79 Matching Annotations
  1. Jan 2024
    1. qPCR’ describes quantitative real-time PCR, which is the PCR amplification of DNA in real time, measured by a fluorescent probe

      Quantified by dyes

    2. Reverse transcription PCR allows the use of RNA as a template to generate complementary DNA

      RNA is our starting product

  2. Nov 2023
    1. However, the additional possibilitythat receptors dimerize at the plasma membrane cannot beexcluded in the absence of additional studies

      Next study ?????????

    2. The D 2Svariant is highly expressed on presynaptic dopaminergicneurons, whereas the D 2L variant is found postsynapti-cally on dopaminergic neuron

      D2short- PREsynaptic D2long-POST synaptic (through the striatum)

    3. CB1R: Striatal GABA neurons -abundant in BG -DS and NAcc

    4. It is thought that the cannabinoid systemnegatively modulates dopamine circuits as activation ofthe CB 1 receptor leads to an attenuation of dopaminesignaling

      CD system downregulates DA circuitry by weakening dopamine signaling (attenuation)

    1. Negative pressure was constantly applied to the U-tubeto prevent agonist leakage and desensitization of receptors

      BIG GAP HERE... THEY BYPASS DESENSITIZATION!!!!

    2. Fura-2 recording of Ca 2 influx was used to show that incubation in 1M nicotine (2– 6 d) upregulates several pharmacological compo-nents of acetylcholine (ACh) responses in ventral midbrain cultures, including a MLA-resistant, DHE-sensitive component that pre-sumably corresponds to42 receptors.

      Fura-2 recordings of calcium influx in ventral midbrain cultures (specifically those that corresponds to a4b2) show increased responses to ACh. Additionally, by tagging a4 or b2 M3-M4 loops with fluorescent proteins, FRET can be measured as a way to directly measure increased assembly of a4b2 neurons.

    3. When cultured transfected midbrain neurons were incubated in 1Mnicotine, there was increased FRET in the cell body, denoting increased assembly of42 receptors. Thus, changes in42 receptorassembly play a role in the regulation of42 levels and responses in both clonal cell lines and midbrain neurons, and the regulation mayresult from Ca 2-stimulated pathways

      Changes in receptor assembly (due to application of nicotine) have a role in the regulation of receptor levels.

    1. From the slope of the regression fit (37 nA /cpm), it is possible tocalculate the number of bound nicotine molecules per f unctional chan-nel.

      what the hell regression line are they talking about?

    2. It is hypothesized that desensitization of neuronal nicotinicacetylcholine receptors (nAChRs) induced by chronic exposureto nicotine initiates upregulation of nAChR number. To test thishypothesis directly, oocytes expressinga4b2 receptors werechronically incubated (24–48 hr) in nicotine, and the resultingchanges in specific [ 3H]nicotine binding to surface receptors onintact oocytes were compared with functional receptor desen-sitization. Four lines of evidence strongly support the hypoth-esis. (1

      Paper 1 Hypothesis: Desensitization of neuronal nACHRs induced by chronic nicotine exposure initiates upregulation of receptor amount. - chronic nicotine -> desensitization -> upregulation of nAChRs amt -4 lines of evidence: 1. 1/2-max [nicotine] necessary to produce BOTH desensitization & upregulation. 2. [nicotine] for 1/2 maximal binding to surface nAChRs also similar. 3. this follows the same pattern, though a higher conc, is needed, for alp3bet4. 4. Mutant a4b2 that do not recover from desensitization are upregulated quickly by acute application of nicotine.

  3. Oct 2023
    1. high unitary conductance

      how many ions can pass through the channel.

    2. Cys-loop receptor

      a family of pentameric ligand gated ion channels

    3. Here, we present structures of the a7 nicotinic receptor in three conformational states: a resting-likeclosed-channel state in complex with the antagonist a-bungarotoxin (a-bgt), an activated, open-channel statestabilized by the agonist epibatidine and the positive allosteric modulator PNU-120596, and a de- sensitized,closed-channel state in complex with epibatidine alone. These structures provide a framework for probing a7’sunique biophysical and pharmacological properties and allow us to compare the gating mechanism of a7 withthose in other Cys-loop receptors. We identify a structural element at the C terminus that we call the ‘‘latch’’and pinpoint residues in this substructure required for coupling ligand binding to channel gating. We furtherdiscover an anionic residue in the extracellular domain required for the high unitary conductance and calciumpermeability seen in a7 nicotinic receptors. Finally, we observe a structural interaction that may contribute torapid desensitization inherent to a7

