Acta histochemica最新文献_第2页

Autophagy markers expression pattern in developing liver of the yotari (dab1 -/-) mice and humans 自噬标志物在yotari （dab1-/-）小鼠和人肝脏发育中的表达模式。

IF 2.3 4区生物学 Q4 CELL BIOLOGY

Acta histochemica

Pub Date : 2024-12-07 DOI: 10.1016/j.acthis.2024.152224

Edita Dražić Maras , Nela Kelam , Anita Racetin , Ejazul Haque , Maja Dražić , Katarina Vukojević , Yu Katsuyama , Mirna Saraga-Babić , Natalija Filipović

Autophagy plays an important role in the physiology and pathology of the liver. Several negative autophagy regulators have been discovered, including epidermal growth factor receptor (EGFR), mediated by activation of the PI3K/Akt/mTOR signaling pathway. Disabled-1 (Dab1) is one of the mediating adaptor factors of PI3K/Akt/mTOR signaling pathways. We investigated the potential impact of Dab1 on autophagy-related markers (LC3B, LAMP2A, HSC70, and GRP78) in the developing liver by using a model of yotari mice and compared it with autophagy marker expression in human liver development. Mouse embryos were obtained at gestation days 13.5 and 15.5 (E13.5 and E15.5), and a total of 5 normal human conceptuses were obtained between gestation days 5 and 10. Histological sections were analyzed by immunohistochemistry. The highest expression of the early endosome-forming factor LC3B and the microautophagy factor LAMP2a was observed at the transition from embryonic to early fetal phase, whereas the expression of the chaperones HSC 70 and GRP78 was highest at embryonic phase. The expression patterns of three of these factors in mouse liver were different from those in human liver: the expression of LC3B was high at E13.5, that of HSC 70 at 15.5, whereas the expression of GRP78 did not change significantly. On the other hand, the expression pattern of LAMP2a was similar to that in human development and was higher at E15.5 than at E13.5. Moreover, knockout of Dab1 resulted in significantly lower expression of LC3B and LAMP2a in mouse embryo livers (at E13.5), indicating a possible role of Dab1 in regulating autophagy during embryonic development in the liver.

自噬在肝脏的生理和病理中起着重要的作用。已经发现了几种负自噬调节因子，包括表皮生长因子受体（EGFR），通过激活PI3K/Akt/mTOR信号通路介导。Disabled-1 （Dab1）是介导PI3K/Akt/mTOR信号通路的适配因子之一。我们通过yotari小鼠模型研究了Dab1对发育中的肝脏中自噬相关标志物（LC3B、LAMP2A、HSC70和GRP78）的潜在影响，并将其与人类肝脏自噬标志物的表达进行了比较。在妊娠第13.5和15.5天获得小鼠胚胎（E13.5和E15.5），在妊娠第5 - 10天共获得5个正常的人胚胎。组织切片采用免疫组织化学分析。早期内体形成因子LC3B和微自噬因子LAMP2a在胚胎期到胎早期表达最高，而伴侣蛋白HSC 70和GRP78在胚胎期表达最高。其中3个因子在小鼠肝脏中的表达模式与人肝脏不同：LC3B在E13.5时高表达，HSC 70在15.5时高表达，而GRP78的表达变化不明显。另一方面，LAMP2a的表达模式与人类发育相似，在E15.5时高于E13.5时。此外，敲除Dab1导致小鼠胚胎肝脏中LC3B和LAMP2a的表达显著降低（E13.5），表明Dab1可能在胚胎发育过程中调节肝脏自噬中发挥作用。

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引用次数: 0

Molecular signature-based labeling techniques for vascular endothelial cells 血管内皮细胞的分子标记技术。

