BACKGROUND & AIMS: :Secretory granules carrying fluorescent cargo proteins are widely used to study granule biogenesis, maturation, and regulated exocytosis. We fused the soluble secretory protein peptidylglycine alpha-hydroxylating monooxygenase (PHM) to green fluorescent protein (GFP) to study granule formation. When expressed in AtT-20 or GH3 cells, the PHM-GFP fusion protein partitioned from endogenous hormone (adrenocorticotropic hormone, growth hormone) into separate secretory granule pools. Both exogenous and endogenous granule proteins were stored and released in response to secretagogue. Importantly, we found that segregation of content proteins is not an artifact of overexpression nor peculiar to GFP-tagged proteins. Neither luminal acidification nor cholesterol-rich membrane microdomains play essential roles in soluble content protein segregation. Our data suggest that intrinsic biophysical properties of cargo proteins govern their differential sorting, with segregation occurring during the process of granule maturation. Proteins that can self-aggregate are likely to partition into separate granules, which can accommodate only a few thousand copies of any content protein; proteins that lack tertiary structure are more likely to distribute homogeneously into secretory granules. Therefore, a simple "self-aggregation default" theory may explain the little acknowledged, but commonly observed, tendency for both naturally occurring and exogenous content proteins to segregate from each other into distinct secretory granules.

背景与目标： ：带有荧光货物蛋白的分泌颗粒被广泛用于研究颗粒的生物发生，成熟和调控的胞吐作用。我们将可溶性分泌蛋白肽基甘氨酸α-羟基化单加氧酶（PHM）与绿色荧光蛋白（GFP）融合，以研究颗粒的形成。当在AtT-20或GH3细胞中表达时，PHM-GFP融合蛋白从内源激素（促肾上腺皮质激素，生长激素）分配到单独的分泌性颗粒库中。外源性和内源性颗粒蛋白均被存储并响应促分泌素而释放。重要的是，我们发现内容蛋白的分离不是过表达的产物，也不是带有GFP标签的蛋白所特有的。内腔酸化或富含胆固醇的膜微区都没有在可溶性蛋白质分离中起重要作用。我们的数据表明，货物蛋白的固有生物物理特性决定了它们的差异分选，在颗粒成熟过程中发生了分离。可以自我聚集的蛋白质很可能会分解成单独的颗粒，这些颗粒只能容纳几千个拷贝的任何含量的蛋白质。缺乏三级结构的蛋白质更有可能均匀地分布到分泌颗粒中。因此，一个简单的“自我聚集默认”理论可以解释很少有人承认但普遍观察到的自然存在和外源性蛋白质彼此分离成不同分泌颗粒的趋势。

BACKGROUND & AIMS: :Six different secretory proteins of molecular weights (15, 26, 30, 41, 55 and 70 kDa) were isolated from 8-day-old culture filtrate of Mycobacterium tuberculosis H37Ra using different column chromatography techniques. These proteins were further examined for their ability to induce cell mediated (T-cell proliferation assay) and humoral immune response (ELISA) in mice immunized with total culture filtrate proteins. Out of six proteins, three proteins showed good reactivity. However, the activity was at a maximum with 30 kDa antigen. The immune response induced by 30 kDa antigen emulsified in Freund's incomplete adjuvant (FIA) was investigated and was found to be dose dependent. The T-cell response induced by this protein was skewed towards T-helper (Th1) cells as determined by the pronounced secretion of interleukin-2 (IL-2) and gamma-interferon (IFN-gamma). The protective activity of the 30 kDa protein was also evaluated and compared with reference to Bacillus Calmette Guerin (BCG) vaccine in the mice challenged with virulent M. tuberculosis H37Rv. The degree of protection afforded by the 30 kDa antigen on the basis of mortality and the significant decrease in c.f.u.'s recovered from different organs (lung, liver, spleen) after 30 days of challenge with LD50 of M. tuberculosis H37Rv was significantly higher in comparison to BCG vaccinated animals. However, the degree of immunity induced by this antigen decreased with time (when challenged 8 and 12 weeks post-immunization) but it was still comparable with BCG. These findings suggest that 30 kDa secretory protein of M. tuberculosis is the key immunoprotective antigen and may be a suitable candidate for the development of an alternative subunit vaccine against tuberculosis.

