Testicular histomorphometry and the proliferative and apoptotic activities of the seminiferous epithelium in Syrian hamster during spontaneous recrudescence after exposure to short photoperiod

Syrian hamsters are photoperiodic rodents in which reproduction, including testicular function, is stimulated by long photoperiod exposure and curtailed by exposure to a short photoperiod. The objectives of this study were to characterize the testis histomorphometrically and to determine the role of the proliferation and apoptosis phenomena in the recovery of the seminiferous epithelium during spontaneous recrudescence after exposure to short photoperiod. The study was performed using conventional light microscopy, proliferating cell nuclear antigen and terminal deoxynucleotidyl transferase (TdT)‐mediated dUTP in situ nick end labelling staining, image analysis software, and transmission electron microscopy in three recrudescence groups: initial recrudescence (IR), advanced recrudescence (AR) and total recrudescence (TR). The results morphometrically pointed to the gradual recovery of the testicular and tubular volumes, as well as of the seminiferous epithelium. Among the IR and AR groups, the increase in testicular and tubular volumes was accompanied by an increase in tubular diameter and length, with an increase in interstitial volume. From AR to TR, there was an increase in the tubular and total volumes, but, in this case, with a gradual increase in tubular diameter. Recovery of the seminiferous epithelium was accompanied by changes in apoptosis and proliferation activities. The first decreased halfway through the process, and the second remained higher than the control levels throughout the recrudescence stage. Alterations in the spermatozoa were ultrastructurally observed, which indicated that spermiogenesis was not yet completely normal. In conclusion, spontaneous testicular recrudescence in Syrian hamster comprises two histomorphometrical phases: the first related to an increase in tubular length and diameter and interstitial volume and the second depending principally on the gradual increase in tubular diameter. The restoration of the seminiferous epithelium is due to apoptosis reaching normal values in the AR group accompanied by higher proliferative activity than that observed in the Control group.     

Testicular dynamics in Syrian hamsters exposed to both short photoperiod and low ambient temperature

The object of this study was to determine the details of morphological dynamics of spermatogenesis in Syrian hamsters exposed to both short photoperiod and low ambient temperature. Eight-week-old male hamsters, kept in a long photoperiod (14 h L, 10 h D), were transferred to a short photoperiod (6 h L, 18 h D) and kept there for 13 weeks to induce testicular regression. Some hamsters were then transferred from the room at 23 degrees C to that at 5 degrees C (5 degrees C group). Remaining hamsters were continuously kept at 23 degrees C (23 degrees C group). Thereafter, the morphology was examined. As a result, it took only 8 weeks until spermatogenesis recovered in the 23 degrees C group. However, it was not until 20 weeks that spermatogenesis was recognized in the 5 degrees C group. As the regulation of seasonal testicular activity is characterized by coordinated shifts in the relationships among mitosis, meiosis, and apoptosis, the changes in the proliferative and apoptotic activities were examined. Although no significant difference in proliferative activity of spermatogonia between the 5 degrees C and the 23 degrees C groups was confirmed, a notable increase in the rate of apoptosis was observed in the 5 degrees C group. Furthermore, this increase was more salient during the hibernation period. These findings suggest that both cold ambient temperature and hibernation caused the delay of testicular recrudescence and this delay arose from the increase of apoptotic activity but not the change in proliferative activity in spermatogonia in the 5 degrees C group.     

Arteriolar nephrosclerosis in the Syrian hamster

Syrian hamsters developed spontaneous renal lesions that resembled those of arteriolar nephrosclerosis in man, and differed from other spontaneously occurring or virus-induced renal diseases in other rodent species. Morphologic changes were mainly degenerative with little cellular exudation and were associated with histologic changes in the intrarenal vasculature. The renal lesions were progressive, often fatal and sometimes were complicated by glomerular amyloidosis with the nephrotic syndrome and uremia. Endstage kidneys often had fibrinoid necrosis of intrarenal arterioles and thus resembled lesions characteristic of the malignant phase of human essential hypertension. Fibrinoid necrosis of small arterioles was common in the uterus, ovaries or testes of affected animals; it was less frequent in mesenteric or coronary vessels. Cardiac thrombosis, often involving the left atrium or left atrioventricular valves, also was common. Changes occurred earlier and often were more severe in females than in males. This disease was a major cause of morbidity and mortality and hampered life-span studies.     

