Reversal of hyperalgesia by transplantation in lateral hypothalamic lesioned rats

Lateral hypothalamus (LHA) plays a very important role in the modulation of nociceptive behaviour. The stimulation of LHA is known to produce analgesia of both tonic and phasic pain. The present study reports hyperalgesia induced by lateral hypothalamic lesions and the effect of fetal (gestation day 16) hypothalamic transplant on the nociceptive response to phasic thermal noxious stimulation [tail flick latency (TFL)] in LHA lesioned rats. The TFL decreased significantly (12.91 +/- 3.91 sec to 10.51+/- 1.23 sec) following LHA lesion. However, after transplantation, the TFL did not change. This is the first report of a hypothalamic transplant inducing recovery of a nociceptive response.     

Amygdalar tissue transplants improve recovery of nociceptive behaviour in rats

PURPOSE: Amygdala plays a very important role in the mediation of emotional and affective components of pain. Bilateral amygdalectomy increases the threshold for vocalization, a measure of emotional reactivity, without any change in the tail flick latency. The present work was designed to study the recovery of emotional nociceptive behaviour following neural tissue transplantation in lesioned rats. METHODS: In a group of adult wistar rats lesions of the central nucleus of amygdala (CeA) were produced electrolytically. In a separate group of rat's amygdalar tissue was transplanted at the lesioned site 2 days thereafter. The vocalization tests, viz. simple vocalization (SV) and vocalization after discharge (VA), which were used to study the emotional nociceptive behaviour were carried out both before and after pro-ducing lesion/transplant. RESULTS: Bilateral CeA lesions increased the thresholds for SV and VA significantly (p < 0.001), indicating analgesia. Following amygdalar tissue transplantation a significant decrease (p < 0.05) in the thresholds were observed when compared with the lesioned group, although when compared with the basal data it showed an increase (p<0.05). CONCLUSIONS: The results indicate a partial recovery of the vocalization response following amygdalar tissue transplantation.     

Recovery of preoptic-anterior hypothalamic functions after transplantation

Kainic acid lesion of the medial preoptic area resulted in the impairment of the thermoregulation and disruption of the reproductive cycle in female rats. The preoptic-anterior hypothalamic area from fetal donor was transplanted into the lesioned area ofthe host. After transplantation, these animals showed signs of estrus cyclicity and a recovery in basal temperature towards normal. They also showed a reduced shift of body temperature on exposure to a hot environment. Thus the female rats, which received transplantation, showed some degree of recovery of functions which were disrupted by kainic acid lesions.     

Activation of the satiety center by auricular acupuncture point stimulation

Stimulation of the rat inner auricular regions that correspond to the human pylorus, lung, trachea, stomach, esophagus, endocrine, and heart acupuncture points evoked potentials in the hypothalamic ventromedial nucleus (HVM), the satiety center. Needle implantation into any of these points reduced the body weight to its initial 290 g after the rat had gained about 410 g in 20 days, and significantly reduced initial 450-g body weights (p less than 0.01, Student's t test) in 14 days. Stimulation of other acupuncture points did not evoke HVM potentials and did not reduce body weight. After the HVM was lesioned, body weight increased and acupuncture point needling had no effect on body weight. Needling of the auricular acupuncture points evoked no potentials in the lateral hypothalamus (LHA), the feeding center, and had almost no influence on weight reduction induced by LHA lesion.     

Differential expression of neurotrophin and neurotrophin receptor mRNAs in and adjacent to fetal midbrain grafts implanted into the dopamine-denervated striatum