背景与目标： ：使用不同的柱色谱技术从结核分枝杆菌H37Ra的8天龄培养滤液中分离出六个分子量分别为15、26、30、41、55和70 kDa的分泌蛋白。在用总培养滤液蛋白免疫的小鼠中，进一步检查了这些蛋白诱导细胞介导的能力（T细胞增殖测定）和体液免疫应答（ELISA）。在六种蛋白质中，三种蛋白质显示出良好的反应性。但是，对于30kDa抗原，活性最大。研究了在弗氏不完全佐剂（FIA）中乳化的30 kDa抗原诱导的免疫反应，发现该反应是剂量依赖性的。该蛋白诱导的T细胞反应偏向T辅助（Th1）细胞，这取决于白介素2（IL-2）和γ-干扰素（IFN-γ）的明显分泌。还评估了30 kDa蛋白的保护活性，并与卡介苗芽孢杆菌（BCG）疫苗进行了比较，比较了在有毒结核分枝杆菌H37Rv攻击的小鼠中的行为。在死亡的基础上，由30 kDa抗原提供的保护程度以及从结核分枝杆菌H37Rv的LD50攻击30天后从不同器官（肺，肝，脾）回收的cfu的显着降低大大高于H37Rv。与接种卡介苗的动物比较。但是，这种抗原诱导的免疫度随时间降低（在免疫后8周和12周激发时），但仍可与BCG相提并论。这些发现表明，结核分枝杆菌的30kDa分泌蛋白是关键的免疫保护抗原，并且可能是开发替代性抗结核亚单位疫苗的合适候选者。

BACKGROUND & AIMS: :Possible function(s) proposed in the literature for Paramecium trichocysts have been controversial for a long time. The oldest hypothesis, defence against protozoan predators, has now obtained support from experiments with Dileptus as well as by our observation of a vigorous lateral displacement of cells due to synchronous, massive local trichocyst expulsion. Recently discovered secretory lectins could also operate in defence or improve the availability of food organisms. Other hypotheses on trichocyst function - even if they appeared unlikely to us a priori-have also been analyzed experimentally, but no convincing evidence in favour of any of them could be found.

背景与目标： ：长期以来，文献中提出的草履虫毛囊的可能功能一直存在争议。最早的假说是防御原生动物的掠食性，现在已经获得了Dileptus实验的支持，以及我们观察到的由于同步，大量局部毛囊虫驱逐而引起的剧烈细胞侧移。最近发现的分泌型凝集素还可以起到防御作用或改善食用生物的利用度。关于毛囊功能的其他假设-即使对我们来说都不是先验的假设，也已经通过实验进行了分析，但是找不到令人信服的证据来支持它们。

BACKGROUND & AIMS: :We herein demonstrate that mast cells express all known members of the group II subfamily of secretory phospholipase A2 (sPLA2) isozymes, and those having heparin affinity markedly enhance the exocytotic response. Rat mastocytoma RBL-2H3 cells transfected with heparin-binding (sPLA2-IIA, -V, and -IID), but not heparin-nonbinding (sPLA2-IIC), enzymes released more granule-associated markers (beta-hexosaminidase and histamine) than mock- or cytosolic PLA2alpha (cPLA2alpha)-transfected cells after stimulation with IgE and Ag. Site-directed mutagenesis of sPLA2-IIA and -V revealed that both the catalytic and heparin-binding domains are essential for this function. Confocal laser and electron microscopic analyses revealed that sPLA2-IIA, which was stored in secretory granules in unstimulated cells, accumulated on the membranous sites where fusion between the plasma membrane and granule membranes occurred in activated cells. These results suggest that the heparin-binding sPLA2s bind to the perigranular membranes through their heparin-binding domain, and lysophospholipids produced in situ by their enzymatic action may facilitate the ongoing membrane fusion. In contrast to the redundant role of sPLA2-IIA, -IID, and -V in the regulation of degranulation, only sPLA2-V had the ability to markedly augment IgE/Ag-stimulated immediate PGD2 production, which reached a level comparable to that elicited by cPLA2alpha. The latter observation reveals an unexplored functional segregation among the three related isozymes expressed in the same cell population.