Inhibin secretion in the golden hamster (Mesocricetus auratus) testis during active and inactive states of spermatogenesis induced by the restriction of photoperiod

Gonadal function in the male golden hamster (Mesocricetus auratus) was investigated during exposure to a short photoperiod condition. Within 3 weeks of exposure to the short photoperiod condition, FSH and testosterone in the plasma significantly decreased, and subsequently immunoreactive (ir)-inhibin significantly decreased. Testicular contents of ir-inhibin and testosterone, and pituitary contents of LH and FSH also significantly decreased by 3 weeks with regression of weight of testes, epididymis and seminal vesicles and sperm head count. Circulating LH varied but not significantly. Thereafter, all reproductive parameters and secretion of LH, FSH, ir-inhibin and testosterone gradually recovered after 17 weeks of exposure even though animals continued to be subjected to the short photoperiod condition. Plasma concentrations of inhibin B and inhibin pro-alphaC were detectable and were significantly decreased after 15 weeks of exposure to the short photoperiod, but their levels were still detectable. Immunopositive reaction of inhibin alpha and betaB subunits was found in Sertoli cells and Leydig cells in the regressed testes of animals subjected to short photoperiod as was also seen in animals before exposure to the short photoperiod. Although the spermatogenic cycle was suppressed like prepubertal animals, the present study showed that the testicular recovery, so-called refractoriness, is functionally different from the developing stage of immature animals, especially with regard to inhibin secretion. The present results showed that changes in FSH preceded changes in inhibin during the regression and recovery phases, indicating that FSH is a major regulatory factor of inhibin secretion in male golden hamsters. The present study also demonstrated that regressed testes still secrete a small amount of bioactive inhibin during exposure to a short-photoperiod condition.     

成年公猪睾丸及附睾中促黄体素受体mRNA表达的研究

试验采用FQ-PCR技术进行了成年公猪睾丸及附睾中促黄体素受体mRNA表达的研究,结果表明:促黄体素受体mRNA不仅在睾丸内表达,在附睾中也有表达;睾丸、附睾中促黄体素受体mRNA的表达量分别为(1.42±0.08)×109 copies/mL和(6.54±0.21)×109copies/mL,睾丸两组织间促黄体素受体mRNA表达量无明显差异(P>0.05).     

钼镉联合诱导对麻鸭肾功能及肾Bcl-2基因mRNA表达的影响

旨在观察钼镉联合诱导对麻鸭肝功能及肝Bcl-2基因表达的影响。选用360羽11日龄健康"江南2号"麻鸭随机分成6组,每组60只(公母各30羽)。以(NH4)6Mo7O24·4H2O和3CdSO4·8H2O分别作为钼源、镉源,在每kg基础日粮中添加Mo、Cd,各组的剂量分别为:对照组(Mo 0mg,Cd 0mg)、低钼组(Mo 15 mg,Cd 0 mg)、高钼组(Mo 100mg,Cd 0mg)、镉组(Mo 0mg,Cd 4mg)、低钼镉组(Mo 15mg,Cd 4mg)、高钼镉组(Mo 100mg,Cd 4mg)。试验期120d,在试验第30、60、90、120天,每组随机选10羽鸭(公母各5羽)采血并分离血清,检测ALT、AST的活性和TP、ALB、GLB、DBIL、IBIL的含量;在试验第60、120天取肝组织检测Bcl-2mRNA表达量。结果显示:随着不同剂量Mo、Cd的添加,ALT、AST的活性除低钼组外均显著高于对照组(P0.05);TP、ALB含量各试验组与对照组差异不显著(P0.05);GLB的含量在第30天各试验组与对照组差异不显著(P0.05),第60、90天低钼镉组、高钼镉组显著高于对照组(P0.01),第120天除低钼组外均显著高于对照组(P0.01);肝组织中Bcl-2mRNA表达量,第120天,除低钼组外均显著低于对照组(P0.01),并且这种变化趋势,钼镉联合组比钼镉单独组更加明显。钼、镉及其联合诱导会导致麻鸭肝功能损伤、肝Bcl-2mRNA表达量下调,钼镉呈协同作用。     