IF 2.3 4区生物学 Q4 CELL BIOLOGY

Acta histochemica

Pub Date : 2024-12-06 DOI: 10.1016/j.acthis.2024.152222

Krutika H. Dobariya, Divya Goyal, Hemant Kumar

Vascular endothelial cells (VECs) play a crucial role in the development and maintenance of vascular biology specific to the tissue types. Molecular signature-based labeling and imaging of VECs help researchers understand potential mechanisms linking VECs to disease pathology, serving as valuable biomarkers in clinical settings and trials. Labeling techniques involve selectively tagging or marking VECs for visualization. Immunolabeled employs antibodies that specifically bind to VECs markers, while fluorescent tracers or dyes can directly label VECs for imaging. Some techniques use specific carbohydrate residues on cell surface, while others employ endothelial-specific promoters to express fluorescent proteins. Additionally, VEC can be labeled with contrast agents, radiolabeled tracers, and nanoparticles. The choice of labeling technique depends on study context, including whether it involves animal models, in vitro cell cultures, or clinical applications. Herein, we discussed the various labeling methods utilized to label VECs and the techniques to visualize them.

血管内皮细胞（VECs）在血管生物学的发展和维持中起着至关重要的作用。基于分子特征的VECs标记和成像帮助研究人员了解将VECs与疾病病理联系起来的潜在机制，在临床环境和试验中作为有价值的生物标志物。标记技术包括选择性地标记或标记VECs以实现可视化。免疫标记使用特异性结合VECs标记的抗体，而荧光示踪剂或染料可以直接标记VECs进行成像。一些技术利用细胞表面特定的碳水化合物残基，而另一些技术则利用内皮特异性启动子来表达荧光蛋白。此外，VEC可以用造影剂、放射性示踪剂和纳米颗粒进行标记。标记技术的选择取决于研究背景，包括是否涉及动物模型、体外细胞培养或临床应用。在此，我们讨论了用于标记VECs的各种标记方法和可视化技术。

引用次数: 0

Corrigendum to “MicroRNA-146b-5p suppresses cholangiocarcinoma cells by targeting TRAF6 and modulating p53 translocation” [Acta Histochem. (2021) 123 7 151793] MicroRNA-146b-5p 通过靶向 TRAF6 和调节 p53 转位抑制胆管癌细胞》[Acta Histochem. (2021) 123 7 151793]的更正。

IF 2.3 4区生物学 Q4 CELL BIOLOGY

Acta histochemica

Pub Date : 2024-11-09 DOI: 10.1016/j.acthis.2024.152214

Yiyue Ren , Xiaoyan Wang , Tong Ji , Xiujun Cai

引用次数: 0

Exploring the oncogenic role of RGS19 in bladder cancer progression and prognosis 探索 RGS19 在膀胱癌进展和预后中的致癌作用

IF 2.3 4区生物学 Q4 CELL BIOLOGY

Acta histochemica

Pub Date : 2024-10-31 DOI: 10.1016/j.acthis.2024.152212

Lei Yan , Guangyue Luo , Chengxiang Han , Jialin Meng , Chaozhao Liang

This study investigates the role of autophagy-related genes (ARGs) in bladder cancer (BLCA), focusing on the regulator of G protein signaling 19 (RGS19). Using data from The Cancer Genome Atlas (TCGA) and the Human Autophagy Database (HADb), we identified RGS19 as significantly upregulated and linked to poor prognosis in BLCA. Kaplan-Meier survival analysis confirmed its association with increased mortality and. In vitro, RGS19 knockdown in BLCA cell lines inhibited proliferation, migration, and invasion, while inducing apoptosis and autophagy. Transmission electron microscopy showed autophagic structures in RGS19-silenced cells. In vivo, a xenograft mouse model demonstrated reduced tumor growth with RGS19 knockdown. Immunohistochemical (IHC) analysis revealed decreased Ki67 and increased autophagy markers in tumors with reduced RGS19. Pathway analysis suggested RGS19 acts through the cGMP-PKG signaling pathway, validated by altered expression of soluble guanylate cyclase (sGC), protein kinase G (PKG1), phosphodiesterase 5 A (PDE5A), vasodilator-stimulated phosphoprotein (VASP), and phosphorylated VASP (p-VASP) upon RGS19 knockdown. These results highlight RGS19 as a potential biomarker and therapeutic target in BLCA.