背景与目标： ：我们在本文中证明肥大细胞表达分泌性磷脂酶A2（sPLA2）同工酶II组的所有已知成员，而具有肝素亲和力的细胞显着增强胞吐反应。用肝素结合（sPLA2-IIA，-V和-IID）转染大鼠肥大细胞瘤RBL-2H3细胞，但未结合肝素不结合（sPLA2-IIC），酶释放的颗粒相关标志物（β-己糖胺酶和组胺）多于用IgE和Ag刺激后，模拟或胞质PLA2alpha（cPLA2alpha）转染的细胞。 sPLA2-IIA和-V的定点诱变表明，催化结构域和肝素结合结构域对于该功能都是必不可少的。共聚焦激光和电子显微镜分析表明，存储在未刺激细胞分泌颗粒中的sPLA2-IIA积累在膜质部位，在质膜中质膜和颗粒膜之间发生融合，并在活化细胞中发生。这些结果表明，与肝素结合的sPLA2s通过其肝素结合结构域与外周膜结合，并且通过其酶促作用原位产生的溶血磷脂可以促进正在进行的膜融合。与sPLA2-IIA，-IID和-V在脱粒调节中的冗余作用相反，只有sPLA2-V具有显着增加IgE / Ag刺激的PGD2立即产生的能力，该水平达到了与诱发的水平相当的水平。由cPLA2alpha。后者的观察结果揭示了在同一细胞群体中表达的三种相关同工酶之间尚未探索的功能分离。

BACKGROUND & AIMS: :FKBPs define a subfamily of peptidyl-prolyl cis/trans isomerases (PPlases). PPlases are known to play roles in cellular protein folding, protein interactions and signal transduction. Here we describe NcFKBP22 from Neurospora crassa, a novel type of FKBP. NcFKBP22 is synthesized as a precursor protein with a cleavable signal sequence. In addition to a typical FKBP domain in the amino-terminal part mature NcFKBP22 contains a novel second domain which is unique amongst all known FKBPs. The amino acid composition of this carboxyterminal domain is highly biased. Secondary structure predictions suggest that this domain may form an amphipathic alpha-helix. The carboxy-terminus of NcFKBP22 is -HNEL, a potential endoplasmic reticulum (ER) retention signal, suggesting that NcFKBP22 is a resident protein of the ER.

背景与目标： ：FKBP定义了肽基-脯氨酰顺/反异构酶（PPlases）的亚家族。已知PP酶在细胞蛋白质折叠，蛋白质相互作用和信号转导中发挥作用。在这里，我们描述了一种新的FKBP类型，来自Neurospora crassa的NcFKBP22。 NcFKBP22被合成为具有可裂解信号序列的前体蛋白。除了氨基末端部分的典型FKBP结构域外，成熟的NcFKBP22还包含一个新颖的第二结构域，该结构域在所有已知的FKBP中都是唯一的。该羧基末端结构域的氨基酸组成高度偏向。二级结构预测表明，该结构域可能形成两亲性α-螺旋。 NcFKBP22的羧基末端是-HNEL，这是潜在的内质网（ER）保留信号，表明NcFKBP22是ER的驻留蛋白。

BACKGROUND & AIMS: :The tubulovesicles of gastric parietal cells sequester H+/K+-ATPase molecules within resting parietal cells. Stimulation of parietal cell secretion elicits delivery of intracellular H+/K+-ATPase to the apically oriented secretory canaliculus. Previous investigations have suggested that this process requires the regulated fusion of intracellular tubulovesicles with the canalicular target membrane. We have sought to investigate the presence of critical putative regulators of vesicle fusion on immunoisolated gastric parietal cell tubulovesicles. Highly purified tubulovesicles were prepared by gradient fractionation and immunoisolation on magnetic beads coated with monoclonal antibodies against the alpha subunit of H+/K+-ATPase. Western blot analysis revealed the presence of Rab11, Rab25, vesicle-associated membrane protein 2 (VAMP-2) and secretory carrier membrane proteins (SCAMPs) on immunoisolated vesicles. The same cohort of proteins was recovered on vesicles immunoisolated with monoclonal antibodies against SCAMPs and VAMP-2. In contrast, whereas immunoreactivities for syntaxin 1A/1B and synaptosome-associated protein (SNAP-25) were present in gradient-isolated vesicles, none of the immunoreactivity was associated with immunoisolated vesicles. The observation of VAMP-2 and two Rab proteins on immunoisolated H+/K+-ATPase-containing tubulovesicles supports the role for tubulovesicles in a regulated vesicle fusion process. In addition, the presence of SCAMPs along with Rab11 and Rab25 implicates the tubulovesicles as a critical apical recycling vesicle population.