      HERE IS OUR RESULTS - we found the structure of the a7 receptor in - closed channel w bungarotoxin agonist - open channel state w agonist and PAM - desensitized closed-channel w agonist alone - identify a structural element at the C terminus, called "latch," and residues (aka amino acid -_-) required for "high unitary conductance (?) and Ca2+ permeability. Important to understand the gating mechanism for a7 and other Cys-loop receptors - Structural interaction that could explain rapid desensitization

    4. The rapid desensitization of a7 prevents excess calcium from entering thecells and allows fine-tuning of neuronal activity

      broad cellular effect

    5. and stimulation of anti-inflammatory pathways in the immunesystem

      receptor calcium influx effect

    6. he influx of calcium through a7 is associated with activation of voltage-dependent calciumchannels in neurons

      receptor effect

    7. desensitizes much faster than other nicotinic receptor

      property

    8. a7 has relatively low affinity for the classic agonistsacetylcholine and nicotine

      property

    9. homopentamer

      five identical subunits

    10. Deficits in a7 function are linked to mental illnesses such as schizophrenia

      Scope

    11. peri-synaptically

      In the context of neuroscience and the study of synaptic transmission, researchers often examine not only the synapse itself but also the peri-synaptic environment, which includes structures and molecules that play a role in the communication between neurons at the synapse

    12. pentameric

      a protein composed of five sub-unit

    Annotators

  4. Sep 2023
    1. the closed E2 statepredominates even in the absence of ATP,

      SERCA pumps are powered by ATP (adenosine triphosphate), which provides the energy needed for active transport of calcium ions against their concentration gradient into the ER. By controlling calcium ion levels, SERCA pumps contribute to the precise regulation of numerous cellular processes and help maintain overall cell health and function.

    2. sarcoplasmic reticulum Ca 2+-ATPase (SERCA) is activated by Ca 2+

      SERCA pump's primary function is to transport calcium ions (Ca2+) from the cytoplasm into the lumen of the ER

    1. presence of tetrodo-toxin (1 mM

      Why?

    Annotators

    1. In addition, VU0409551increased pERK1/2 in the absence of added glutamate

      It can also activate this pathway on its own (agonist activity)????

      Its okay bc other mGlu5 PAMS do this... why?

    2. ational model of allosterism (Gregory et al., 2012) allowed forquantification of VU0409551 agonist efficacy (t B : 1.04), affinity(KB : 89 nM), and cooperativity (b: 1.43) with glutamate.

      What?

    3. VU0409551 also potentiated glutamate-induced phosphoryla-tion of extracellular signal-regulated kinase 1/2 (pERK1/2) inmGlu 5 -expressing cells (Figure 1E)

      In Figure 1E, VU0409551 is shown to potentiate the phosphorylation of extracellular signal-regulated kinase 1/2 (pERK1/2) in mGlu5-expressing cells in response to glutamate stimulation.

      Figure 1E shows that when cells are exposed to different concentrations of the novel PAM along with varying levels of glutamate, the PAM enhances the strength of the cellular response related to ERK1/2 phosphorylation. This indicates that VU0409551 effectively boosts the activation of the ERK1/2 pathway in response to glutamate stimulation.

    4. shifts the glutamate concentration response curve with a maximum fold shift of 11 at 30 mM.

      The "maximum fold shift of 11" mentioned in the caption likely refers to the highest fold shift observed among all the tested concentrations of the novel PAM. In this case, at 30 microM of the novel PAM, the response to glutamate was enhanced 11-fold compared to when glutamate was administered alone!!!!!!!!

      This indicates that the novel PAM at 30 microM was particularly effective in increasing the potency of the glutamate response.

    5. ncreasing concentrations ofVU0409551 resulted in progressive leftward shifts in the gluta-mate concentration-response curve with a maximum fold shiftof 11 at a 30 mM concentration of VU0409551 (Figure 1C).

      Figure 1C: Increasing the conc. of VU leads to leftward shifts i the dose-response curve. Shifting left means smaller amount of glutamate is needed to reach a EC50.

