Interplay between stress response genes associated with attention‐deficit hyperactivity disorder and brain volume

The glucocorticoid receptor plays a pivotal role in the brain's response to stress; a haplotype of functional polymorphisms in the NR3C1 gene encoding this receptor has been associated with attention‐deficit hyperactivity disorder (ADHD). The serotonin transporter (5‐HTT) gene polymorphism 5‐HTTLPR is known to influence the relation between stress exposure and ADHD severity, which may be partly because of its reported effects on glucocorticoid levels. We therefore investigated if NR3C1 moderates the relation of stress exposure with ADHD severity and brain structure, and the potential role of 5‐HTTLPR. Neuroimaging, genetic and stress exposure questionnaire data were available for 539 adolescents and young adults participating in the multicenter ADHD cohort study NeuroIMAGE (average age: 17.2 years). We estimated the effects of genetic variation in NR3C1 and 5‐HTT, stress exposure and their interactions on ADHD symptom count and gray matter volume. We found that individuals carrying the ADHD risk haplotype of NR3C1 showed significantly more positive relation between stress exposure and ADHD severity than non‐carriers. This gene–environment interaction was significantly stronger for 5‐HTTLPR L‐allele homozygotes than for S‐allele carriers. These two‐ and three‐way interactions were reflected in the gray matter volume of the cerebellum, parahippocampal gyrus, intracalcarine cortex and angular gyrus. Our findings illustrate how genetic variation in the stress response pathway may influence the effects of stress exposure on ADHD severity and brain structure. The reported interplay between NR3C1 and 5‐HTT may further explain some of the heterogeneity between studies regarding the role of these genes and hypothalamic–pituitary–adrenal axis activity in ADHD.     

刺激大鼠中脑导水管周围灰质诱发的脊髓背根电位及其传出途径的分析

电刺激大鼠中脑导水管周围灰质(PAG),在腰5(L_5)背根可记录到—稳定的负性背根电位(DRP),简称 PAG-DRP。PAG-DRP 具有空间和时间总和性质,沿背根作电紧张性扩布,且能被 GABA 能拮抗剂印防己毒素(Picrotoxin)所抑制。电解损毁中缝大核(NRM)对刺激背侧 PAG 诱发的 PAG-DRP 无明显影响,而可使刺激腹侧 PAG 诱发的 PAG-DRP电位幅值降低40%左右。结果表明,PAG 下行抑制作用中有突触前抑制参与;NRM 参与腹侧 PAG-DRP 的产生,背侧 PAG-DRP 则可能由 NRM 以外的其他核团中继。     

猫下丘脑腹内侧核与旁中央中脑区神经联系的电生理学观察

用电脉冲刺激戊巴比妥钠麻醉和三碘季铵酚麻痹的31只猫的下丘脑腹內侧核(HVM),记录旁中央中脑区的平均诱发电位(AEP)。HVM 刺激点46个,旁中央中脑区引导点1046个,其中阳性反应点103个,快波反应151个,大多数为“峰电位式”快波。用逐渐增强的电脉冲刺激 HVM 可得等级式和“全或无”式两类反应。用不同频率的双刺激(PST)测试了111个 AEP 的反应性,其中有38个能跟随500Hz 以上的 PST,27个能跟随200—499Hz 的PST,46个仅能跟随199Hz 以下的 PST。给予短串刺激时,得到跟随500Hz 以上及500Hz以下的两类反应。这些结果提示 HVM 与旁中央中脑区之间既有经突触的投射又有直接的纤维连接。据计算,这些直接纤维的传导速度约为0.29—4.1m/s,相当于细有髓鞘和无髓鞘两类纤维。另外还描述了可能属于中枢轴突的某些传导特性。反映直接纤维连接的 HVM 刺激点与中脑中央灰质反应点的位置关系提示两者之间可能具有背腹侧方向的对应关系。     

电针对青霉素所致大脑皮层痫样放电的抑制作用

青霉素置于大鼠大脑皮层感觉运动区所诱发的皮层痫样放电,可被一些“穴位”电针所抑制,表现为放电频率、振幅及时程之减少。青霉素使逆向刺激锥体束所产生的感觉运动皮层表面负波减小,而电针则促进其恢复。因此电针制痫与它促进回返抑制的恢复有关。将微量纳洛酮注入腹腔、脑室及中脑导水管周围灰质、伏隔核均能翻转电针的作用。将纳洛酮直接注入放置青霉素处的皮层感觉运动区,针效亦被阻断。这表明电针制痛中有内啡肽参与。电刺激中脑导水管周围灰质及伏隔核有显著制痫作用。     