This study examined the expression of neurotrophins and neurotrophin receptors in the lesion/transplanted striatum at four different time points after transplantation. The ventral mesencephalic region was dissected from a single rat fetus at embryonic day 14 (E14) and implanted into the denervated striatum of rats with unilateral 6-hydroxydopamine lesions. Transplanted rats were killed at 1, 2, 3, or 4 weeks after transplantation surgery and the brains subsequently prepared for semiquantitative in situ hybridization analysis of neurotrophin and neurotrophin trk receptors. Hybridization of cRNA probes for trkB or trkC showed a time-dependent reduction within the transplant during the first 4 weeks after transplantation; hybridization of brain-derived neurotrophic factor or tyrosine hydroxylase mRNA probes within the transplant did not change significantly during the same posttransplantation period. Hybridization of the trkB mRNA probe in host striatum adjacent to the transplant was significantly higher than probe hybridization in the corresponding region of the intact striatum during the first 2 weeks after transplantation, but by the 3rd and 4th week, probe hybridization in the denervated/transplanted and intact striatum were the same. Lesioned animals without transplants maintained higher trkB mRNA probe hybridization in the denervated striatum than in the intact striatum at the same postlesion time points suggesting that lesioned/transplanted animals show a normalization of trkB mRNA probe hybridization. Hybridization of the trkC mRNA probe in the lesioned/transplanted striatum was significantly lower than that observed in the intact striatum 4 weeks after transplantation; however, at this same time point we observed a similar reduction of trkC probed hybridization in lesioned animals without transplants. The results of the study show dynamic neurotrophic activity occurring within the transplant and host tissue during the first month of transplant development.     

Intracerebroventricular Transplantation of Embryonic Neuronal Tissue from Inflammatory Resistant into Inflammatory Susceptible Rats Suppresses Specific Components of Inflammation

To more directly define the role of central nervous system factors in susceptibility to peripheral inflammatory disease, we examined the effect of intracerebroventricular transplantation of neuronal tissue from inflammatory resistant into inflammatory susceptible rats on subcutaneous carrageenan-induced inflammation (a measure of innate immunity), and on the relative percentage of naive and memory T helper cells in peripheral blood (a measure of the anamnestic immune response). Female inflammatory disease susceptible Lewis (LEW/N) rats transplanted with hypothalamic tissue from inflammatory resistant Fischer (F344/N) rats exhibited >85% decrease in carrageenan inflammation compared to naive LEW/N rats, LEW/N rats transplanted with F344/N spinal cord, or sham-operated animals. LEW/N rats transplanted with LEW/N hypothalamic tissue exhibited >50% decrease in carrageenan inflammation. In contrast, intracerebroventricular transplantation of neuronal tissue did not affect the characteristically twofold higher percentage of naive versus memory T helper cells in LEW/N rats, suggesting that the central nervous system (CNS) and hypothalamus play a greater role in the innate inflammatory response than in the acquired immune processes. Grafted tissue survived well and did not show extensive gliosis or inflammation. Compared to naive LEW/N rats, LEW/N rats transplanted with F344/N or LEW/N hypothalamic tissue expressed significantly greater hypothalamic corticotropin releasing hormone mRNA. LEW/N rats transplanted with F344/N hypothalamic tissue also showed significant increases in plasma corticosterone responses to lipopolysaccharide. These data indicate that intracerebroventricular transplantation of fetal hypothalamic tissue from inflammatory resistant into inflammatory susceptible rats suppresses peripheral inflammation partially through hypothalamic factors. These findings have implications for understanding the contribution of specific neuronal tissue in regulation of components of the immune/inflammatory response and in susceptibility to inflammatory disease. Furthermore, this model could be used in the development of potential new treatments for inflammatory/autoimmune diseases aimed specifically at sites within the CNS.     

Effect of lateral hypothalamic area lesions on serotonin-containing epithelial cells in rat duodenum

The aphagic effect of lateral hypothalamic area (LHA) lesions induced by kainic acid injection on serotonin (5HT)-containing epithelial cells in rat duodenum was studied in comparison with food deprived rats. The densities of 5HT cells were evaluated by quantitative morphometry using fluorescent histochemical longitudinal sections. The levels of 5HT/mg tissue were determined by HPLC on acid extracts of isolated villous epithelium. Body and duodenal weight losses, as well as duodenal mucosal atrophy, were comparable in the two groups of starved animals, i.e., one is food deprived and the other is lesioned animals. The numbers of 5HT cells in these starved groups were similar to those in controls so that the densities were increased. The 5HT/mg tissue in food-deprived animals was similar to that in controls but significantly higher in the LHA-lesioned group. These results are interpreted in terms of a possible central regulatory mechanism that is intact in the food-deprived animals but destroyed in the lesioned animals.     

