The roles of intraflagellar transport (IFT) protein 25 in mammalian signaling transduction and flagellogenesis / 亚洲男科学杂志(英文版)

Cilium, an organelle with a unique proteome and organization, protruding from the cell surface, generally serves as a force generator and signaling compartment. During ciliogenesis, ciliary proteins are synthesized in cytoplasm and transported into cilia by intraflagellar transport (IFT) particles, where the inner counterparts undergo reverse trafficking. The homeostasis of IFT plays a key role in cilial structure assembly and signaling transduction. Much progress has been made on the mechanisms and functions of IFT; however, recent studies have revealed the involvement of IFT particle subunits in organogenesis and spermatogenesis. In this review, we discuss new concepts concerning the molecular functions of IFT protein IFT25 and how its interactions with other IFT particle subunits are involved in mammalian development and fertility.

Research progress on the distribution of primary cilia and related signaling pathways involved in odontogenesis / 口腔疾病防治

@#Primary cilia are organelles present on most mammalian cells that sense environmental changes and transduce signaling, and they are the key coordinators of various signaling pathways during tissue development. This article reviews the progress of research on the distribution of primary cilia in tooth development and the related signaling pathways. A literature review shows that in odontogenesis, primary cilia play an important role in the mutual induction of the epithelium and mesenchyme; during the continuous proliferation and differentiation of cells, the distribution of primary cilia is temporally and spatially dependent. Although the reason for this distribution is still unclear, some experimental evidence indicates that this phenomenon is compatible with the function of cells and tissues in which primary cilia are distributed. Primary cilia are involved in the regulation of two important signaling pathways, Hedgehog and Wnt, in odontogenesis. Genes encoding cilia (such as Kif3a, Evc/Evc2 and Ift) can affect the development of teeth by regulating these two signaling pathways, and there is an interaction between the two signaling pathways. Deletion of related genes (such as Ofd1 and Bbs) can damage the transmission of upstream and downstream signals by damaging the structure or function of cilia, thereby causing various types of dental dysplasia, including small teeth, enamel hypoplasia, missing teeth, or craniofacial deformities.

Comparison Of The Effectiveness Of Transcutaneous Electrical Nerve Stimulation (TENS) Vs. Interferential Therapy (IFT) For Relief Of Pain In Primary Dysmenorrhea

Background: Dysmenorrhea or pain during menstruation is commonly seen in young women and largely impacts their quality of life. Pain medications are largely used to tackle this situation; however, they have side effects with regular use. Safe, effective pain-relieving modalities are crucial to help women cope with this situation. There is extensive literature proving the effectiveness of TENS for relieving the pain in primary dysmenorrhea; however, literature proving the same for IFT is sparse. Thus this research aims to investigate and compare the effectiveness of both the modalities in primary dysmenorrhea. Methods: 30 samples(17-25) were included in this study. Using the Simple Random Sampling method, the subjects were divided into two groups. One group received therapy through TENS(80-120 Hz, 20mins, Intensity as per tolerance) and the other through IFT(4PV,80-150HZ, 20 mins, Intensity as per tolerance). Electrical stimulation was given in the region of maximal pain. The Intensity of pain before and after treatment was recorded using the Numerical Pain Scale. The duration of pain relief and the need for analgesic post the therapy was also noted. Results: TENS and IFT are both extremely effective for the relief of pain in primary dysmenorrhea (P<0.0001). There is no significant difference in their effect on the comparison (P=0.422). And most of the subjects in both groups did not need analgesics after therapy. Conclusion: Both TENS and IFT are equally effective in the relief of pain in primary dysmenorrhea. However, tolerance to IFT currents is better among young women as they are medium frequency currents.

