scholarly journals L1 Retrotransposons Are Transcriptionally Active in Hippocampus of Rat Brain

2016 ◽  
Vol 117 (1) ◽  
pp. 42-53 ◽  
Author(s):  
Somnath Mukherjee ◽  
Deepak Sharma ◽  
Kailash C. Upadhyaya

LINE1 (L1) is an autonomous, non-LTR retrotransposon and the L1 family of retrotransposons constitute around 17%, 20% and 23% in the human, mouse and rat genomes respectively. Under normal physiological conditions, the retroelements remain by and large transcriptionally silent but are activated in response to biotic and abiotic stress conditions and during perturbation in cellular metabolism. They have also been shown to be transiently activated under certain developmental programs. Using RT-PCR, we show that the L1 elements are transcriptionally active in the hippocampus region of the brain of four-month-old rat under normal conditions without any apparent stress. Twenty non-redundant LINE1-specific reverse transcriptase (RTase) sequences form ORF2 region were isolated, cloned and sequenced. Full length L1 element sequences complementary to the isolated sequences were retrieved from the L1 database. In silico analysis was used to determine the presence of these retroelements proximal (up to 10 kb) to the genes transcriptionally active in the hippocampus. Many important genes were found to be in close proximity of the transcriptionally active L1 elements. Transcriptional activation of the elements possibly affects the expression of the neighbouring genes.

2020 ◽  
Vol 6 (2) ◽  
pp. 205511692094147
Author(s):  
Christopher Hoey ◽  
George Nye ◽  
Angela Fadda ◽  
Janet Bradshaw ◽  
Emi N Barker

Case summary A 7-month-old Siberian cat was presented for investigation of acute onset multifocal neurological deficits. Neurological examination documented dull mental status and an ambulatory left hemiparesis. Serum biochemistry documented marked hyperglobulinaemia. MRI of the brain identified marked leptomeningeal contrast enhancement extending along the brainstem caudally to involve the cranial cervical spinal cord. MRI of the cervical spine further identified a subarachnoid diverticulum that extended from the level of the obex to the C2–C3 vertebrae. Cerebrospinal fluid quantitative RT-PCR was positive for the presence of feline coronavirus. Histopathology revealed pyogranulomatous meningitis and choroid plexitis, uveitis and nephritis. Relevance and novel information This article describes the first reported case of a subarachnoid diverticulum associated with feline infectious peritonitis.


Genetics ◽  
2000 ◽  
Vol 155 (3) ◽  
pp. 1105-1117 ◽  
Author(s):  
W John Haynes ◽  
Kit-Yin Ling ◽  
Robin R Preston ◽  
Yoshiro Saimi ◽  
Ching Kung

Abstract Pawn mutants of Paramecium tetraurelia lack a depolarization-activated Ca2+ current and do not swim backward. Using the method of microinjection and sorting a genomic library, we have cloned a DNA fragment that complements pawn-B (pwB/pwB). The minimal complementing fragment is a 798-bp open reading frame (ORF) that restores the Ca2+ current and the backward swimming when expressed. This ORF contains a 29-bp intron and is transcribed and translated. The translated product has two putative transmembrane domains but no clear matches in current databases. Mutations in the available pwB alleles were found within this ORF. The d4-95 and d4-96 alleles are single base substitutions, while d4-662 (previously pawn-D) harbors a 44-bp insertion that matches an internal eliminated sequence (IES) found in the wild-type germline DNA except for a single C-to-T transition. Northern hybridizations and RT-PCR indicate that d4-662 transcripts are rapidly degraded or not produced. A second 155-bp IES in the wild-type germline ORF excises at two alternative sites spanning three asparagine codons. The pwB ORF appears to be separated from a 5′ neighboring ORF by only 36 bp. The close proximity of the two ORFs and the location of the pwB protein as indicated by GFP-fusion constructs are discussed.


2017 ◽  
Vol 123 (6) ◽  
pp. 1532-1544 ◽  
Author(s):  
Thomas M. Langer ◽  
Suzanne E. Neumueller ◽  
Emma Crumley ◽  
Nicholas J. Burgraff ◽  
Sawan Talwar ◽  
...  