钼镉联合诱导对麻鸭肾功能及肾Bcl-2基因mRNA表达的影响

旨在研究钼镉联合诱导对麻鸭肾功能及肾脏中Bcl-2基因mRNA表达量的影响。选用360羽健康的11日龄"江南2号"麻鸭并随机分成6组,每组60只(公母各30羽)。(NH_4)_6Mo_7O_(24)·4H_2O和3CdSO_4·8H_2O分别作为本试验的钼源和镉源,在每千克基础日粮中添加不同剂量的Mo、Cd,各组的处理情况分别为:对照组(Mo 0 mg,Cd 0mg)、低钼组(Mo 15 mg,Cd 0 mg)、高钼组(Mo 100 mg,Cd 0 mg)、镉组(Mo 0 mg,Cd 4 mg)、低钼镉组(Mo 15mg,Cd 4mg)、高钼镉组(Mo 100mg,Cd 4mg)。试验期120d。在试验期第30、60、90、120天时采集血样分离血清用于检测血清中Cr、UA、BUN、Na~+、K~+、Cl-、Mg~(2+)的含量;于60、120d采集鸭肾组织检测Bcl-2 mRNA的表达量。结果显示,120d高钼镉组Cr和BUN显著高于对照组(P0.05);120d试验组血清中UA低于对照组但差异不显著(P0.05);120d试验组血清Na~+、Cl-显著低于对照组(P0.05);试验组血清K~+低于对照组但差异不显著(P0.05);90、120d高钼组和高钼镉组的血清Mg~(2+)显著低于对照组(P0.05);在60、120d,除低钼组外,其余试验组鸭肾中Bcl-2mRNA表达量均极显著低于对照组(P0.01),高钼镉组极显著高于其他组(P0.01);钼镉联合组变化比单独组更显著。此结果表明钼、镉及其联合诱导可导致麻鸭肾功能损伤、肾脏中Bcl-2 mRNA表达量下调,且钼镉呈协同作用。     

小花棘豆中毒对家兔睾丸细胞凋亡及Bcl-2、Bax表达的影响

将24只家兔随机分为对照组和试验Ⅰ、Ⅱ、Ⅲ组,试验组饲粮分别按照Ⅰ组15%(苦马豆素含量30mg/kg)、Ⅱ组30%(苦马豆素含量60mg/kg)、Ⅲ组45%(苦马豆素含量90mg/kg)的比例添加小花棘豆,对照组仅饲喂青干草,试验期70d;分别于攻毒后第14,35,70天每次每组随机采集2只家兔的睾丸,通过TUNEL法检测细胞凋亡,real-time PCR检测Bcl-2、Bax mRNA的表达,免疫组化检测Bcl-2、Bax蛋白的表达。结果显示:从第35天开始,试验Ⅱ组和Ⅲ组家兔睾丸细胞凋亡指数、Bcl-2和Bax mRNA的表达均与对照差异极显著(P<0.01),试验Ⅰ组家兔睾丸细胞凋亡指数、Bcl-2和Bax mRNA的表达均与对照差异显著(P<0.05);试验组家兔睾丸组织Bcl-2蛋白表达均明显低于对照,Bax蛋白表达均明显高于对照,其差异性随中毒时间的延长而变化。结果表明:小花棘豆中毒可导致家兔睾丸组织生精细胞凋亡,且与小花棘豆中毒呈现一定的时间-剂量效应,这种作用可能与Bcl-2和Bax基因表达有关。     

Effects of short photoperiod on energy intake,thermogenesis, and reproduction in desert hamsters (Phodopus roborovskii)

Desert hamsters (Phodopus roborovskii) are the least known species in the genus Phodopus with respect to ecology and physiology, and deserve scientific attention, particularly because of their small body size. Here, the responses of energy metabolism and reproductive function to short photoperiods in desert hamsters were investigated. Male and female desert hamsters were acclimated to either long day (LD) (L:D 16:8 h) or short day (SD) photoperiods (L:D 8:16 h) for three months, and then the females were transferred back to an LD photoperiod for a further five months, while at the end of the SD acclimation the males were killed and measurements were taken for serum leptin as well as molecular markers for thermogenesis. We found that like the other two species from the genus Phodopus, the desert hamsters under SD decreased body mass, increased adaptive thermogenesis as indicated by elevated mitochondrial protein content and uncoupling protein‐1 content in brown adipose tissue, and suppressed reproduction compared to those under LD. However, different from the other two species, desert hamsters did not show any differences in energy intake or serum leptin concentration between LD and SD. These data suggest that different species from the same genus respond in different ways to the environmental signals, and the desert adapted species are not as sensitive to change in photoperiod as the other two species.     

