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Modulation of Neurogenesis by Novel Natural or Synthetic Compound S

Thesis Info

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Author

Majeed, Saba

Program

PhD

Institute

University of Karachi

City

Karachi

Province

Sindh

Country

Pakistan

Thesis Completing Year

2018

Thesis Completion Status

Completed

Subject

Pharmacology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/11208/1/Saba%20Majeed_Pharmacology_2018_UoK_PRR.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726667773

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The discovery of novel molecules capable of modulating neurogenesis will contribute to explore novel therapeutic approaches and to clarify the physiological role of neurogenesis. Development of small molecules can be targeted to the regulation of progenitor cell proliferation, survival, migration or maturation and might be applied to augmenting physiological neurogenesis already present in the dentate gyrus or subventricular zone/olfactory bulb or to normally non-neurogenic regions important for neuropathological conditions. In the present study, in vitro model for neurogenesis was developed by isolation of neonatal rat hippocampal and cortical cells. These cells were both cultured independently and with bone marrow stromal cells (BMSCs). The effect of Isoxylitones (ISOX) during proliferation, differentiation and survival of the cultured cells under normal and hypoxic condition was studied. A dose dependent increase in cell viability of treated cells was observed in alamarBlue and MTT assays for proliferation. BrdU cell proliferation assay for growth of newly born cells was also performed and significantly higher cell population was recorded in the treated groups. To study the effect of ISOX on recovery of hypoxic cells, the cortical and hippocampal cells were cultured and given hypoxic shock followed by ISOX treatment. A significant recovery effect was seen which ensures its capability to modulate neuronal cell survival. The markers associated with neuronal lineage were analyzed to study the stage-specific neuronal differentiation and functional maturation of ISOX treated cultures. Themarkers included in the present study were nestin, β-tubulin III, GFAP and calretinin. In addition, transcription factors markers, NeuroD1 and Neurogenin-2, were used to analyze the induction of differentiation and activating factors for neurogenesis. The immunocytochemical analysis of above mentioned markers suggests that ISOX is involved in enhancement of neural progenitor cells as nestin expression was increased. The ISOX also supported the neuronal differentiation through inducing NeuroD1 and Neurogenin-2 transcription factors. The cocultured hippocampal and cortical cells with or without ISOX treatment demonstrated the generation of neuronal cells with marked increase of nestin, tubulin and calretinin expression. Additionally a significant induction of transcription factors NeuroD1 and Neurogenin-2 was also observed in cocultured treated groups. Based on our findings, we conclude that ISOX have a potential to support cortical and hippocampal cells survival and induction of transcription factors NeuroD1 and Neurogenin-2 and neuronal protein markers (nestin tubulin, calretinin) may provide cell stage-specific molecular and cellular mechanistic clues for understanding the potential effects of ISOX on the neurodevelopment. Our results regarding cocultured cells demonstrated the successful differentiation of BMSCs into neuronal like cells by co-culturing with hippocampal and cortical cells. Additionally the ISOX have capability to potentiate this differentiation of neuronal cells in cocultures. In short its cell survival and differential effects can be utilized for therapeutic rationale in neurodegenerative disorders.
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مولانا حسرت موہانی