本研究调查了自噬相关基因（ARGs）在膀胱癌（BLCA）中的作用，重点研究了G蛋白信号转导调节因子19（RGS19）。利用癌症基因组图谱（TCGA）和人类自噬数据库（HADb）的数据，我们发现RGS19在膀胱癌中显著上调并与不良预后相关。Kaplan-Meier生存分析证实了RGS19与死亡率增加和预后不良有关。在体外，敲除 BLCA 细胞系中的 RGS19 可抑制细胞增殖、迁移和侵袭，同时诱导细胞凋亡和自噬。透射电子显微镜显示了 RGS19 沉默细胞中的自噬结构。在体内，异种移植小鼠模型显示，RGS19 基因敲除可减少肿瘤生长。免疫组化（IHC）分析表明，在 RGS19 被敲除的肿瘤中，Ki67 降低，自噬标记物增加。通路分析表明，RGS19 是通过 cGMP-PKG 信号通路发挥作用的，可溶性鸟苷酸环化酶（sGC）、蛋白激酶 G（PKG1）、磷酸二酯酶 5 A（PDE5A）、血管扩张剂刺激的磷蛋白（VASP）和磷酸化 VASP（p-VASP）的表达在 RGS19 基因敲除后发生改变也验证了这一点。这些结果突出表明，RGS19 是 BLCA 的潜在生物标记物和治疗靶点。

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引用次数: 0

miR-103–3p attenuates liver injury with severe acute pancreatitis by inhibiting pyroptosis through miR-103–3p/NLRP1 axis miR-103-3p通过miR-103-3p/NLRP1轴抑制热蛋白沉积，从而减轻重症急性胰腺炎的肝损伤。

IF 2.3 4区生物学 Q4 CELL BIOLOGY

Acta histochemica

Pub Date : 2024-10-30 DOI: 10.1016/j.acthis.2024.152211

Wenquan Zhang , Min Du , Yingjian Jiang , Jiang Wang , Yue Yu , DianLiang Zhang

Background

Severe acute pancreatitis (SAP) is a common digestive system disorder in clinical practice, and it is often associated with liver damage in patients with severe acute pancreatitis. Several studies have indicated that pyroptosis plays a role in liver damage following severe acute pancreatitis (SAP). However, the precise mechanisms remain unclear. This study aims to elucidate the association and specific mechanisms between liver injury following SAP and pyroptosis, providing theoretical support for research on SAP-induced liver injury.

Methods

A rat model of SAP with concomitant liver injury was successfully established. The expression levels of miR-103–3p across different liver tissue groups were quantified using quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). Bioinformatic analyses and dual-luciferase reporter assays confirmed that NLRP1 is a direct target of miR-103–3p. In vivo assessments of miR-103–3p levels were performed, and the extent of cell pyroptosis during liver injury post-SAP was evaluated through western blotting, qRT-PCR, scanning electron microscopy, histopathology, immunofluorescence, and immunohistochemistry. The role of miR-103–3p in regulating NLRP1-mediated pyroptosis and its impact on SAP-induced liver injury were validated.

Results

This study reports that following SAP-induced liver injury, the expression of miR-103–3p in liver tissue was significantly decreased, and cell pyroptosis was involved in the process of liver injury. Experimental validation indicated that NLRP1 was a downstream target of miR-103–3p. Overexpression of miR-103–3p in vitro significantly alleviated the severity of liver injury following SAP, while simultaneously inhibiting cell pyroptosis.

Conclusion

These findings indicate that pyroptosis may be linked to SAP-induced liver injury and that miR-103–3p mitigates hepatocyte pyroptosis by reducing liver injury through the suppression of NLRP1 expression.