Transplanted adult neural progenitor cells survive, differentiate and reduce motor function impairment in a rodent model of Huntington's disease

The present study investigated the ability for adult rat neural progenitor cells to survive transplantation, structurally repopulate the striatum and improve motor function in the quinolinic acid (QA) lesion rat model of Huntington's disease. Neural progenitor cells were isolated from the subventricular zone of adult Wistar rats, propagated in culture and labeled with BrdU (50 microM). Fourteen days following QA lesioning, one group of rats (n = 12) received a unilateral injection of adult neural progenitor cells ( approximately 180,000 cells total) in the lesioned striatum, while a second group of rats (n = 10) received a unilateral injection of vehicle only (sham transplant). At the time of transplantation adult neural progenitor cells were phenotypically immature, as demonstrated by SOX2 immunocytochemistry. Eight weeks following transplantation, approximately 12% of BrdU-labeled cells had survived and migrated extensively throughout the lesioned striatum. Double-label immunocytochemical analysis demonstrated that transplanted BrdU-labeled progenitor cells differentiated into either astrocytes, as visualized by GFAP immunocytochemistry, or mature neurons, demonstrated with NeuN. A proportion of BrdU-labeled cells also expressed DARPP-32 and GAD67, specific markers for striatal medium spiny projection neurons and interneurons. Rats transplanted with adult neural progenitor cells also demonstrated a significant reduction in motor function impairment as determined by apomorphine-induced rotational asymmetry and spontaneous exploratory forelimb use when compared to sham transplanted animals. These results demonstrate that adult neural progenitor cells survive transplantation, undergo neuronal differentiation with a proportion of newly generated cells expressing markers characteristic of striatal neurons and reduce functional impairment in the QA lesion model of Huntington's disease.     

Partial recovery of gustatory function after neural tissue transplantation to the lesioned gustatory neocortex

The ability of homotypic cortical tissue grafts to induce recovery of function after a gustatory neocortex (GN) lesion was studied using the conditioned taste aversion (CTA) paradigm. On acquisition day, 26 GN-lesioned and 8 sham-lesioned rats were presented with a saccharin solution, followed by an injection of the illness-inducing agent lithium chloride (LiCl). On the test day, 2 days later, saccharin was presented again. The GN-lesioned rats showed significantly less aversion to saccharin on the test day, indicating that the lesion impaired their ability to form taste-illness association. Nine of the lesioned rats were then bilaterally transplanted with fetal GN tissue. Nine weeks after the transplantation, the rats were presented with a LiCl solution, which served as both a tastant and an illness-inducing agent. An NaCl solution, which tasted very similar to the LiCl solution, was used to test the CTA to salt 3 days later. The nontransplanted rats consumed significantly more LiCl than the transplanted and sham-operated rats on the acquisition day, but both transplanted and nontransplanted rats consumed more NaCl than sham-operated rats on the test day. Nissl and Golgi stainings showed numerous somata and extensive arborization of neurons within the grafts. The results indicate that fetal GN grafts can restore the ability to integrate gustatory and visceral inputs but not to form long-lasting taste-illness associations.     

Partial recovery of gustatory function after neurol tissue transplantation to the lesioned gustatory neocortex

The ability of homotypic cortical tissue grafts to induce recovery of function after a gustatory neocortex (GN) lesion was studied using the conditioned taste aversion (CTA) paradigm. On acquisition day, 26 GN-lesioned and 8 sham-lesioned rats were presented with a saccharin solution, followed by an injection of the illness-inducing agent lithium chloride (LiCl). On the test day, 2 days later, saccharin was presented again. The GN-lesioned rats showed significantly less aversion to saccharin on the test day, indicating that the lesion impaired their ability to form taste-illness association. Nine of the lesioned rats were then bilaterally transplanted with fetal GN tissue. Nine weeks after the transplantation, the rats were presented with a LiCl solution, which served as both a tastant and an illness-inducing agent. An NaCl solution, which tasted very similar to the LiCl solution, was used to test the CTA to salt 3 days later. The nontransplanted rats consumed significantly more LiCl than the transplanted and sham-operated rats on the acquisition day, but both transplanted and nontransplanted rats consumed more NaCl than sham-operated rats on the test day. Nissl and Golgi stainings showed numerous somata and extensive arborization of neurons within the grafts. The results indicate that fetal GN grafts can restore the ability to integrate gustatory and visceral inputs but not to form long-lasting taste-illness associations.     