Neuromodulator interdependence posits that changes in one or more neuromodulators are compensated by changes in other modulators to maintain stability in the respiratory control network. Herein, we studied compensatory neuromodulation in the hypoglossal motor nucleus (HMN) after chronic implantation of microtubules unilaterally ( n = 5) or bilaterally ( n = 5) into the HMN. After recovery, receptor agonists or antagonists in mock cerebrospinal fluid (mCSF) were dialyzed during the awake and non-rapid eye movement (NREM) sleep states. During day studies, dialysis of the µ-opioid inhibitory receptor agonist [d-Ala2, N-MePhe4, Gly-ol]enkephalin (DAMGO; 100 µM) decreased pulmonary ventilation (V̇i), breathing frequency ( f), and genioglossus (GG) muscle activity but did not alter neuromodulators measured in the effluent mCSF. However, neither unilateral dialysis of a broad spectrum muscarinic receptor antagonist (atropine; 50 mM) nor unilateral or bilateral dialysis of a mixture of excitatory receptor antagonists altered V̇i or GG activity, but all of these did increase HMN serotonin (5-HT) levels. Finally, during night studies, DAMGO and excitatory receptor antagonist decreased ventilatory variables during NREM sleep but not during wakefulness. These findings contrast with previous dialysis studies in the ventral respiratory column (VRC) where unilateral DAMGO or atropine dialysis had no effects on breathing and bilateral DAMGO or unilateral atropine increased V̇i and f and decreased GABA or increased 5-HT, respectively. Thus we conclude that the mechanisms of compensatory neuromodulation are less robust in the HMN than in the VRC under physiological conditions in adult goats, possibly because of site differences in the underlying mechanisms governing neuromodulator release and consequently neuronal activity, and/or responsiveness of receptors to compensatory neuromodulators. NEW & NOTEWORTHY Activation of inhibitory µ-opioid receptors in the hypoglossal motor nucleus decreased ventilation under physiological conditions and did not affect neurochemicals in effluent dialyzed mock cerebral spinal fluid. These findings contrast with studies in the ventral respiratory column where unilateral [d-Ala2, N-MePhe4, Gly-ol]enkephalin (DAMGO) had no effects on ventilation and bilateral DAMGO or unilateral atropine increased ventilation and decreased GABA or increased serotonin, respectively. Our data support the hypothesis that mechanisms that govern local compensatory neuromodulation within the brain stem are site specific under physiological conditions.


2011 ◽  
Vol 14 (4) ◽  
pp. 573-577 ◽  
Author(s):  
J. Niczyporuk ◽  
E. Samorek-Salamonowicz ◽  
W. Kozdruń ◽  
Z. Mizak

The survey of wild birds for West Nile virus in PolandTwo thousand one hundred and forty birds belonging to 39 different species from different locations in Poland were examined. The study has taken place from the early spring till late autumn 2007-2010 when the activity of the mosquitoes was the highest. The brain samples were taken from the birds and whole cellular RNA was isolated, then the RT-PCR and NRT-PCR were performed to detect the presence of West Nile virus (WNV). The obtained results were confirmed by the commercial WNV Kit. No genetic material of WNV was found in the examined samples.


1999 ◽  
Vol 12 (7) ◽  
pp. 592-603 ◽  
Author(s):  
Corinne Mhiri ◽  
Pierre J. G. M. De Wit ◽  
Marie-Angèle Grandbastien

The copia-like Tnt1 element of tobacco is one of the few active plant retrotransposons and is transcriptionally activated, in tobacco and in heterologous species, by biotic and abiotic stress factors. In order to establish more precisely the link between Tnt1 activation and plant defense responses, the expression of the Tnt1 promoter was studied in a gene-for-gene pathosystem, the interaction between tomato and the fungal pathogen Cladosporium fulvum. In compatible interactions, Tnt1 expression is highly induced throughout the leaf regions colonized by the fungus, while in incompatible interactions Tnt1 induction is transient and localized in distinct foci. Tnt1 expression after fungal inoculation parallels the differential activation of tomato defense genes. Tnt1 expression is induced by nonspecific factors of plant or fungal origin present in apoplastic fluids of leaf tissues infected by virulent races of C. fulvum, but is also activated by specific factors resulting from the interaction between fungal avirulence peptides and plant resistance genes. Tnt1 activation by apoplastic fluids containing avirulence peptides of C. fulvum is detected soon after elicitation. These results demonstrate that Tnt1 transcriptional activation correlates with biological responses of tomato to infection by C. fulvum and is mediated through signals originating from both race-specific and non-race-specific perception pathways.


Stroke ◽  
2021 ◽  
Vol 52 (Suppl_1) ◽  
Author(s):  
tao wang ◽  
Guokun Zhou ◽  
mingdi he ◽  
yuanyuan xu ◽  
w.g. Rusyniak ◽  
...  

Introduction: Acidosis is one prevalent phenomenon in ischemic stroke. The literature has shown that protons directly gate acid-sensing ion channels (ASICs) and proton-activated chloride channel, both lead to neuronal injury However, it is unclear whether protons activate metabotropic pathways in brain neurons. There are four proton-sensitive G-protein coupled receptors (GPCRs): GPR4, GPR65, GPR68 and GPR132. It remains unknown whether any of these GPCRs mediate acid-induced signals in brain neurons or whether they contribute to ischemia-induced brain injury. Methods: Total RNA from human cortical tissue or mouse brain was isolated using TRIzol and an RNase Kit. Standard RT-PCR was performed to determine the expression of these GPCRs in the brain. An in vitro slice injury model was used for functional screening. To determine the effect of ischemia, WT and knockout male mice were subjected to MCAO. To study brain injury, brains were sectioned coronally at 1 mm intervals and stained by vital dye immersion: (2%) 2,3,5-triphenyltetrazolium hydrochloride (TTC). Locomotor analysis and corner test were used to assess behavior outcome. Adeno-associated virus (AAV) -mediated gene delivery was used to determine the outcome of GPR68 overexpression. Results: RT-PCR showed that brain tissue expressed GPR4, -65, and -68. The expression of GPR68 was evident at 30 cycles. In organotypic slices, compared to the WT, deleting GPR4 or GPR65 had no effect while deleting GPR68 significantly increased acidosis-induced neuronal injury. At both 24 hour and 72 hour after 45 minutes MCAO, GPR68 deletion increased brain injury (p=0.0020 for 24hour, p=0.0392 for 72hour, Mann-Whitney U test). WT and GPR68-/- mice did not differ in baseline locomotor activities or corner test. On the third day following MCAO, GPR68-/- exhibited significantly more left rotations (p=0.0287, Mann-Whitney U test) than WT animals. Lastly, mice receiving AAV-GPR68 exhibited an average infarct of 21.97 ± 12.4%, significantly (p = 0.0022, Mann-Whitney U test) smaller than those receiving AAV-GFP (37.2 ± 6.8%). Conclusion: These data showed that GPR68 functions as a neuroprotective proton receptor in the brain.