背景：重症急性胰腺炎（SAP）是临床上常见的消化系统疾病，重症急性胰腺炎患者往往伴有肝损伤。一些研究表明，热蛋白沉积在重症急性胰腺炎（SAP）后的肝损伤中起一定作用。然而，其确切机制仍不清楚。本研究旨在阐明 SAP 后肝损伤与热蛋白沉积之间的关联和具体机制，为 SAP 诱导肝损伤的研究提供理论支持：方法：成功建立了伴有肝损伤的 SAP 大鼠模型。方法：成功建立了伴有肝损伤的 SAP 大鼠模型，并利用定量反转录聚合酶链反应（qRT-PCR）对不同肝组织组 miR-103-3p 的表达水平进行了定量分析。生物信息学分析和双荧光素酶报告实验证实，NLRP1 是 miR-103-3p 的直接靶标。研究人员对 miR-103-3p 的水平进行了体内评估，并通过 Western 印迹、qRT-PCR、扫描电子显微镜、组织病理学、免疫荧光和免疫组织化学等方法评估了 SAP 后肝损伤过程中细胞热解的程度。研究验证了 miR-103-3p 在调控 NLRP1 介导的热蛋白沉积中的作用及其对 SAP 诱导的肝损伤的影响：结果：本研究发现，SAP 诱导的肝损伤后，肝组织中 miR-103-3p 的表达明显降低，细胞裂解参与了肝损伤的过程。实验验证表明，NLRP1是miR-103-3p的下游靶标。在体外过表达 miR-103-3p 能明显减轻 SAP 损伤后肝损伤的严重程度，同时抑制细胞嗜热：这些研究结果表明，肝细胞脓毒症可能与 SAP 诱导的肝损伤有关，而 miR-103-3p 可通过抑制 NLRP1 的表达减轻肝损伤，从而缓解肝细胞脓毒症。

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引用次数: 0

Ultrastructure and immunohistochemistry of apteric skin in ratites and its epidermal soft cornification 大鼠皮肤的超微结构和免疫组织化学及其表皮软角化。

IF 2.3 4区生物学 Q4 CELL BIOLOGY

Acta histochemica

Pub Date : 2024-10-30 DOI: 10.1016/j.acthis.2024.152213

Lorenzo Alibardi

An electron microscopy and immunohistochemistry study has been conducted to acquire comparative information on the structure of apteric skin in ratites, ostrich and emu. The epidermis is thin in the neck of both species and thicker in the dorsal region where acidic and neutral keratins are present in the viable epidermis and stratum corneum. The dermis in both species is mostly occupied by collagen fibrils that form large bundles, often organized in alternated layers in the deeper part of the dermis. Numerous collagen fibrils contact the basement membrane of the epidermis. Sparse tactile Meissner or Krause sensilli are present among the thick collagen bundles. The ostrich epidermis in the dorsal skin is thicker than in the neck, with a columnar basal layer, 3–5 intermediate suprabasal layers and a thick corneous layer. The epidermis of the neck in emu is very thin, featuring two-three narrow cell layers above a flat basal layer and a relatively thick corneous layer. Basal and suprabasal keratinocytes contain lipid droplets and small keratin bundles but no keratohyalin accumulates in pre-corneous cells. The thin corneocytes form a multilayered corneous layer. Loricrine is present in pre-corneous and corneous layers while CBPs, formerly indicated as beta-keratins, are absent in apteric epidermis.

通过电子显微镜和免疫组织化学研究，我们获得了大鼠、鸵鸟和鸸鹋皮肤结构的比较信息。这两种动物颈部的表皮较薄，背部较厚，酸性和中性角蛋白存在于有活力的表皮和角质层中。两种动物的真皮层大部分被胶原纤维占据，胶原纤维形成大束，通常在真皮深层交替排列。许多胶原纤维与表皮的基底膜接触。在粗大的胶原纤维束中有稀疏的触觉梅斯纳（Meissner）或克劳斯（Krause）感觉器。鸵鸟背部表皮比颈部表皮厚，有一个柱状基底层、3-5个中间基底上层和一个厚角质层。鸸鹋颈部的表皮非常薄，在平坦的基底层和相对较厚的角质层上有两三层狭窄的细胞层。基底层和上基底层角质细胞含有脂滴和小的角质蛋白束，但角质层前细胞中不积聚角质纤维蛋白。薄角质细胞形成多层角质层。角质层前细胞和角质层中含有 Loricrine，而凋亡表皮层中则没有 CBPs（以前称为 beta-角蛋白）。

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引用次数: 0

Section thickness is identical for the sliding microtome and rotary microtome under the continuous cooling device condition 在连续冷却装置条件下，滑动切片机和旋转切片机的切片厚度相同。