背景与目标： ：胃壁细胞的微管壁螯合静息壁细胞内的H / K -ATPase分子。刺激壁细胞分泌引起将细胞内H / K -ATP酶递送至根尖定向的分泌小管。先前的研究表明，该过程需要细胞内微管小泡与小管靶膜的受控融合。我们试图研究免疫隔离的胃壁细胞微管小泡上囊泡融合的关键假定调节剂的存在。通过梯度分级分离和在磁珠上免疫分离制备高度纯化的微管小球，所述磁珠上涂有针对H / K -ATPaseα亚基的单克隆抗体。蛋白质印迹分析显示免疫分离的囊泡上存在Rab11，Rab25，囊泡相关膜蛋白2（VAMP-2）和分泌性载体膜蛋白（SCAMPs）。在针对SCAMPs和VAMP-2的单克隆抗体免疫分离的囊泡上回收了同一批蛋白质。相反，尽管在梯度分离的囊泡中存在语法1A / 1B和突触体相关蛋白（SNAP-25）的免疫反应性，但免疫反应性均与免疫分离的囊泡无关。在含有免疫隔离的H / K -ATPase的微管小泡上观察到VAMP-2和两种Rab蛋白支持了微管小泡在调节的囊泡融合过程中的作用。此外，SCAMPs与Rab11和Rab25的存在暗示着微管小泡是一个重要的根尖再生小泡种群。

BACKGROUND & AIMS: :Here we present a patient with a parotid secretory carcinoma (SC) with high-grade transformation. A 65-year-old female was referred to our hospital due to a gradually growing right parotid tumor discovered initially about 4 years earlier. MRI imaging detected a right parotid tumor 50 mm in the longer axis. Fine needle aspiration cytology indicated a class III tumor. Nine months after her initial visit, she revisited our department because of pain, trismus and facial paralysis. MRI detected a tumor 69 mm in the longer axis and 64 mm in the shorter axis and a biopsy specimen revealed parotid cancer. Furthermore, positron emission tomography revealed a synchronous small cell lung cancer (SCLC). Chemoradiotherapy for the SCLC was performed followed by an extended total parotidectomy for the parotid SC. Histological findings and ETV6-FISH analysis confirmed a parotid SC with high-grade transformation. Two months after the surgery, CT revealed a loco-regional recurrence and proton beam therapy (70.2 GyE/26 Fr) was performed. Three months after the proton beam therapy, CT indicated pleural effusion and lung metastasis, and fine needle aspiration cytology revealed the metastatic SC. Eight months after the surgery, the patient died due to the lung metastasis of SC.

背景与目标： ：这里我们介绍了一名患有高度转化性腮腺分泌癌（SC）的患者。由于最初约4年前发现的逐渐增长的右腮腺肿瘤，一名65岁的女性被转诊到我们医院。 MRI成像在长轴上检测到右腮腺肿瘤50毫米。细针穿刺细胞学检查显示为III类肿瘤。初诊九个月后，由于疼痛，三头肌和面部瘫痪，她重新访问了我们的科室。 MRI在长轴上检测出一个69mm的肿瘤，在短轴上检测到一个64mm的肿瘤，活检标本显示腮腺癌。此外，正电子发射断层扫描显示同步性小细胞肺癌（SCLC）。对SCLC进行化学放射治疗，随后对腮腺SC进行全面的腮腺切除术。组织学发现和ETV6-FISH分析证实腮腺SC具有高度转化。手术两个月后，CT显示局部复发，并进行了质子束治疗（70.2 GyE / 26 Fr）。质子束治疗三个月后，CT显示胸腔积液和肺转移，细针穿刺细胞学检查显示有转移性SC。手术后八个月，患者因SC的肺转移而死亡。

BACKGROUND & AIMS: :Fc receptor-like (FCRL) 4 is an immunoregulatory receptor expressed on a subpopulation of human memory B cells of mucosa-associated lymphoid tissue. Fc receptor function of FCRL4 was demonstrated by binding of IgA to FCRL4 following heat aggregation of the Ig. In this study, we demonstrate that FCRL4 recognizes J chain-linked systemic IgA in the absence of heat aggregation. We further demonstrate that mucosal secretory IgA is not recognized by FCRL4 and that systemic IgA binding can be competitively inhibited by recombinant secretory component protein. Finally, we provide evidence that primary FCRL4-bearing human memory B cells are constitutively bound to IgA. Our study provides a mechanism for the negative regulatory activity of FCRL4 on AgR-mediated B cell activation.