维生素E缺乏对雏鸡脾脏淋巴细胞Caspase-3活性及Caspase-3mRNA表达的影响

用低维生素E(VE)和添加不饱和脂肪酸的日粮饲喂雏鸡建立VE缺乏雏鸡模型,采用比色法和半定量RT-PCR法分别检测脾脏淋巴细胞Caspase-3活性及Caspase-3 mRNA的表达水平。结果显示,VE缺乏组雏鸡脾脏淋巴细胞Caspase-3活性和Caspase-3 mRNA丰度高于对照组,组间差异均极显著(P<0.01)。结果表明,Caspase-3活性及Caspase-3 mRNA丰度的改变,是VE缺乏雏鸡脾脏淋巴细胞凋亡的调控机制之一。     

丁酸钠对体外培养猪肠黏膜上皮细胞寡肽转运载体及钠氢交换载体mRNA表达的影响

为探讨丁酸钠对肠道寡肽转运载体(PepT1)及钠氢交换载体(NHE2、NHE3)mRNA表达丰度的调控,取刚出生未吮乳的仔猪小肠黏膜组织,建立猪小肠上皮细胞(IEC)分离及原代培养方法。分别用0,2,4,8 mmol/L的丁酸钠处理体外培养的猪小肠黏膜上皮细胞96 h后,提取细胞总RNA,以18S rRNA为内标基因,用实时荧光定量RT-PCR法(SYBR GreenⅠ试剂盒)检测PepT1、NHE2及NHE3 mRNA在不同浓度丁酸钠处理细胞中的表达丰度。结果表明,体外培养的猪小肠黏膜上皮细胞用丁酸钠处理96 h后,PepT1和NHE2 mRNA的表达丰度均显著增加(P0.05),且呈现剂量依赖效应;NHE3 mRNA表达丰度的剂量效应不明显,只有在高浓度丁酸钠(8 mmol/L)组时才显著高于对照组(P0.05)。     

Embryonic and postnatal differentiation of olfactory epithelium and vomeronasal organ in the Syrian hamster

The details of the embryonic and postnatal differentiation of the olfactory epithelium (OE) and vomeronasal organ (VNO) were examined by light and electron microscopy in the Syrian hamster. At 10 days of gestation, the nasal placode is invaginated to form the olfactory pit on either side at the rostral end of the embryo. Abundant mitotic figures are observed near the free surface of the epithelium lining the olfactory pit. At 11 days of gestation, the mass of the epithelium lining a recess is separated from the medial wall of the olfactory pit to form the VNO. At 13 days of gestation, mitotic figures become observable in the basal layer of the vomeronasal sensory epithelium (VSE) in addition to the superficial to middle layers, while in the OE mitotic figures are observed mainly in the middle to basal layer. At 1 day after birth, the OE is almost complete in differentiation. On the other hand, the VSE differentiate slowly to retain some immature properties even at 10 days after birth. These findings suggest that the olfactory function seems to be solely ascribed to the OE for a while after birth. The significance of mitotic figures are discussed in the course of development with special reference to the origin of the nasal placode from the central nervous system.     

不同光照时长对鸡睾丸褪黑素受体及合成酶表达的影响

旨在研究不同光照时长对鸡睾丸组织中褪黑素受体及合成酶表达规律的影响。选用20周龄体质量相近的AA(爱拔益加)种公鸡共120只,随机分为3组,分别使用8.0,11.5和16.0h光照处理,饲喂4周,饲喂结束后颈动脉放血致死,采取睾丸。采用实时荧光定量PCR和Western blot方法检测鸡睾丸组织中褪黑素受体及合成酶mRNA和蛋白表达量。结果表明,鸡睾丸组织中褪黑素受体MEL-1A、MEL-1B和MEL-1C,褪黑素合成酶AANAT的mR-NA及蛋白表达量随光照时长增加而显著降低(P0.01);但合成酶ASMT表达量无显著变化(P0.05)。综上,褪黑素受体MEL-1A、MEL-1B和MEL-1C及合成酶AANAT的表达与光照时长呈负相关。     

Pathology of acute Leptospira interrogans serotype icterohaemorrhagiae infection in the Syrian hamster

The pathology of acute Leptospira interrogans serotype icterohaemorrhagiae infection in the Syrian hamster was investigated up to 7 days after infection using histology, electron microscopy and an indirect fluorescence test for leptospires. The disease was characterized by the presence of many leptospires in the tissues, jaundice, leukocytosis, haemorrhages, endothelial alteration and thrombotic glomerulopathy. The leptospires were present intravascularly, in the interstitium penetrating between liver cells and tubular epithelial cells and in the tubular lumina. The presence of leptospires was not necessarily associated with lesions. These findings support both pathogenetic mechanisms suggested in the literature, namely: the ability of leptospires to penetrate actively between cells with detachment of tight junctions, without obvious lesions to the cells, and an immune-mediated process with immune complex formation and binding and activation of complement resulting in leukocytosis, thrombotic glomerulopathy, endothelial alteration and haemorrhages.     