Phocanema decipiens: Growth,reproduction, and survival in seals

Captive harbor seals (Phoca vitulina) and gray seals (Halichoerus grypus) were fed infective larvae of Phocanema decipiens, an anisakine nematode from the flesh of Atlantic cod (Gadus morhus). Ova of P. decipiens were first detected in the feces of harbor seals 21(17–30) days after exposure; the patency period was 15 to 45 days. In gray seals, the prepatent period was 19(16–23) days; patency 20–60 days. By the sixth week of infection in harbor seals, mean body lengths of adult females and males of P. decipiens were 60.8(40.8–76.2) and 54.3(45.5–60.8) mm, respectively; mean fecundity of female nematodes was 156,000 ova. In infections of similar duration in gray seals, females and males of P. decipiens were 82.1(69.7–104.3) and 64.4(53.8–72.7) mm in length, respectively; mean fecundity of females was 366,000 ova. In sensitizing infections in harbor seals, 28% of P. decipiens survived to early patency (Days 25–30) while only 9% of the nematodes survived to midpatency (Days 35–45). In sensitizing infections in gray seals, 56% of P. decipiens survived to early patency (Days 20–30) and 48% survived to midpatency (Days 35–50). Seals with existing or recent P. decipiens infections resisted reinfection; <50% of the nematodes in challenge infections in gray seals survived to Day 3 and <10% survived to patency. Growth of the nematodes, however, was not retarded in the challenge infections and resistence to reinfection subsided when seals were maintained anisakinefree for 2–6 months after loss of prior natural or experimental infections. Natural anisakine infections were surveyed in 16 harbor and 53 gray seals from the Nova Scotia mainland. The mean incidence of P. decipiens was 62(5–177) in harbor seals and 577(11–1694) in gray seals; incidence varied seasonally and with age of host. Adult females of P. decipiens from harbor seals were 64.0(49.2–79.8) mm in length and contained 1.68(0.87–2.73) × 10⁵ ova; females from gray seals were 78.3(62.3–92.1) mm in length and contained 2.39(0.69–4.39) × 10⁵ ova.     

Hypothermia elicited by the intracerebral microinjection of neurotensin

Changes in rectal temperature were measured after the intracerebral microinjection of neurotensin (2.5 μg/0.5 μl) at 135 sites in the rat. At 63 of the 135 microinjection sites, the tridecapeptide produced a rapid onset of hypothermia ranging in magnitude from 0.8 to 2.3°C below the baseline rectal temperature. The drop in rectal temperature persisted for 2–4 hours following the microinjection. The greatest concentrations of neurotensin-sensitive sites were found in the medial preoptic region of the hypothalamus and in the periaqueductal gray area, both of which contain relatively large amounts of endogenous neurotensin. Other active sites were found in the ventral thalamus, the dorsomedial hypothalamus, and in the diagonal band of Broca. At no injection site did neurotensin evoke an increase in rectal temperature. These data support the proposition that neurotensin may act endogenously to mediate heat-loss mechanisms in the rat. The data provide further evidence indicating a potent neuromodulatory role for neurotensin.     

A METHOD TO COLLECT AND PROCESS SKIN BIOPSIES FOR CELL CULTURE FROM FREE-RANGING GRAY WHALES (ESCHRICHTIUS ROBUSTUS)

For researchers studying mysticere whales few methods for determining gender or for collecting biochemical and genetic information from unrestrained animals are available. The objective of this study was to develop a reliable method for collecting viable tissue samples for establishing continuous cell cultures from skin biopsies of free-ranging whales. A method to collect and process these samples is presented. Six of seven skin biopsies from gray whales were established in cell culture. Our results suggest that the viability of the samples is improved by (1) sterile processing in the field, (2) minimizing the time between collection and delivery to the cell culture facility, (3) reducing the concentration of antifungal agent, and (4) placing tissue explants under a coverslip. While the results reported in this paper are based on a small sample size, we believe that if the procedures are followed, they will increase the probability of successfully culturing cetacean tissue. Established cell lines can supply replenishable material from identified whales still living in the wild. These cultures can then be used for determination of sex from karyotypes, and for assessing genetic relationships of cetaceans from inherited protein, chromosomal and DNA polymorphisms. These much needed analytical tools can be used to determine familial and populational relationships, leading to a better understanding of mating systems, stock identification and effective population sizes of wild cetaceans.     

中央灰质内注射去甲肾上腺素对刺激兔下丘脑诱发室性期前收缩的影响

家兔用乌拉坦(700mg/kg)和氯醛糖(35mg/kg)静脉麻醉,用三碘季铵酚制动,在人工呼吸下进行实验。用电刺激下丘脑近中线区的方法诱发室性期前收缩(HVE)。中脑中央灰质(CG)内微量注射去甲肾上腺素(NA,4μg/2μl)对 HVE 有易化效应,使 HVE 次数增多。微量注射β-肾上腺素能受体阻断剂心得宁(2μg/2μl)使 HVE 次数明显减少,而α-肾上腺素能受体阻断剂酚妥拉明(2μg/2μl)则对 HVE 次数无明显减少作用。事先在 CG 内微量注射心得宁也可阻断 CG 内注入 NA 对 HVE 的易化效应。在 CG 内注射 NA 后,再在 CG 内微量注射吗啡,后者对 HVE 的抑制效应仍然存在。但在 GG 内注射 NA 可减少或消除刺激腓深神经对 HVE 的抑制作用。上述结果提示,在 CG 内β-受体的激活可增加 HVE 次数。刺激腓深神经对 HVE 的抑制作用可能部份是通过内源性吗啡样物质抑制 CG 内 NA 的释放而实现的。     

脊髓中的甲啡肽和强啡肽参与吗啡下行镇痛

在大鼠中脑导水管周围灰质(PAG)注射吗啡10μg 可引起明显的镇痛作用,并有部位特异性。在脊髓蛛网膜下腔注射抗甲啡肽 IgG20μg 或抗强啡肽 IgG20μg,均能大部分对抗 PAG内注射吗啡引起的镇痛作用,这提示甲啡肽和强啡肽参与自 PAG 到脊随的下行抑制作用。本实验利用蛋白质 A-琼脂糖 CL-4B 亲和层析柱从血清中纯化 IgG,这种方法简单,提纯速度快,且 IgG 纯度高,为中枢微量注射抗体研究神经肽的生理功能提供了方便。