背景与目标： ：Fc受体样（FCRL）4是在与黏膜相关的淋巴组织的人类记忆B细胞亚群上表达的免疫调节受体。在Ig热聚集后，IgA与FCRL4的结合证明了FCRL4的Fc受体功能。在这项研究中，我们证明了FCRL4在不存在热聚集的情况下识别J链连接的全身IgA。我们进一步证明，粘膜分泌型IgA不被FCRL4识别，全身性IgA结合可被重组分泌成分蛋白竞争性抑制。最后，我们提供的证据表明，带有原代FCRL4的人类记忆B细胞与IgA组成性结合。我们的研究为FCRL4对AgR介导的B细胞活化的负调控活性提供了一种机制。

BACKGROUND & AIMS: :The secretory pathway Ca(2+) ATPase (SPCA) provides the Golgi apparatus with a Ca(2+) supply essential for Ca(2+)-dependent enzymes involved in the post-translational modification of proteins in transit through the secretory pathway. Ca(2+) in the Golgi apparatus is also agonist-releasable and plays a role in hormone-induced Ca(2+) transients. Although the Ca(2+) ATPase inhibitors thapsigargin is more selective for the sarcoplasmic-endoplasmic reticulum Ca(2+) ATPase (SERCA) than for SPCA, no inhibitor has been characterised that selectively inhibits SPCA. A number of inhibitors were assessed for their selectivity to the human SPCA1d compared to the more ubiquitous human SERCA2b. Each isoform was over-expressed in COS-7 cells and the Ca(2+)-dependent ATPase activity measured in their microsomal membranes. Both bis(2-hydroxy-3-tert-butyl-5-methyl-phenyl)methane(bis-phenol) and 2-aminoethoxydiphenylborate (2-APB) selectively inhibited SPCA1d (with IC(50) values of 0.13 μM and 0.18 mM, respectively), which were of 62- and 8.3-fold greater potency than the values for hSERCA2b (IC(50) values; 8.1 μM and 1.5mM, respectively). Other inhibitors tested such as bis-phenol-A, tetrabromobisphenol-A and trifluoperazine inhibited both Ca(2+) ATPases similarly. Furthermore, bis-phenol was able to mobilize Ca(2+) in cells that had been pre-treated with thapsigargin. Therefore we conclude that given the potency and selectivity of bis-phenol it may prove a valuable tool in further understanding the role of SPCA in cellular processes.

背景与目标： ：分泌途径Ca（2）ATPase（SPCA）为高尔基体提供了Ca（2）供应，这对于参与通过分泌途径转运的蛋白质的翻译后修饰的Ca（2）依赖性酶必不可少。高尔基体中的Ca（2）也是激动剂可释放的，并在激素诱导的Ca（2）瞬变中起作用。尽管Ca（2）ATPase抑制剂thapsigargin对肌浆网-内质网Ca（2）ATPase（SERCA）的选择性比对SPCA的选择性更高，但尚无抑制剂选择性抑制SPCA的特征。与更普遍的人SERCA2b相比，评估了许多抑制剂对人SPCA1d的选择性。每个同工型在COS-7细胞中过表达，并在其微粒体膜中测量Ca（2）依赖的ATPase活性。双（2-羟基-3-叔丁基-5-甲基-苯基）甲烷（双酚）和2-氨基乙氧基二苯基硼酸酯（2-APB）选择性抑制SPCA1d（IC（50）值为0.13μM和0.18 mM分别比hSERCA2b的值高62倍和8.3倍（IC（50）值；分别为8.1μM和1.5mM）。测试的其他抑制剂，如双酚A，四溴双酚A和三氟哌嗪类似地抑制了两个Ca（2）ATPases。此外，双酚能够动员已经用毒胡萝卜素预处理的细胞中的Ca（2）。因此，我们得出结论，鉴于双酚的效力和选择性，它可能被证明是进一步了解SPCA在细胞过程中的作用的有价值的工具。