مولانا حسرتؔ موہانی
افسوس کہ آخر مولانا حسرت موہانی بھی چل بسے۔مولانا کی شخصیت کا پیکر دو چیزوں سے بنا تھاایک شعر وسخن اوردوسری سیاست۔سیاست اس پیکر کے ساتھ جسم کی نسبت رکھتی تھی، اس بناپر جب جسم مٹی میں ملاتوسیاست بھی فناہوگئی لیکن شعر و سخن اس پیکر کی روح تھی جو مرنے کے بعد باقی رہتی ہے اس لیے حسرت کی شاعری اب بھی زندہ ہے اورزندہ رہے گی۔
مرحوم سیاست میں کبھی ایک روش میں قایم نہیں رہے وہ کبھی کسی پارٹی میں شریک ہوئے کبھی کسی میں،ان کی سیاست کاآغاز کانگریس میں شرکت سے ہوا اور اس کاخاتمہ لیگ کے پُرجوش کارکن ہونے پر ہوگیا۔ان دونوں کی درمیانی مدت میں سیاسی اعتبار سے وہ کبھی کسی روپ میں نظر آئے اورکبھی کسی جامہ میں وہ دیکھے گئے لیکن ہرجگہ اورہرمقام پربیباک خلوص ان کا امتیازی وصف رہا۔ یہی وجہ ہے کہ جن لوگوں سے وہ سیاسی اختلاف رائے رکھتے تھے وہ بھی ان کی قدر کرتے اور ان کا احترام ملحوظ رکھتے تھے، وہ خواہ کسی رنگ اورکسی بھیس میں ہوتے ان کا اندازِ قدالگ سے الگ پہچان لیا جاتاتھا۔ ملک کی جدوجہد آزادی میں ان کا اتنا بڑا حصہ ہے کہ اس جدوجہد کی کوئی تاریخ مرحوم کے شاندار تذکرہ کے بغیر کامل نہیں ہوسکتی۔ ایک زمانہ تھا کہ حسرت کانام بچہ بچہ کی زبان پر تھا اورلوگ ان کے ایثار و قربانی،محنت وجفاکشی،برطانوی حکومت سے نفرت اور اس سلسلہ میں ان کی سخت ضداورہٹ کی داستانیں مزے لے لے کر اورجوش ومسرت کے ساتھ بیان کرتے تھے، لیکن مرحوم کے یہ وہ اوصاف وکمالات ہیں جن کو لوگوں نے خود ان کی زندگی میں ہی بھلا دیاتھا اوروہ آخر میں’’یوسف بے کارواں‘‘ہوکررہ گئے تھے۔
حسرت کی شاعری جوانمٹ اورزوال ناآشنا ہے اس کا اصل جوہر حسن تغزل ہے۔ انھوں نے اپنے...

Concept of Tolerance in Islam for Promotion of Human Unity

Islam is a divine religion. It is based on divine revelation and Sunnah of the Holy prophet (SAW). Tolerance, patience, forbearing and broadmindedness are the hallmarks of the last divine religion, Islam. In the article the ideology of co-existence has been discussed. Verses of the Holy book indicate importance of the tolerance. Similarly some events out of the Seerah of the Holy prophet (SAW) have been mentioned as references. In these pages it is impossible to explain scope of the subject; however efforts have been made to elaborate the Islamic perspective of tolerance. May Allah Almighty enable all of us to follow teachings of Islam in true spirit.

Studies on Regulation of Intronic Polyadenylation by Rna Helicase P68/Ddx5 and U1snrnp in Mcf7 Cell Line

In eukaryotes, pre-mRNA has to undergo different processing steps; addition of cap at 5′ end, removal of introns to join functional exons, and addition of poly(A) tail at 3′ end to become fully functional mRNA. These processing events are linked to each other; one process effects efficiency of the other. Addition of polyA tail is not only limited at 3` end but poly A sequences are also found in genes (within exons and introns). The use of one of these intronic PAS results in primary transcript with different 3`UTR. DDX5 (p68) and DDX17 (p72) are RNA helicases performing different cellular functions; mi-RNA processing, transcription, mRNA processing, cell proliferation/transformation, cellular development and cancer. Four human genes MET, BCCIP, TGM2, and SMAD2 were selected to determine the relationship between p68, U1 snRNP, and activation of intronic polyadenylation. Genes were cloned into pGL3-TK vector having Firefly luciferase reporter gene. Mutation was introduced in 5`splice site to block U1. MCF7 cells were transfected with si-RNAp68/p72. After 24 hours cells were co-transfected with WT and Mut plasmids and pRL-SV40 control vector. Expression level of short isoform was determined by Dual Luciferase Reporter assay. The results suggest the role of p68 in IPA activation. Quantitative PCR was performed on uncleaved/total mRNA that confirmed the role of p68 RNA helicase in IPA acting through U1snRNP. To exclude the possibility that IPA is activated by splicing inhibition, si-RNAs against two splicing factors were used. If competitive inhibition of splicing result in IPA activation then should get the same expression level of short isoform with both si-RNAs, but it was not the case. IPA activation was seen only after si-U1 70k treatment while no or little short isoform was observed after si-U2AF65. These results clearly prove that IPA activation is not related to splicing inhibition. Overexpression of p68 and p72 enhanced the IPA event, again confirming the role of p68 in IPA. All experimental results prove that p68 activates intronic polyadenylation by removing U1 from 5`ss. Influence of p68 on IPA is not direct but it is acting through U1.