NEFA和BHBA对体外培养的脂肪细胞HSL、ADPN mRNA表达的影响

在体外培养的牛脂肪组织和脂肪细胞中,分别添加5个浓度梯度的NEFA和BHBA,均设3个重复,通过荧光定量PCR技术,观测不同浓度的NEFA和BHBA对牛脂肪细胞ADPN mRNA与HSL mRNA丰度的影响。结果表明:NEFA在一定浓度范围内(0.2~0.8 nmol/L)对ADPN mRNA表达有显著促进作用并呈剂量依赖性,而高浓度(&gt;1.6 nmol/L)时又显著下调其表达;0.2~0.8 nmol/L NEFA对HSL mRNA的表达有抑制作用,呈剂量依赖性,但在高浓度时(&gt;0.8 nmol/L)反而促进了其表达(P&lt;0.05)。低浓度的BHBA对HSL mRNA的表达无显著抑制作用,高浓度(1.2 mmol/L)的BHBA显著下调HSL mRNA的表达,并呈剂量依赖性;低浓度(&lt;0.6mmol/L)的BHBA对ADPN mRNA的表达无明显作用,但高浓度的BHBA(&gt;0.6 mmol/L)对ADPN mRNA的表达具有明显的抑制作用(P&lt;0.05)。结论:代谢中间产物可通过促进ADPN mRNA或抑制HSL mRNA的表达来调节脂肪代谢。     

缺Se对雏鸡血清组胺水平及空肠H2R mRNA表达的影响

将40只1日龄雏鸡随机分成低Se试验组(日粮Se含量0.032 mg/kg)和正常对照组(日粮Se含量0.229mg/kg),分别在30、45、60和75 d断头处死取样,用ELISA法检测组胺浓度及半定量RT-PCR法测定空肠2型组胺受体(H2R)mRNA表达水平.结果表明,血清组胺水平在对照组与缺Se组及缺Se组组间比较差异均极显著(P<0.01);Se缺乏时鸡空肠H2R mRNA表达水平呈升高趋势,对照组与缺Se组及缺se组组间比较差异极显著(P<0.01);在整个试验期间,缺Se组血清Se含量与组胺浓度及H2R mRNA表达水平呈时间-效应关系.缺se刺激肥大细胞脱颗粒,使血清中组胺浓度和空肠H2R mRNA的表达水平升高,可能对硒缺乏引起的空肠组织损伤具有保护作用.     

缺Se对雏鸡血清组胺水平及空肠H_2R mRNA表达的影响

将40只1日龄雏鸡随机分成低Se试验组（日粮Se含量0.032 mg/kg）和正常对照组（日粮Se含量0.229mg/kg）,分别在30、45、60和75 d断头处死取样,用ELISA法检测组胺浓度及半定量RT-PCR法测定空肠2型组胺受体（H2R）mRNA表达水平。结果表明,血清组胺水平在对照组与缺Se组及缺Se组组间比较差异均极显著（P〈0.01）;Se缺乏时鸡空肠H2R mRNA表达水平呈升高趋势,对照组与缺Se组及缺Se组组间比较差异极显著（P〈0.01）;在整个试验期间,缺Se组血清Se含量与组胺浓度及H2R mRNA表达水平呈时间-效应关系。缺Se刺激肥大细胞脱颗粒,使血清中组胺浓度和空肠H2R mRNA的表达水平升高,可能对硒缺乏引起的空肠组织损伤具有保护作用。     

Allometric analysis of the postnatal development of the Syrian golden hamster pancreas

The objective of the present study was to analyse allometrically the growth of the Syrian golden hamster pancreas during days 2 to 70 of postnatal development. Body and pancreatic mass were determined, followed by stereological determination of the absolute volume of each morphological compartment of the pancreas. The marked pancreatic growth, by 4360%, was due to an increase in the absolute volume of all morphological compartments, mainly the acini which showed an increase of 10 431%. Bivariate allometric analysis of pancreatic mass and morphological compartmental volume in relation to body mass gain showed: (1) a biphasic pattern for pancreatic mass, acinar volume, excretory duct volume and stromal volume, with the first phase being observed from 2 to 21 days of age and the second from 21 to 70 days of age, with allometry coefficients of 1.537-0.513, 1.770-0.543, 1.651-0.506 and 0.967-0.258, respectively, and (2) a monophasic pattern from 2 to 70 days for intercalated duct volume and islet volume, with allometry coefficients of 0.913 and 1.727, respectively. These results show that during the growth of the pancreas in relation to that of the body some structures - acini and excretory ducts - follow the growth pattern of the organ, while others - intercalated ducts and islets - show a different pattern. This may be related to the genetic growth characteristics of each compartment itself or to some relationship between compartments during some stage of the ontogenetic development of this organ.