BACKGROUND & AIMS: :Intestinal secretion was evoked in periarterially denervated jejunal segments of anesthetized rats and cats by exposing the intestines to the heat stable (ST) toxins from a strain of Escherichia coli producing both STa and STb toxins. The secretion was significantly inhibited and to about the same relative extent by the addition of each one of the three following drugs: hexamethonium (i.v., rats), lidocaine (applied on the serosal surface, rats) and tetrodotoxin (intra-arterial, cats). Atropine inhibited fluid secretion in some experiments. It is proposed that a nervous mechanism is mediating part of the secretory response to Escherichia coli heat stable toxins, since three different drugs, which influence nervous activity in different ways, significantly diminished the secretory response. A model for the secretory nervous reflex(es) within the enteric nervous system is proposed; Escherichia coli heat stable toxins activate a "receptor cell" in the epithelium, which then stimulates surrounding dendritic nerve endings via the release of unknown substance(s). A nicotinic receptor is involved but further characteristics of the nervous reflex(es) remain to be elucidated.

背景与目标： ：通过将肠暴露于产生STa和STb毒素的大肠杆菌菌株的热稳定（ST）毒素，在麻醉的大鼠和猫的动脉周围神经支配的空肠段中引起肠道分泌。通过添加以下三种药物中的每一种，分泌被显着抑制，并且相对抑制程度大致相同：六甲铵（静脉注射，大鼠），利多卡因（应用于浆膜表面，大鼠）和河豚毒素（动脉内，猫） 。在某些实验中，阿托品抑制体液分泌。提出神经机制介导了对大肠杆菌热稳定毒素的分泌反应的一部分，因为以不同方式影响神经活动的三种不同药物显着降低了分泌反应。提出了肠道神经系统内分泌神经反射的模型。大肠杆菌的热稳定毒素会激活上皮细胞中的“受体细胞”，然后通过释放未知物质刺激周围的树突神经末梢。涉及烟碱样受体，但神经反射的进一步特征尚待阐明。

BACKGROUND & AIMS: :Chronic administration of a high tolbutamide dose to rats induces islet hypertrophy associated with a decreased insulin content per islet and with a diminished insulin release in response to a glucose or leucine stimulus. These changes are reversible after discontinuation of tolbutamide. Chronic administration of a low tolbutamide dose (effective on islet size, on insulin content per islet, or on leucine-induced insulin release is normal in the presence of glucagon (5 mug/ml) or theophylline (5 mM). Since islet hypertrophy occurs following administration of high tolbutamide doses only and is associated with hypofunction rather than with hyperfunction, it seems hardly conceivable that the therapeutic principle of tolbutamide is based on a beta-cytotrophic effect. B-cell hypofunction seems to be due to at least three factors: the decrease in the insulin content per islet, an impairement in secretory signal recognition, and an interference with the process of signal transmission.

背景与目标： ：对大鼠长期给予高剂量的甲苯磺丁酰胺会诱发胰岛肥大，这与每个胰岛的胰岛素含量降低以及对葡萄糖或亮氨酸刺激的胰岛素释放减少有关。停用甲苯磺丁酰胺后，这些变化是可逆的。在存在胰高血糖素（5杯/毫升）或茶碱（5毫米）的情况下，长期给予低剂量的甲苯磺丁酰胺（对胰岛大小，每个胰岛胰岛素含量或亮氨酸诱导的胰岛素释放有效）是正常的。仅在服用高剂量的甲苯磺丁酰胺后，它与功能减退而不是功能亢进有关，因此很难想象甲苯磺丁酰胺的治疗原理是基于β-细胞营养作用，而B细胞功能减退似乎是由于至少三个因素引起的：每个胰岛中胰岛素含量的减少，分泌信号识别的障碍以及对信号传输过程的干扰。

BACKGROUND & AIMS: :The dynamic activities of mitochondria and lysosomes, which play important roles in maintaining cellular homeostasis, were observed without labeling by using highly sensitive photothermal (PT) microscopy. This imaging modality allows for the direct observation of cellular organelles that contain endogenous chromophores, with high temporal and spatial resolution. We identified mitochondria and lysosomes inside living mammalian cells via simultaneous dual-color imaging. Moreover, dynamic imaging revealed that the lysosomes make contact with mitochondria and move between sites within the dynamic mitochondrial network. Since mitochondrial and lysosomal functions are intricately connected, PT microscopy should provide in-depth understanding of cellular functions associated with mitochondria-lysosome communication as well as insights into various human diseases caused by dysfunction of these organelles.