      Proves that there is a dose-response dependent nature of the drug (aka its the drug thats modifying glutamate). Leftward shifts in concentration-response curves are often used to characterize the potency of drugs or compounds. In this case, it helps establish that VU0409551 is a potent positive allosteric modulator (PAM) of the mGlu5 receptor.

    6. but potentiatedthe response to an EC 20 concentration of glutamate with anEC 50 of 235 nM (Figure 1B).

      Description of Figure 1B

      Figure 1B, along with its title, shows that the novel PAM VU0409551 potentiates the response to an EC20 (effective concentration at which 20% of the maximum response is achieved) concentration of glutamate with a potency of 235 nM in mGlu5-expressing R10A rat cells.

      The figure's main point is to show us what the EC50 is.

      Demonstrating that VU0409551 potentiates the response to glutamate is important because it confirms that this compound has the desired pharmacological effect as a positive allosteric modulator (PAM) of the mGlu5 receptor. This information is crucial for understanding the compound's potential therapeutic applications and its suitability for further development as a drug candidate. It also helps in characterizing the compound's dose-response relationship, which is vital for dosing in experimental and clinical settings.

    7. U0409551 behaved as a classic mGlu 5 PAM (Conn et al.,2014) in rat mGlu5 -expressing HEK293A cells and did notpossess intrinsic agonist activity (Figure 1A

      Figure 1A: VU behaves as a classic mGlu5 PAM, but is not an agonist s A key characteristic of a classic PAM is that it does not directly activate the receptor on its own (i.e., it lacks intrinsic agonist activity). In the graph, this would be indicated by the absence of a calcium mobilization response when the novel PAM is applied alone (in the absence of the endogenous agonist, such as glutamate). If the PAM had intrinsic agonist activity, you would see a calcium response even in the absence of an agonist.

    8. Based on these studies, we initi-ated an effort to optimize novel mGlu5 PAMs that potentiatemGlu 5 -mediated Gaq signaling and calcium mobilization

      Statement on what they did. They are trying to optimize a novel PAM that well potentiate mGlu5 mediated Galpha-q signaling and Ca2+ mobilization. What they DONT want is a PAM that potentiates coupling of mGlu5 to modulation of NDMAr currents.

      Why dont they want this? The key point here is that they are trying to selectively enhance certain aspects of mGlu5 receptor function while avoiding the enhancement of its interaction with NMDAR currents. This suggests that there may be some undesired side effects or interactions when mGlu5 and NMDAR currents are potentiated together. By designing PAMs that specifically target one aspect of mGlu5 function (GaQ signaling and calcium mobilization) without affecting another (NMDAR currents), they hope to achieve a more targeted and potentially safer therapeutic effect.

    Tags

    Annotators

    1. poten-tiates NMDAR-independent long-term depression (LTD) butdoes not potentiate NMDAR-dependent long-term potentiation(LTP) in the rat hippocampus

      Where does it potentiate LTD and LTP? where? LTP experiments are NDMA specific Does this change current going thru NDMA receptors?

    2. characterization of a novel mGlu 5 PAM

      Does this PAM target mGlu5

    3. induces a robust potentiation of mGlu 5 couplingto Gaq -mediated calcium mobilization

      Does it induce Calcium?

    Annotators

    1. n additionalproperty commonly encountered with signal transduction proteins: the occurrence of cooperativeinteractions between multiple ligand-binding sites

      For GPCRS, this means that different configurations lead to different pathways. The combination of GPCRs can also cause allosteric change. Don't always think of GPCR has homologs, they can be heterologous

    2. As presented in this review, several well-established examples can be fitted better by thisscheme than by the KNF induced-fit mechanism.

      Whereas MWC says the the protein can be bound or unbound, however, the ligand being around increases the chance of a conformational change

    Annotators

  5. Aug 2023
    1. y contrast, the allosteric scheme implies that agonists and antagonists, as well as positive andnegative allosteric modulators, select and stabilize structurally different conformations that maybe modeled for the active, resting, and eventually desensitized states, exploiting the considerableamount of data rapidly emerging from structural and molecular dynamics studies

      Agonists, Antagonists, PAMs, and NAMs select and stabilize different conformations that may be in the active, resting, or desensitized states.