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Dongjiao Wang ◽  
Ling Wang ◽  
Weihua Su ◽  
Yongjuan Ren ◽  
Chuihuai You ◽  
...  

AbstractWRKY transcription factors play significant roles in plant stress responses. In this study, a class III WRKY gene ScWRKY5, was successfully isolated from sugarcane variety ROC22. The ScWRKY5 was a nucleus protein with transcriptional activation activity. The ScWRKY5 gene was constitutively expressed in all the sugarcane tissues, with the highest expression level in the stem epidermis and the lowest in the root. After inoculation with Sporisorium scitamineum for 1 d, the expression level of ScWRKY5 was significantly increased in two smut-resistant varieties (YZ01-1413 and LC05-136), while it was decreased in three smut-susceptible varieties (ROC22, YZ03-103, and FN40). Besides, the expression level of ScWRKY5 was increased by the plant hormones salicylic acid (SA) and abscisic acid (ABA), as well as the abiotic factors polyethylene glycol (PEG) and sodium chloride (NaCl). Transient overexpression of the ScWRKY5 gene enhanced the resistance of Nicotiana benthamiana to the tobacco bacterial pathogen Ralstonia solanacearum, however the transiently overexpressed N. benthamiana was more sensitive to the tobacco fungal pathogen Fusarium solani var. coeruleum. These results provide a reference for further research on the resistance function of sugarcane WRKY genes.


1999 ◽  
Vol 81 (4) ◽  
pp. 1974-1977 ◽  
Author(s):  
Daniel Dilks ◽  
Huai-Ping Ling ◽  
Mark Cockett ◽  
Patricia Sokol ◽  
Randy Numann

Cloning and expression of the human Kv4.3 potassium channel. We report on the cloning and expression of hKv4.3, a fast inactivating, transient, A-type potassium channel found in both heart and brain that is 91% homologous to the rat Kv4.3 channel. Two isoforms of hKv4.3 were cloned. One is full length (hKv4.3 long), and the other has a 19 amino acid deletion (hKv4.3 short). RT-PCR shows that the brain contains both forms of the channel RNA, whereas the heart predominantly has the longer version. Both versions of the channel were expressed in Xenopus oocytes, and both contain a significant window or noninactivating current seen near potentials of −30 to −40 mV. The inactivation curve for hKv4.3 short is shifted 10 mV positive relative to hKv4.3 long. This causes the peak window current for the short version to occur near −30 mV and the peak for the longer version to be at −40 mV. There was little difference in the recovery from inactivation or in the kinetics of inactivation between the two isoforms of the channel.


2019 ◽  
Vol 36 ◽  
pp. 98-104
Author(s):  
B. K C ◽  
K. Kaphle

 Taenia solium is a zoonotic cestode parasite which causes cysticercosis in human and porcine cysticercosis in pigs. The infective stage of T. solium develops in pig while that of T. saginata develop in cattle and buffalo. The adult stages of T. solium are obligatory intestinal parasites for man. In humans the cysticercus larvae commonly encyst in the brain, causing neurocysticercosis. The disease was first described in pig by Aristophanes and Aristotle in 3rd century B.C. In Nepal, Taenia cysts were observed for first time in pork meat slaughtered in Kangeswari, Kathmandu more than 30 years ago. T. solium is the cause of 30% of epilepsy cases in many endemic areas where people and roaming pigs live in close proximity. More than 50 million of the world’s populations are infected worldwide and 50000 die from Neurocysticercosis yearly. In Nepal, the prevalence ranges from 0.002-0.1% in general population. Accurate diagnosis required detailed post mortem examination involving slicing of affected tissue to determine the viability of cysts. Cysticerci occur most commonly in striated muscles tissues and brain of pigs. The application of vaccination with TSOL18 recombinant vaccine along with oral medication of oxfendazole can reduces the transmission by pigs whereas use of niclosamide is found effective in humans. Hence the use of medication eventually reduces the incidence of neurocysticercosis in human and procine cysticercosis in pigs.


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