背景与目标： ：通过使用高灵敏度的光热（PT）显微镜，无需标记即可观察到线粒体和溶酶体的动态活动，它们在维持细胞稳态中起着重要作用。这种成像方式可以以高的时间和空间分辨率直接观察包含内生发色团的细胞器。我们通过同步双色成像鉴定了活的哺乳动物细胞内的线粒体和溶酶体。此外，动态成像显示溶酶体与线粒体接触并在动态线粒体网络内的位点之间移动。由于线粒体功能和溶酶体功能错综复杂地联系在一起，因此，PT显微镜应能深入了解与线粒体-溶酶体通讯相关的细胞功能，并深入了解由这些细胞器功能障碍引起的各种人类疾病。

BACKGROUND & AIMS: :Carcinomas develop in complex environments that include a diverse spectrum of cell types that influence tumor cell behavior. These microenvironments represent dynamic systems that contribute to pathologic processes. Damage to DNA is a notable inducer of both transient and permanent alterations in cellular phenotypes. Induction of a DNA damage secretory program is known to promote adverse tumor cell behaviors such as proliferation, invasion, metastasis, and treatment resistance. However, prior studies designed to identify genotoxic stress-induced factors evaluated actively proliferating in vitro cultures of cells such as fibroblasts as experimental models. Conversely, the vast majority of benign cells in a typical tumor microenvironment (TME) are not proliferating but rather exist in quiescent (i.e., G0) or in terminally differentiated states. In this study, the diversity and magnitude of transcriptional responses to genotoxic damage in quiescent prostate fibroblasts were assessed using gene expression profiling. The secretory damage response in quiescent cells was highly concordant with that of actively dividing cells. Quiescent human prostate stroma exposed to genotoxic agents (e.g., mitoxantrone) in vivo resulted in significant upregulation (2.7- to 5.7-fold; P ≤ 0.01) of growth factors and cytokines including IL1β, MMP3, IL6, and IL8. The paracrine effects of damaged quiescent cells consistently increased the proliferation and invasion of prostate cancer cells and promoted cell survival and resistance to apoptosis following exposure to chemotherapy.Implications: Benign quiescent cells in the TME respond to genotoxic stress by inducing a secretory program capable of promoting therapy resistance. Developing approaches to suppress the secretory program may improve treatment responses. Mol Cancer Res; 15(7); 842-51. ©2017 AACR.

背景与目标： 癌在复杂的环境中发展，其中包括影响肿瘤细胞行为的多种细胞类型。这些微环境代表了有助于病理过程的动态系统。 DNA损伤是细胞表型瞬时和永久性变化的明显诱因。已知DNA损伤分泌程序的诱导可促进不良的肿瘤细胞行为，例如增殖，侵袭，转移和治疗抗性。然而，旨在鉴定遗传毒性应激诱导因素的先前研究评估了诸如成纤维细胞等细胞的体外培养中活跃增殖的实验模型。相反，在典型的肿瘤微环境（TME）中，绝大多数良性细胞并未增殖，而是以静止状态（即G0）或终末分化状态存在。在这项研究中，使用基因表达谱评估了静态前列腺成纤维细胞中对遗传毒性损伤的转录反应的多样性和强度。静止细胞的分泌损伤反应与主动分裂细胞的分泌损伤反应高度一致。体内暴露于遗传毒性剂（例如米托蒽醌）的静态人前列腺基质导致生长因子和包括IL1β，MMP3，IL6和IL8在内的细胞因子显着上调（2.7-至5.7倍; P≤0.01）。受损的静态细胞的旁分泌作用持续增加前列腺癌细胞的增殖和侵袭，并促进化疗后的细胞存活和对凋亡的抗性。意义：TME中的良性静态细胞通过诱导能够促进分泌的分泌程序来对遗传毒性应激作出反应。治疗抵抗力。开发抑制分泌程序的方法可以改善治疗反应。分子癌症研究； 15（7）； 842-51。 ©2017 AACR。