    2. As a result of symmetry properties,hetero-oligomers may occur more frequently in heterologous oligomers

      ?

    Annotators

    1. looked at what mightcause burst firing in depression

      What causes burst firing in depression

    2. The team also provoked LHb neuronsinto burst firing using optogenetics, a tech-nology that allows neurons to be activatedwith light.

      optogenetics

    3. discovered a dis-tinctive pattern of rapid bursts in the LHbof rats that display depressionlike behav-iors.

      evidence connecting depression and LHb... specifically the "burst" activity?

    4. LHb, a small, central brain region thatacts like the dark twin of the brain’s rewardcenters by processing unexpectedly un-pleasant events

      definition of LHb

    5. ather works via interaction with anotherchemical, glutamate

      works with GLUTAMARE

    6. though there are multiple theories,researchers do not quite know howketamine combats depression.17

      gap of knowledge to be explored?

    Annotators

    1. whereas mGlu5 -induced potentiation of NMDAR currentsin forebrain neurons is mediated by interactions of mGlu 5 withNMDARs through adaptor proteins that are independent ofsignaling through G a

      NMDAr potentiation induced by mGlu5 in FOREBRAIN neurons are mediated by interactions between mGlu5 & NMDAr via ADAPTOR proteins, INDEPENDENT of G-alpha-q

    2. GTP-binding protein subuni

      what is this?

    3. the hypothesis that mGlu5 -induced potentiation of NMDARcurrents is required for in vivo efficacy of mGlu 5 PAMs has notbeen directly teste

      Is mGlu5 induced potentiation of NMDAr currents necessary for effective mGlu5 PAMs?

    4. deletion or blockade of mGlu5 exacerbates psychotomi-metic-like and cognition-disrupting effects of NMDAR antago-nists

      inhibition of signaling buddy makes cognitive symptoms of NMDA antagonists (elicits NMDA hypofunction) WORSE

    5. potentiates

      A persistent increase in synaptic strength following high-frequency stimulation of a chemical synapse.

    Annotators

    1. Innate immunity is elicited earlier thanadaptive immunity in response to primary infection

      Innate, then adaptive.

  6. Jul 2023
    1. "Don't deny me my denial.'' ''Oh, that's brilliant. As long as he claims to be in denial, he doesn'teven have to appear to suffer,' I said. 'One of these days, all this grief he'savoiding is going to knock him on his ass.

      Great quote on Perry

    2. 'You could say, "I've got some good news and some bad news."''What's the good news?''You don't have AIDS.'

      More dialoguje between the POV and Perry showing responses

    3. AIDS to them was whathunger is to the fed, something we think we can imagine because we'vebeen on a diet

      Wow

    4. Because if it came out positive, he'd commit suicide. Anyway, Noah'scorning to take you to tea. Where's Enzo?'

      Dialogue highlighting response

    5. We pin our hopes onantivirals that work in vitro. What should we fear? What should we drawhope from? What is reasonable? I worry: How much fear is choice of fearin my case

      Again we see the fear response from POV.

    6. 'I haven't been of much help to you,' Perry said. 'I never even calledto see how you were doing when you had shingles.''I didn't need help,' I said, although I had mentioned this very fact inmy journal: 'March 30. I am blistered from navel to spine. My guts rise

      Perry's response: to be absolved, snarky

    7. Tm very distrustful of this sentimentality, this tendency toward willfulpathos,' I said. 'A kid I met on the train was going to a bereavementcounselor.''You've become a cynic since this all began.''That's not cynicism, that's despair

      POV guy's response to things. Seriousness, cynicism

    1. seemed Sylvia had been looking a long time for something like this,something heavy enough to leave an impression, yet so fragile it couldmake you sorry

      Feels aptly like a description of him in his sickness

    2. "Ah, you live and learn.""You live," Theo said.Sylvia blinked

      woof

    1. inally he said: Somethings you just let happen

      juicy callback

    2. n the distance a ways. Someone burning a field maybe.But it's the wrong time of year. People are still harvesting corn.

      nature

  7. Jun 2023
    1. If the newcommon wisdom that hotly overt homophobes are men who are "insecureabout their masculinity" supplements the implausible, necessary illusionthat there could be a secure version of masculinity (known, presumably, bythe coolness of its homophobic enforcement) and a staJ.,le, intelligible wayfor men to feel about other men in modern heterosexual capitalist patriarchy, what tighter turn could there be to the screw of an. already offcenter, always at fault, endlessly blackmailable male identity ready to bemanipulated into any labor of channeled violence? 1

