侧,腹内侧核背侧。本文首次观察到在PVN和SON之间有VP阳性的神经纤维相联系。ICC和免疫电镜研究进一步证明在正中隆起外带存在VP阳性纤维,含大颗粒囊泡的VP末梢紧邻门脉毛细血管。将HRP注入第四脑室用HRP逆行追踪与ICC方法相结合,在PVN中观察到双标细胞,与直接用ICC法所见VP阳性轴突伸入第三脑室腔的结果一致。说明PVN中存在接触脑脊液神经元。干渴动物正中隆起的内、外带中的VP免疫反应明显减弱。     

LHRH pathways in rat brain: 'deafferentation' spares a sub-chiasmatic LHRH projection to the median eminence

Localization of luteotrophic hormone-releasing hormone (LHRH) was examined by immunocytochemistry in untreated male rats and rats that received an anterior hypothalamic deafferentation. Gonadotrophic function was assessed by examining testicular weight and morphology. All of the antisera used in this study were able to reveal LHRH cell bodies and fibers. Cells from the medial preoptic area, (particularly the preoptic periventricular or median preoptic nuclei) and lateral anterior hypothalamus sent axons to the median eminence. The fiber tracts were loosely organized into (1) a tract that coursed through the organum vasculum of the lamina terminalis between the optic nerves and along the ventral surface of the optic chiasm; (2) a tract that coursed to the organum vasculum from the preoptic area and then traversed along the floor of the third ventricle; (3) a tract that coursed from the medial preoptic area to the median eminence along the lateral walls of the third ventricle; and (4) a tract that arose from the more caudal portions of the LHRH cell field in the lateral preoptic and lateral anterior hypothalamus, coursed along with the fibers of the medial forebrain bundle and turned medially at the caudal hypothalamus to enter the median eminence. An anterior cut which served most of the connections between the medial preoptic area and hypothalamus but did not penetrate through the optic chiasm served tract 3 and most of tracts 2 and 4, but spared the subchiasmatic projection (tract 1). The fibers that remained in the median eminence were sufficient to retain gonadotrophic function. This study provides an explanation for the variable effects of deafferentation and lesions of the anterior hypothalamus and preoptic area on gonadotrophic function.     

Effects of corticotropin releasing factor on genetically obese (fatty) rats

Corticotropin releasing factor (CRF) has been administered into the third ventricle of lean and genetically obese Zucker fatty rats in both acute and chronic experiments. Following a single injection of CRF (5 micrograms or approximately 1 nmole) there was an acute reduction of food intake in both the lean and obese animals, but the effect was greater in the obese. This effect persisted for the first three hours but was no longer detectable in either lean or genetically obese animals at 6 hours. Binding of GDP to mitochondria from interscapular brown adipose tissue in 21-hour deprived animals was lower in the fatty rats than in the lean controls. The injection of CRF significantly increased GDP binding in both the lean and fatty rats. During chronic infusion of CRF into the third ventricle of fatty rats, there was a significant decrease in food intake in the obese rats and fall of body weight in both groups. The basal levels of GDP binding were significantly lower in the saline-infused fatty rats than in the saline-infused lean controls. The chronic infusion of CRF increased GDP binding in the fatty rats but not in the lean animals. The CRF-treated values for GDP binding in fatty rats however, remained significantly below the baseline values in the control animals. Chronic CRF infusion also significantly lowered glucose levels in the fatty rat. These studies are consistent with the hypothesis that CRF may be involved in the decreased food intake and increased sympathetic activity observed in genetically obese fatty rats following adrenalectomy.     

烫伤大鼠第三脑室伸长细胞的免疫组化研究

目的探讨伸长细胞在烫伤应激中是否参与转运促肾上腺皮质释放因子(CRF)、加压素(VP)和胆囊收缩素(CCK)等应激激素。方法在大鼠清醒状态下给予标准的25％Ⅲ°皮肤烫伤,用免疫细胞化学ABC法显示第三脑室室管膜内的伸长细胞。结果大鼠烫伤后24h大量CRF免疫反应阳性的伸长细胞出现在第三脑室底壁和外侧壁,其顶部与脑脊液接触,基部伸出一条细长的突起走向正中隆起的毛细血管附近;位于第三脑室侧壁伸长细胞的突起似与深面脑组织的毛细血管接触。未观察到含VP和CCK的伸长细胞。结论结果为伸长细胞转运CRF提供了形态学依据,提示伸长细胞可能参与应激状态下CRF超短正反馈回路的调节。     

Effect of ovariectomy on the hypothalamic content of immunoreactive gonadotropin-releasing hormone in the female mouse as revealed by quantitative immunocytochemistry and radioimmunoassay

Our radioimmunoassay data reveal that ovariectomy for either one or two months resulted in a significant decrease in the GnRH content of the mouse basal hypothalamus, while in mice ovariectomized for two weeks the GnRH levels did not differ significantly from controls. Parallel immunocytochemical analyses gave similar results. Tissue sections from specific regions of the median eminence in intact and ovariectomized mice were immunolabeled for GnRH with the peroxidase-antiperoxidase procedure of Sternberger. Differences in area and average unit density of the reaction product were measured with a Quantiment 720 image analyzer. At two weeks after ovariectomy, a majority of matched sections from intact and ovariectomized animals, taken in each sampled region of the median eminence, showed no visually apparent difference in GnRH immunoreactivity. Quantitative image analysis of these preparations revealed no statistically significant difference in either the area of neural tissue covered by deposit or the average unit density of this material in any given region. In contrast, in mice ovariectomized for either one or two months, immunolabeled GnRH was decreased throughout the cephalo-caudal extent of the median eminence. In each region, a reduction in reaction deposit was observed over most of the area in which GnRH is localized, especially in the medial aspects of this distribution. Image analysis of these preparations revealed a significant reduction of both specimen area immunolabeled for GnRH and the average unit density of reaction product in each region of the median eminence. While the decrease in density was uniform throughout the median eminence, the greatest reduction in area covered by deposit occurred in the infundibular region. The demonstration of a decrease in basal hypothalamic GnRH in the chronically ovariectomized mouse suggests that GnRH neurosecretion is increased in response to the absence of the ovaries and supports the premise that tonic secretion of this neurohormone, in the mouse, may be subject to regulation by the gonads. At two weeks after ovariectomy, it is possible that changes in GnRH secretion occur without a concomitant alteration in hypothalamic hormone content. In addition, the quantitative immunocytochemical data suggest that GnRH neurons responsive to ovariectomy terminate throughout the rostro-caudal extent of the mouse median eminence, especially in the medial sites of localization. A proportionally greater number of these endings appear to be present within the infundibular region of the median eminence, relative to the other regions sampled.