BACKGROUND & AIMS: :Specialization of many cells, including the acinar cells of the salivary glands and pancreas, milk-producing cells of mammary glands, mucus-secreting goblet cells, antibody-producing plasma cells, and cells that generate the dense extracellular matrices of bone and cartilage, requires scaling up both secretory machinery and cell-type specific secretory cargo. Using tissue-specific genome-scale analyses, we determine how increases in secretory capacity are coordinated with increases in secretory load in the Drosophila salivary gland (SG), an ideal model for gaining mechanistic insight into the functional specialization of secretory organs. Our findings show that CrebA, a bZIP transcription factor, directly binds genes encoding the core secretory machinery, including protein components of the signal recognition particle and receptor, ER cargo translocators, Cop I and Cop II vesicles, as well as the structural proteins and enzymes of these organelles. CrebA directly binds a subset of SG cargo genes and CrebA binds and boosts expression of Sage, a SG-specific transcription factor essential for cargo expression. To further enhance secretory output, CrebA binds and activates Xbp1 and Tudor-SN. Thus, CrebA directly upregulates the machinery of secretion and additional factors to increase overall secretory capacity in professional secretory cells; concomitant increases in cargo are achieved both directly and indirectly.

背景与目标： ：许多细胞的专业化，包括唾液腺和胰腺的腺泡细胞，乳腺产生乳汁的细胞，粘液分泌的杯状细胞，产生抗体的浆细胞以及产生密集的骨和软骨细胞外基质的细胞，需要扩大分泌机器和特定于细胞类型的分泌货物的规模。使用组织特异性基因组规模的分析，我们确定果蝇唾液腺（SG）中分泌能力的增加如何与分泌负荷的增加协调起来，果蝇唾液腺（SG）是获得有关分泌器官功能专业化的机制的理想模型。我们的发现表明，bZIP转录因子CrebA直接结合编码核心分泌机制的基因，包括信号识别颗粒和受体的蛋白质成分，ER货物转运子，Cop I和Cop II囊泡以及结构蛋白和酶。这些细胞器。 CrebA直接结合SG货物基因的一个子集，而CrebA结合并增强Sage的表达，Sage是货物表达必不可少的SG特异性转录因子。为了进一步增强分泌输出，CrebA结合并激活Xbp1和Tudor-SN。因此，CrebA直接上调分泌机制和其他因素，以增加专业分泌细胞的总体分泌能力。直接和间接实现了货运量的同时增加。

BACKGROUND & AIMS: :Rapid actions of T3 on TSH synthesis in posttranscriptional steps, such as polyadenylation and translation rate, have already been described. The focus of this paper was to characterize rapid actions of T3 on TSH secretion and the involvement of actin and microtubule cytoskeleton in this process. For that, sham-operated (SO) and thyroidectomized (Tx) rats were subjected to acute or chronic treatment with T3. We observed a disarrangement in microtubule and actin cytoskeletons and an increase in Tshb mRNA levels in Tx rats, whereas the total TSH protein content was reduced in the pituitary gland as a whole, but increased in the secretory granules close to the plasma membrane of thyrotrophs, as well as in the extracellular space. The acute T3 dose promoted a rapid increase and redistribution of TSH secretory granules throughout the cytoplasm, as well as a rearrangement in actin and microtubule cytoskeletons. The T3 chronic treatment outcome reinforces the acute effects observed and, additionally, evinces an increase in the α-tubulin content and a rearrangement in microtubule cytoskeleton. Thus, T3 is able to rapidly suppress TSH secretion and, in parallel, to promote a rearrangement in actin and microtubules assembly throughout the pituitary gland, effects that seem to be independent from each other.

背景与目标： ：已经描述了T3在转录后步骤中对TSH合成的快速作用，例如聚腺苷酸化和翻译速率。本文的重点是表征T3对TSH分泌的快速作用以及肌动蛋白和微管细胞骨架在此过程中的参与。为此，对假手术（SO）和甲状腺切除（Tx）大鼠进行T3急性或慢性治疗。我们在Tx大鼠中观察到微管和肌动蛋白细胞骨架的紊乱以及Tshb mRNA水平的增加，而垂体的总TSH蛋白含量总体上减少了，但在靠近甲状腺营养细胞质膜的分泌颗粒中增加了，以及在细胞外空间急性T3剂量促进了TSH分泌颗粒在整个细胞质中的快速增加和重新分布，以及肌动蛋白和微管细胞骨架的重排。 T3慢性治疗的结果可增强观察到的急性作用，此外，还需要增加α-微管蛋白含量和微管细胞骨架的重排。因此，T3能够迅速抑制TSH分泌，并同时促进整个垂体的肌动蛋白和微管装配的重排，效果似乎彼此独立。