      The author is suggesting that the notion that overtly homophobic men are "insecure about their masculinity" perpetuates the false belief that there is a fixed and secure form of masculinity that men should adhere to. This idea creates an illusion that there is a stable and acceptable way for men to feel about other men within a heterosexual patriarchal society. By labeling homophobia as a result of insecurity about masculinity, it implies that there is a "correct" way for men to express their masculinity, which often involves conforming to heteronormative expectations. This reinforces the existing power dynamics and norms within society that privilege heterosexual relationships and marginalize other forms of sexual and gender expression.

    2. efinitional barriers between "the homosexual" (minority) and "the heterosexual" (majority) are fortified, in this century, by nonhomosexuals,and especially by men against men, that most saps one's ability to believein "the homosexual" as an unproblematically discrete category of persons. Even the homophobic fifties folk wisdom of Tea and Sympathydetects that the man who most electrifies those barriers is the one whoseown current is at most intermittently direct.

      The author is discussing the reinforcement of definitional barriers between "the homosexual" (minority) and "the heterosexual" (majority) in the present century, particularly by nonhomosexuals and men against men. This insistence on maintaining these barriers undermines the belief in "the homosexual" as a clearly defined category of individuals. The author refers to the homophobic attitudes of the 1950s and how even then, there was an awareness that those who most vehemently enforced these barriers were often individuals whose own sexual desires were not consistently directed. During the period when the concept of the "homosexual" was being developed, Freud provided a countervalent perspective that emphasized the fluidity of sexual desire and the potential bisexuality of all individuals. This perspective challenged the rigid definitions and enforcement of homophobic attitudes by offering a psychologically nuanced understanding of the motives behind such behaviors. However, Freud's account was influenced by heterosexist and masculinist ethical norms, which were effectively concealed within a developmental narrative. The idea that overt homophobes are men who are "insecure about their masculinity" contributes to the illusion that there is a secure version of masculinity and a stable way for men to feel about other men within a heterosexual patriarchal society. This further reinforces an already skewed and fault-ridden male identity that can easily be manipulated into acts of violence.

    3. ocates in about the nineteenth century a shift in European thought fromviewing same-sex sexuality as a matter of prohibited and isolated genitalacts (acts to which, in that view, anyone might be liable who did not havetheir appetites in general under close control) to viewing it as a function ofstable definitions of identity (so that one's personality structure mightmark one as a homosexual, even, perhaps, in the absence of any genitalactivity at all). T hus, according to Alan Bray, "To talk of an individual [inthe Renaissance] as being or not being 'a homosexual' is an anachronismand ruinously misleading,"14 whereas th�period stretching roughly '2._e_tween Wilde and Proust was prodigally productive of atte�E_ts to name,explain, and define.this new kind of creature, the homosexual person-aproject SO urgent tfiat It spawne in its rage of distinction- ane�en ne�rcategory, that of the heterosexual persoriJ 5

      Foucault and other historians point out a change in European thinking during the 19th century regarding same-sex sexuality. Previously, it was seen as a matter of prohibited acts, where anyone who didn't have strict control over their desires could engage in such acts. However, a shift occurred where same-sex sexuality came to be seen as part of a person's identity. This means that one's personality traits or characteristics could identify them as a homosexual, even if they didn't engage in any sexual activity. In contrast, during the period from Wilde to Proust, there was a strong desire to name, explain, and define this new category of individuals, the homosexual person. This urgency to distinguish and define led to the emergence of another category, that of the heterosexual person.

    4. This is true first and generallybecause erotic identity, of all things, is never to be circumscribed simply asitself, can never not be relational

      . This is because our own understanding of our sexual identity is not just about ourselves; it is always connected to others and influenced by our relationships with them. Furthermore, the challenges and conflicts surrounding being gay in our society reflect and bring attention to the challenges and conflicts within the broader framework of compulsory heterosexuality.

    5. Afterall, the position of those who think they know something about one thatone may not know oneself is an excited and empowered on

      has literally anything changed also self-callout

    6. ssuerus does not suggest that Esther is going through a phase, oris just angry at Gentiles

      !!!!!