Cultured human foetal cerebral cortex, transfected with tyrosine hydroxylase cDNA, as a source of neural transplant material

Summary. Obtaining an adequate supply of foetal dopaminergic tissue to treat Parkinson's disease by neural transplantation can be difficult. In this study primary cultures of human foetal cerebral cortex cells were transfected, using cationic lipids, with a eukaryotic expression vector (pCIneo-THI) containing the cDNA for human tyrosine hydroxylase isoform I (TH). Cortical cells from human (10–14 week) foetuses were cultured for 11 days in vitro and transfected twice with pCIneo-TH1 during this time. The double transfection process resulted in 3–4% of the cells becoming TH positive. When grafted into the striatum of 6-OHDA lesioned rats the transfected foetal cerebral cortex cells reduced amphetamine-induced circling behaviour by 75%, while grafts of untransfected cells had no significant effect on turning. TH transfected foetal cerebral cortex cells may therefore be a useful alternative supply of tissue for use in neural transplants to treat Parkinson's disease. Received April 4, 2000; accepted February 5, 2001     

Feeding pattern in obese Zucker rats after dopaminergic and serotonergic LHA grafts.

To study the role of the lateral hypothalamic area (LHA) dopamine and serotonin in the regulation of feeding pattern during obesity, embryonic dopaminergic and serotonergic neurons from mesencephalon and rombencephalon of lean rats were grafted into the LHA of adult obese Zucker rats. Compared to the pregrafting period, a smaller increase in meal size occurred in both serotonin-grafted (9%) and dopamine-grafted (31%) rats vs control rats (51%). There was also a smaller decrease in meal number in both serotonin-grafted (3%) and dopamine-grafted (13%) rats vs control rats (28%). Although the changes in feeding pattern resulted in a decrease in total food intake in serotonin-grafted rats (5%) vs control rats, no differences in body weight gain were observed in grafted vs control rats for the duration of the study. Since adult obese Zucker rats are known to have an increased meal size and decreased meal number relative to lean rats, the data indicate the involvement of LHA dopamine and serotonin in the regulation of feeding pattern during obesity.     

Behavioural recovery following transplantation of substantia nigra in rats subjected to 6-OHDA lesions of the nigrostriatal pathway. I. Unilateral lesions

The ability of embryonic substantia nigra transplants to compensate for behavioural deficits induced by unilateral destruction of the nigrostriatal dopamine pathway has been investigated in adult rats. Six days following unilateral 6-OHDA lesions of the nigrostriatal pathway, the adequacy of the lesion was assessed by measurement of the intensity of ipsilateral amphetamine-induced rotation. All rats then received surgical cavities in the cortex overlying the head of the caudate-putamen on the lesioned side. In 51 rats, transplants of embryonic substantia nigra were placed on the dorsal surface of the caudate-putamen, and the remaining 19 rats served as unilateral lesioned controls. Behavioural testing was conducted approximately 3 months after transplantation: (a) the transplant animals alone showed a marked reduction in ipsilateral rotation induced by 5 mg/kg amphetamine (‘compensation’); (b) although both transplanted and control rats expressed equal contralateral rotation at a dose of 0.25 mg/kg apomorphine, the transplant animals alone showed a marked reduction in rotation at a lower dose of 0.05 mg/kg; (c) the transplanted rats showed less asymmetry in spontaneous rotational behaviour than controls, and the asymmetry was further reduced by mild tailpinch; (d) when tested for spontaneous choice behaviour in a T-maze, control rats showed 97% selection of the arm ipsilateral to the 6-OHDA lesion, whereas the transplanted rats that were well compensated on the amphetamine rotation test turned to the contralateral side on 30–40% of choices; (e) no transplant-induced changes were found in contralateral sensory inattention on a sensorimotor test battery, whether tested spontaneously or under mild tailpinch-induced activation. The results support the conclusion that dopaminergic reinnervation of the dorsal neostriatum is capable of inducing functional recovery in many, but not all, behavioural tests which involve side choice or bias, not only after pharmacological activation but also in the spontaneous behaving animal.     