IF 2.3 4区生物学 Q4 CELL BIOLOGY

Acta histochemica

Pub Date : 2024-10-24 DOI: 10.1016/j.acthis.2024.152208

Mayuna Fukuzawa, Raia Kushibiki, Yuki Kanehira, Akifumi Ishizawa, Moe Kameda, Sayaka Kobayashi, Yoshimi Nishijima, Masanao Saio

To date, no report has compared section thickness (ST) between the sliding microtome (SM) and rotary microtome (RM). We used the ice pack (IP) condition, in which the paraffin block was not cooled during slicing, and a continuous cooling device (CCD) for continuous cooling during slicing. The ST was greater for the SM than the RM in the IP condition, but it was identical between the devices under the CCD condition. Thus, we used the CCD condition for subsequent studies. In formalin-fixed, paraffin-embedded (FFPE) fish sausage blocks, the ST of the tissue surface (T-surface) was significantly concaved compared to that of the paraffin surface (P-surface) for both microtomes. On the contrary, in FFPE human kidney blocks, ST did not differ between the T-surface and P-surface. Furthermore, the eosin-positive area of PAM-stained specimens was affected by ST, and the color tone of the thickest sample differed from that of the median or thinnest sample. Our data indicated that we should use CCD conditions to ensure that ST is uniform regardless of the type of microtome. In addition, for quantitative analysis, we should utilize ST measure equipment and specimens with a constant ST.

迄今为止，还没有报告比较过滑动切片机（SM）和旋转切片机（RM）的切片厚度（ST）。我们使用了冰袋（IP）条件，即在切片过程中不冷却石蜡块，并使用持续冷却装置（CCD）在切片过程中持续冷却。在 IP 条件下，SM 的 ST 值大于 RM，但在 CCD 条件下，两种装置的 ST 值相同。因此，我们在后续研究中使用了 CCD 条件。在福尔马林固定、石蜡包埋（FFPE）的鱼肠块中，两种显微切片的组织表面（T 表面）的 ST 都比石蜡表面（P 表面）的 ST 明显凹陷。相反，在 FFPE 人肾组织块中，T 表面和 P 表面的 ST 没有差异。此外，PAM 染色样本的伊红阳性面积也受 ST 的影响，最厚样本的色调与中位样本或最薄样本的色调不同。我们的数据表明，无论使用哪种显微切片机，我们都应使用 CCD 条件来确保 ST 的一致性。此外，为了进行定量分析，我们应该使用 ST 测量设备和具有恒定 ST 的样本。

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引用次数: 0

Histological changes in skeletal muscle induced by heart failure in human patients and animal models: A scoping review 人类患者和动物模型心力衰竭引起的骨骼肌组织学变化：范围综述。

IF 2.3 4区生物学 Q4 CELL BIOLOGY

Acta histochemica

Pub Date : 2024-10-23 DOI: 10.1016/j.acthis.2024.152210

Akinori Kaneguchi , Naoyoshi Sakitani , Takuya Umehara

Objective

This scoping review aimed to characterize the histological changes in skeletal muscle after heart failure (HF) and to identify gaps in knowledge.

Methods

On April 03, 2024, systematic searches were performed for papers in which histological analyses were conducted on skeletal muscle sampled from patients with HF or animal models of HF. Screening and data extraction were conducted by two independent authors.

Results and conclusion

A total of 118 papers were selected, including 33 human and 85 animal studies. Despite some disagreements among studies, some trends were observed. These trends included a slow-to-fast transition, a decrease in muscle fiber size, capillary to muscle fiber ratio, and mitochondrial activity and content, and an increase in apoptosis. These changes may contribute to the fatigability and decrease in muscle strength observed after HF. Although there were some disagreements between the results of human and animal studies, the results were generally similar. Animal models of HF will therefore be useful in elucidating the histological changes in skeletal muscle that occur in human patients with HF. Because the muscles subjected to histological analysis were mostly thigh muscles in humans and mostly lower leg muscles in animals, it remains uncertain whether changes similar to those seen in lower limb (hindlimb) muscles after HF also occur in upper limb (forelimb) muscles. The results of this review will consolidate the current knowledge on HF-induced histological changes in skeletal muscle and consequently aid in the rehabilitation of patients with HF and future studies.