    7. The suggested closeted Supreme Court clerk who struggled with thepossibility of a self-revelation that might perceptibly strengthen gay sistersand brothers, but would radically endanger at least the foreseen course ofher or his own life, would have an imagination filled with possibilitiesbeyond those foreseen by Esther in her moment of risk

      note /beyond/ those foreseen

    8. Proust, in fact, insistently suggests as a sort of limit-case of one kind ofcoming out precisely the drama of Jewish self-identification, embodied inthe Book of Esther and in Racine's recasting of it that is quoted throughoutthe "Sodom and Gomorrah" books of A la recherche.

      The story goes, "Proust, in his work "A la recherche," explores the theme of coming out through the story of Esther, as depicted in the Book of Esther and Racine's adaptation of it. Esther, a Jewish queen, conceals her Jewish identity from her husband, King Assuerus, due to the belief that her people are considered unclean and abominations against nature. The king's advisor, Aman, fuels his hatred towards Jews and envisions a world without them.

      Esther's cousin Mardochee advises her to reveal her true identity at a critical moment, similar to how gay individuals may approach coming out to their homophobic parents. Esther decides to take the risk, saying, "And if I perish, I perish." The revelation of her secret identity carries immense power and challenges the king's political animosity towards her people with his personal love for her. The question arises whether the king will reject her or accept her as she is.

      The biblical story and Racine's play depict a dream or fantasy of coming out, where Esther's eloquence and self-disclosure have a transformative effect. In just a few lines, both the king and Aman realize the futility of their anti-Semitic views. The revelation of identity within the intimate space of love disrupts the public notions of natural and unnatural, purity and impurity. Esther's willingness to risk losing the love and support of her husband has the potential to save not only her own life but also her people."

    9. There is no question that to fixate, as I have done, on the scenariosketched here more than flirts with sentimentality. This is true for quiteexplicable reason

      In this passage, Sedgwick is reflecting on the limitations and potential sentimentalism that can arise from fixating on individual acts of revelation, such as coming out as LGBTQ+. She acknowledges that there is a tendency to attribute significant transformative power to individual acts of disclosure, believing that they can single-handedly challenge and dismantle systemic oppressions.

      However, Sedgwick argues that it is important to recognize the limited leverage that individual revelations actually have in the face of large-scale and institutionalized forms of oppression. While she acknowledges that acts of coming out can be powerful and disruptive, she emphasizes that they cannot completely circumscribe the consequences within predetermined boundaries, such as distinguishing between the "personal" and the "political" realms.

      Moreover, Sedgwick cautions against expecting transformative potential solely from theatrical displays of ignorance that are already deeply ingrained in societal institutions. She suggests that merely exposing or performing individual acts of revelation within a system of institutionalized ignorance is unlikely to bring about substantial transformation on its own.

    10. it isindicative for homophobia in a way it cannot be for other oppressions.

      damn

    11. that a lot of the energy of attention and demarcation that has swirled £,around issues of homosexuality since the end of the nineteenth century, inEurope and the United States, has been impelled by the distinctivelyindicative relation of homosexuality to wider mappings of secrecy anddisclosure, and of the private and the public, that were and are criticallyproblematical for the gender, sexual, and economic structures of theheterosexist culture at large, mappings whose enabling but dangerousincoherence has become oppressively, durably condensed in certain figures of homosexualit

      This is her thesis.

    12. first place, on the grounds that he had failed to note on his originalemployment application that he had been, in college, an officer of astudent homophile organization- a notation that would, as school officials admitted in court, have prevented his ever being hired. The rationalefor keeping Acanfora out of his classroom was thus no longer that he haddisclosed too much about his homosexuality, but quite the opposite, thathe had not disclosed enough.

      damn

    13. Ameditation that lacks that particular utopian organization

      She argues that without a utopian framework that envisions a future where the closet no longer exists, there is a danger of inadvertently glamorizing or justifying the negative aspects of the closet, such as its oppressive effects, distortions, disempowerment, and suffering.

    14. will risk presenting as inevitableor somehow valuable its exactions, its deformations, its disempowermentand sheer pain

      Talking about this phenomena normalizes if not glamorizes that conflicts that entails being closeted.