Neurobehavioural, neurochemical and electrophysiological studies in 6-hydroxydopamine lesioned and neural transplanted rats

Unilateral injection of 6-hydroxydopamine (6-OHDA) into the caudate nucleus of rat caused degeneration of dopaminergic terminals, evidenced by significant (P⊂0.05) elevation of spontaneous and drug-induced motor behaviour, enhanced DA receptor binding and significant increase in the neuronal firing rate of caudate neurons, suggesting supersensitivity of dopaminergic receptors. Eight weeks following the transplantation of embryonic cell suspensions from caudate at the lesioned site, a significant restoration of the enhanced ³H spiperone binding and neuronal activity of caudate neurons was observed in comparison with lesioned rats.These results clearly demonstrate that transplanted embryonic neuronal tissue at the lesioned site is capable of restoring the neuronal deficits caused by 6-OHDA as evidenced by significant amelioration in neurochemical, behavioral and electrophysiological alterations.     

Glucocorticoids are required for meal-induced changes in the expression of hypothalamic neuropeptides

Glucocorticoid deficiency is associated with a decrease of food intake. Orexigenic peptides, neuropeptide Y (NPY) and agouti related protein (AgRP), and the anorexigenic peptide proopiomelanocortin (POMC), expressed in the arcuate nucleus of the hypothalamus (ARC), are regulated by meal-induced signals. Orexigenic neuropeptides, melanin-concentrating hormone (MCH) and orexin, expressed in the lateral hypothalamic area (LHA), also control food intake. Thus, the present study was designed to test the hypothesis that glucocorticoids are required for changes in the expression of hypothalamic neuropeptides induced by feeding. Male Wistar rats (230-280 g) were subjected to ADX or sham surgery. ADX animals received 0.9% NaCl in the drinking water, and half of them received corticosterone in the drinking water (B: 25 mg/L, ADX+B). Six days after surgery, animals were fasted for 16 h and they were decapitated before or 2 h after refeeding for brain tissue and blood collections. Adrenalectomy decreased NPY/AgRP and POMC expression in the ARC in fasted and refed animals, respectively. Refeeding decreased NPY/AgRP and increased POMC mRNA expression in the ARC of sham and ADX+B groups, with no effects in ADX animals. The expression of MCH and orexin mRNA expression in the LHA was increased in ADX and ADX+B groups in fasted condition, however there was no effect of refeeding on the expression of MCH and orexin in the LHA in the three experimental groups. Refeeding increased plasma leptin and insulin levels in sham and ADX+B animals, with no changes in leptin concentrations in ADX group, and insulin response to feeding was lower in this group. Taken together, these data demonstrated that circulating glucocorticoids are required for meal-induced changes in NPY, AgRP and POMC mRNA expression in the ARC. The lower leptin and insulin responses to feeding may contribute to the altered hypothalamic neuropeptide expression after adrenalectomy.     