Bardet-Biedl syndrome protein-8 is involved in flagellar membrane protein transport in Chlamydomonas reinhardtii / 生物工程学报

Cilia and flagella on eukaryotic cells are polarized organelles extending from the surfaces of cells, which participate not only in cell motility, but also in signal transduction and other processes. Structural or functional abnormalities of cilia can cause various human diseases, termed ciliopathies. Bardet-Biedl syndrome (BBS) is a ciliopathic human genetic disorder, and the pathogenesis is that mutated BBS genes result in abnormal cilia function. In order to study the pathogenic genes BBS8, we screened bbs8 mutant in Chlamydomonas reinhardtii and did a lot of physiology and biochemistry experiments. We affirmed that BBS8 protein was a cilia protein and had specific localization in the basal body by immunofluorescence (IF). The bbs8 mutant lost photokinesis, and it was defective in flagella shortening with drug induction. The results of silver staining and mass spectrometric analysis showed aberrant accumulation of flagellar proteins in the mutant flagella. We concluded that the BBS8 protein plays a significant role in flagellar membrane proteins transport, and the BBS8 protein might mediate retrograde transport to exert physiological function in the process.

Prokaryotic expression and purification of Chlamydomonas reinhardtii intraflagellar transport protein 46(IFT46) and preparation of polyclonal antibody / 生物工程学报

IFT46 is one of the important components of intraflagellar transport complex B in Chlamydomonas reinhardtii, and plays important roles in the assembly, movement and perception of ciliary. To study its functional mechanism, a GST-tagged and an MBP-tagged prokaryotic expression plasmid, pGEX-2T-ift46 and pMAL-C2X-ift46 were constructed, respectively, by inserting ift46 into the pGEX-2T and pMAL-C2X vector, and then transformed into Escherichia coli BL21 (DE3) for protein expression. SDS-PAGE (15%) analysis results showed that the molecular weights of the fusion protein GST-IFT46 and MBP-IFT46 were 70 kDa and 86 kDa, respectively. We used the fusion protein GST-IFT46 purified by affinity adsorption purification (more than 95% purity) for immunity to New Zealand white rabbits. The 5th immune serum was collected and the antibody titer was determined to be 256 000 by ELISA. The antiserum was purified by Protein A affinity adsorption purification and immobilized MBP-IFT46 purification, and the specificity of polyclonal antibodies was evaluated by Western blotting and immunofluorescence. Results showed that the polyclonal antibody prepared could specifically and precisely bind IFT46 in C. reinhardtii, and IFT46 was mainly concentrated at basal body regions and few localized along the entire length of the flagellum as punctuated dots, which will make a foundation to further study the mechanism of IFT46 in cilia related diseases such as obesity, diabetes and polycystic kidney disease.

The effects of IFT80 proteins on tumors / 中国肿瘤临床

Objective:To investigate Intraflagellar Transport 80 (IFT80) protein expression in bone, lung, pancreatic, stomach, in-testinal, prostate, breast, and ovarian cancers to explore its mechanism in cancer cell proliferation and to diagnose and identify new tar-gets in cancer treatment. Methods:Immunohistochemistry was used to investigate the expression of IFT80 in gastric cancer tissue of different stages and in eight other kinds of human cancer tissues. We studied the relationship between cancer cell proliferation and inhi-bition of IFT80. Immunofluorescence method and cell culture were used to study the cilia and IFT80. Results:Results showed the fol-lowing:a) the expression of IFT80 was high in gastric and lung carcinoma tissues, moderate in breast and colorectal cancers, low in bone and ovarian cancers, and nearly absent in prostate and pancreatic cancers;b) inhibition of IFT80 in the A549 cancer cell line accel-erated cell proliferation and resulted in shorter, lower quality cilia;and c) IFT80 was abundantly expressed in cancer tissues of well-dif-ferentiated stage-IIA gastric cancer and normal gastric tissues, but was hardly expressed in late-stage, poorly differentiated gastric can-cer. IFT80 could have various degrees of expression in gastric carcinoma of other stages and differentiation. Conclusions:Different can-cer organs showed variation in IFT80 expression. IFT80 can be distributed in the organs with mechanical motion function, such as lungs and stomach. IFT80 is distributed on the cell cilia and can adjust the number and length of the cilia by reducing IFT80 protein ex-pression. Through a variety of ways, IFT80 directly or indirectly participates in the proliferation of cancer cells. Thus, the lowest or nearly zero expression of IFT80 can be seen in cancer tissues of high-grade malignancy, such as advanced cancers with poor differentia-tion.