目的本综述旨在描述心力衰竭（HF）后骨骼肌组织学变化的特征，并找出知识空白：2024年4月3日，系统检索了对心力衰竭患者或心力衰竭动物模型骨骼肌进行组织学分析的论文。筛选和数据提取由两位独立作者完成：共筛选出 118 篇论文，包括 33 项人体研究和 85 项动物研究。尽管各研究之间存在一些分歧，但还是观察到了一些趋势。这些趋势包括从慢到快的转变，肌肉纤维尺寸、毛细血管与肌肉纤维比率、线粒体活性和含量的减少，以及细胞凋亡的增加。这些变化可能是导致高频后易疲劳和肌力下降的原因。尽管人类和动物研究结果之间存在一些分歧，但总体上结果相似。因此，心房颤动动物模型将有助于阐明人类心房颤动患者骨骼肌组织学上的变化。由于接受组织学分析的肌肉在人类中大多是大腿肌肉，而在动物中大多是小腿肌肉，因此目前仍不确定高血脂后下肢（后肢）肌肉是否也会发生类似于上肢（前肢）肌肉的变化。本综述的结果将巩固目前关于高频诱导的骨骼肌组织学变化的知识，从而有助于高频患者的康复和未来的研究。

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引用次数: 0

The elastic system: A review of elastin-related techniques and hematoxylin-eosin/phloxine applicability for normal and pathological tissue description 弹性系统：弹性蛋白相关技术及苏木精-伊红/荧光素在正常和病理组织描述中的适用性综述。

IF 2.3 4区生物学 Q4 CELL BIOLOGY

Acta histochemica

Pub Date : 2024-10-22 DOI: 10.1016/j.acthis.2024.152209

Thalles Fernando Rocha Ruiz , Luara Jesus Ferrato , Lorena Gabriela de Souza , Gervásio Evangelista Brito-Filho , Ellen Cristina Rivas Leonel , Sebastião Roberto Taboga

The elastic system is one of the most developed interstitial elements in connective tissue. With diverse functions, pre-elastic and elastic fibers contribute to the distensibility and malleability of several organs. Also, microanalyses of the elastic system were obtained by different histological techniques that were employed over years to describe normal and pathological conditions. Compared to conventional stains, hematoxylin-eosin/phloxine (HE/P) under fluorescence and confocal microscopy presented a highly detailed observation of the elastic system in different organs and scenarios. This technique provides a better demarcation of the elastic fibers, favoring their description in relation to their deposition and aggregation in different organs. Also, fibrils with low aggregation or loss of this characteristic are observed in an optimal view in the skin, heart valves, and large-caliber blood vessels. Degradation, fragmentation, and rupture were also well described by the HE/P technique. Several organs, such as the mammary gland, prostate, skin, aorta, and lung, could be described with precision under this technique. In association with non-linear microscopy, the results of the research presented in this paper improved and detailed characteristics of precise pathogenesis. Thus, the HE/P technique presented an interesting efficiency to demonstrate alterations and structures in which the elastic system showed a relevant role, and when compared to other techniques it demonstrated a similar or better result. In addition, it is expected that future studies can reveal more information about the elastin and interactions with specific dyes, thus allowing a greater understanding of the great efficiency of this technique.

弹性系统是结缔组织中最发达的间隙元素之一。前弹力纤维和弹力纤维具有多种功能，对多个器官的伸缩性和延展性做出了贡献。多年来，人们采用不同的组织学技术对弹性系统进行微观分析，以描述正常和病理状态。与传统染色法相比，荧光显微镜和共聚焦显微镜下的苏木精-伊红/荧光素（HE/P）可对不同器官和情况下的弹性系统进行非常详细的观察。这种技术能更好地划分弹性纤维，有利于描述它们在不同器官中的沉积和聚集情况。此外，在皮肤、心脏瓣膜和大口径血管中，还能以最佳视角观察到低聚集或失去这种特性的纤维。HE/P 技术对降解、碎裂和断裂也有很好的描述。乳腺、前列腺、皮肤、主动脉和肺等多个器官都能通过该技术精确描述。结合非线性显微镜，本文介绍的研究结果改进了精确发病机制的详细特征。因此，HE/P 技术在展示弹性系统在其中发挥相关作用的改变和结构方面具有令人感兴趣的效率，与其他技术相比，它显示出类似或更好的结果。此外，未来的研究有望揭示更多有关弹性蛋白以及与特定染料相互作用的信息，从而让人们更深入地了解这种技术的巨大功效。