Direct effects of 3,4-dihydroxybutanoic acid gamma-lactone and 2,4,5-trihydroxypentanoic acid gamma-lactone on lateral and ventromedial hypothalamic neurons

The mechanism of central actions of endogenous sugar acids, 3,4-dihydroxybutanoic acid gamma-lactone (3,4-DB) and 2,4,5-trihydroxypentanoic acid gamma-lactone (2,4,5-TP) which have been newly identified as satiety and hunger substances respectively, was investigated. Intracellular recordings were made from neurons in the lateral hypothalamic area (LHA) of rats and ventromedial hypothalamic nucleus (VMH) of guinea pigs, brain areas referred to as feeding and satiety centers, respectively. The LHA neurons hyperpolarized by 3,4-DB show no change in membrane input resistance while the depolarized VMH neurons are associated with an increase in membrane resistance. The mechanisms related to the action of 3,4-DB on these hypothalamic neurons are similar to those in case of glucose on the glucose-sensitive neuron in the LHA and the glucoreceptor neuron in the VMH. 2,4,5-TP depolarized LHA neurons but hyperpolarized VMH neurons with a decrease in the membrane resistance. Our findings indicate that 3,4-DB and 2,4,5-TP have reciprocal effects on each LHA and VMH neurons, with regard to neuronal excitability.     

Noninvasive method of immortalized neural stem-like cell transplantation in an experimental model of Huntington's disease

A loss of neostriatal neurons is a characteristic of Huntington's disease (HD), and neural tissue transplantation has been performed directly into the striatum. Since the neural stem cells have ability to migrate into the lesion site, we administered immortalized neural stem-like cells (NSC) into the ventricle or via a tail vein following unilateral intrastriatal quinolinic acid lesioning in Sprague-Dawley rats. To identify transplanted NSC, cells were encoded with lac Z and beta-galactosidase histochemistry was performed. lac Z+ cells were detected in the lesioned striatum but tissue damage or tumor formation was not observed. This study shows that NSC migrate into the striatum, either from ventricle or from the systemic circulation, providing less invasive routes for stem cell application in HD.     

神经营养因子NGF、BDNF对AD模型鼠海马移植的影响

目的:研究神经营养因子NGF、BDNF对AD模型鼠海马移植后行为和形态学变化。方法:24.只AD模型鼠随机分成4组:单纯胚基底前脑细胞悬液移植组(ST组)、含NGF或BDNF胚基底前脑细胞悬液移植组(NGF组)、(BD-NF组)和模型对照组(M组),移植后3月进行行为测试并比较移植区AchE细胞数和纤维密度,运用方差分析和SNK检验进行组间比较。结果:行为测试移植3组明显优于模型M组,含因子组又较ST组效果好(P<0.01),两因子组间差异无显著性(P>0.05);因子组存活细胞数均高于ST组,NGF组细胞数多于BDNF组(P<0.05),纤维密度两组相似(P>0.05)。结论:海马内存活胆碱能神经元能代偿受损胆碱能神经元的功能,改善动物的学习记忆功能;NGF、BDNF均能促进胆碱能神经元存活,增加AchE细胞数目和突起,但BDNF促进神经元突起延伸作用较好,而NGF则对神经元保护作用较强。     

Fibroblast Growth Factor Receptor-1 in the Lateral Hypothalamic Area Regulates Food Intake

Previous studies have shown that acidic and basic fibroblast growth factor (aFGF and bFGF) and certain fragments of the aFGF N-terminal suppress food intake in rats due to their inhibitory actions on the glucose-sensitive neurons in the lateral hypothalamic area (LHA). The present study was planned to determine the role of FGF receptor-1 (FGFR-1), which was found in the LHA neurons of rats, on feeding regulation. The structure–activity relationship of aFGF fragments in feeding suppression was also investigated. An injection of anti-FGFR-1 antibody (250 and 350 ng) into the bilateral LHA significantly increased food intake. Synthesized aFGF fragments were infused into the III ventricle to elucidate the structure–activity relationship on the inhibition of feeding. Although aFGF-(1–29) did not affect food intake, [Ser¹⁶]aFGF-(1–29) (400 ng) and [Glu¹⁶]aFGF-(1–29) (400 ng), in which the cysteine residue at position 16 of aFGF-(1–29) was replaced with structurally similar serine and glutamic acid, were observed to significantly inhibit food intake. These findings suggest that endogenous FGFR-1 in the LHA plays an important role in FGF-induced feeding suppression, while, in addition, the dissolving disulfide bond formation in aFGF fragments enhances their inhibitory effects on feeding.