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引用次数: 0

Neuronal splicing regulator RBFOX3 (NeuN) distribution and organization are modified in response to monosodium glutamate in rat brain at postnatal day 14 出生后第14天大鼠脑内神经元剪接调节因子RBFOX3（NeuN）的分布和组织在谷氨酸钠的作用下发生改变。

IF 2.3 4区生物学 Q4 CELL BIOLOGY

Acta histochemica

Pub Date : 2024-10-19 DOI: 10.1016/j.acthis.2024.152207

Anaís Monzerrat García Juárez, Nidia Jannette Carrillo González, Tania Campos-Ordoñez, Yadira Gasca Martínez, Graciela Gudiño-Cabrera

Neuronal splicing regulator RNA binding protein, fox-1 homolog 3 (NeuN/RbFox3), is expressed in postmitotic neurons and distributed heterogeneously in the cell. During excitotoxicity events caused by the excess glutamate, several alterations that culminate in neuronal death have been described. However, NeuN/RbFox3 organization and distribution are still unknown. Therefore, our objective was to analyze the nucleocytoplasmic distribution and organization of NeuN/RbFox3 in hippocampal and cortical neurons using an excitotoxicity model with monosodium glutamate salt (MSG). We used neonatal Wistar rats administered subcutaneously with 4 MSG mg/kg during the postnatal day (PND) 1, 3, 5, and 7. The control group was rats without MSG administration. On 14 PND, the brain was removed, and coronal sections were used for immunodetection with the antibody NeuN, DAPI, and the propidium iodide staining for histological evaluation. The results indicate that in the control group, NeuN/RbFox3 was organized into macromolecular condensates inside and outside the nucleus, forming defined nuclear compartments. Additionally, NeuN/RbFox3 was distributed proximal to the nucleus in the cytoplasm. In contrast, in the group treated with MSG, the distribution was diffuse and dispersed in the nucleus and cytoplasm without the formation of compartments in the nucleus. Our findings, which highlight the significant impact of MSG administration in the neonatal period on the distribution and organization of NeuN/RbFox3 of neurons in the hippocampus and cerebral cortex, offer a new perspective to investigate MSG alterations in the developmental brain.

神经元剪接调节器 RNA 结合蛋白 fox-1 同源物 3（NeuN/RbFox3）在有丝分裂后的神经元中表达，并在细胞中异质性分布。在由过量谷氨酸引起的兴奋性中毒事件中，已描述了几种最终导致神经元死亡的改变。然而，NeuN/RbFox3 的组织和分布仍不为人知。因此，我们的目的是利用谷氨酸一钠（MSG）兴奋毒性模型，分析 NeuN/RbFox3 在海马和皮层神经元中的核细胞质分布和组织。我们使用新生 Wistar 大鼠，在出生后第 1、3、5 和 7 天皮下注射 4 毫克/千克 MSG。对照组为未注射味精的大鼠。出生后第 14 天，取出大鼠大脑，用 NeuN 抗体、DAPI 和碘化丙啶染色对冠状切片进行免疫检测，并进行组织学评估。结果表明，在对照组中，NeuN/RbFox3在细胞核内外组织成大分子凝聚体，形成明确的核区。此外，NeuN/RbFox3 还分布在细胞质中核的近端。相比之下，在用味精处理的组中，NeuN/RbFox3呈弥漫性分布，分散在细胞核和细胞质中，没有在细胞核中形成小室。我们的研究结果突显了新生儿期服用味精对海马和大脑皮层神经元NeuN/RbFox3的分布和组织的重要影响，为研究味精在大脑发育过程中的改变提供了一个新的视角。

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引用次数: 0